457 selected compound then co-infused in the microfluidic device, in mice or in 1500 the human spleens along with the same gametocyte population exposed to the solvent control. Normal RBCs, asexual ring-IRBCs and heated RBCs IMPLEMENTING ENHANCED HIGH-RESOLUTION were used as controls. We first confirmed that unlike stage I-IV, stage SURVEILLANCE USING SPATIAL DECISION SUPPORT SYSTEMS V gametocytes from an in vitro culture were not markedly retained in TO GUIDE TARGETED RAPID RESPONSE IN MULTI-DRUG microsphere-based microplate filters and in human spleen perfused ex- RESISTANT AREAS OF VIETNAM vivo, consistent with the hypothesis that deformability of gametocytes is a major determinant of their circulation in peripheral vessels. Using Gerard C. Kelly1, Colin Ohrt1, Hieu M. Truong2, Hien T. Nguyen3, the microfluidic device, we showed that stage V exposed to a recently Nicholas Martin4, Xa X. Nguyen2, Quy Anh Nguyen2, Roland identified stiffening compound C were enriched to 74.9% (vs. 25.08% for Gosling1, Jennifer Wegbreit1, Duong T. Tran2, Duong T. Tran2, unexposed controls, p=0.0001 paired t test) in narrow 2 μm-wide spaces Thang D. Ngo2 mimicking inter-endothelial slits in the spleen. In macrophage-depleted 1University of California San Francisco, San Francisco, CA, United States, C57 Bl/6 mice, immature gametocytes (10 mice) and heated RBCs (4 2National Institute of Malariology, Parasitology and Entomology, Hanoi, mice) were cleared by 86% or 75% in 3 hours, respectively. By contrast, Vietnam, 3U.S. Embassy, Hanoi, Vietnam, 4Naval Medical Research Center- a majority of mature gametocytes (5 mice) or normal RBCs (4 mice) were Asia, Singapore, Singapore still circulating 3 hours after infusion (Retention rates: 44% and 30%, Emerging artemisinin resistant in the Greater Mekong Subregion respectively (p=0.0058, p=0.0002). Similar results were observed in human (GMS) has important implications for . A project was spleens. Mature circulating gametocytes can be stiffened to induce their established to research, develop and implement enhanced surveillance and mechanical retention, thereby interrupting transmission. The stiffening targeted appropriate intervention measures to stop the spread of multi- effect can now be validated in a biomimetic microfluidic device and in a drug resistant malaria through elimination of the disease in the region. simple rodent model as a prerequisite before further development. The aims of this project are to pilot a spatial decision support system (SDSS) approach to conduct high-resolution surveillance to guide swift 1499 and targeted responses. Pilot sites were established in selected communes in Vietnam with associated customised SDSS developed. Publically THE USE OF RESPONDENT DRIVEN SAMPLING METHODS available topographic geographic information system data were uploaded TO IDENTIFY MALARIA PREVENTION KNOWLEDGE AND into the SDSS to provide baseline information. Household and forest BEHAVIORS BY MIGRANT AND MOBILE POPULATIONS IN transmission location data were located and enumerated through field- WESTERN CAMBODIA based geographical reconnaissance using handheld computers. Passively John Hustedt1, Chandary Rang1, Sophal Uth1, Sara Canavati1, detected malaria cases were geo-referenced to the suspected transmission John Macom1, Po Ly2, Siv Sovannaroth2, Arantxa Roca-Feltrer1, location sites upon diagnosis. Using case location data in the SDSS, active Tarekegn Abeku1, Sylvia Meek1 transmission foci were automatically classified and response areas-of- interest (AOI) generated. Supporting data (including population, location 1 2 Malaria Consortium, Phnom Penh, Cambodia, National Center for and number of sleeping locations within the AOI) were automatically Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia produced in the SDSS and sent to village health workers and district Mobile and migrant populations (MMPs) along the Thai-Cambodian level units to mobilize appropriate responses. Complete pilot data for border are at high-risk for malaria infection and have been found with presentation are expected in September 2014. This new approach utilizing artermisinin resistant parasites. However, the mobile nature of this novel geo-spatial tools to support targeted, appropriate and aggressive population makes it difficult to adequately measure malaria infection response measures to support malaria elimination in areas of global and risk behaviors, which is vital as we move to elimination in the region. significance will be presented. Utilizing respondent driven sampling methods, MMPs residing within two villages in Palin province (Pang Rolim and Sala Krau) were recruited 1501 in two independent rounds of sampling (602 in 2013 and 604 in 2014). All responses were adjusted for network size and recruitment patterns RESTRATIFICATION OF MALARIA EPIDEMIOLOGY IN allowing for calculation of population-adjusted statistics. While the VIETNAM FOR MORE EFFECTIVE APPLICATION OF LIMITED prevalence of Plasmodium vivax is estimated to be 0.2% among the RESOURCES general population, this study found 2.0% and 1.3% of MMPs in these 1 1 1 networks to be infected with P. vivax in 2013 and 2014 respectively, and Thang D. Ngo , Hung X. Le , Hung M. Nguyen , Thieu Q. 1 1 1 1 an absence of P. falciparum. Most respondents from Pang Rolim, from Nguyen , Trung D. Ho , Xa X. Nguyen , Anh Q. Nguyen , Ha S. 1 2 3 4 both rounds, identified having seen malaria messages within the previous Dinh , Roland Gosling , Bruce Lee , Hien T. Nguyen , Gerard C. 3 5 3 1 three months (99.7%, 95% CI: 97.6-100 in 2013 and 99.0%, 95% CI: Kelly , Nicholas Martin , Colin Ohrt , Duong T. Tran , The NIMPE 1 95.9-99.8 in 2014). However, in Sala Krau, the percentage of respondents Advisory Groups answering similarly decreased from 97.0% (95% CI: 94.1-98.4) in 2013 to 1National Institute of Malariology, Parasitology and Entomology, Hanoi, 59.1% (95% CI: 51.3-66.4) in 2014. While knowledge related to malaria Viet Nam, 2University of California San Francisco, San Francisco, CA, transmission, symptoms and prevention increased noticeably in Pang United States, 3U.S. Embassy, Hanoi, Viet Nam, 4Naval Medical Research Rolim, similar knowledge remained low in Sala Krau across both rounds. Centre - Asia, Singapore, Singapore Furthermore, while the percentage of respondents from Pang Rolim who The National Malaria Control Program in Vietnam is updating malaria didn’t use a net the previous night remained the same across both rounds epidemiology in order to more effectively apply limited malaria diagnosis, (2.4%), there was a slight increase in non-users in Sala Krau from 6.1% prevention and treatment resources. The most recent prior restratification (95% CI: 0.9-6.7) to 9.5% (95% CI: 5.3-16.4). These findings correlate was conducted in 2009. This on-going 2014 restratificaton effort (2009- with the fact that there were increased efforts on malaria prevention in 2013 data) is using the same methods of 2009 to collect all malaria case Pang Rolim (eg. concerts and videos with prevention messaging) and not data to the commune (county) level. Indicators for classification are based in Sala Krau; suggesting that as MMPs change frequently there is a need on the average number of confirmed cases per 1000 population over the for sustained public health efforts to reach this population, especially 5 year period, the presence of at least one of the three malaria vectors, within an elimination context. socioeconomic disadvantaged or border commune, poor health system, drug resistant parasites, chemically resistant mosquitoes, and migratory populations. Each indicator has a score, with the sum of the scores used to define the level of endemicity and priority for interventions. This score will be used to characterize each commune into one of five zones (no

astmh.org 458 malaria transmission, area at risk for reintroduction of malaria, low (>0- 1503 1/1000), medium (1-5/1000), or high (>5/1000)). The current levels of malaria endemicity using historical passive case detection data will be BASELINE EPIDEMIOLOGICAL CHARACTERISTICS OF determined by September 2014. Using available data, greater precision of PARTICIPANTS ENROLLED IN A TRIAL OF INTERMITTENT where malaria transmission is occurring and populations at risk will also MASS SCREENING AND TREATMENT FOR MALARIA IN be estimated. Enhanced methods to collect these data prospectively will WESTERN KENYA be developed. Additionally, data on all prior malaria interventions for the last 5 years will also be collected and entered in to a database. These will Aaron M. Samuels1, Norbert A. Odero2, Wycliffe Onyango2, be analyzed to estimate the impact of prior malaria control interventions Kephas Otieno2, Peter Otieno2, Ya Ping Shi1, Mary J. Hamel1, Kim in an operations research model to help select which methods should Lindblade1, Simon Kariuki2, Meghna Desai1 be continued or reassessed. Methods to prospectively assess the impact 1Centers for Disease Control and Prevention, Atlanta, GA, United States, of new interventions in an on-going and iterative fashion will also be 2Kenya Medical Research Institute, Kisumu, Kenya developed. We will present the new 2014 restratification data and The prevalence of malaria infection remains high in western Kenya despite compare and contrast it with the 2009 data. We will illustrate how these over 30 years of control efforts. Community parasite prevalence in children new data will be used to better target interventions. The plan to collect <5 years of age was estimated at >80% in the early 1980s, and dropped more precise prospective information, as well as the status of the analysis to 26% by 2008. Prevalence rose to 43% in 2011. Individuals with of intervention impact, will also be presented. asymptomatic parasitemia account for 90% of infections in specific age groups and may be sustaining the continued high level of transmission. 1502 Mathematical models suggest that strategies targeting the asymptomatic population could reduce malaria transmission. We have initiated a multi- LAMP AS DIAGNOSTIC TOOL FOR DETECTION OF SUB- year two-arm cluster randomized controlled trial to evaluate the impact PATENT ASYMPTOMATIC MALARIA INFECTIONS IN PRE- of intermittent mass screen and treat (iMSaT) campaigns for malaria in ELIMINATION SETTINGS IN NORTHERN NAMIBIA Siaya County, western Kenya. We describe baseline cross-sectional survey Davis R. Mumbengegwi1, Munyaradzi Tambo1, Ronnie A. Bock1, results. Twenty compounds were randomly selected from each of 20 Roland D. Gosling2, Hugh J. Sturrock2 clusters during peak malaria transmission season in July-August, 2013. All consenting individuals within houses of selected compounds were asked 1University of Namibia, Windhoek, Namibia, 2University of California San demographic, behavioral and symptom-based questions by community Francisco, San Francisco, CA, United States health workers using personal digital assistants. Blood samples were tested The global map of malaria is shrinking with 34 out of 99 malaria endemic for malaria using a combination HRP2/pLDH rapid diagnostic test (RDT), countries embarking on eliminating the disease. Transmission in Namibia light microscopy (LM), and polymerase chain reaction (PCR). A total of has declined dramatically from 477,786 cases in 2000 to 1546 in the 1,987 persons living in 605 households from 359 selected compounds 2013 malaria season. Namibia is now in the pre-elimination phase of were interviewed. Of these, 1,402 consented for both RDTs and LM. malaria and is targeting elimination by 2020. A new challenge facing the Baseline malaria infection prevalence was 47.1% (95% Confidence elimination campaign is the detection of asymptomatic malaria cases with interval [CI] 43.9-50.2) and 36.6% (CI: 33.3-39.8) by RDT and LM, low levels of parasitaemia. These infections are difficult to detect as they respectively. RDT positivity was strongly associated with age; 64.9% (CI: are below the threshold of routinely used Rapid Diagnostic Test (RDT) 57.6-72.2), 70.8% (CI: 65.1-76.4), and 27.8% (CI: 24.5-31.1) of persons kits, yet can result in onward transmission. Molecular tools based on DNA aged <5 years, 5-15 years, and older than 15 years were RDT positive (P < amplification such as PCR are sensitive and specific enough to detect low 0.0001), respectively. Only 25% who were RDT positive reported a fever in parasitaemia but their routine use requires expensive, highly technical the prior 24 hours, and 45% reported a fever in the previous two weeks. equipment and expertise. Loop-mediated isothermal amplification (LAMP) Overall, history of fever in the previous 2 weeks was not associated with is a tool based on DNA amplification and has the advantages of PCR RDT positivity, PR 0.94 (CI: 0.84-1.04). Of persons reporting a fever in the yet is requires less expertise and equipment. This study was conducted previous 2 weeks, RDT positive individuals were as likely to seek care as to determine the usefulness of LAMP as a diagnostic tool to detect those who were not, PR 1.04 (CI: 0.93-1.16). PCR results and multivariable asymptomatic, sub-patent infections found during reactive case detection analyses are pending. The large proportion of infections that were not in Engela district in Northern Namibia. All RDT confirmed malaria cases associated with fever or care-seeking behavior suggests that strategies reported in the Engela district and members of their households as well targeting the asymptomatic population may be beneficial for reducing as occupants of the four surrounding households were recruited into the malaria transmission in western Kenya. study. RDTs and dried blood spots (DBS) of all subjects were collected and DNA was extracted from both using the chelex method. LAMP was run 1504 using DNA extracted from all the collected samples and results detected as fluorescence under a UV light. Preliminary results from 416 RDTs and COMMUNITY-LEVEL MALARIA SURVEILLANCE IN SOUTHERN DBS collected during follow up of 11 index cases, showed 11 individuals PROVINCE, ZAMBIA - AN ANALYSIS OF PERCEPTIONS, positive by RDT and 18 positive by LAMP. Thus 7 additional secondary PRACTICE AND PROGRESS IN AN AREA TARGETING malaria cases associated with index cases (a 1.6 fold increase) were ELIMINATION detected by LAMP over RDTs. This shows LAMP could be a useful tool to detect sub-patent asymptomatic malaria infections at low transmission and Lynne Lohfeld1, Tokozile Kangome-Ngwenya2, Anna M. may be a suitable diagnostic tool for use in pre-elimination settings. Winters2, Zunda Chisha2, Busiku Hamainza3, Mulakwa Kamuliwo3, John Miller4, Matthew Burns2, Daniel J. Bridges2 1McMaster University, Hamilton, ON, Canada, 2Akros, Lusaka, Zambia, 3Zambia National Malaria Control Center, Lusaka, Zambia, 4PATH-MACEPA, Lusaka, Zambia Since its inception in 2011, the training techniques and personnel associated with the program have evolved and matured using anecdotal feedback from community health workers (CHWs) trained to deliver the program, their supervisors and district-level staff. We have recently conducted a systematic qualitative program review to determine how malaria is perceived by the program implementers and beneficiaries, how the community received the surveillance platform, and solutions to key

astmh.org 459 operational challenges that could improve implementation. A criterion- antimalarial drugs and host immunity in clearing both asexual parasites based sampling framework based on training regime and performance and transmission-stage gametocytes. We modeled the pharmacokinetics level of facilities offering the program was used to select six (PK) and pharmacodynamics of two artemisinin-based combination posts, three per district, and a representative from associated stakeholder (ACTs), artemether-lumefantrine (AL) and dihydroartemisinin- groups (community health workers delivering the program, their clinic- piperaquine (DP), and one gametocytocidal drug, primaquine, using based supervisors, regional-level staff, and community members). age-based dosing and weight-dependent PK. We show that current Individual interviews and focus group discussions were then held in these dosing regimens, especially current fixed-dose recommendations for selected sites. Service providers, supervisors, program administrators and DP, significantly underdose children. We also find that asexual-stage community members all credited the program with helping to reduce the immunity alone is insufficient to explain low gametocyte prevalence in number of malaria cases. Barriers to fuller implementation of the program populations with endemic malaria; host physiological responses are likely included transportation (e.g. ensuring all CHWs had working bicycles), to modulate prevalence of the sexual stage. We identify a maximum EIR communication (e.g. providing CHWs with working cell phones and “talk above which co-dosing ACTs with primaquine has little effect on reducing time” to transmit data by phone) and supplies (e.g. ensuring adequate prevalence, and we demonstrate that a minimal level of individual number of RDT kits to test for malaria in clinics and communities, compliance is necessary for mass drug treatments to impact transmission. artemether-lumefantrine to treat uncomplicated malaria cases, and Pharmacological modeling of antimalarials can guide community decisions antipyretics for malaria-negative patients to encourage future visits to rural in drug administrations and alert administrators to the most likely and health centers). Results from this review will be used when developing deleterious modes of drug failure. plans to scale-up the program for delivery in other parts of Zambia. 1507 1505 SIMULATION OF MALARIA PARASITE RESERVOIR REACHING MIGRANT AND MOBILE POPULATIONS THROUGH COVERAGE USING REACTIVE CASE DETECTION AND ACTIVE A PRIVATE SECTOR INITIATIVE: MALARIA BED NET LENDING COMMUNITY FEVER SCREENING FROM CENSUS DATA IN SCHEME IN CAMBODIA SOUTHERN ZAMBIA Soy Ty Kheang Joshua O. Yukich1, Adam Bennett1, Rudy K. Yukich2, Busiku University Research Co. LLC, Phnom Penh, Cambodia Hamainza3, Kafula Silumbe4, John M. Miller4, Thomas P. Eisele1 While tremendous strides have been made toward eliminating malaria 1Tulane University School of Public Health and Tropical , New in Cambodia, pockets of risk persist in border areas that are remote and Orleans, LA, United States, 2Sensorstar, Inc., Ellicott City, MD, United have high population mobility. Lacking knowledge of malaria, migrant States, 3National Malaria Control Centre, Ministry of Health, Lusaka, and mobile populations (MMPs) are particularly at risk. The PMI/USAID Zambia, 4PATH-MACEPA, Lusaka, Zambia Control and Prevention of Malaria (CAP-Malaria) Project in the Greater There is need for malaria elimination programs to determine when and Mekong Sub-region works with private sector employers to increase where reactive case detection (RCD) is most effective and feasible. Geo- access to and use of long-lasting insecticidal nets (LLINs) by their migrant referenced census data on over 80,000 individuals from 6 rounds of a workers. Employers range from small farm owners to large companies that mass test and treatment (MTAT) intervention with rapid diagnostic tests manage rubber, cassava plantations, and hydroelectric dam construction. (RDT) in southern Zambia (2012-2013) were analyzed using a Monte-Carlo Employers receive a stock of LLINs and malaria educational materials for simulation algorithm to assess the coverage, sensitivity and specificity of their employees. The employers then lend the LLINs to their workers, potential RCD systems. Data on household location and composition, fever retrieving them prior to their departure for reuse with other migrant history, treatment seeking, and RDT results for all individuals were included employees. An evaluation of the lending model was conducted during the in MTAT census data. Simulations were conducted within 25 health facility harvesting season in late 2013. The study assessed access to and utilization catchments, and the following parameters were varied in sensitivity of LLINs by migrant workers, and explored reasons for non-use. Interviews analysis: RCD search radius or number of households searched, sensitivity were conducted with 207 farm owners and 712 workers. Results showed and specificity of diagnostics used to identify index cases, treatment that farm owners were generally satisfied with the LLIN lending model. seeking probability, household and individual RCD participation level, Some employers (28%) ran out of nets. LLIN uptake among the workers sensitivity and specificity of diagnostic used during RCD search. Results was high, most (93%) had a bed net at their residence, and almost all indicate that RCD and active community fever screening are potentially (96%) reported sleeping under a bed net the previous night. Half of the efficient ways of identifying the parasite reservoir. However, substantial workers (58%) had received an LLIN from their employer. The main barrier resources are required before meaningful fractions of the parasite reservoir for not using a LLIN was that it was considered too stiff (29%). A fifth of are found in a single search round. Treatment seeking for fevers and respondents also said they were allergic to the insecticides. Half of the access to care are key limiting factors to the sensitivity of an RCD system farm workers said they would be willing to pay a small amount for their for identifying the parasite reservoir in the community. A shift from RCD own net, suggesting an opportunity for subsidized vouchers for LLINs. to active community fever screening would improve the fraction of the parasite reservoir identified, especially in areas with poor access to care. 1506 However, the fraction of the parasite reservoir identified remains small given feasible search criteria in both systems. Multiple RCD rounds may MODELING PHARMACOKINETICS AND improve the fraction identified over a given period of time. PHARMACODYNAMICS OF ANTIMALARIAL DRUGS IN THE EPIDEMIOLOGICAL MODELING (EMOD) MODEL WITH IMPLICATIONS FOR TRANSMISSION REDUCTION Jaline Gerardin, Edward Wenger, Philip Eckhoff Intellectual Ventures, Bellevue, WA, United States Modeling approaches can predict the effect of large-scale drug deployments on reducing morbidity over several years of campaigns, leading to better understanding of the role of antimalarial drugs in eradication. The Epidemiological Modeling (EMOD) simulation program includes individual-level modeling of vectors and within-host dynamics, providing an ideal environment in which to study the interplay of

astmh.org 460 1508 days after. Infectivity to mosquitoes was measured by the presence of oocysts 7 days post infected feeding. There was no severe or serious MAPPING GLOBAL MALARIA CONNECTIVITY FOR STRATEGIC adverse event. Preliminary analysis on the first 30 participants enrolled ELIMINATION PLANNING showed no differences among the three groups in mean change in hemoglobin following treatment on day 1 (p=0.89), day 2 (p=0.77), day 3 1 2 3 Andres Garcia , Alessandro Sorichetta , David L. Smith , Deepa K. (p=0.10), day 7 (p=0.61), day 14 (p=0.66), and day 28 (p=0.81). The mean 4 2 Pindolia , Andrew Tatem hemoglobin was 13.8 g/dL (range: 11.5, 16.2) at day 0, 13.9 g/dL (range: 1Bill & Melinda Gates Foundation, Seattle, WA, United States, 2University 11.3, 17.9) at day 7, and 14.0 g/dL (range: 11.6, 17.3) at day 28. In the of Southampton, Southampton, United Kingdom, 3Johns Hopkins School control group, compared to day 0 there was no reduction in infectivity of Public Health, Baltimore, MD, United States, 4Clinton Health Access on day 2 or day 7, -22.8% (95% CI -100%,100%) and 40.0% (95% CI Initiative, Boston, MA, United States -86%, 100%), respectively. In the 0.125 mg/kg dose group, compared Calls for the eradication of malaria require the development of global to day 0, there was 86.3% (95% CI 39.4%, 100%) reduction on day2 and regional strategies based on a strong and consistent evidence base. and 100% reduction in day 7. In the 0.5 mg/kg dose group, compared to Evidence from the previous global malaria eradication program and day 0 there was a 100% reduction in day 2 and 90.1% (95% CI 64.7%, more recent transborder control campaigns have shown the importance 100%) reduction on day 7. In summary our preliminary results indicate a of accounting for human movement in introducing infections to areas higher reduction in infectivity in the 0.5 mg dose group (100% reduction targeted for elimination. Here, (micro)census, survey and cellphone-based at day 2) without safety concerns. human movement data and models were analysed with network analysis tools to map globally the connectivity of both countries and subnational 1510 administrative units through population movements. These data were also combined with and P. vivax malaria transmission GENOME-SCALE PROTEIN MICROARRAY ANALYSIS OF maps and a global population dataset to identify the likely principal PLASMODIUM FALCIPARUM AND P. VIVAX SYMPTOMATIC sources and destinations of imported cases. Results indicate that certain AND ASYMPTOMATIC INFECTIONS IN A REMOTE VILLAGE groups of countries and regions within countries are much more strongly OF THE PERUVIAN AMAZON connected by high levels of population movement than others. The 1 2 3 mapping here of both communities of subnational regions and countries Raul Chuquiyauri , Douglas Molina , Huw Davies , Hugo Rodriguez4, Alejandro Llanos-Cuentas5, Philip Felgner3, Joseph M. linked by high levels of population exchange, and ‘natural’ migration 1 boundaries that display reduced movement of people and infections Vinetz between regions has practical utility. These inform the design of malaria 1University of California San Diego, San Diego, CA, United States, 2Antigen elimination strategies by identifying regions afforded protection from Discovery Inc., Irvine, CA, United States, 3Division of Infectious Diseases, re-colonisation by natural ‘firebreak’ regions of reduced connectivity. For Department of Medicine, University of California Irvine, Irvine, CA, United more isolated areas, a regionally-focussed control or elimination program States, 4Directorate of Health, Iquitos, Peru, 5Tropical Medicine Institute is likely to stand a better chance of success than those receiving high “Alexander von Humboldt”, Universidad Peruana Cayetano Heredia, Lima, levels of visitors and migrants from high transmission regions. Moreover, Peru we demonstrate how the mobility and malaria connectivity framework The Peruvian Amazon is hypoendemic for Plasmodium falciparum (Pf) provides an evidence base for informing the design and simulation of and P. vivax (Pv) malaria. To assess, at a population level, the proportion malaria elimination strategies globally. of people infected by the endemic malaria parasites, a prospective cohort study was done in Santa Emilia, a remote-rural village in the Peruvian 1509 Amazon in 2013 where the main seasonality for malaria relates to river height rather than rainfall. Light microscopy and an aldolase gene- PHASE 2A DOSE ESCALATION STUDY OF SAFETY AND specific qPCR assay that detects both Pv and Pf were used to assess the EFFICACY OF LOW SINGLE-DOSE PRIMAQUINE FOR presence of parasitemia. Plasmas were obtained for genome-scale protein GAMETOCYTOCIDAL ACTIVITY AGAINST P. FALCIPARUM IN microarray analysis using a combined Pf500/Pv500 chip containing a SUB-SAHARAN AFRICA down selected list of the 500 of the top most sero-reactive antigens for each Pf and Pv. At baseline, 33(22%) and 77(51%) of 151 subjects 1 2 1 Halimatou Diawara , Joelle Brown , Ibrahima Baber , were positive by microscopy and q-PCR, respectively. On the last survey, 1 1 1 Almahamoudou Mahamar , Koualy Sanogo , Harouna Soumare , 3(2%) and 8(5%) of 157 subjects were positive by microscopy and 1 2 2 1 Fanta Koita , Eugenie Poirot , Jimee Hwang , Sekou Traore , q-PCR, respectively. Asymptomatic parasitemia detected by microscopy 3 4 1 Francois Nosten , Teun Bousema , Alassane Dicko , Roland ranged from 22% to 40%, while for q-PCR ranged from 41% to 60%. A 2 Gosling significant proportion of infections detected by q-PCR, 39% and 47% for 1Malaria Research and Training Center, Bamako, Mali, 2Global Health Pf and Pv, respectively, were undetected using microscopy. Comparison Group, University of California San Francisco, San Francisco, CA, United of proportions of negative subjects at each of the eleven surveys revealed States, 3Mahidol Oxford University Research Unit in Bangkok, Oxford, there was a significant increase of negative subjects during September United Kingdom, 4University of Nijmegen, Nijmegen, Netherlands to December surveys in comparison to March surveys. Protein microarray Primaquine is the only currently available drug with strong gametocytocidal analysis was done with 324 plasma samples: 132 matched paired samples properties against the more mature gametocytes and known to behighly from the two time points and 60 unpaired samples. The top 200 most effective in reducing gametocyte carriage and infectivity to mosquitoes. sero-reactive antigens were selected for comparison. Seroreactivity However its deployment hasbeen limited because of the safety concerns. increased with age and in response to documented malaria infection. To identify the lowest efficacious dose of PQ, we conducted a Phase 2a Seroreactivity was lower in September than in March for all age groups, dose escalation study of safety and efficacy of low single-dose primaquine paralleling mosquito abundances (related to river height, not rainfall). in non deficient G6PD male in Ouelessebougou Mali. The first 50 While there was a gradual increase in seroreactivity associated with participants aged 5 to 50 years with Plasmodium falciparum gametocyte at age, the differential seroreactivity between March (high) and September blood smear, were randomly allocated to one of the following treatment (low) was greatest in youngest and this difference decreased with age. groups with primaquine at 0, 0.125 mg/kg and 0.5 mg/kg. All participants Prior to embarking on an elimination strategy, monitoring changes in received standard dose dihydroartemisinin-piperaquine. Subjects were transmission intensity and identification of malaria foci is mandatory seen at days 0, 1, 2, 3, 7, 14 and 28 for hemoglobin measurement and for best intervention efforts. This population-based study of a malaria- assessment adverse events. Mosquitoes were fed on blood meal using endemic population identified new serological markers of infection using membrane feeding assay before administration of drug and 1, 2 and 7

astmh.org 461 genome-scale protein microarray. This new tool has important potential often need to perform their own data analysis, which is time-consuming for providing key control and elimination data for national surveillance and resource-intensive. In an attempt to address this problem, we have programs. developed the “Free Surveillance App” (FREESAPP), an online application that converts time-series data from national surveillance systems 1511 into interactive graphs similar to those produced by the Gapminder Foundation. The applet can output burden distribution and decision- INVESTIGATING OPERATIONAL STRATEGIES FOR making trees for cost-effectiveness assessments as well as utilize satellite ANTIMALARIAL DRUG ADMINISTRATION IN ZAMBIA’S maps for background illustration. The overall goal of this application is to SOUTHERN PROVINCE: A SIMULATION STUDY allow public health officers to efficiently and accurately use the surveillance data to guide their decision-making practice at each of the levels of the 1 2 2 1 Erin M. Stuckey , John M. Miller , Megan Littrell , Nakul Chitnis , local health system hierarchy. To achieve this goal, FREESAPP allows users 2 Rick W. Steketee to plot the trends in malaria burden using a variety of interactive displays 1Swiss Tropical and Public Health Institute, Basel, Switzerland, 2PATH (i.e., bubble, bar, and line charts). FREESAP allows users to include and Malaria Control and Evaluation Partnership (MACEPA), Seattle, WA, United adjust for various covariates of interest (i.e., incident rate, population States size, P. falciparum proportion, time, etc.) using different mathematical Malaria elimination requires reducing both the potential of mosquitoes transformations. Users can contrast trends against epidemiological to transmit parasites and the infectious reservoir of parasites in humans, thresholds for each reporting level, which are automatically updated with including asymptomatic infections. To achieve this goal in Southern each week of data entry. Furthermore, FREESAPP users are able to estimate Province, Zambia a mass test and treat (MTAT) campaign using artemether- costs of implementing interventions by using a cost-effectiveness algorithm lumefantrine was conducted from 2011-2013 to complement high that is adjustable by population size, distance, and coverage rates. In order coverage of long lasting insecticide-treated nets. In order to identify to allow data managers to update the system without altering their current factors likely to increase campaign effectiveness, a modeling approach reporting protocol, FREESAPP was developed using a combination of four was applied to investigate the simulated effect of alternative operational free tools: Motion Chart, Google Earth, Google maps, and R software. strategies for MTAT in Southern Province. OpenMalaria, a discrete-time, Given its open source free format, FREESAPP may contribute to enhancing individual-based stochastic model of malaria, was parameterized for the the local readiness and response capacity at each of the levels of the range of transmission intensities observed in the study area to simulate Health System hierarchy in most malaria endemic countries. FREESAPP may antimalarial administration for interruption of transmission. Simulations also potentially be used for other reportable diseases, therefore facilitating were run for scenarios with a range of artemisinin-combination therapies improved public health decision-making. (ACTs), proportion of the population reached by the campaign, targeted age groups, frequency of campaign rounds, Plasmodium falciparum 1513 test protocols, and the addition of drugs aimed at preventing onward transmission. Scenarios were evaluated based on the reduction in all-age EVALUATING REACTIVE CASE DETECTION ACTIVITIES IN parasite prevalence during the peak transmission month following the RANONG PROVINCE, THAILAND campaign, compared to the currently-implemented strategy. Simulation Prayuth Sudathip1, Jennifer Koonings2, Chris Cotter3, Roly results suggest that the most important determinant of success in reducing Gosling3 prevalence is the coverage of the population achieved in the campaign. 1 However, even with high coverage with mass drug administration (MDA) Bureau of Vector-Borne Diseases, Department of Disease Control, 2 in areas with a pre-intervention all-age parasite prevalence of less than Bangkok, Thailand, Global Health Sciences, University of California San 3 10%, simulations suggest that elimination would require more than one Francisco, San Francisco, CA, United States, Global Health Group at the year of campaign implementation. Including single low-dose primaquine, University of California San Francisco, San Francisco, CA, United States which acts as a gametocide, to the drug regimen did not further reduce Active case detection (ACD) strategies are recommended by the World prevalence. The addition of an endectocide, such as ivermectin, resulted in Health Organization and are implemented in many low transmission a lower simulated parasite prevalence and warrants further investigation. settings worldwide as a critical part of malaria elimination programs. ACD Simulation results indicate a high proportion of low density infections strategies include determining the origin of infection, case investigation, were missed by rapid diagnostic tests that would be treated and cleared and responding to locally acquired cases of malaria, known as reactive with MDA. The optimal implementation strategy for MTAT/MDA will vary case detection (RACD). Effectively implementing RACD requires substantial by background level of prevalence and rate of infections imported to the programmatic and human resources. Thailand is pursuing a spatially- area. Success of the campaign depends on continued coverage of vector progressive approach to eliminate malaria from 80 percent of the country control interventions to ensure sustained gains in reduction of disease by 2020. Between April and June 2014, the Thailand Bureau of Vector- burden. Borne Diseases (BVBD) conducted an evaluation in Ranong Province to identify best practices and inform RACD efforts in the province and across 1512 Thailand. Using a standardized monitoring and evaluation (M&E) tool, five districts within the province were evaluated, and included a mix of high, A FREE SURVEILLANCE APP FOR PLANNING MALARIA medium and low transmission settings. Case investigation and RACD rates ELIMINATION INTERVENTIONS AND OUTBREAK RESPONSES and reporting timeliness were analyzed through secondary data extraction AT THE COMMUNITY LEVEL IN MALARIA ENDEMIC from the national malaria information system and district-level malaria COUNTRIES clinics and measured against defined indicators. Questionnaires were administered to 60 malaria clinic staff regarding RACD operations and 1 2 1 Antonio M. Quispe , Josiah L. Kephart , Andres G. Lescano , procedures. A financial analysis of RACD-related expenditures was also 2 2 Timothy M. Shields , Clive J. Shiff collected and analyzed to determine the primary cost drivers and operating 1U.S. Naval Medical Research Unit - 6, Lima, Peru, 2Johns Hopkins costs for RACD. Findings from the evaluation will inform the BVBD on Bloomberg School of Public Health, Baltimore, MD, United States program efficiency within Ranong Province, identify best practices and Planning malaria elimination interventions and outbreak responses at gaps in RACD activities, and will assist the BVBD in optimizing RACD the community level represents an operational challenge in most malaria program effectiveness. endemic countries. This happens primarily because most health systems report surveillance data at the regional and country levels only, despite data collection at the community level. Therefore, when planning interventions and responses at the community level, decision makers

astmh.org 462 1514 safe and well tolerated. An international consortium was established to facilitate development of PfSPZ Vaccine for use in geographically focused LAMP FOR THE DETECTION OF SUB-MICROSCOPIC MALARIA Pf malaria elimination campaigns, and a 4-stage clinical development INFECTIONS IN THE CONTEXT OF MALARIA ELIMINATION IN plan (CDP) delineated. In Stage 1, which is in progress, trials at 3 sites CAMBODIA in the US, and in Mali, Tanzania, Equatorial Guinea, and Germany are assessing the reproducibility of the VRC 312 trial, and optimizing 1 2 1 1 Nimol Khim , Heidi Hopkins , Saorin Kim , Dany Dourng , durability, heterologous protection, and dosage regimens. These trials 3 4 1 Vincent Sluydts , Somony Heng , Lydie Canier , Sovanaroth are intended to establish: (1) the reproducibility of the findings from the 4 4 3 3 Siv , Tho Sochantha , Lies Durnez , Marc Coosemans , Vincent study conducted at VRC; (2) protection against heterologous Pf, including 1 5 2 1 Deubel , David Bell , Iveth J. Gonzalez , Didier Menard naturally acquired Pf; (3) durability of protection; (4) protective efficacy 1Institut Pasteur in Cambodia, Phnom Penh, Cambodia, 2Foundation of different dosage regimens of PfSPZ Vaccine - regimens of 5, 4, 3, 2 or for Innovative New Diagnostics, Geneva, Switzerland, 3Institute of 1 doses of 1.35x105 to 2.2x106 PfSPZ/dose by direct venous inoculation , Antwerp, Belgium, 4National Center for Parasitology, (DVI) or IM routes; (5) an assay/biomarker that predicts protection; and Entomology and Malaria Control, Phnom Penh, Cambodia, 5Global Good/ (6) optimal approaches for DVI administration. >400 doses of 2.7x105 to Intellectual Ventures Lab, Seattle, WA, United States 2.2x106 PfSPZ/dose have been administered in the US and Africa, and the In the context of government’s commitment to eliminate malaria in vaccine has been safe and exceptionally well tolerated. Stage 2 will include Cambodia in 2025, additional efforts and new methodologies to detect age de-escalation and escalation and regimen optimization trials; Stage 3 malaria parasites in asymptomatic individuals are needed. Indeed, in very will be pivotal phase 3 trials; and Stage 4 will include mass administration low transmission settings such as Cambodia, asymptomatic infections campaigns to halt transmission and eliminate Pf malaria from populations remain the major reservoir of malaria parasites contributing to maintain of > 200,000 individuals. Progress and plans will be explained. disease transmission. As a consequence, the detection and treatment of the asymptomatic carriers is a crucial step in progress towards 1516 malaria elimination. This represents a new challenge as the proportion of asymptomatic parasite carriers is unknown. To date, although PCR SPECTRUM OF INFECTIOUS DISEASES IN RURAL CLINIC FOR methods show lower detection threshold compared to microscopy, Nested REFUGEES AND DISPLACED POPULATION ON RWANDA-DR or Real time PCR assays required fully equipped laboratory and trained CONGO BORDER: ANALYSIS OF 10,051 PATIENTS technicians. These approaches are not suitable as point-of-care in the Katarina Molnarova1, Dominik Hes1, Gertruda Mikolasova2, field, contrarily to LAMP, which can be done on a simple bench top in Lenka Michalikova3, Jaroslava Brnova3, Nada Kulkova2, Vladimir a clinic, with basic reagents and equipment, by personnel with only a Krcmery2 few days’ training in the technique. To assess the performance of this 1 2 promising tool, we have conducted a retrospective study on 516 samples Bl. Clémentine-Anuarita Healthcare Centre, Bigugu, Rwanda, Laboratory 3 from asymptomatic individuals collected in Rattanakiri province, eastern of Molecular Microbiology, Bratislava, Slovakia, Faculty of Health Sciences Cambodia. Ten microliters of DNA extracted by Instagene matrix from and Social Work, Trnava, Slovakia dried blood spots were used for both LAMP (Pan detection) and Real time Migrants and internally displaced in sub-Saharan Africa are subject PCR. Positive specimens with Pan LAMP were screened for falciparum of increasing threat of infectious diseases (IDs) due to contaminated species (P. falciparum LAMP reaction). The results between the two water supplies (cholera, typhoid fever), food (salmonellosis, shigellosis), techniques were compared to calculate the diagnostic accuracy. Based malnutrition (tuberculosis-TB, HIV) and absence of housing (pneumonia, on the LAMP detection, the prevalence of malaria infection was 19.8%. upper respiratory tract infections - RTI). Cross sectional study in area Compared to the Real time PCR, the specificity and the sensitivity of the close to DRC border (Sud-Kiwu) and Rwanda (Bisesero) in two clinics malaria LAMP kit was 93.3% (95% CI: 90.5% - 95.4%) and 86.4% (95% serving for 50 000 population (of them 25 000 internally displaced and CI: 76.6% - 92.7%), respectively. We concluded that LAMP detection has refugees in UNHCR camps) was performed to asses occurrence of major similar performances with the Real time PCR. In addition, LAMP results ID in 2013. Bigugu clinic is located in altitude of 2350 m and Bisesero are available just one hour after sample processing begins for 14 samples. United Nations High Commissioner for Refugees (UNHCR) camp is in We suggest that high throughput LAMP assay for a large-scale screening 1150 m above sea level.Of 10 051 patients, only 31 (0,3%) had malaria, would be developed for a step forward to malaria eradication. and 26 of them (0,26%) had true highland malaria (without down country travelling history), confirmed both microscopically and with 1515 rapid diagnostic test (RDT). Commonest IDs were upper RTI representing (72-89%) of all visits, followed by diarrheal and gastro-enteric diseases STAGE 1 TRIALS OF THE CLINICAL DEVELOPMENT PLAN FOR (13-19%). Also, 26-77% of all children were infected by geohelmints. PFSPZ VACCINE FOR GEOGRAPHICALLY FOCUSED MALARIA Only one case of neuroinfection was recorded. Urinary tract infections ELIMINATION CAMPAIGNS and sexually transmitted diseases were rare as well (1-4%).Among 10 051 outpatient visits in two rural clinics, serving for UNHCR registered Stephen L. Hoffman refugees from DRC in Rwanda and internally displaced population near International PfSPZ Vaccine Clinical Consortium; VRC, Laboratory of Sud-Kiwu Province. Malaria was extremely rare due to high altitude, and Malaria and Vaccinology, DMID of National Institute of diarrheal and gastrointestinal infections were relatively rare, too. Of all ID, and Infectious Diseases, UMB CVD, Bamako Malaria Research and upper RTI were the commonest, while neuroinfections (such as bacterial Training Center, IHI Bagamoyo, Swiss Tropical and Public Health Institute, or viral meningitis and sleeping sickness) were only exceptional. Very Naval Medical Research Center, Walter Reed Army Institute of Research, high proportion of RTI was associated with malnutrition and very low University of Tubingen, MoH Equatorial Guinea, CRESIB, Centers for socio-economic status in areas of high altitudes above sea level with low Disease Control and Prevention, , Rockville, MD, United States temperature. A vaccine for geographically focused malaria elimination campaigns must provide sustained sterile protection against infection. The Sanaria® PfSPZ Vaccine was developed to address this need. It is composed of aseptic, purified, cryopreserved, vialed PfSPZ. In a recent clinical trial at the Vaccine Research Center (VRC), NIAID, NIH, PfSPZ Vaccine protected 6/6 (100%) subjects against controlled human malaria infection at the highest dosage regimen administered (5 IV doses of 1.35x105 PfSPZ), There was a dose response in regard to antibody and T cell responses, and the vaccine was

astmh.org 463 1517 microscopically, plus rapid diagnostic testing (RDT) has been used since 2013. Seasonality of severe malaria was observed among majority of cases SPECTRUM OF INFECTION DISEASES IN BURUNDIAN RURAL in period from April to November, with 113 to 221 cases per month with HOSPITAL IN GASURA IN DRY VERSUS RAINY SEASON - up to 9 deaths on severe malaria (monthly among 3707 admissions, in 2012/2013 2013). Altogether, 1438 patients (38.8 %) had severe malaria clinically confirmed as fever plus severe anemia, or respiratory distress syndrome, 1 1 1 1 M. Kolibab , Jana Trilisinska , Eva Smrekova , Tomas Razsony , or cerebral malaria, or liver, or kidney failure, or severe hypoglycemia with 1 1 1 Eliska Maradova , Alexandra Mamova , Miriam Balazova , George acidosis. Of 1438 severe malaria cases, 76 died (5.3 %). Relatively low 2 1 2 Benca , Bianca Voinescu , Vladimir Krcmery mortality may be explained with: (i) good access to the hospital, (ii) pre- 1Health Centre in Gasura, Burundi, Gasura, Burundi, 2Slovak Tropical referral administration of antimalarial drugs due to education campaign Institute, Bratislava, Slovakia with the in 2010 - 2013, and (iii) use of artemisinin-based combination Seasonal cycles of infectious diseases have been variously attributed to (ACT) since 2010 in Marial Lou. Severe malaria in travelers changes in atmospheric conditions, the prevalence or virulence of the returning to Europe is associated with up to 20 % mortality. But it can by pathogen, or the behaviour of the host. Also malaria has been observed moresuccessfully treated on site in tropics due to semi-immune population, during last 10 years with increasing frequency in areas above sea level early pre-referral administration of antimalarial drugs and early empiric (highlands malaria). The aim of this research was to asses if rainy/dry intramuscular and venous administration of antimalarial drugs; resulting to seasons are related with seasonal variation of infectious diseases similar to 5 % mortality even in more severe cases. Kenya or Uganda highlands (western Kenya and south-western Uganda). Monthly incidences of malaria versus respiratory infections in June to 1519 November (rainy seasons) versus January to April (dray seasons) have been compared in 2012/2013 in community hospital in Gasura located in GENETIC DIVERSITY AND POPULATION STRUCTURE 1283 m.a.s. (Burundi). This hospital had about 50 beds and outpatients OF PLASMODIUM VIVAX INFECTIONS AFTER RADICAL department. Hospital staffs was composed of 2 doctors, 8 nurses, 2 lab TREATMENT IN A RURAL COMMUNITY OF CENTRAL technicians and pharmacist, with a patient flow of 40-120 patients daily VIETNAM in the outpatients department and 2-10 inpatients daily. Malaria diagnosis Nguyen Van Hong1, Christopher Delgado-Ratto2, Pham was made microscopically (according to WHO guidelines) and was Vinh Thanh1, Peter Van Den Eede3, Nguyen Thi Huong Binh1, confirmed with rapid diagnostic test (RDT; according to manufacturer’s Umberto D’Alessandro3, Tran Thanh Duong1, Jean – Pierre Van instructions). Malaria was responsible for approximately 42,5 - 48,8% of Geertruyden2, Annette Erhart3, Anna Rosanas-Urgell3 all admission or consultations in rainy seasons but only 12,5% -29,7% 1 in dry seasons. In dry season, proportion of respiratory tract infections National Institute of Malariology, Parasitology and Entomology, Hanoi, 2 increased from 19% in June to 42% in November and replaced malaria. Vietnam, Faculty of Medicine, University of Antwerp, Antwerp, Belgium, 3 Both malaria and pneumonia showed significant seasonal variations in Institute of Tropical Medicine, Antwerp, Belgium occurrence despite of attitude of community health care centre in Gasura In Vietnam the malaria burden has been drastically reduced over the past (1283 m.a.s.). In Burundi highlands health care centre in Gasura seasonal two decades but Plasmodium vivax is becoming increasingly important variations despite of high attitude (1283 m.a.s.) was observed with mainly due to its relapsing nature and the difficulty to radically cure increasing proportion of malaria from 29,7 to 42% during rainy season dormant parasites from the liver. A two-year cohort study was conducted replaced respiratory tract infection increased from 19 to 42% in dry season in Central Vietnam to assess the efficacy of the radical cure regimen based vice versa. Malaria in the Burundi highlands represented growing problem on a 10-day Primaquine (0.5mg/kg/d) in combination with the standard with variations in prevalence in rainy versus dry seasons. 3-day chloroquine (total 25mg/kg) regimen. We report the genetic diversity and population structure of P. vivax infections before and after radical 1518 treatment. All day 0 (n=247) and post treatment P. vivax infections (n=788) detected by microscopy and PCR during the 2-year monthly follow-up SEVERE MALARIA AMONG 3,707 ADMISSIONS IN SOUTH were genotyped using 16 previously described microsatellites. Genetic SUDANESE HOSPITAL FOR INTERNALLY DISPLACED diversity, linkage disequilibrium, population structure and haplotype POPULATION clustering were analyzed in post treatment samples and compared to Day 0 samples. All markers were highly polymorphic with 3 to 30 alleles 1 1 1 Alexandra Mamova , Miriam Balazova , Lenka Dibusova , Veronika per marker and heterozigosity (He) values ranging from 0.35 to 0.90. 1 1 1 1 Sladeckova , Marian Bartkovjak , Juraj Benca , John Mutuku Muli , Overall He values were not significantly different between day 0 and post- 2 3 2 Lenka Michalikova , Nada Kulkova , Jaroslava Brnova , Gertruda treatment samples (He = 0.64 and 0.66 respectively). In addition, 71.0% 3 1 1 Mikolasova , Maria Raucinova , Jirina Kafkova , Vladimir of all infections were polyclonal (76.9% at day 0 vs. 69.2% post-treatment 4 Krcmery samples) and the average multiplicity of infection (MOI) was 1.9 parasites/ 1St. Francesis of D´Assisi Mission Hospital, St. Elizabeth Tropical Program person (MOI = 2.1 at D0, MOI =1.8 at recurrences). Genetic diversity Marial Lou, Mapuordit, South Sudan, 2Trnava University, Faculty of of parasite population experimented significant changes when parasite Health Sciences and Social Work, Department of Theoretical Disciplines population before treatment was compared with parasite population and Laboratory Investigation Methods, Trnava, Slovakia, 3Laboratory of in the second year follow up (FST= 0.21). which may suggest a delayed Molecular Microbiology, St. Elisabeth University College of Health and effect of the intervention or may reflect the intense follow up (with Social Science, Bratislava, Slovakia, 4St. Elisabeth University College of treatment of all cases) study design. In order to estimate multilocus linkage Health and Social Science, Bratislava, Slovakia disequilibrium (LD) changes between day 0 and post-treatment samples, Severe malaria is responsible for 1.2 million deaths worldwide, 90% of we calculated the index of association IsA (which is zero for LD). We them in Sub-Saharan Africa, mainly in children below 5 years of age. observed higher LD in post-treatment than day 0 parasite population (IsA The aim of this study was to assess proportion of severe malaria among = 0.093, P=0.0001 and IsA = 0.039, P=0.0001, respectively), suggesting all admissions hospital in area with internally displaced population in inbreeding and a clonal population structure. Overall parasite population south Sudan. Data on infectious diseases were analyzed at admission in the study is genetically diverse, and has a low effective recombination within 12 months 1.1. 2013 - 31.12.2013 at St. Francesis of D´Assisi rate that contrasts with the high number of polyclonal infections. mission Hospital located in Marial Lou, South Sudan, built for internally displaced refugees coming from north to South Sudan due to civil war (1982 - 2005) and Darfour Conflict (2002 - 2012)and for about 50 000 Dinka population. Diagnosis of severe malaria was done clinically, plus

astmh.org 464 1520 targeted to foci of infection that do not accurately reflect the true nature of transmission in the community, ensuing interventions may incorrectly SEASONALITY IN MALARIA TRANSMISSION - IMPLICATIONS be perceived as being ineffective. To determine the impact of sample size FOR CASE-MANAGEMENT WITH LONG-ACTING ARTEMISININ and choice of malaria metric, data for 17,500 individuals residing in 3,200 COMBINATION THERAPIES compounds, approximately one third of the population, collected during a cross-sectional survey in the western Kenyan highlands were used to 1 2 2 2 Matt Cairns , Patrick Walker , Lucy Okell , Jamie Griffin , Tini identify foci of transmission in the community. All structures in the area 2 3 3 Garske , Kwaku Poku Asante , Seth Owusu-Agyei , Diadier were digitized to provide a total census of the area and several structures 4 5 6 1 Diallo , Alassane Dicko , Badara Cisse , Brian Greenwood , Daniel can comprise a compound. Model-based geostatistical methods were 1 2 1 Chandramohan , Azra Ghani , Paul Milligan used to analyze the spatial variation of parasite prevalence, as determined 1London School of Hygiene & Tropical Medicine, London, United Kingdom, by polymerase chain reaction (PCR), a measure for current infection, and 2MRC Centre for Outbreak Analysis and Modelling, Imperial College by seropositivity, a measure of malaria exposure. Informative thresholds London, London, United Kingdom, 3Kintampo Health Research Centre, of risk were defined in order to identify foci in the spatial distribution of Kintampo, Ghana, 4London School of Hygiene & Tropical Medicine, now the two outcomes. The impact of the sample size on both the accuracy at PATH- Initiative, Dakar, Senegal, 5Malaria Research of prevalence estimates and the ability of the model to identify foci was and Training Center, Bamako, Mali, 6Universite Cheikh Anta Diop, Dakar, assessed through a simulation study. Preliminary findings suggest that Senegal foci defined by the two outcome measures were measures were only Seasonality in malaria transmission is a key driver of malaria epidemiology, moderately correlated (r=0.43) with only 36% of structures identified and has important implications for the effectiveness of interventions. One by both outcome measures. Among 14 discrete foci identified as having aspect that is not well understood is how seasonality affects the number increased risk by one or both outcomes 6 clusters were identified by both of repeat malaria episodes that occur soon after a previous clinical attack, metrics, although only 3 had good overlap. Five clusters (592 structures) and which might be prevented if a long-acting artemisinin combination were identified based on PCR but missed using seroprevalence, and 3 therapy (LACT) was used to manage the initial episode. We estimated the clusters (1214 structures) were missed using PCR but identified using separate and combined effects of transmission intensity and seasonality on seroprevalence. In terms of sample size, initial findings indicate that the timing and concentration of repeat malaria episodes, using data from halving the sample size would have a minimal impact on model efficiency six cohort studies in West Africa, and then used an individual-based model for generating the predicted surface for both outcome measures. of malaria transmission across sub-Saharan Africa to extrapolate these results across a range of settings. Seasonality was quantified using the 1522 Markham seasonality index (MSI), taking account of areas with bimodal seasonality patterns, and the concentration of malaria episodes in time UTILITY OF PCR-BASED SURVEILLANCE METHODS RELATIVE was quantified using a modified version of the Gini index. We explored TO AGE IN RAINY AND DRY SEASONS IN SOUTHERN 10% intervals of the MSI, and simulated transmission intensity that MALAWI equates to prevalence in 2-10 year olds ranging from 5-60%. In settings Jenny A. Walldorf1, Jenna E. Coalson2, Andy Bauleni3, Kondwani where prevalence is less than 10%, repeat malaria episodes constitute a Nkanaunena3, Clarissa Valim4, Lauren M. Cohee1, Nelson small fraction of the total burden, and few repeat episodes occur within Chimbiya3, Jacqueline Fiore5, Atupele Kapito-Tembo3, Karl Seydel6, the window of protection provided by currently available drugs. However, Don Mathanga3, Terrie E. Taylor6, Miriam K. Laufer1 in higher transmission settings, and particularly in highly seasonal settings, 1 repeat malaria becomes increasingly important, with up to 20% of the University of Maryland Baltimore, Baltimore, MD, United States, 2 total clinical burden in children estimated to be due to repeat episodes University of Michigan School of Public Health, Ann Arbor, MI, United 3 4 within four weeks of a prior attack. At a given level of transmission States, Malaria Alert Center, Blantyre, Malawi, Harvard School of Public 5 intensity and annual incidence, the concentration of repeat malaria Health, Boston, MA, United States, College of Osteopathic Medicine, East 6 episodes in time, and consequently the protection from LACTs, is always Lansing, MI, United States, Blantyre Malaria Project, Blantyre, Malawi highest in the most seasonal areas. As a result, the degree of seasonality, Routine malaria surveillance often relies on microscopic or rapid diagnostic in addition to the overall intensity of transmission, should be considered test (RDT) parasite detection, which misses low density infections. We have by policy makers when deciding between ACTs that differ in terms of the previously shown in cross-sectional surveys that school age children are duration of post-treatment prophylaxis they provide. at high risk of PCR-detectable asymptomatic infections in dry and rainy seasons. Using the same survey data, we now assess risk factors for low 1521 density infection to determine the utility of PCR by age and transmission setting. Submicroscopic infection was defined as Plasmodium falciparum DETECTING FOCI OF MALARIA TRANSMISSION: detected by PCR but with a negative malaria smear read by quality- IMPLICATIONS OF SAMPLE SIZE AND CHOICE OF MALARIA controlled microscopy. Sub-RDT infection was defined as PCR detection METRIC with microscopy negative or <200 parasites per microliter. Among all PCR-detected infections, infections were more likely to be submicroscopic 1 2 3 Gillian H. Stresman , Emanuele Giorgi , Amrish Baidjoe , Philip in the dry (56%,180/319) vs. rainy season (38%, 208/544, p<.0001). The 4 5 5 5 Knight , Wycliffe Odongo , Chrispin Owaga , Shehu Shagari , proportion of infections that were submicroscopic among adults (≥16 5 6 1 Euniah Makori , Jennifer Stevenson , Chris Drakeley , Jonathan years), school age (6-15 years), and young children (≤5 years), was 72% 1 2 1 Cox , Peter Diggle , Teun Bousema (69/96), 55% (93/168), and 33% (18/54) in the dry season and 56% 1London School of Hygiene & Tropical Medicine, London, United Kingdom, (99/178), 30% (81/274), and 30% (27/91) in the rainy season, respectively 2Lancaster University, Lancaster, United Kingdom, 3Radboud University (p<.0001 both seasons). In mixed modeling, relationships between age Nijmegen Medical Centre, Nijmegen, Netherlands, 4University of the West and low density infections varied by season. In the rainy season, adults of England, Bristol, United Kingdom, 5KEMRI/Centers for Disease Control had 3.0 [95% CI: 1.7, 5.2] increased odds of submicroscopic infection, and Prevention, Kisumu, Kenya, 6Johns Hopkins Bloomberg School of while school age children did not differ from young children. In the same Public Health, Macha, Zambia communities in the dry season, both adults (OR 5.3 [2.6, 11.0]) and school Identifying malaria foci in endemic settings can be challenging. Many age children (OR 2.5 [1.3, 5.0]) had increased odds of submicroscopic pragmatic decisions must be made, including the sample size to employ infection relative to young children. Associations were independent of and which malaria metric to use. However, it is not known how these district, net use, house materials, and gender. The relationship between decisions affect the ability to detect foci of infection, particularly the age and sub-RDT infections followed a similar pattern. Microscopy and boundaries of foci identified. If control or elimination programs are RDTs have inconsistent performance across age groups and seasons. While

astmh.org 465 both microscopy and molecular detection have demonstrated the highest transmission study stratified by ecological zone. We followed a cohort malaria burden among school age children, surveillance with microscopy of 2145 participants of all ages(715 per zone, selected by multistage or RDT alone fails to detect most infections among adults and half of the cluster sampling)once a month from April 2011 to March 2012.A history infections in school age children. of fever within the previous 2weeks was elicited at each visit and further information obtained by questionnaire from those with a positive history, 1523 after which a finger-prick blood sample was taken for the preparation of blood smears. Data was analyzed in STATA12.We completed 77% ESTIMATING THE MALARIA ATTACK RATE OF A TANZANIAN of all planned visits;8% of participants reported fever,3% had used an MILITARY COHORT IN THE SEARCH FOR NON-IMMUNE artemisinin combination therapy (ACT) for treatment of perceived fever POPULATIONS FOR MALARIA PROPHYLAXIS, VACCINE AND and 6% had used an insecticide treated bed-net (ITN)the night before TREATMENT STUDIES visits.The incidence of slide confirmed malaria per 1000 person years was 85 in the Forest,41 in the Coastal and 13 in the Lakeside zones.Verified 1 2 3 Charles E. Mwanziva , Shekalaghe Seif , Lucky Temu , Sarah ITN use the night before visits in each of the zones was 3%,4% and 9% 4 5 6 6 Chiduo , George Amoo , David Schnabel , Lalaine Anova , Colin respectively.The absence of a ceiling in a room was associated with an 6 6 7 1 Ohrt , Eyako Wurapa , Akili Kalinga , Christopher Mswanya , excess risk of malaria of 15%. Malaria incidence per 1000 person years 1 1 Yadon M. Kohi , Adam S. Mwambulanga was 119 in those aged 0-4years,136 in those aged 5-9,50 in those aged 1Tanzania People’s Defence Forces, Dar es Salaam, United Republic 10-19, 9 in those aged 20-29,18 in those aged 30-39 and 24 in those of Tanzania, 2Ifakara Health Institute, Bagamoyo, United Republic of over 40 years of age.Overall rates had decreased by 40% from the 1994 Tanzania, 3Tanzania People’s Defence Forces, Walter Reed Program - levels.The Lakeside zone had the lowest incidence despite vast irrigated Tanzania, United Republic of Tanzania, 4Walter Reed Program - Tanzania, fields and the lowest access to ACTs.The Forest zone with the lowest Dar es Salaam, United Republic of Tanzania, 5Amethyst Technologies LLC, verified ITN use and ownership(25%) and highest access to ACTs bore the Baltimore, MD, United States, 6Walter Reed Army Institute of Research, brunt of morbidity.The data suggest that in an area of declining malaria Silver Spring, MD, United States, 7National Institute for Medical Research, transmission,efficient surveillance is required to promptly determine Tukuyu, United Republic of Tanzania levels and patterns of morbidity to identify remaining foci for targeted In Tanzania, malaria is the leading cause of outpatient and inpatient health interventions service attendance, and accounts for about 32% of hospital deaths. The disease is unevenly distributed in the country. Some areas of Tanzania have 1525 little to no malaria transmission as a result of anti-malarial interventions (e.g., use of treated bednets and ACT) as well as differences in climate CONTRIBUTIONS OF WOMEN WITH CHILDREN AND YOUTH and geography, while other areas are highly endemic for malaria. Exposure WORKERS TO SPATIAL MALARIA TRANSMISSION IN SUB- to the malaria parasite gives individuals the ability to develop immunity SAHARAN AFRICA and asymptomatic infection. However, naive populations from low-risk John M. Marshall1, Mahamoudou B. Touré2, André Lin areas have no protective immunity and are at high risk of developing the Ouédraogo3, Micky Ndhlovu4, Samson S. Kiware5, Ashley Rezai1, disease. This study aims at determining the malaria attack rate of proposed Emmy Nkhama4, Jamie T. Griffin1, T. Deirdre Hollingsworth6, non-immune individuals from non-malarious areas when entering a Seydou Doumbia2, Nico Govella5, Neil M. Ferguson1, Azra C. training camp in a highly endemic area. 500 recruits from Tanzania Ghani1 People’s Defence Forces (TPDF) from non-endemic areas were selected 1 2 by multistage random sampling; consenting, eligible participants were Imperial College London, London, United Kingdom, Malaria Research 3 followed for six months. Malaria smears were collected every fortnight and Training Center, Bamako, Mali, Centre National de Recherche et 4 by active and passive detection of infection at the camp health facility. de Formation sur le Paludisme, Ouagadougou, Burkina Faso, Chainama 5 Blood samples for PCR and serological tests were collected. Malaria College of Health Sciences, Lusaka, Zambia, Ifakara Health Institute, 6 diagnosis was confirmed by malaria microscopy. There was a high rate Dar es Salaam, United Republic of Tanzania, Liverpool School of Tropical of study subject follow-up; 98.1% (491/500) individuals participated in Medicine, Liverpool, United Kingdom all activities, while 8 withdrew their participation for personal reasons. Recent investment in global malaria control has led to malaria prevalence 21% (102/491) were terminated after confirmed malaria infection by dropping in many parts of the world. As people play a dominant role in clinical laboratory test (study end point), and one participant died from a parasite dispersal, a quantitative understanding of human movement non-malarial infection. The malaria attack rate was found to be as low as patterns is relevant to determining how best to maintain gains made 18% to above 24%. Plasmodium falciparum was the predominant species through these efforts. We conducted a survey of human movement detected. This study revealed one of very few non-immune populations patterns in four countries throughout sub-Saharan Africa - Mali, Burkina with sufficient malaria exposure to conduct malaria prevention studies of Faso, Zambia and Tanzania - with additional questions on malaria risk a reasonable size, made possible by the heterogenous disease distribution factors and cell phone usage behavior, the latter to enable anonymous cell in the country. TPDF recruits from non-endemic areas may be an ideal non- phone signal data to be better-correlated with movement patterns. A total immune population for future malaria prophylaxis, vaccine and treatment of 4,352 individuals were interviewed and 6,141 trips recorded. A cluster trials. analysis highlighted two distinct traveler groups of relevance to malaria transmission - women traveling with children (all four countries) and youth 1524 workers (Mali). Women with children were predominantly between the ages of 16 and 45 and were more likely to travel to areas of relatively high MALARIA INCIDENCE IN A DISTRICT WITH THREE malaria prevalence in Mali (p<0.001) and Zambia (p=0.035) compared to ECOLOGICAL ZONES IN SOUTHERN GHANA other travelers. They were also more likely to own bed nets in Burkina Faso (p=0.001) and Zambia (p<0.001), to use bed nets in Zambia (p<0.001) and Alberta A. Quartey Tanzania (p=0.046), and to own a cell phone in Mali (p<0.001), Burkina Kwame Nkrumah University of Science and Technology, Kumasi, Ghana Faso (p<0.001) and Zambia (p<0.001). Taking into account that children Information on malaria incidence,age patterns of morbidity,mortality are especially receptive to malaria parasites, women with children were and serological exposure can be combined with entomological data estimated to account for the majority of spatial malaria transmission in to help target enhanced malaria control. Although the Dangme West Mali, Burkina Faso and Zambia. Malian youth workers were predominantly District in Ghana hosts a research centre and has conducted many between the ages of 16 and 29 and were more likely to travel to areas malaria interventions,the last malaria transmission measurement was of relatively high malaria prevalence (p<0.001) and for longer durations undertaken in1994.We have therefore undertaken a new detailed malaria (p<0.001) compared to other travelers. They were estimated to make a

astmh.org 466 significant contribution to spatial malaria transmission in Mali. Knowledge we determine the presence and the prevalence of these sub-species in of the spatial patterns of malaria transmission and the contributions of Mali using molecular analysis of blood blotted onto filter papers. Between key traveler groups to this spread will assist in the design of control and 2011 and 2013, 7044 volunteers were screened by light microscopy surveillance programs targeting “hot spots” of malaria transmission. in the context of various clinical studies conducted in five sites of Mali with different malaria epidemiology (Bougoula-Hameau, Faladje, Kolle, 1526 Pongonon and Sotuba). Thick smears were made and read onsite by experienced microscopists. Genomic DNA was extracted using Qiagen EPIDEMIOLOGY OF MALARIA INFECTION AMONG SCHOOL- kits. First, ssrRNA-based PCR methods detecting the P. ovale specy were AGED CHILDREN IN KINTAMPO NORTH DISTRICT, GHANA: performed. Second, nested PCR of P. ovale tryptophan-rich antigen (potra) AN EVALUATION OF BEHAVIOR, NUTRITIONAL STATUS, gene designed to distinguish the sub-species P. ovale. curtisi and P. ovale. HOOKWORM CO-INFECTION AND ANTIBODY RESPONSES wallikeri were run. Overall, 84/7044 (1.2%) of slides were positive for P. ovale. To date, 483 dried blood spots were analyzed by PCR. ssrRNA 1 1 2 Jennifer Du Mond , Benjamin T. Simms , Dylan Davey , Joseph analysis revealed 12 (2.5%) cases of P. ovale. Potra analysis showed that 3 1 3 Otchere , Debbie Humphries , Josephine Quagraine , Samuel 6/12 (50%) were P. ovale curtisi, 5/12 (41.7%) were P. ovale wallikeri and 4 2 3 2 Newton , Lisa Harrison , Daniel Boakye , Michael Wilson , Michael 1 sample was not sub-typable. We show the two recently described P. 2 Cappello ovale sub-species were both present in Mali. 1Yale University School of Public Health, New Haven, CT, United States, 2Yale University School of Medicine, New Haven, CT, United States, 1528 3Noguchi Memorial Institute for Medical Research, Accra, Ghana, 4Kintampo Health Research Center, Brong Ahafo, Ghana USING DHIS2 FOR ROUTINE MONITORING OF QUALITY OF A cross-sectional study was conducted in June 2010 in the Kintampo HEALTH SERVICES IN THE PRIVATE SECTOR North Municipality, Ghana. Children (n=286) from 16 schools were Purity Njagi1, Martin Dale1, Victor Lara1, Preston Izulla2, enrolled after screening (n=844) if they had HAZ Z-score below -1.80 or Stephanie Dolan1, Nikki Charman1 above -0.10, with no more than one child from each household. Serum, 1 2 fecal samples, and household surveys were used to assess the associations Population Services International, Nairobi, Kenya, PS Kenya, Nairobi, between the presence of malaria parasites, malaria parasitemia, and Kenya individual and household risk factors including nutritional status, District Health Information System (DHIS2) has increasingly become hookworm co-infection, household risk prevention behaviors, and serum the preferred Health information system for effective management of measures of parasite-specific immunoglobulin G (IgG). The primary factors health data in many countries, mostly for collection and management associated with reduced risk of malaria infection included spraying the of routine service data at various levels of the health sector. However, house in the past year (OR=0.04, p<0.001), the child having a health DHIS2 capabilities can also be optimised for routine monitoring of quality care visit in the past year (OR=0.39, p<0.001), household malaria in of health services in the private sector: Population Services International the past year (OR=0.37, p=0.001), higher hookworm antibody levels, (PSI) is implementing a multi-country project aiming to create a market and geographic location, while greater household food insecurity was for malaria Rapid Diagnostic Tests (mRDTs) to improve quality of care associated with reduced risk of high levels of parasitemia. Primary risk in the private health sector by enabling effective treatment based on factors for elevated parasite density included the house being sprayed diagnosis. This project is being implemented by PSI in Kenya, Madagascar in the past year (OR=9.83, p<0.001), household bednet usage (higher and Tanzania. The use of DHIS2 for the purpose of tracking provider proportion of use associated with greater parasitemia), household and level case management will be presented, with specific focus the power child history of malaria in the past year (OR=2.80, p=0.039; OR=0.15, of DHIS2 to convert routine data into decision-making. DHIS2-enabled p<0.001, respectively), frequency of consumption of protein-rich food tablets can be used to (i) undertake provider quality of care and service groups, and geographic location, while those with the highest hookworm preparedness assessments, (ii) automatically score and benchmark the antibody levels showed reduced parasite density. Hookworm infection provider’s performance on site, and (iii) provide effective on the spot was associated with increased risk of malaria infection (OR=2.65, p=.10) feedback for continuous improvement. The power of DHIS2 dashboards and higher density of malaria parasites among those infected (OR=2.81, to manage and give feedback to providers will be highlighted, bridging p=0.001). These risk factors highlight areas of programmatic interest, information on provider quality of service, productivity based on caseloads, particularly the elevated risks of malaria infection and higher density of and behaviour change based on the adoption stairway. In addition, an parasites among those infected with hookworm. Further research should innovative adaptation of DHIS2 to allow effective allocation of resources elucidate the mechanism of this interaction, and treatment measures through automated supervision planning, taking into account provider should focus on reducing the burden of hookworm in malaria endemic quality of service benchmarks will be discussed (frequency and scheduling areas. of provider assessments in particular). The presentation will further demonstrate how DHIS2 can enable program managers to effectively track 1527 service provision and make informed decisions leading to program quality improvement through the use of tailored dashboards. Potential future links PREVALENCE OF TWO NEWLY RECOGNIZED HUMAN with national systems will also be discussed, given the widespread use of MALARIA SPECIES IN MALI: PLASMODIUM OVALE CURTISI DHIS2 by governments in the countries we work. AND P. OVALE WALLIKERI 1529 Nouhoum Diallo1, Sangare P. Cheick1, Antoine Dara1, Aboubacrine Haidara1, Bakari Fofana1, Bakari Sidibe1, Souleymane USE OF SATELLITE IMAGERY TO ESTABLISH A SAMPLING Dama1, Omar B. Traore1, Aliou Traore1, Sekou Toure1, Amadou FRAME AND MEASURE HOUSEHOLD MOVEMENT IN Togo1, Niawalou Dara1, Moctar Coulibaly1, Issaka Sagara1, Khalid SOUTHERN ZAMBIA BETWEEN 2007 AND 2011 Bechir2, Colin Sutherland2, Ogobara K. Doumbo1, Abdoulaye Djimde1 Timothy M. Shields1, Jessie Pinchoff1, William Moss1, Jailos 2 2 1 1University of Sciences, Techniques and Technologies of Bamako, Bamako, Lubinda , Harry Hamapumbu , Frank Curriero Mali, 2London School of Hygiene & Tropical Medicine, London, United 1Johns Hopkins School of Public Health, Baltimore, MD, United States, Kingdom 2Johns Hopkins School of Public Health, Macha, Zambia Ovale malaria is caused by two sympatric Plasmodium ovale sub-species; High-resolution satellite imagery can be used to establish a sampling their biology and morbidity need to be investigated. In the present study frame for epidemiologic research and to describe patterns of household

astmh.org 467 distribution and movement. Assessing the frequency and geographic 1531 distribution of household movement by comparing satellite images taken over time may suggest a time period for satellite image accuracy MALARIA ATTRIBUTABLE FRACTION TO FEVER IN A COHORT and utility for epidemiological research. All households in a 575 km2 OF PAPUA NEW GUINEAN CHILDREN region of southern Zambia were enumerated based on satellite images 1 2 2 taken in 2007 and in 2011. Movement of households in the study area Anna Rosanas-Urgell , Inoni Betuela , Benson Kiniboro , Danielle 2 2 2 2 was assessed by comparing the images to calculate the percentage of I. Stanisic , Lorna Samol , Leanne J. Robinson , Peter Siba , Ivo 3 households that were built, removed or stayed the same. We created a Mueller spatial intensity map to identify geographic areas of household movement, 1Institute of Tropical Medicine, Antwerp, Belgium, 2Papua New Guinea and to describe the spatial variation in household movement. There were Institute of Medical Research, Madang, Papua New Guinea, 3Walter and a total of 3,287 household enumerated in 2007 and 3,721 in 2011. 970 Eliza Hall Institute, Parkville, Australia households were newly observed in 2011 and 536 were no longer present. In areas of high malaria endemicity, a high proportion of individuals is Reporting a net change of 434 households occurring over the four year asymptomatic, thus are infected with malaria parasites without developing period does not adequately describe the population movement within clinical symptoms. In addition, those who develop symptoms in the this region. Spatial variation around key features, such as around the new presence of malaria parasites, may be ill due to non-malaria diseases. sealed road, points to non-uniform dynamics in population movement. Therefore, a case definition able to distinguish between malaria and non- These population dynamics may have implications for field studies working malaria morbidity is essential to correctly estimate the burden of disease, in this area over this time period. design adequate control strategies -including case management-, and measure the effect of malaria interventions. In a cohort of 500 Papua 1530 New Guinean children aged 1 to 5 years and followed up for 9 months, logistic regression methods were used to estimate the risk of fever by REDEFINING THE URBAN-RURAL CONTINUUM FOR MALARIA parasite density above specific cut-offs. Plasmodium falciparum and P. RISK: NEW APPROACHES TO CHARACTERIZING PATTERNS IN vivax attributable fraction (AF) and population attributable fraction (PAF) MALAWI to fever were calculated from odds ratio estimates for each case definition. Sensitivity and specificity of case definitions were also evaluated. P. Peter S. Larson1, Joseph N. Eisenberg1, Carl H. Campbell2, Don P. falciparum AF ranges from 79% to 94% when all parasite densities and Mathanga3, Mark L. Wilson1 densities higher than 50000 P/µL are used as cut-off values, respectively. 1 University of Michigan School of Public Health, Ann Arbor, MI, United Overall, the PAF of fever to P. falciparum was 16% when all parasite 2 3 States, University of Georgia, Athens, GA, United States, University of densities in the presence of fever were considered. On the other hand, P. Malawi College of Medicine, Blantyre, Malawi vivax AF increased from 9% (all parasite densities) to 85% when parasites Urban and rural setting is often included in the analysis of infectious densities higher than 10000 P/µL were used. P. vivax PAF exhibit the disease risk, as it may be an important determinant of pathogen highest values (5-7%) when a cut-off of >1500 P/µL was used. Estimates transmission. This dichotomous designation, however, is politically defined, of the sensitivity and specificity of case definitions cut-off by parasite and masks the true biological and social causal determinants of risk. To density show that a low P. falciparum cut-off (<2500 P/µL) achieves high improve understanding of disease patterns and targeting of interventions, sensitivity (80-100%), while when only using high parasite density cut-off transmission-specific characterization of the urban-rural continuum is values high specificity (90%) is obtained. 80% sensitivity and specificity needed. We developed and tested a composite measure for malaria in is achieved with P. vivax cut-off value > 10000 P/µL.Approximately 80% Malawi that included features such as health facilities, roads, rivers, lakes to 94% of fevers with P. falciparum infections occurring in Papua New and electricity, along with census-derived population density. Analysis was Guinean children aged 1 to 5 years are attributable to malaria regardless based on a household-level survey of infection and various risk factors of parasite densities. On the other hand, only 9% of fevers occurring in among 6-59 month old children. Geographic distances to features were the presence of P. vivax infections, at any parasite density, are attributable estimated and a principal component analysis (PCA)-based composite to malaria, suggesting that clinical tolerance against low density P. vivax measure for each household was produced for all points on a fine-scale infections is already acquired at this young age. grid throughout Malawi. Statistical relationships of all factors were tested against Plasmodium parasitemia status using multivariate regression 1532 based methods, including potential household-level confounding factors such as treated net use and material wealth. Urban-like and rural-like PRIVATE SECTOR READINESS FOR MALARIA CASE areas existed throughout Malawi, even within areas classified as ”urban” MANAGEMENT AND MALARIA MARKET COMPOSITION and ”rural” by the Malawi Government. Individual factors associated BEFORE AND AFTER THE AFFORDABLE FACILITY - with urban and rural divides, including proximity to health services and MALARIA (AMFM): RESULTS FROM THREE PILOT COUNTRIES roads, as well as population density, were associated with Plasmodium 1 2 2 2 infection. Community-level factors associated with human settlements Megan Littrell , Julius Njogu , Raymond Sudoi , Kevin Duff , 3 4 5 2 and urban development were predictive of decreased malaria risk, even Hana Bilak , Vamsi Vasireddy , Kate O’Connell , Stephen Poyer in the presence of more traditional household-level prevention methods 1Population Services International, Washington, DC, United States, such as ITN use. Infection probability was similar for most “rural” areas, 2Population Services International, Nairobi, Kenya, 3Population Services but declined linearly after a breakpoint with increasing urbanicity, as International, Chevry, France, 4Independent Consultant, Chicago, IL, United measured by the PCA based composite. Dichotomized measures of States, 5Independent Consultant, Yangon, Myanmar urban and rural spaces fail to adequately characterize environments People living in malaria endemic countries often turn to the private which might be associated with risks for infectious disease transmission. sector for fever case management. However, private sector markets are Politically determined destinations may ignore “urbanized” pockets within typically characterized by low levels of readiness for appropriate malaria traditionally “rural” areas, and rural-like spaces within areas classified as case management, particularly in comparison with public health facilities. “urban.” A composite measure which analyzes many factors associated to Efforts to improve this have included the Affordable Medicines Facility - varying degrees with spaces roughly defined as rural and urban presents malaria (AMFm), which aimed to improve availability and affordability of an opportunity to refine places of disease risk. quality assured Artemisinin Combination Therapy (ACT) in both public and private sectors. The pilot demonstrated favorable improvements in private sector readiness for malaria case management in most countries. The private sector comprises a diverse set of actors, including regulated and health facilities, and unregulated drug shops and general

astmh.org 468 retailers. We examine the level of private sector readiness post-AFMm, 1534 and compare private sector market composition before, during, and after the pilot in Madagascar, Nigeria and Uganda. Trends in key readiness EFFECTS OF AGE AND CONTROL INTERVENTIONS indicators are examined, including availability of blood testing and ACT, ON PREVALENCE OF PLASMODIUM FALCIPARUM and provider knowledge. Favorable trends in private sector readiness may GAMETOCYTEMIA DURING PEAK MALARIA TRANSMISSION be driven by improvements across existing market actors, or may be an SEASON IN WESTERN KENYA indirect effect of shifting market composition towards a market dominated by regulated market actors. In the context of varying degrees of Zhiyong Zhou1, Rebecca M. Mitchell1, Simon Kariuki2, improvement, we examine the private sector malaria market composition Christopher Odero2, Peter Otieno2, Kephas Otieno2, Vincent over time. Multiple nationally representative outlet surveys were conducted Were2, Mary Hamel1, John Gimnig1, Edward D. Walker3, Meghna between 2009 and 2013 by the ACTwatch project in the 3 countries. Desai1, Ya Ping Shi1 During each survey, a census of all outlets with the potential to sell 1Malaria Branch, Division of Parasitic Diseases and Malaria, Centers for antimalarials was conducted, allowing examination of relative market Disease Control and Prevention, Atlanta, GA, United States, 2Kenya composition and antimalarial market share between and within sectors. Medical Research Institute, Kisumu, Kenya, 3Department of Microbiology Private sector readiness improved over time in all 3 countries although to and Molecular Genetics, East Lansing, MI, United States varying degrees across contexts and measures of readiness. We examine Gametocytes are the sexual stage of Plasmodium parasites responsible for readiness trends over time in relation to shifts in market composition, and malaria transmission from human to mosquito host. However, risk factors discuss implications for improving private sector readiness for malaria case for gametocytemia such as age and ameliorating effects of interventions management. (antimalarial and ITN use) are poorly understood. We measured the prevalence and density of P. falciparum gametocytes by pfs25 gametocyte 1533 mature stage V marker using real-time quantitative nucleic acid sequence- based amplification assay (QT-NASBA) and by pfg377 female gametocyte TRENDS IN AVAILABILITY OF MALARIA MEDICINES AND marker using qRT-PCR on samples from a cross sectional survey conducted DIAGNOSTICS IN KINSHASA, DR CONGO FROM 2009-2013 at peak transmission season in 2012 in Siaya County of western Kenya. Julie Archer1, Godéfroid Mpanya2, Stephen Poyer1, Antoinette We also measured parasitemia using 18S QT-NASBA. Total of 446 samples Tshefu3, Willy Onema2, Jamie Ciesla2, Joris Losimba Likwela4, (225 malaria smear-positive and 221 randomly selected smear-negative) Vamsi Vasireddy5, Megan Littrell6 from 832 individuals were used to determine gametocyte carriage. Overall, 18S-NASBA detected 354 positives, of which 129 infections were from 1Population Services International, Nairobi, Kenya, 2Association de Santé smear-negative individuals. Pfs25 was detected in 78.7% of smear positive Familiale/PSI, Kinshasa, Democratic Republic of the Congo, 3Ecole de Santé and 10.9% of smear negative samples while pfg377 was found in 55.1% Public de Kinshasa, Kinshasa, Democratic Republic of the Congo, 4National of smear positive and 2.7% of smear negative samples. In multivariable Malarial Control Program, Kinshasa, Democratic Republic of the Congo, analysis, children (<5 and 5-15 years old) were more likely positive with 5Independent Consultant, Chicago, IL, United States, 6Population Services pfs25 and pfg377 than adults >15 years old (pfs25: OR 3.4, CI 2.0-6.0 International, Washington, DC, United States and OR 4.0, CI 2.1-7.7; pfg377: OR 15.3, CI 6.3-37.1 and OR 7.6, CI 3.0- Malaria is a leading cause of illness and death in the Democratic Republic 19.3, respectively). Children <5 years were more likely pfg377 positive of the Congo (DRC) where the malaria disease burden is estimated to than children 5-15 years old (OR 2.0, CI 1.2-3.4); however, gametocyte be the second highest in the world. Malaria is endemic throughout the density detected by pfg377 or pfs25 did not differ between these two age country, including in the large urban agglomeration of Kinshasa. National groups of children. Anemia (Hb < 11 g/dl) was associated with higher 18S policy has recommended diagnostic testing since 2006, and the national density (1.45x per log10, CI 1.18-1.80, p=0.0005). Importantly, anemia treatment protocol includes two artemisinin-based combination therapies was also associated with pfs25 and pfg377 positive status (OR 2.1, CI (ACT) for first-line treatment. However, according to past surveys, access 1.4-3.4 and OR 2.4, CI 1.4-3.9). The odds of being pfs25 positive were to and use of malaria diagnostics and ACT remains low. A 2009 survey lower in individuals using ITNs (OR 0.41, CI 0.23-0.71). No differences conducted by ACTwatch found that 60% of antimalarial-stocking public were seen for pfs25 density between individuals with and without ITN and private sector outlets in Kinshasa had ACT in stock, but a 2010 use. Antimalarial use (90% artemether-lumefantrine) during the two ACTwatch household survey found that just 15% of children with fever weeks prior to the survey was associated with fewer pfs25 carriers (OR had received a diagnostic test for malaria and only 4.5% of those with 0.32, CI 0.17-0.62), but not with pfs25 density. These results show that fever had received an ACT. In a late 2013 follow-up to the 2009 survey, children provided the highest gametocyte reservoir compared to adults ACTwatch conducted a representative cross-sectional survey of 3,654 and anemia was associated with an increased risk of gametocytemia. ITN public and private sector health facilities and retail outlets in Kinshasa and antimalarial use decreased the gametocyte prevalence, but not the to assess the availability, price and market share of antimalarials and gametocyte density in the study population. malaria diagnostics. Findings from this survey and a trend analysis will be presented. Preliminary results show that availability of malaria diagnostics 1535 remains low and the antimalarial market is still dominated by quinine in the private sector. Results will be examined in the context of qualitative IDENTIFYING FACTORS ASSOCIATED WITH MALARIA research findings from in-depth interviews conducted in 2014 in Kinshasa PARASITEMIA IN MOZAMBIQUE USING A GEOSTATISTICAL with private doctors, pharmacists, drug shop owners and clients. There are MODEL renewed efforts in DRC to increase the quality of fever case management following World Health Organization recommendations calling for James Colborn1, Kathryn Colborn2, Edward Wenger2, Guidion universal parasitological confirmation before treatment. This survey of the Mathe3, Graca Matsinhe3 landscape of availability of ACT and malaria diagnostics, as well as ACT 1Centers for Disease Control and Prevention, Maputo, Mozambique, market share will inform renewed case management efforts in Kinshasa 2Intellectual Ventures, Seattle, WA, United States, 3National Malaria that can then be scaled up throughout the country. Control Program, Maputo, Mozambique Malaria is among the largest contributors to child mortality in Mozambique. Donors such as the Global Fund and the US President’s Malaria Initiative have been successful in controlling malaria by supporting national scale-up of interventions, though transmission remains high in many parts of the country. Malaria Indicator Surveys (MIS) and Demographic Health Surveys (DHS) are intermittent household surveys

astmh.org 469 aimed at collecting nationally representative demographic and health 1537 related data, including socio-economic status (SES), insecticide treated net (ITN) ownership and use, coverage with indoor residual spraying (IRS), and COLLABORATIVE DATA MANAGEMENT FRAMEWORK FOR malaria parasitemia prevalence by rapid diagnostic test (RDT) for children BORDER MALARIA RESEARCH IN SOUTHEAST ASIA ICEMR 6-59 months of age. When examined independently, these surveys may CENTER not capture annual variation in malaria transmission. To overcome this, 1 2 3 1 we combined both the 2007 MIS and the 2011 DHS data to capture Ming-Chieh Lee , Ying Wang , Kirakorn Kiattibutr , Guofa Zhou , 4 3 5 the inherent spatial and temporal variation in environmental factors and Jeeraphat Sirichaisinthop , Jetsumon Prachumsri , Liwang Cui , 1 malaria transmission. Using these data, we estimated the associations Guiyun Yan between relevant factors and interventions and cluster-level parasitemia 1University of California Irvine, Irvine, CA, United States, 2Third Military prevalence using a geostatistical model. This model included a fixed Medical University, Chongqing, China, 3Faculty of Tropical Medicine, factor for survey year, IRS coverage, ITN use, age, SES (wealth quintiles), Mahidol University, Bangkok, Thailand, 4Vector Borne Disease Training environmental factors, and a spatial random effect. Our results suggested Center, Saraburi, Thailand, 5Pennsylvania State University, University Park, that higher SES (OR=0.79; 95% CI 0.73, 0.85) and coverage of IRS up PA, United States to 40% (OR=0.93; 95% CI 0.88, 1.00) were associated with decreased Document and data archiving for the research activities are the tedious odds of parasitemia, and higher monthly average enhanced vegetation but important tasks through the whole scientific research project. It is a index (increased greenness; OR=5.10; 95% CI 1.83, 16.91) was associated challenge for the scientist to maintain a solid, flawless and well organized with increased odds of parasitemia; ITN use was not associated with data structure and data entry procedures especially for the project included parasitemia, although this could be related to the low levels of ITN use international collaboration within multi-cultures and languages. Southeast in both survey years (cluster medians=0% and 33%, respectively). These Asia ICEMR center is focused on bolder malaria transmission between findings suggest that socioeconomic status and vegetation are important several Southeast Asia countries. We collaborated with several Universities, factors to consider for understanding malaria transmission. Additionally, government agencies, local hospital and clinics, and field research labs the lack of evidence using this analytic method for additional protection by in the remote border area of China, Myanmar, and Thailand. How to IRS coverage above 40% bears further investigation. standardize the field survey and data entry procedures and semi-real-time to share research results is the goal for data manager and coordinators to 1536 conquer. Therefore, we design multi-language survey forms and data entry system as the tools to provide consistent interface and user experience for ASSESSING THE IMPACT OF CLINICAL MENTORSHIP ON all research staffs. In order to expedite the data sharing and secured data MALARIA DIAGNOSIS AND TREATMENT PRACTICES IN services, we utilized several open source applications and cloud computing UGANDA services to sustain our database system. To minimize the catastrophe of Deepa K. Pindolia1, Kathleen Maloney1, Alex Ogwal2, Chelsea computer hardware/software collapse or network traffic congestion, the Solmo1, Vicky Abenakyo2, Albert P. Okui3, Anthony Mbonye3, failover and load balancing features were setup in main server and several Bosco B. Agaba3, Lindsay Bryson1, Caitlin Dolkart1, Justin M. mirror servers and offsite remote backup scheme were implemented. To Cohen1, Arnaud Le Menach1 maintain the maximum data quality and structure, the standard operating procedures (SOPs) for field survey, data entry, data QA/QC were developed 1 2 Clinton Health Access Initiative, Boston, MA, United States, Clinton and documented. Currently the database system is online since July 2012. Health Access Initiative, Kampala, Uganda, 3Ministry of Health, Kampala, Uganda 1538 To address problems associated with malaria misdiagnosis and inappropriate treatment, Uganda has implemented nationwide case MALARIA PREVALENCE AND SPOROZOITIC INDEX IN management trainings, and procured Rapid Diagnostic Tests (RDTs) SUBURBAN AREA IN KINSHASA BEFORE ITN MASS widely in the public sector since 2012. Despite these efforts, only 60% DISTRIBUTION, DR CONGO of suspected malaria cases were tested and 35% of negative tests were provided with anti-malarials in early 2014. In this study, a clinical Aimée Kahindo, Betricia Mavuzi, Freddy Kabasele, Serge M. mentorship for health workers, as a potential tool to overcome barriers Kayembe, Tommy M. Nseka, Solange E. Umesumbu, Thierry L. of testing and adherence, and to improve overall fever case management Bobanga is evaluated. This is a cluster randomized control trial, with one control University of Kinshasa, Kinshasa, Democratic Republic of the Congo and two intervention arms: one with district-selected peer mentors and Malaria is a parasitic disease due to Plasmodium transmitted by a female one with facility in-charges as mentors. Mentorships occur on a monthly mosquito of genus. It is a public health problem which causes basis over 6 months, starting in April 2014 in 150 public health facilities a high mortality in less 5 years children. Estimated deaths by malaria word distributed in 17 pilot medium endemicity districts. Diagnosis and report are around 60000. This study interest is to update data on malaria treatment patient-level data are collected at monthly intervals in all study prevalence in Kinshasa suburb area, in order to provide data for the facilities. Multivariate logistic regressions will be performed to assess the stratification and control of the disease. The objectives of the study were impact of clinical mentorship on confirmatory diagnosis and on adherence to assess malaria prevalence into sub urban Kinshasa area population, to to test results adjusted for covariates such as age, health facility level determine the most concerned population group, to identify plasmodial and district locations. At baseline, there were no significant differences species and to determine sporozoitic index. An analytical cross-sectional in clinical diagnosis and adherence between any of study arms. Over 6 survey was conducted in the village of LUZIZILA to 329 people whose months, it is expected that clinical mentorship improves confirmatory age ranged between 6 months and 76 years for the period from August diagnosis and adherence to test results in both intervention arms, and that 10 to 25, 2013 Blood sample for a thick, a thin smear was conducted differences of at least 10% are identified pre and post intervention and to determine the prevalence of malaria and determining plasmodial between each intervention arm and the control group. Clinical mentorship species. The sporozoitic index was determined by ELISA Results The overall can work as an effective method to increase confirmatory diagnosis and prevalence of positive thick smears was 53.2%. In the age group under adherence to test results, and improve overall fever case management, 5 years, prevalence was higher with 68.8%. Between 6 and 15 years it providing countries with a novel and effective tool to meet their national was 55% beyond 15 years the prevalence stood at 37.3%. P. falciparum malaria case management targets. was found in 97.7% of cases in thin smear slides The sporozoitic index was 11%. In conclusion, the prevalence found place Luzizila in an hyperendemic area. After the distribution we can expect a reduction of these indices

astmh.org 470 1539 out to the risk of transfusional malaria, even in areas of low endemicity, since asymptomatic donors could be accepted based on clinical screening. MALARIA INFECTION IS ASSOCIATED WITH PREGNANCY Moreover, the lack of knowledge about this silent malaria outside the LOSS IN OUELESSEBOUGOU, MALI Amazon Region increases the risk of transmission. The use of platforms with different approaches could minimize this invisible risk. Almahamoudou Mahamar1, Naissem Andemel2, Youssoufa Sidibe1, Adama Dembele1, Moussa Traore1, Oumar Attaher1, 1541 Moctar Tounkara1, Djibrilla Issiaka1, Kadidia B. Cisse1, Bakary S. Diarra1, Moussa B. Kanoute1, Amadou Barry1, Patrick E. Duffy2, SUB PATENT INFECTION OF PLASMODIUM FALCIPARUM IN 1 2 Alassane Dicko , Michal Fried NORTHWESTERN PERU 1Malaria Research and Training Center, Faculty of Medicine, and 1 2 , University of Sciences Techniques and Technologies of Bamako, Juan Jose Contreras-Mancilla , Dionicia Gamboa , Angel Rosas- 1 1 3 Bamako, Mali, 2Laboratory of Malaria Immunology and Vaccinology, Aguirre , Alejandro Llanos-Cuentas , Socrates Herrera National Institute of Allergy and Infectious Diseases, National Institutes of 1Instituto de Medicina Tropical Alexander von Humboldt, Lima, Peru, Health, Rockville, MD, United States 2Instituto de Medicina Tropical Alexander von Humboldt, Departamento In malaria endemic areas, pregnant women are more susceptible to de Ciencias Celulares y Moleculares, Facultad de Ciencias, Universidad 3 malaria infection compared to their non-pregnant counterparts. While Peruana Cayetano Heredia, Lima, Peru, Latin American Center for Malaria the relationships between pregnancy malaria (PM) and outcomes such as Research, Cali, Colombia severe maternal anemia and low birth weight are well established, there The North-western region in Peru, is an area categorized as low endemicity have been limited studies on the relationship between PM and pregnancy for malaria after the El Niño Southern Oscillation (ENSO) phenomenon loss particularly in areas of high malaria transmission. We evaluated the that increased the number of malaria cases to a peak with more than 200 relationship of fetal loss to malaria infection among pregnant women in 000 cases in 1998 for both species, Plasmodium vivax and P. falciparum. Ouelessebougou Mali from November 2010 to January 2014. Pregnant The new treatment scheme implement in 2000 in this area decreased the women were enrolled during the antenatal consultation visits and number of P. falciparum cases in this area and since 2008, no cases have followed up to delivery. Malaria infection in peripheral blood was detected been reported by Ministry of Health. In Peru, microscopy is the diagnostic by bloodsmear, and submicroscopic infection by PCR when the BS was test used as routine by the Ministry of Health; but there are other technics negative. The proportion of women with submicroscopic malaria infection as the Polymerase Chain Reaction (PCR), which is more sensitive to detect at delivery was 25.5% and pregnancy loss occurred in 5.8% of the cohort and identified correctly the species of Plasmodium, under the microscopy (80/ 1,377). Submicroscopic infection at delivery was associated with detection limit. In June of 2013, a total of 750 individuals from 3 urban increased odds of fetal loss (unadjusted OR = 3.26, 95% confidence areas were enrolled in a surveillance study in Piura, a malaria endemic interval (CI) 1.35 - 7.89; and adjusted OR = 3.35, 1.37 - 8.16). A recent region North-western of Peru: 350 from Bellavista, 329 from Obrero and positive bloodsmear also increased the odds of fetal loss. In summary, 71 from Querecotillo. From each individual, a blood sample was taken preliminary analysis indicates that a submicroscopic malaria infection is to prepare 2 slides for microscopy and a filter paper for PCR diagnostic. associated with four times increase in odds of the pregnancy loss. Microscopy diagnosis was performed twice, one a local level and the second one by an expert microscopist as quality control. The DNA 1540 extraction from the filter paper was done by the Chelex-100 method and the PCR was based in a Real time protocol using specific probes to detect ASYMPTOMATIC INFECTIONS AND MALARIA TRANSMITTED P. falciparum and/or P. vivax. No malaria cases was detected by microscopy; BY BLOOD TRANSFUSION: AN INVISIBLE RISK but PCR detected two positive cases for P. falciparum only, one case was located in Obrero and the other one in Bellavista. The parasitaemia level 1 1 2 1 Giselle F. Lima , José E. Levi , Maria E. Carvalho , Juliana Inoue , in both cases was lower than 450 parasites/µL and no symptom was 1 2 1 Angélica D. Hristov , Luis G. Nascimento , Arianni R. Sanchez , present at moment when the sample was taken. These results showed the 1 2 Eduardo M. Ramos-Sanchez , Maria J. Costa-Nascimento , Silvia presences of P. falciparum in the North-western region of Peru and stress 1 1 M. Di Santi , Maria Carmen A. Sanchez out the need to implement more sensitive tools for malaria diagnostic in 1São Paulo University, São Paulo, Brazil, 2SUCEN, São Paulo, Brazil areas of low endemicity where microscopy cannot detect if the country Transfusion transmitted malaria represents a major challenge, essentially aims to improve control measures looking into malaria elimination. due to the occurrence of asymptomatic infections. The vector transmission in Brazil mainly occurs in the Amazon Region, where 166,864 cases 1542 were notified in 2013. Outside the endemic region sporadic cases of autochthonous malaria are reported, including asymptomatic carriers of IMPLICATIONS AND EFFECTS OF DIVERSE PLASMODIUM Plasmodium. In the state of São Paulo, transfusional cases were detected, VIVAX RELAPSE DISTRIBUTIONS IN SIMULATIONS OF due to asymptomatic donors harboring P. malariae, one of them leading to VARYING TRANSMISSION SETTINGS the death of the recipient. The occurrence of parasitemia without clinical Philip A. Eckhoff, Caitlin A. Bever, Jaline Gerardin, Edward A. symptoms in addition to the fact that Plasmodium can survive in stored Wenger red blood cells between 2 and 6º C for up to three weeks, increases the risk of transmission. In order to minimize the possibility of transfusional Institute for Disease Modeling, Bellevue, WA, United States cases, the use of platforms including molecular and serological tests might Control and elimination of Plasmodium vivax is complicated by recurring point out donors suspected of harboring Plasmodium. We tested samples relapses from hypnozoites in the liver of infected individuals. Different from 56 candidates for blood donation living in an area of São Paulo State strains of vivax exhibit different patterns of relapse, ranging from the where asymptomatic infections are reported. Thick blood smear, PCR, Chesson strain with an initial infection followed by early relapses, to ELISA with recombinant P. vivax MSP119 antigen and SD Bioline Malaria strains with mixes of short and longer relapses, to North Korean strains Pf/Pv immunochromatographic test were used. Two samples (3.5% ) (0.98 that have infrequent early infections and longer latencies to relapse. -12.1) were positive by thick blood film for Plasmodium, in a very low Which strains predominate in a given geographic region depends on the parasitemia. One of them was also positive by PCR, indicating the presence local transmission setting, and earlier observational and modeling studies of P. malariae. ELISA detected 53.6% (40.7- 65.9) of samples reagent for by various groups have allowed classification of different malaria zones. P. vivax, with Reactivity Index ≥1.0. SD Bioline detected antibodies against We present a new model for P. vivax transmission, host interactions, and P. vivax MSP and CSP recombinant antigens in 48.2% (35.6 - 60.9) of the relapse distributions and incorporate it into the EMOD model for malaria samples. The frequency of positive samples in the serological tests pointed transmission. The broad diversity of relapse patterns is recreated with

astmh.org 471 a simple set of biological and immunological mechanisms, providing a 1545 mechanistic mathematical framework for comparing different strains. The fitness and population-level effects of different relapse patterns are then ANTIBODIES TO PLASMODIUM VIVAX MSP1-19 explored for a variety of transmission settings, with reference to earlier RECOMBINANT ANTIGEN IN BLOOD DONORS FROM work by others on classification of malaria zones. Finally, implications BRAZILIAN LOW ENDEMIC AREAS of different relapse patterns for control and elimination efforts scaling 1 1 up in the Solomon Islands and other settings are explored in simulation. Arianni R. Sanchez , Eduardo M. Ramos-Sanchez , Silvano 2 1 1 Interventions simulated include primaquine and chloroquine combinations Wendel , Silvia M. Di Santi , Hiro Goto , Maria Carmen A. 1 and vector control. Sanchez 1São Paulo University, São Paulo, Brazil, 2Sírio Libanês Hospital, São Paulo, 1543 Brazil In non-endemic and low endemic areas, transfusion-transmitted malaria NONINVASIVE SURVEILLANCE OF ZOONOTIC MALARIA (TTM) is a rarely reported event. However, four TTM were detected in Josephine Siregar1, Christina Faust2, Lydia Murdiyarso1, Andrew São Paulo State, in Southeastern Brazil, including one death. Infected Dobson2 donors were identified as asymptomatic carriers with displacements to the Atlantic forest biome in São Paulo State. Due to the immune status of 1Eijkman Institute of Molecular Biology, Jakarta, Indonesia, 2Princeton these donors, the parasite densities are low and undetectable in the thick University, Princeton, NJ, United States blood smear or rapid diagnostic tests requiring the use of other methods The fifth human malaria parasite, Plasmodium knowlesi, is a novel for detection in blood banks outside the endemic areas. In this study, since public health threat in Southeast Asia. The parasite is primarily found in Plasmodim vivax is the most prevalent species in Brazil, we assessed the macaques, but within the last decade it has been increasingly recorded in prevalence of anti- P. vivax MSP1-19 IgG antibodies among blood donors humans, particularly on the island of Borneo. Human malaria treatment is from Southeastern Brazil. Initially, for validation, ELISA-PvMSP1-19 was effective for the parasite in humans, but to prevent transmission in the first assayed with 197 sera from patients with positive thick-blood smear for P. place a better understanding of prevalence in its natural hosts is necessary. vivax yielding 96.95% sensitivity. A specificity of 100.0% was achieved in The objectives of this research are to develop and optimize noninvasive serum specimens from 101 normal individuals and 98.21% in 168 serum sampling methods for macaque malaria in wild populations. By using specimens from other diseases patients. After validation, 1,974 blood bank naturally infected macaques, we will compare blood and fecal samples to serum samples were tested: 1,309 from São Paulo and 665 from Rio de determine if noninvasive samples offer a logistical solution to widespread Janeiro. These samples were collected after the donors had been screened surveillance of macaque . Results of this project will be applied to by clinical parameters, provided they were considered fit to donate and field collected specimens and inform experimental designs for surveillance had signed the informed consent form. Regarding samples from São of this pathogen in Borneo and elsewhere. Results from this work are Paulo, 1.15% (N=15) positivity was achieved. In Rio de Janeiro samples, essential for understanding malaria prevalence in macaque hosts and the positivity was 1.65% (N=11). The reactivity index (RI) of the positive controlling emergence of the pathogen in new human populations. samples ranged from 8.98 to 1.16 (Sao Paulo) and 13.03 to 1.08 (Rio de Janeiro). The detection of specific antibodies is not necessarily a marker of 1544 parasitemia or disease, but the detection of anti- P. vivax IgG antibodies in blood bank donors in non-endemic areas constitutes an alert that impel us A COMPARATIVE CASE CONTROL STUDY OF THE to review the adopted criteria for screening of the donors aiming to reduce DETERMINANTS OF CLINICAL MALARIA IN THE GAMBIA the risk of TTM in these areas without losing donations. Kalifa A. Bojang1, Julia Mwesigwa1, Eugene L. Kama Kama2, Serign J. Ceesay1, Serign J. Ceesay1, Fanta Njie1 1546 1 2 MRC Unit, Banjul, Gambia, National Institute of Public Health, Conakry, HOUSEHOLD-LEVEL SOCIAL AND ENVIRONMENTAL Guinea FACTORS ASSOCIATED WITH BED NET OWNERSHIP AND The massive deployment of life saving malaria interventions has not DIFFERING MALARIA PREVALENCE IN SOUTHERN MALAWI only resulted in a decline in disease burden but a change in the risk of infection and disease. The study reassesses the importance of known risk Lindsay R. Townes1, Dyson Mwandama2, Don P. Mathanga2, factors and reviews socio-demographic determinants of malaria risk in Mark L. Wilson1 the population. We conducted a case-control study involving 150 children 1University of Michigan, Ann Arbor, MI, United States, 2Malaria Alert aged 6 months to 12 years with slide-confirmed malaria recruited from Centre, Blantyre, Malawi the outpatient clinics of three health facilities (cases) in the Greater Banjul Since 2002, Malawi’s primary malaria prevention program has been a area, The Gambia. One hundred and fifty controls were matched on age, nationwide, health facility-based distribution of insecticide-treated nets residence, and were negative for malaria. We collected information on (ITNs). Despite these efforts, ITN ownership in households with under-5 the use of long lasting insecticidal nets, occupation of parents, housing children remains sub-optimal. Knowing what characterizes such low- structure, knowledge of malaria and socio-demographic factors. The mean ownership households may improve targeting of ITN distribution. From age of study participants was 6.8 (SD 3.3) years with 147 (49%) being June to August 2011, data on household ITN ownership, child malaria males. Significant determinants of malaria risk were parent’s occupation: status, house location, building materials, and nearby environmental mother as trader (OR 0.18, 95% CI 0.04 - 0.73, p = 0.017), father as characteristics were collected by cross-sectional survey from 398 trader (OR 0.02, 95% CI 0.002- 0.193, p = 0.001), civil servants (OR households in two rural Traditional Authorities (TAs) of southern Malawi, 0.04, 95% CI 0.008- 0.257, p =0.001) or handyman (OR 0.03, 95% CI Sitola and Nsamala, which were in the catchment area of Machinga 0.005- 0.182, p < 0.001). Children sleeping in rooms with windowpanes District Hospital (MDH). The proportion of households in Sitola reporting had a 76% reduction in their odds of malaria (OR 0.24, 95%CI 0.07- bed net ownership was significantly lower (OR 0.59, 95% CI 0.39-0.89), 0.82, p = <0.022. Household socio-economic status plays an important and the prevalence of malaria among under-5s significantly higher (OR role in management of illnesses. The ability of mothers to engage in an 4.20, 95% CI 2.74-6.46) than in Nsamala, this despite higher bed net occupation increases household resources to access healthcare and on use among owners in Sitola (OR 2.56, 95% CI 1.00-6.57). Households time. The balance between the type of mother’s occupation and her time in Sitola were also much more likely to be located within 50 m of active available to supervise the child is an interesting emerging issue that needs agriculture (OR 9.60, 95% CI 4.93-18.68), of brick-making sites (OR 4.59, further investigation. 95% CI 2.34-9.01), and of water sources (OR 11.87, 95% CI 3.49-40.40). Households in the two TAs did not differ with respect to housing materials,

astmh.org 472 recent or current maternal pregnancy, number of children, or number of previously reported anemia and parasitemia surveys were conducted household residents. Among those with a bed net, there was no difference serially in 3 rainy seasons (RS) and 3 dry seasons (DS) in 2010-13, covering in whether they had received their bed net at MDH or whether the net 11,945 children under five from 179 communities. In spite of high was insecticide-treated. Curiously, within the higher malaria prevalence coverage for insecticide-treated bed nets (>75% use in each RS) and context of Sitola, ITN ownership was not significantly associated with child indoor residual spraying (IRS) with pyrethroid pesticides in years 2 and 3 malaria status, land use/land cover, quality of housing materials, nor recent (>98% households sprayed), investigators found unexpectedly high and maternal pregnancy; however, current maternal pregnancy was inversely geographically heterogeneous malaria prevalence. To better define and associated with net ownership (OR 0.35, 95% CI 0.13-0.95). In contrast, explain this heterogeneity, this study enhanced the survey dataset with current maternal pregnancy had no association with net ownership in remotely sensed data, then analyzed by ecologic zones, delineated as Nsamala (OR 0.69, 95% CI 0.31-1.50), while houses built with higher urban (Zone 1, n=1131); rocky uplands (Zone 2, n=5234, >750 ft altitude); quality materials were more likely to own at least one bed net (OR 3.31, transition (Zone 3, n=3456), and riverine plains (Zone 4, n=2124, <550 95% CI 1.20-9.12). These results suggest a geographical disparity in ITN ft). The RS odds ratios for microscopic malaria parasitemia in children distribution between Nsamala and Sitola, which may be reduced through living in Zones 2, 3, and 4, as compared with Zone 1, were respectively improved targeting of pregnant women in Sitola. 3.9 (95% CI: 2.8-5.4), 7.6 (95% CI:5.7-10.3), and 11.1 (95% CI: 8.0- 15.6; all p values here and in the following <0.0001). Zone 4 parasitemia 1547 prevalence across the 3 years was 65.6-72.1% in the RS and 39.8-54.9% in DS. Among 17 variables with statistically significant odds ratios (OR) for MALARIA TRANSMISSION IN BOLENGE HEALTH ZONE, malaria risk, 12 exhibited a zonal gradient favoring reduced risk in Zone EQUATORIAL SETTING, DR CONGO 1 vs. Zone 4, with Zone 2 intermediate. In the RS these included lower wealth quintile (OR=3.6; 95% CI:2.7-4.7), caregiver’s lack of education Thierry L. Bobanga1, Billy S. Kunyu2, Solange E. Umesumbu3, (OR = 2.7; CI: 2.1-3.3), ethnicity (OR = 3.9; CI: 3.2-4.8), lack of health Olivier A. Fataki1, Prosper Abanda4, Dieudonné N. Mumba1 insurance coverage (OR 3.0; CI=2.4-3.6), higher vegetation index (OR=1.6; 1University of Kinshasa, Kinshasa, Democratic Republic of the Congo, CI: 1.1-2.3), lower human influence index (OR 5.2; CI:3.8-7.1); and >3 2National Institute of Biomedical Research, Kinshasa, Democratic Republic km distance to nearest health facility (OR=2.4; CI:1.9-3.1), among others. of the Congo, 3National Malaria Control Program, Kinshasa, Democratic DS findings were similar. No consistent zonal gradient was found for the Republic of the Congo, 4Bolenge Health Zone, Mbandaka, Democratic malaria control measures (ITN use, ACT use, IRS). Findings suggest that, Republic of the Congo in spite of high coverage with ITNs and pyrethroid-based IRS, high malaria Malaria constitutes a public health major problem in Democratic Republic prevalence in northern Ghana may be found in locations where reduced of Congo (DRC). For malaria control , one of adopted approach is socioeconomic status and isolation coincide with low-lying terrain. Such Insecticide treated Net(ITN). In order to evaluate the malaria transmission, areas may require additional and/or modified methods for vector and level in a area , different parameters are therefore used. One of them is parasite control. the parasitic parameter which includes plasmodic index (PI), gametocytic index (GI), parasitemia density (PD) and the plasmodial species. Another 1549 parameter is the sporozoitic index (SI). This study aimed to assess the level of transmission in a stable Heath Zone where ITN have been partially DEVELOPING A PASSIVE MALARIA CASE DETECTION distributed. A transversal study has been conducted from 11th of October STRATEGY IN TANZANIAN MILITARY HEALTH FACILITIES AND to 17th of November 2011 in Bolenge Heath Zone where in, there are 3 MALARIA EPIDEMIOLOGY DATA COLLECTED TO DATE Health Areas( Bolenge, Wendji Secli and Bongonde) separated between 1 2 3 themselves by the distance of at least 10 kilometers, have been selected. Akili Kalinga , Christopher Mswanya , George Amoo , Charles 4 5 5 6 Thick blood smear and thin blood smear have been done in all members of Mwanziva , Sarah Chiduo , Lucky Temu , Lalaine Anova , Colin 6 the households which remount to 185 in total, which include by the way Ohrt 1066 subjects. Anopheles were captured in household for determining 1National Institute of Medical Research, Dar es Salaam, United Republic SI Results The global PI in Bolenge Health zone was of 41.8 %. The rate of Tanzania, 2Tanzania Peoples Defense Forces, Dar es Salaam, United of mosquisto bednet utilization was 95 %, 13 % and 23 %, respectively Republic of Tanzania, 3Amethyst Technologies LLC, Baltimore, MD, United in Bolenge health area, Bongonde and wendji-Secli and in the same States, 4Tanzania Peoples Defense Force, Dar es Salaam, United Republic way, the PI was of 32.7 %, 50.4 % and 42.2 %; p<0.01. The global of Tanzania, 5Walter Reed Malaria Programme-Tanzania, Dar es Salaam, average parasitemia of 3 Heath areas was of 2213±354 trophozoites/ µl United Republic of Tanzania, 6Walter Reed Army Institute of Research, (2326.±54; 3182±603 and 965.±194 respectively in Bolenge, Bongonde Silver Spring, MD, United States and Wendji-Secli health areas and in the same way, GI was of 3.7 %, The capability to accurately track malaria incidence is essential to 10.4 % and 4.4 % SI was respectively 5, 7 et 10 in Bolenge , Wendji measuring the true impact and efficacy of malaria control interventions or secli and Bongonde and..Plasmodium falciparum was found at 99.9 %. field evaluations of malaria therapeutics, vaccines, etc. However, gathering All anopheles were An.gambiae s.s M molecular form. In conclusion, reliable malaria epidemiology data in rural and resource-challenged transmission was high in Bolenge Health Zone, it was very raised in settings is often a daunting and difficult endeavor. In Tanzania, malaria Bongonde Health Area, where the rate of the ITN use was low. persists as a major cause of morbidity and mortality. The US Army has partnered with the Tanzania People’s Defence Forces (TPDF) and the 1548 Tanzania National Service Program (JKT) to support the TPDF’s efforts to improve malaria management in a number of TPDF and JKT camps. DETERMINANTS OF RISK OF MALARIA PARASITEMIA IN The TPDF is an important provider of health services for both military BUNKPURUGU-YUNYOO DISTRICT, NORTHERN GHANA, and civilian populations, especially in remote areas where TPDF camps INCORPORATING REMOTE SENSING AND SURVEY DATA are based. Our initial efforts to perform passive malaria case detection relied on collecting microscopy slides from sites for cross-checking. Paul J. Psychas1, Brooke Eckman1, Benjamin Abuaku2, Collins These efforts were resource intensive and resulted in dubious success Ahorlu2, Kwadwo Koram2 and questionable data. This led us to transition our focus to the use of 1University of Florida Emerging Pathogens Institute, Gainesville, FL, United malaria rapid diagnostic tests (RDTs) with RDT readers. In our approach, States, 2Noguchi Memorial Institute of Medical Research, University of we deployed the Deki Reader, a rugged, mobile in vitro diagnostic device Ghana, Legon, Ghana which interprets commercially available RDTs. Several advantages provided Ghana’s malaria control strategy prioritizes the northern savannah regions by the Deki system include real-time quality control measures, the ability due to persistent hyperendemicity. In Bunkpurugu-Yunyoo district, for remote quality assurance (QA), and the automatic organization of

astmh.org 473 the data in a centralized web-based portal. After deploying devices to 1551 each site, training is provided to site staff regarding use of RDTs and Deki Readers and sites are allowed a period of practice. After the sites become CAN HEALTH MANAGEMENT INFORMATION SYSTEM (HMIS) active, QA monitors review the mRDT database and conduct QA quarterly DATA BE USED TO MAKE INDOOR RESIDUAL SPRAY POLICY visits for trouble-shooting, to cross-check mRDT results against laboratory, DECISIONS IN BENIN? , and pharmacy records, and technician compliance and accuracy 1 2 1 for data transmission. In our first year of implementation, our approach Evelyne S. Alyko Chaffa , Elana Fiekowsky , Joel Sodjinou , 2 3 3 has undergone several adjustments to adapt to a number of challenges, Benjamin Johns , Allison Belemvire , Kristen George , Christen 3 however we have made great strides improving the reliability of the Fornadel data. We will present the challenges and successes experienced with 1Abt Associates, Cotonou, Benin, 2Abt Associates, Bethesda, MD, United implementation of our approach and the data collected to date. States, 3United States Agency for International Development, Washington, DC, United States 1550 The President’s Malaria Initiative (PMI) has supported Indoor Residual Spraying (IRS) in Benin as part of its malaria control strategy. The PMI/ PRECISE GENOTYPING TOOLS FOR INVESTIGATING USAID-funded Africa Indoor Residual Spraying (AIRS) Project assessed TRANSMISSION DYNAMICS OF PLASMODIUM FALCIPARUM the feasibility of using health facility data to inform targeted spray GAMETOCYTES decision-making, and assess IRS’s effect on malaria caseloads. AIRS Rahel Wampfler1, Lincoln Timinao2, Hans-Peter Beck1, Issiaka accessed routinely HMIS data from nine IRS intervention districts and Soulama3, Alfred B. Tiono3, Peter Siba2, Ivo Mueller4, Ingrid Felger1 five neighboring comparison districts, covering the period 2006 - 2013, including the period of IRS implementation from 2011-2013. We then 1 2 Swiss Tropical and Public Health Institute, Basel, Switzerland, Papua New assessed the association between IRS and the number of malaria cases 3 Guinea Institute of Medical Research, Goroka, Papua New Guinea, Centre reported by health facilities for both the total population and children National de Recherche et de Formation sur le Paludisme, Ouagadougou, under five years of age. Two models were developed for this assessment. 4 Burkina Faso, Walter and Eliza Hall Institute, Parkville, Australia In the first model, we first assessed, for a subset of areas with available Differentiation between gametocyte-producing Plasmodium falciparum data, the association of IRS on the Entomological Inoculation Rate clones depends on high stage-specific expression and high genetic diversity (EIR), we then analyzed the association between EIR and the number of of a genotyping marker in the study area. High-resolution typing methods malaria cases as reported in the HMIS data. Finally, we combined these are crucial for longitudinal tracking of gametocyte producing clones in two steps to evaluate the association of IRS on reported malaria cases multiple infections. Pfs230 and pfg377 are classical length-polymorphic through the EIR. The second model measured the association of IRS markers for differentiation of gametocytes. We have evaluated capillary directly on reported malaria cases in the HMIS data. The sample size for electrophoresis-based differentiation of 6 length-polymorphic gametocyte the first model was small, yet it serves to assess the validity of the second genes. These assays were applied to asexual parasites by targeting model. For the analyses, we used a difference-in-difference approach genomic DNA from field samples and in parallel to gametocytes from that controls for rainfall, other malaria and health interventions, and the same blood samples by targeting gametocyte-specific RNA. Highest time trends. We assessed the internal validity of the data assessing the diversity was found for pfs230 with 18 alleles and for pfg377 with 15 association of other malaria interventions on health facility utilization alleles in 111 samples from PNG. When assays were performed in parallel and putting the treatment on a year before IRS took place to measure on RNA and DNA from 46 samples from Burkina Faso, 85.7% of all pfs230 the association of a “false” treatment in the second model. We assessed samples and 59.5% of all pfg377 samples contained at least one matching the association separately for two classes of insecticides, carbamates genotype in DNA and RNA. Out of the 93 PCR fragments amplified from and organophosphates. As estimated against the comparison group, DNA of all samples by pfs230, 41 (44.1%) were not observed in the initial results show a 20 to 30 percent decrease in the number of corresponding RNA sample. Vice versa we found that 42.9% (39/91) confirmed cases per person per month associated with carbamates of pfs230 fragments detected in RNA failed to be amplified from the and a 5 to 20 percent decrease per person per month associated with corresponding DNA samples. The imperfect detection in both, DNA and organophosphates. The sensitivity tests and the falsification tests make RNA, was identified as major limitation for investigating transmission us question the internal validity of the second model; the falsification test dynamics, owing primarily to the volume of blood processed and the showed a statistically significant effect in a year that IRS did not take place. incomplete representation of all clones in the sample tested. This finding Based on this research, we cannot recommend using HMIS data to make emphasises the importance of expression levels of gametocyte-specific IRS targeting policy decisions in Benin without further data validation. markers as well as optimal sampling and preservation of DNA and RNA. Larger volumes may improve clone detectability of abundant low-density gametocyte carriers and of initially sequestered gametocyte clones in follow-up samples. Application of these methods to samples from cohort studies will help to explain additional factors influencing detectability of gametocyte clones, e.g. the dynamics of gametocytogenesis of a specific parasite clone over the duration of its infection.

astmh.org 474 1552 years manifests as severe malarial anemia [Hemoglobin (Hb) <6.0g/dL; any density parasitemia]. High levels of Immunoglobulin (Ig)-G to a number of INVESTIGATING THE SPECIFICITY AND KINETICS OF surface proteins and invasion ligands have been associated with protection

PLASMODIUM FALCIPARUM-SPECIFIC IGG RESPONSES from malaria. Morever, recent studies suggest that MSP1-42 interacts THAT ASSOCIATE WITH PROTECTION FROM MALARIA: A with heparin-like molecules on the RBC. Adhesion is mediated by the P. LONGITUDINAL STUDY IN MALI falciparum erythrocyte membrane protein 1 (PfEMP-1), expressed at the surface of infected erythrocytes and is linked to both antigenic variation Eugene Liu1, Jeff Skinner1, Tuan M. Tran1, Aissata Ongoiba2, and cytoadherence. The role of antibodies against these antigens in the Shanping Li1, Safiatou Doumbo2, Didier Doumtabe2, Younoussou pathogenesis of SMA remains largely unknown. We therefore sought to Kone2, Aboudramane Bathily2, Seydou Dia2, Moussa Niangaly2, elucidate the role of these antibodies by measuring circulating IgG levels Jules Sangala2, Chiung-Yu Huang3, Huw Davies4, Aarti Jain4, Arlo against these antigens in children (n=117) presenting with acute malaria at Randall5, Ogobara K. Doumbo2, Kassoum Kayentao2, Boubacar Siaya County Hospital, western Kenya. Complete hematological measures Traore2, Philip L. Felgner4, Peter D. Crompton1 were obtained with a Beckman Coulter Counter®, and Giemsa-stained 1National Institutes of Health, Rockville, MD, United States, 2Mali slides were used to determine parasitemia. Participants were stratified International Center of Excellence in Research, University of Sciences, based on Hb status as non-SMA (n=91), Hb≥6.0 g/dL and SMA (n=26), Technique and Technology of Bamako, Bamako, Mali, 3Research Program Hb<6.0 g/dL. Results presented here show that circulating IgG against in Quantitative Sciences, Division of Biostatistics/Bioinformatics, MSP1-42 were comparable between non-SMA and SMA groups [Median The Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD, United (Interquartile range) non-SMA 128.83 (295) and SMA 151.19 (321); 4 5 States, University of California Irvine, Irvine, CA, United States, Antigen P=0.963]. In addition, the IgG levels against MSP1-42 did not correlate Discovery Inc., Irvine, CA, United States with parasite density (ρ=0.065; P=0.487). Similarly, circulating IgG against Antibodies play a key role in malaria immunity, but antibody-mediated PfEMP-1 was comparable between the groups [Median (Interquartile protection is only acquired after years of Plasmodium falciparum exposure, range) non-SMA 420.0 (590) and SMA 377.97 (710); P=0.632] and was leaving children vulnerable to repeated bouts of symptomatic malaria. not associated with parasite density (ρ=-0.006; P=0.952). These results suggest that the levels of circulating IgG against MSP1- and PfEMP-1 are Our previous work in Mali suggests P. falciparum protein microarrays 42 can be used in population-based studies to investigate the antigen not correlated with malaria disease severity in acutely infected children specificity of protective antibodies and the kinetics of their acquisition; from this holoendemic region. however, our prior work was limited by small samples size, a lack of active surveillance for clinical malaria and P. falciparum infection, and few time 1554 points. To address these limitations we conducted an independent two year cohort study in the rural village of Kalifabougou, Mali where intense THE ROLE OF THE INHIBITORY FC RECEPTOR, FCΓRIIB malaria transmission occurs from June to December. Active surveillance IN HOST IMMUNE RESPONSE AND SUSCEPTIBILITY TO for clinical malaria and P. falciparum infection was done weekly and bi- MALARIA IN PAPUA NEW GUINEA weekly, respectively. Of the 695 enrollees in the cohort study (aged 2 Alexander M. Barron1, Gloria Tavera1, Arlene E. Dent1, Peter M. months to 25 years), we focused the present analysis on the 268 subjects Siba2, James W. Kazura1, Christopher L. King1 who were P. falciparum PCR negative at enrollment before the malaria 1Case Western Reserve University, Cleveland, OH, United States, 2Papua season. A microarray with 1024 P. falciparum proteins was probed with New Guinea Instititute of Medical Research, Goroka, Papua New Guinea plasma collected from these 268 subjects at four time points: before the six-month malaria season, during the first episode of febrile malaria Binding of human Fragment crystalizable gamma receptor 2b (FcγRIIb) of the ensuing malaria season (if it did occur), after the malaria season, on B cells to antigen (Ag)-containing immune complexes (ICs) mediates a and after the subsequent 6-month dry season. In ongoing analyses critical inhibition of host immune responses. Homozygosity for a FcγRIIb that we expect to complete by mid 2014, we are comparing antibody missense mutation (p.Ile232Thr) that reduces inhibition is associated with profiles of children who were prospectively classified as clinically immune protection against severe malaria in Kenyan children. Protection against (documented infection not followed by fever) or susceptible to malaria, as severe malaria in p.232Thr/Thr (TT) Kenyan children may result from a well as individuals who showed evidence of sterile protection. We are also more robust humoral immune response and/or enhanced phagocytosis of modeling the breadth, magnitude and kinetics of P. falciparum-specific malaria-infected erythrocytes (iRBC), which may select for this mutation. antibody responses from 2 months to 25 years--the age range over which This hypothesis is supported by observations that TT frequency is high in clinical immunity to malaria is acquired in this population. This rich dataset malaria endemic areas (5-11%) and low elsewhere (~1-3%). In Papua New is shedding light on fundamental properties of the human antibody Guinea, the frequency of I232T phenotypes did not significantly differ response to malaria and may help identify novel malaria vaccine targets. between groups living in different malaria endemic versus non-endemic areas (p = 0.3794), however preliminary analysis suggests that individuals 1553 with TT phenotype were less likely to be infected with P. vivax malaria (p = 0.0319, N = 544). Studies are underway to correlate TT phenotype CIRCULATING IMMUNOGLOBULIN (IGG) AGAINST with risk of malaria disease. To understand how the TT phenotype may mediate protection from malaria infection and disease, we examined MSP1-42 AND PFEMP-1 ARE NOT ASSOCIATED WITH PEDIATRIC MALARIA SEVERITY IN WESTERN KENYA malaria-specific antibody (Ab) levels among different FCGR2B genotypes since TT polymorphism reduces repression of B cell activating pathways. Evans Raballah1, Prakasha Kempaiah2, Samuel B. Anyona1, Unexpectedly, TT Papuans had smaller repertoires and lower levels of Zachary Karim2, Mark Webale1, George Orinda3, Michael F. P. falciparum- and P. vivax-specific Ab than II Papuans (i.e. p = 0.0045 Otieno4, James B. Hittner5, Douglas J. Perkins2, John M. Ong’echa1 for Duffy Binding Protein). Studies are underway to determine the role 1University of New Mexico Laboratories of Parasitic and Viral Diseases, of FcγRIIb in modulating phagocytosis, further elucidating its role in Kisumu, Kenya, 2Center for Global Health, Albuquerque, NM, United regulating host immune responses and susceptibility to malaria. States, 3Department of Biochemistry and Biotechnology, Nairobi, Kenya, 4Department of Medical Laboratory Sciences, Nairobi, Kenya, 5Department of Psychology, Charleston, SC, United States Malaria remains a major cause of morbidity and mortality among naïve immune children and pregnant women of Africa. The majority of cases are caused by Plasmodium falciparum. In holoendemic regions such as western Kenya, severe malaria cases in children under the age of five

astmh.org 475 1555 are carried out in a controlled environment, they allow unprecedented, detailed evaluation of parasite growth dynamics and immunological UNRAVELING THE HUMAN IMMUNE RESPONSE TO responses to infection. However, to date CHMI studies have not been SYMPTOMATIC AND ASYMPTOMATIC PLASMODIUM VIVAX used to investigate mechanisms of naturally-acquired immunity (NAI) INFECTIONS THROUGH SYSTEMS IMMUNOLOGY to P. falciparum infection. We conducted an open label, randomized CHMI study using aseptic, cryopreserved P. falciparum sporozoites (PfSPZ 1 2 2 Vitor R. Mendonca , Andrea Kuhn , Marcus V. Lacerda , Steve Challenge) administered intramuscularly to evaluate infectivity and parasite 3 4 1 Porcella , Manoel Barral-Netto , Peter D. Crompton growth dynamics in healthy Kenyan adults (n=28) with varying degrees 1National Institute of Allergy and Infectious Diseases, Rockville, MD, United of prior exposure to P. falciparum. All participants developed blood-stage States, 2FMT-HVD, Manaus, Brazil, 3National Institutes of Health, Hamilton, infection, however one volunteer remained asymptomatic and blood film MT, United States, 4FIOCRUZ-BA, Salvador, Brazil negative until day 21 post injection of PfSPZ Challenge, despite developing Approximately 2-3 billion people are at risk of Plasmodium vivax infection confirmed blood-stage infection by quantitative polymerase chain reaction worldwide. The nature of the immune response during symptomatic (qPCR). A significant correlation was seen between parasite multiplication and asymptomatic P. vivax infection remains elusive. We are applying rate (PMR) and anti-schizont ELISA OD at screening (p=0.044; r=-0.384). systems biology tools such as genome-wide expression profiling by Our study has shown that CHMI using PfSPZ Challenge is safe in African RNA-seq and multi-parameter flow cytometry to gain insight into host adults who have varying degrees of prior exposure to malaria and that NAI immune responses that associate with protection from disease during P. can impact on PMR post-CHMI, providing a novel method to investigate vivax infection. In the Brazilian Amazon we enrolled three groups: 1) P. the dynamics and mechanisms of blood-stage immunity. vivax-infected adults with fever (n=19), 2) P. vivax-infected adults with no fever or symptoms for 30 days despite persistent parasitemia (n=17), 1557 and 3) age-matched uninfected controls (n=17). Blood samples were collected at enrollment from the first and third groups, whereas blood CYTOKINE RESPONSES TO THE VAR2CSA VACCINE was collected from the second group after the 30-day period without CANDIDATE IN PREGNANT BENINESE fever or symptoms. Standard and chemistry labs were done; Komi Gbedande1, Gilles Cottrell2, Samad Ibitokou1, Bertin and plasma, peripheral blood mononuclear cells (PBMCs) and RNA were Vianou1, Carine Agbowaï1, Marita Troye-Blomberg3, Thor isolated from whole blood. Analysis of the hematologic data showed Theander4, Ali Salanti4, Achille Massougbodji1, Kabirou that symptomatic subjects had a higher percentage of neutrophils Moutairou5, Nicaise Tuikue Ndam2, Philippe Deloron2, Adrian (median 74.1%, p=0.0002) and a lower percentage of lymphocytes Luty2, Nadine Fievet2 (median 21.0%, p=0.0007) compared to asymptomatic subjects. 1Centre d’Etude et de Recherche sur le Paludisme Associé à la Symptomatic subjects also had higher levels of total bilirubin, creatinine Grossesse et à l’Enfance, Cotonou, Benin, 2Institut de Recherche pour le and glucose (p<0.0001 for each comparison) versus asymptomatic Développement, UMR D216 Mère et enfant face aux infections tropicales, subjects. Purified RNA was converted to cDNA and sequenced by Paris, France, 3Department of Immunology, Wenner-Gren Institute, next generation sequencing. In ongoing analysis of the RNA-seq data, Stockholm University, Stockholm, Sweden, 4Department of International differentially expressed pathways and gene sets in immune and susceptible Health, Institute of International Health, Immunology and Microbiology, individuals will be confirmed at the protein level and functionally using University of Copenhagen, Copenhagen, Denmark, 5Faculté des sciences et contemporaneous PBMCs and plasma samples. Molecular and cellular techniques, Université d’Abomey-Calavi, Cotonou, Benin signatures that correlate with protection from malaria fever may yield new hypotheses regarding the biological mechanisms by which malaria The STOPPAM consortium conducted 2 longitudinal cohort studies immunity is induced by P. vivax infection. The resulting datasets may be of in pregnant women in Benin and Tanzania in order to evaluate the considerable value in the urgent effort to develop a malaria vaccine. immunopathological consequences of infections with Plasmodium falciparum during pregnancy. In this context, parasite antigen-specific 1556 cellular responses, in particular to the vaccine candidate antigen VAR2CSA, have received little attention. Here we evaluated both, cytokine (IL10, IL13, EVALUATING CONTROLLED HUMAN MALARIA INFECTION IL17, IFN- γ, TNF-α) responses and the T cells IFN-γ specific responses to the IN KENYAN ADULTS WITH VARYING DEGREES OF PRIOR DBL5 domain of VAR2CSA. In Come, southwestern Benin, we conducted EXPOSURE TO PLASMODIUM FALCIPARUM USING a longitudinal prospective study of ~1000 pregnant women. Women at SPOROZOITES ADMINISTERED BY INTRAMUSCULAR ≤24 weeks of pregnancy were enrolled and followed at each antenatal INJECTION visit until delivery. Peripheral blood mononuclear cellular (PBMC) responses to VAR2CSA-DBL5 in vitro were determined in subgroups of 150 women Susanne H. Hodgson1, Elizabeth A. Juma2, Amina Salim3, at inclusion and 100 at delivery. In each subgroup those harbouring P. Charles Magiri2, Domtila Kimani3, Daniel Njenga2, Alfred Muia2, falciparum infections were matched by gravidity and gestational age Andrew O. Cole2, Caroline Ogwang3, Ken Awuondo3, Brett Lowe3, with mothers with no infection at inclusion and those with no history of Marianne Munene3, Peter F. Billingsley4, Eric R. James4, Anusha infection earlier in the pregnancy. The amounts of IL10, IL13, IL17, IFN-γ Gunasekera4, B. K. Sim4, Patricia Njuguna3, Thomas Rampling1, and TNF-α produced in response to mitogen (PHA) and to VAR2CSA- Adam Richman4, Yonas Abebe4, Gathoni Kamuyu3, Michelle DBL5 were quantified in supernatants of stimulated PBMC. The ex vivo Muthui3, Sean C. Elias1, Sassy Molyneux3, Stephen Gerry5, Alex frequencies of IFN-γ secreting CD4 and CD8 T cells in response to PHA and Macharia3, Thomas N. Williams3, Peter C. Bull3, Adrian V. Hill1, VAR2CSA domains were evaluated in the same maternal PBMC groups. At Faith H. Osier3, Simon J. Draper1, Philip Bejon3, Stephen L. the time of writing, all data have been collected, cytokine concentrations Hoffman4, Bernhards Ogutu3, Kevin Marsh3 have been evaluated and multivariate analyses are under way. Results 1University of Oxford, The Jenner Institute, Oxford, United Kingdom, will be discussed firstly in the context of cytokine profiles that reflect the 2Centre for Clinical Research, Kenya Medical Research Institute, Nairobi, acquisition of a specific cellular memory response to the vaccine candidate Kenya, 3Kenya Medical Research Institute - Wellcome Trust, Centre according to gravidity or to previous P. falciparum infection. Secondly, for Geographical Medical Research (Coast), Kilifi, Kenya,4 Sanaria Inc., we will discuss cytokine profiles as potential markers of protection in the Rockville, MD, United States, 5University of Oxford, Centre for Statistics in context of infection, anemia and birth weight. Medicine, Oxford, United Kingdom Controlled human malaria infection (CHMI) studies, where healthy volunteers are infected with Plasmodium falciparum have become a vital tool to accelerate vaccine and drug development. As CHMI trials

astmh.org 476 1558 to immobilized anti-T Cell Receptor antibody was inhibited by immobilized PD1-ligand in a dose-dependent manner. Our results suggest that VΔ2 cell MFERA (MALAWI) COHORT STUDY: COMMUNITY-BASED function in the fetus is regulated by PD1 in order to limit inflammatory LONGITUDINAL STUDY OF MALARIA IMMUNOLOGY, responses. Prenatal VΔ2 cell stimulation caused by maternal infection may EPIDEMIOLOGY AND GENOMICS induce long-term PD1 up-regulation, and hinder VΔ2 cell responses to pathogens and to BCG vaccination, affecting infant immunity. Esther N. Gondwe College of Medicine, Malawi, Blantyre, Malawi 1560 Malaria is endemic in Malawi, with climatic factors supporting transmission in most of the country except at high elevation. Individuals at risk of STATISTICAL MODELING METHODS REVEAL VARIABLE malaria infection are mostly young children and pregnant women. IMPORTANCE OF ANTI-MALARIAL ANTIBODIES IN KENYAN The risk of severe malarial disease and clinical symptoms declines in CHILDREN children residing in endemic regions when they experience continuing Arlene E. Dent1, Denise Babineau1, Peter Sumba Odada2, John exposure to infectious mosquito bites. Thus older children sustain only Vulule3, Ann Moormann4, James Kazura1 mild symptoms or are asymptomatic. To understand host responses that 1 2 develop with repeated exposures and are associated with clinical disease Case Western Reserve University, Cleveland, OH, United States, Kenya 3 immunity, we undertook a longitudinal cohort study at Mfera Health Medical Research Institute, Kisumu, Kenya, Centers for Disease Control 4 Centre, Chikhwawa district in Malawi. 120 subjects with uncomplicated and Prevention, Kisumu, Kenya, University of Massachusetts, Worcester, malaria are being followed for two years to capture host responses MA, United States during repeated infections. We will also examine host responses in three Naturally acquired immunity (NAI) to Plasmodium falciparum is age groups (1-5 years, 6-12 years and 13-50 years), as age serves as a characterized by age-related control of parasitemia and protection from proxy for clinical immunity. We will present data by age group to include clinical malaria. With the goal of advancing knowledge of how the clinical symptoms, rate of recurrence, host response profiling and parasite magnitude and breadth of anti-malaria IgG antibodies contribute to genotypes. We will also analyse these features in individuals who have NAI, we used plasma from 97 children (1-14 years) who participated in repeated infections to identify changes over time using repeated measures a treatment time to infection study in western Kenya. IgG antibodies to design. A primary focus of the host response studies is the role of type I 24 recombinant merozoite and pre-erythrocytic proteins were measured IFN in the development of clinical immunity. The role of type I IFN to confer by multiplex microsphere assay. A global test had a p value of 0.0601 protection or susceptibility to clinical malaria remains controversial and indicating that there is evidence that antibodies against at least one of thus we will focus on type I IFN responses (cytokine, interferon responsive the antigens is associated with delayed time to infection. Traditionally we genes, type I IFN receptor genetic variants, immunophenotyping of have used only Kaplan-Meier analysis to examine the relationship between relevant cells) in association with disease markers and age. The data antibody responses and time to infection. Here we developed and generated will provide an unprecedented opportunity to understand how compared 6 prediction models: 1) Kaplan-Meier, 2) Univariate Screening, residents of malaria endemic areas develop less severe clinical disease with 3) Backward Elimination, 4) Penalized Regression Models--Least Absolute repeated parasite exposures and these data may inform vaccine strategies. Shrinkage and Selection Operator (lasso) 5) Penalized Regression Model- -Elastic net, and 6) Random Survival Forests. Each method has benefits 1559 and limitations. By comparing the results of all analyses and evaluating the performance of each using the time-dependent Brier score, several PD1 EXPRESSION ON NEONATAL VΔ2 T CELLS MODULATES promising antigen targets that appear to contribute to protection from FUNCTIONAL RESPONSES TO MICROBIAL ANTIGENS infection such as AMA1(FVO), MSP3, MSP1 (3D7), and EBA181 would have been missed if we only used Kaplan-Meier analysis. We conclude that 1 2 1 Sarah E. Boudova , Suchita Chaudhry , Christopher Harman , high level statistical models offer insights into targets of NAI and identify 1 2 2 Miriam Laufer , David Pauza , Cristiana Cairo potential candidates for inclusion in multi-antigen malaria vaccines. 1University of Maryland, School of Medicine, Baltimore, MD, United States, 2University of Maryland, Institute of Human Virology, Baltimore, MD, 1561 United States In utero exposure to microbial antigens primes the fetal immune system, TRACKING THE DEVELOPMENT OF IMMUNITY TO often with negative consequences for infant immunity and responses PLASMODIUM FALCIPARUM AFTER IMMUNIZATION WITH to pediatric vaccines. VΔ2 T cells, a subset of γΔ T cells, play important IRRADIATED SPOROZOITES IN A MALARIA ENDEMIC roles in antimicrobial immunity and participate in responses to Bacille SETTING Calmette-Guérin (BCG) vaccination. We observed that prenatal exposure Irfan Zaidi1, Hama Diallo2, Yeya Dit S. Sarro3, Bourahima Kone3, to plasmodium antigens primes fetal VΔ2 cells, potentially causing Abdoulaye Katile2, Amadou Kone3, Ousmane Koita3, Freda dysregulation of infant VΔ2 cells. We are defining the impact of prenatal 4 1 2 Plasmodium falciparum exposure on neonatal and infant immunity by Omaswa , Sharon Wong-Madden , Mahamdou S. Sissoko , Stephen L. Hoffman4, Ogobara Doumbo2, Sara A. Healy1, Michael measuring changes in VΔ2 cells. In this context, we want to identify Walther1, Patrick Duffy1 inhibitory and activating receptors that modulate VΔ2 responses and differ in expression on fetal versus adult cells. These differences in regulation are 1Laboratory of Malaria Immunology and Vaccinology, National Institute important for normal immune function and may be altered by maternal of Allergy and Infectious Diseases, Rockville, MD, United States, 2Malaria infections. PD1 is a key negative regulator of immune responses and Research and Training Center, Mali-NIAID ICER, University of Science, a marker of T cell functional exhaustion during chronic viral infections Techniques and Technologies of Bamako, Bamako, Mali, 3SEREFO Core and malaria. We compared PD1 expression on healthy North American Immunology Laboratory, ICER, University of Science, Techniques and 4 adult and neonatal VΔ2 cells after stimulation. For adult VΔ2 cells, PD1 Technologies of Bamako, Bamako, Mali, Sanaria Inc., Rockville, MD, expression peaked by day 4 and in most individuals returned to baseline United States by day 14. For neonatal (cord blood) VΔ2 cells, PD1 expression peaked Vaccination with irradiated Plasmodium falciparum (Pf) sporozoites (SPZ) between days 4 and 7, and in most subjects was still elevated at day has been shown to induce sterilizing protection against malaria infection 14, yielding a PD1+ fraction significantly larger than in adults (43.6% in naïve volunteers, but as yet this has not been tested in subjects living versus 8.5%, p<0.0001). PD1 expression on neonatal VΔ2 cells remained in malaria endemic regions who would have pre-existing immunity. In stable up to day 35. The ability of neonatal VΔ2 cells to produce the pro- collaboration with Sanaria Inc. and the Malaria Research Training Centre inflammatory cytokine TNFα and mobilize cytotoxic granules in response (MRTC), the first double-blinded randomized phase 1b trial of the Sanaria®

astmh.org 477 PfSPZ Vaccine (radiation attenuated, aseptic, purified PfSPZ) in a malaria 1563 endemic region is being conducted in Mali. Ninety-three volunteers were randomized to receive five vaccinations of 2.7x105 PfSPZ or normal KIR3DS1 HOST GENOTYPE, IL17 SERUM CONCENTRATION saline placebo. In addition twelve volunteers received two vaccinations AND PLASMODIUM VIVAX CSP GENOTYPES MODULATION to ascertain safety of the vaccine, and nine of these volunteers will OF VIVAX MALARIA PARASITEMIA receive 5 immunizations. Vaccinations began in January 2014, and will 1 2 be completed in July 2014. CD8 T cells may be the key mediators of Giselle M. Viana , Andréa Luciana da Silva , Erian de Almeida 3 1 4 protection against liver stages of P. falciparum, but other cellular subsets Santos , Danielle Regina Barbosa , Gustavo Capatti Cassiano , 5 5 5 may also play a significant role. In earlier studies, CD38 and CD11a on Priscila Vianna , Nadine Glesse , José Artur Chies , Valéria Suenya 2 2 3 CD8 T cells have been used to measure the development of immunity Leal , Lilian de Araújo Mendes , Janaina Mota de Vasconcelos , 1 2 against malaria antigens after vaccination in mice and humans. Whole Ricardo Luiz Machado , João Farias Guerreiro , Marinete Marins 1 2 blood samples are being collected at baseline, 3, 7 and 27 days after each Póvoa , Eduardo José dos Santos vaccination, and used to measure the percentages of CD8, CD4, T and 1Instituto Evandro Chagas - IEC/SVS/MS, Ananindeua, Brazil, 2Universidade NK cells expressing CD38 and CD11a using flow cytometry. The results Federal do Pará - UFPA, Belém, Brazil, 3Universidade Federal do Pará after each of the 5 vaccine doses will be reported, and may be useful in - UFPA, Instituto de Ciências Biológicas, Belém, Brazil, 4Faculdade de discriminating the roles of various immune subsets in conferring protection Medicina de São José do Rio Preto - FAMERP, São José do Rio Preto, Brazil, after PfSPZ vaccination. 5Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, Brazil Malaria is the most important arthropod borne disease in Brazil, occurring 1562 mainly in North region, being a serious Public Health concern, in terms of its high morbidity and mortality rates. Both host and parasite genetic ANTIBODY PROFILING BY PROTEIN MICROARRAY IN NAÏVE features, as well as host immune profile, modulate resistance to infection AND SEMI-IMMUNE INDIVIDUALS IN COLOMBIA AFTER and heterogeneity of clinical/laboratorial manifestations. This study EXPERIMENTAL CHALLENGE WITH PLASMODIUM VIVAX approached: i) the Plasmodium vivax genotypes in Circumsporozoite Huw Davies1, Myriam Arévalo-Herrera2, Mary López-Pérez2, Protein (CSP) region; ii) host KIR genes polymorphisms and; iii) immune Aarti Jain1, Kelly Rubiano2, José Millán Oñate3, Philip L. Felgner1, profile during the infection (concentration of six circulating cytokines: IL- Socrates Herrera2 17, INF-g, TNF-a, IL-10, IL-6, IL-4, IL-2). Furthermore, we evaluated how these factors modulate the parasitemia and how host KIR polymorphism 1 2 University of California Irvine, Irvine, CA, United States, Caucaseco influences the number of CSP genotypes in infected individuals. Fourteen 3 Scientific Research Center, Cali, Colombia, Centro Médico Imbanaco, Cali, KIR genes and their ligands were genotyped by PCR-SSP on 62 P. vivax- Colombia infected individuals living in the town of Goianesia do Pará (Pará, Brazil). Acquisition of naturally acquired immunity to malaria in low transmission Cytokine levels were quantified using a Becton Dickinson cytometric bead intensity regions is often accomplished from relatively few exposures array. CSP genotypes (Vk210, Vk247 and P.vivax-like) were determined by and can occur at any age. This stands in contrast to high transmission PCR-RFLP. Among the 14 KIR genes only KIR3DS1 presence was associated areas where protection is acquired from repeated exposures during the with higher parasitemia (Mann-Whitney test; p=0.01). Moreover, KIR3DS1 first two decades of life. In this study we have evaluated serological presence associates with P. vivax CSP multiple genotypes (Fisher Exact differences in response to live Plasmodium vivax challenge in two groups test; 0.0084). Interestingly, individuals presenting multiple genotypes of P. of individuals from Colombia. These comprised 7 individuals with no vivax CSP showed also higher parasitemia (Mann-Whitney test; p=0.028). history of malaria (“naïve”), and 9 individuals naturally exposed to P. Fulfilling this scenario IL-17 concentration correlated negatively with vivax previously (“semi-immune”). Each individual was infected with parasitemia (r=-0.6702; p=0.024), suggesting a protective role in the P. vivax sporozoites via mosquito bites. As a result, 6 naïve (86%) and parasitemia control. Noteworthy, KIR3DS1 is a key stimulatory receptor 6 semi-immune (67%) individuals developed clinical symptoms in the of Natural Killer cells that produces many cytokines related to immune second week post-challenge. Fever and headache were significantly more response to P.vivax infection being the results suggestive of a role of this frequent or severe in naïve compared to semi-immune individuals, while gene in control of the number of different circulating CSP genotypes blood alanine aminotransferase, aspartate aminotransferase (markers as well as parasitemia and highlights KIR3DS1 role’s in malaria immune of liver function) and C-reactive protein were also significantly higher in response in an Amazonian population. naïve. Overall, the clinical data indicated previous exposure to P. vivax is associated with protection against clinical symptoms in response to P. vivax 1564 challenge. To test whether protection might also be associated with IgG profiles, serum from d0, d5, d11, 3 weeks, and 4 months post-challenge INDUCTION OF HOST CELL AUTOPHAGY PROMOTES THE were probed on a protein microarray displaying 500 P. vivax and 500 P. DEVELOPMENT OF MALARIA LIVER STAGE falciparum sero-reactive exon products. The array did not reveal strong serological differences between naïve and semi-immune individuals at the Wenyue Xu, Chenghao Zhao, Taiping Liu, Taoli Zhou time of challenge. However, a difference was observed in the response to Third Military Medical University, Chongqing, China challenge with P. vivax sporozoites. In both groups, the response peaked Host cell autophagy has been reported to be involved in the restriction of at week 3 and declined thereafter, with the response by the naïve group the growth of a variety of microorganisms, but its role in the development being noticeably stronger and broader in comparison with the semi- of malaria pre-erythrocytic stage is still unknown. Here, we found that immune group. Interestingly, the bulk of the serological response seen in sporozoite infection induced LC3 containing vesicle to surround the exo- the semi-immune group was also associated with those individuals with erythrocytic forms in hepatocyte, indicating the hepatocyte autophagy fever or headache, while those that were asymptomatic had an attenuated of the malaria parasite. Rapamycin treatment significantly increased response. Thus the association between previous exposure and protection the number of the autophagy of exo-erythrocytic form by hepatocyte, against clinical malaria is also associated with lower serological reactivity as and promoted the fusion of autophagy containing malaria parasite measured by protein array, possibly reflecting activity of the memory pool with lysosome. However, host cell autophagy induced by rapamycin in previously-exposed individuals. could significantly promote the development of exo-erythrocytic form in vitro. Further study showed that parasites inside the autophagosome could still survive and replicate normally as same as those in the parasitphorous vacuole, and the acidification of autolysosome was greatly inhibited. Therefore, we firstly provide evidence that sporozoite infection could induce host cell autophagy of malaria parasite, and the

astmh.org 478 induction of hepatocyte autophagy promoted the development of pre- based on three standard deviations above the normal distribution of erythrocytic stage, which might be associated with its ability to suppress our negative serum absorbances (OD of 0.37 and 0.48 for AMA-1 and the acidification of autolysosome. This data indicated the induction of MSP-1 respectively). Between February and May 2013, sample collection autophagy as a novel escape strategy of exo-erythrocytic stage, and shed occurred at four different sites; a rural community, two schools and a new light on the prophylatic therapy against liver stage. clinic in the Ouest and Sud-Est departments of Haiti. Of the 770 samples screened, 170 (22.1%) had been exposed to malaria at one point in their 1565 life time. Age was highly associated with the likelihood of having been exposed (p-value <0.001). After adjusting for age, the sero-conversion rate TRANSCRIPTOME ANALYSIS OF ATYPICAL MEMORY B CELLS calculations indicated that the annual malaria transmission in the Ouest IN THE SETTING OF NATURAL PLASMODIUM FALCIPARUM and Sud-Est department is roughly 1.03%. This data suggests that despite INFECTION the absence of sustained malaria control efforts in Haiti, transmission has remained relatively low over multiple decades. Our results are further 1 1 2 Richard T. Sullivan , Mary Fontana , Isaac Ssewanyana , Harriet supported by passive hospital based surveillance conducted by the Haiti 3 1 1 Mayanja , Prasanna Jagannathan , Margaret Feeney , Charlie Health Surveillance system, which found low country-wide transmission 1 1 Kim , Bryan Greenhouse (<1%). These findings provide valuable information that can be used to 1University of California San Francisco, San Francsico, CA, United States, make a case for the elimination of malaria on the Island of Hispaniola. 2London School of Hygiene & Tropical Medicine, London, United Kingdom, 3Makerere University , Kampala, Uganda 1567 Higher frequencies of an “atypical” phenotype of memory B cells have been associated with chronic exposure to Plasmodium falciparum and TOWARDS FULL PLASMODIUM FALCIPARUM PROTEOME other pathogens, but the function of these cells in malaria pathogenesis AND REACTOME ANTIBODY SCREENING ASSAYS USING and the development of immunity is not understood. Atypical memory REPRODUCIBLE HIGH-THROUGHPUT PROTEIN ARRAYS B cells have been hypothesized to be dysfunctional based on their poor Joseph J. Campo1, Douglas M. Molina1, Chris Hung1, Arlo Z. production of antibodies in vitro, while others have hypothesized that Randall1, Li Liang2, Arti Jain2, D. Huw Davies2, Elizabeth Baum2, these cells actively produce antibodies. However, antibody secretion Peter D. Crompton3, Robert W. Sauerwein4, Robert Seder5, is not the sole function of B cells. To better understand the functional Stephen L. Hoffman6, Denise L. Doolan7, John J. Aponte8, Carlota nature of atypical memory B cells, we performed a systematic evaluation Dobano8, Angela Yee1, Xiaowu Liang1, Philip L. Felgner2 of the differences between atypical and classical memory B cells using transcriptome analysis of both subsets in the presence of naturally 1Antigen Discovery, Inc., Irvine, CA, United States, 2University of California occurring P. falciparum infection. B cells subsets were isolated from 6 Irvine, Irvine, CA, United States, 3Laboratory of Immunogenetics, National parasitemic but asymptomatic children aged 8-10 years old and analyzed Institute of Allergy and Infectious Diseases, National Institutes of Health, on whole genome microarrays. Expression of select genes was confirmed Rockville, MD, United States, 4Department of , by qPCR and/or flow cytometry. Consistent with previously hypothesized Nijmegen, Netherlands, 5Vaccine Research Center, National Institute of atypical memory B cell dysfunction, we found a number of inhibitory genes Allergy and Infectious Diseases, National Institutes of Health, Bethesda, elevated compared to classical memory B cells. However, atypical memory MD, United States, 6Sanaria, Inc., Rockville, MD, United States, 7QIMR B cells were not dormant, but clearly metabolically active. In addition Berghofer Medical Research Institute, Herston, Australia, 8Barcelona to elevated inhibitory genes, atypical memory B cells also upregulated Centre for International Health Research (CRESIB), Barcelona, Spain multiple genes associated with activation, migration, and secretion High throughput (HT) technologies have rapidly advanced discovery of pathways. Upregulation of genes in these pathways suggests a more novel targets for disease diagnostic tests and vaccines and is especially complex function for atypical memory B cells beyond antibody secretion. useful for pathogens with large, complex genomes, such as Plasmodium Indeed, we performed functional assays confirming that these cells did not spp. We have developed a full proteome antibody screening assay for spontaneously secrete IgG ex vivo. We are currently performing additional P. falciparum proteins expressed via an HT in vitro transcription and functional assays to help define other roles atypical memory B cells have in translation (IVTT) system and printed onto nitrocellulose-coated microarray modulating cellular responses. In summary, the transcriptome data suggest chips. Here, we demonstrate the reproducibility of this protein array a functional role for atypical B cells, which, while as yet unknown, may be platform, the design and reactogenicity of the full proteome array, independent of antibody production. and the development of a down-selected P. falciparum “reactome” array. Comparisons included variation in assay dates, sample position 1566 on microarray chips and print batches. Agreement between replicate protein array assays was good. Mean difference and limits of agreement, AGE-SPECIFIC MALARIA SERO-CONVERSION RATES IN HAITI: demonstrated graphically with Bland-Altman dotplots, was 0.03 +/- 0.6 AN ANALYSIS OF MALARIA TRANSMISSION IN THE OUEST normalized intensity (range -6 to 8 on log scale). Correlation between AND SUD-EST DEPARTMENTS OF HAITI replicate measurements of antibody breadth and magnitude were 0.91 and 0.95, respectively. Classification of antigen reactivity showed high 1 1 Michael E. von Fricken , Brandon Lam , Thomas A. concordance (Cohen’s kappa: 0.95, p<0.01). The final down-selection 1 1 2 3 Weppelmann , Will Eaton , Laura Schick , Roseline Masse , and fabrication of a 1,000-feature “Pf1000 reactome” chip is underway 1 1 Madsen V. Beau De Rochars , Benjamin Anderson , Bernard A. and expected completion is in summer of 2014. We are down-selecting 1 Okech based on a tiered antigen reactivity scoring algorithm using antibody 1University of Florida, Gainesville, FL, United States, 2Community Coalition responses from serum/plasma samples of highly exposed individuals for Haiti, Jacmel, Haiti, 3Christianville Foundation, Inc., Gressier, Haiti from sub-Saharan Africa and Papua New Guinea and samples from Malaria transmission continues to occur in Haiti, with 32,000 confirmed experimental models of pre-erythrocytic immunity. Reactive antigens cases of Plasmodium falciparum reported in 2011. As rates of malaria are cross-referenced for confirmation with previously published data decrease, passive surveillance measures become less sensitive for capturing from other regions, including South America and Southeast Asia. transmission intensity. By implementing highly sensitive antibody detection The latest generation of down-selected chips will be a useful tool for methods we aimed to quantifying malaria transmission intensity over time. seroepidemiological studies and screening for antibody immune correlates A total of 770 serum samples were screened for malaria antibodies using of protection, and full proteome chips can be used for identifying novel indirect enzyme-linked immunosorbant assay (ELISA) coated with vaccine targets of monoclonal therapeutic antibodies or potential vaccine target candidates, apical membrane antigen (AMA-1) and merozoite surface antigens. protein-11-19 (MSP-1). The “exposed” cut off value was established

astmh.org 479 1568 sampling. In both 2 and 4 year old children, we found that higher prior malaria incidence was strongly associated with lower frequencies of Tregs IMMUNE MECHANISMS OF CROSS-STAGE PROTECTION (2 yo cohort rho = -0.28, p=0.011; 4yo cohort rho = -0.35, p=0.005)

BY VACCINATION WITH A LATE LIVER STAGE-ARRESTING However, there was no difference in frequencies of Tregs seen in children GENETICALLY ATTENUATED PARASITE with or without current or recent parasitemia. Functional differences

between Tregs from children with high or low prior incidences were Brandon K. Sack, Gladys Keitany, Jessica Miller, Ruobing Wang, investigated in ex vivo microarray analysis, specific and global suppression Ashley Vaughan, Stefan Kappe assays, and during the time course of acute infection. Our data suggests

Seattle Biomedical Research Institute, Seattle, WA, United States that repeated infection results in a loss of functionally suppressive Tregs

Immunizations targeting the pre-erythrocytic stages of Plasmodium have from the peripheral blood. These data may indicate that although Tregs demonstrated sterile protection against malaria by blocking the sporozoite are induced following malaria infection in naïve individuals, this process stages and/or eliminating liver stage parasites. Strategies such as irradiated becomes blunted after chronic repeated malaria exposure, which may have sporozoite immunizations however face a formidable challenge in that implications for the development of effective immune responses. they only confer stage-specific protection, and in consequence, less than complete protection can result in full-blown blood stage infection. 1570 However, we have demonstrated that immunization with sporozoites of P. yoelii genetically attenuated parasites (GAP), which arrest late in the INTERLEUKIN 8 AND TRANSFORMING GROWTH FACTOR- liver stage development and do not progress to blood stage infection, BETA (TGFΒ) AMONG MALARIA PATIENTS IN LAGOS, engenders protective cross-stage immunity against a direct lethal blood NIGERIA stage challenge. Here, we demonstrate that immune mechanisms Wellington A. Oyibo, Uche Igbasi, Ikechuckwu A. Ofili conferring cross-stage immunity are diverse. In C57BL/6 mice, antibodies are both sufficient and necessary for this protection as GAP-immunized College of Medicine of the University of Lagos, Nigeria, Lagos, Nigeria mice depleted of T cells completely control blood stage parasitemia The interaction between pro- and anti-inflammatory cytokines such as whereas immunized mice lacking antibodies succumb to uncontrolled interleukin-8 (IL-8) and transforming growth factor beta (TGF-β) plays an blood stage infection. Conversely, BALB/c mice depend on T cells for important role in malaria pathogenesis and outcome.TGFβ, produced by cross-stage protection. C57BL/6 antibodies recognize antigens in late a wide range of cells, has a pivotal role in the control of the transition liver stages as well as on the surface of blood-stage merozoites but are between pro-inflammatory (Th1-type) and anti-inflammatory (Th2-type) not specific for the well-characterized merozoite surface protein (MSP)- response during the acute and resolving phases of malaria infection. The 1. In contrast, immunization of BALB/c mice engenders anti-blood stage role of IL-8 in Plasmodium falciparum malaria is unknown although studies antibodies but they are lower in quantity and do not recognize the surface indicate it’s likely use as a biomarker of intensity of malaria.The aim of of merozoites. Cross-stage protection is unique to late-liver stage arresting this study was to measure the plasma levels of IL-8 and TGF- β in 136 GAP as animals immunized with an early liver stage-arresting GAP are not individuals with malaria and correlate the production of these cytokines protected from a lethal BS challenge and fail to generate antibodies which with the severity of the disease. IL-8 and TGF- βlevels were determined recognize late-liver stage/blood stage antigens. Therefore, immunization using enzyme-linked immunosorbent assay. The severity of malaria with a late liver stage-arresting GAP induces T and B cell responses capable was established by parasitemia, clinical symptoms and haematological of protecting against multiple stages of Plasmodium infection and thus parameters. The level of IL-8 was found to be substantially elevated constitutes the most potent among vaccination strategies. This unique (508.8±755.1pg/ml) in malaria infected individuals and its value was system also offers an opportunity to identify novel protective antigens, significant in parasitemia levels (43200.0, p<0.05). In contrast TGF-β which are shared with between both the liver stage and blood stage levels were found to be lower in malaria patients (23,672±30,703.8pg/ parasites. ml) compared to non-malaria patients, the mean difference in levels of IL-8 between malaria positive and malaria negative individuals was statistically 1569 significant (p<0.05).The relationship between TGF-β levels and packed cell volume was negatively correlated (r = -0.27) . These findings suggest PERIPHERAL BLOOD FOXP3+ REGULATORY CD4 T CELLS that fine mechanisms regulate the interaction between TGF-β andIL-8 DECLINE WITH INCREASING MALARIA EPISODES IN YOUNG in the immune response to Plasmodium falciparum infection, seemingly CHILDREN directingin vivo modulations in red cell population, and indicating a likely role in susceptibility to malaria Michelle Boyle1, Samuel Wamala2, Prasanna Jagannathan1, Iejoma Eccles-James1, Katherine Bowen1, Tara McIntyre1, Lila 1571 Farrington1, Felistas Nankya2, Charles Ebusu2, Mayimuna Nalubega2, Kate Naluwu2, Esther Sikomyu2, Victor Bigira2, James IMPACT OF ACUTE MALARIA ON PRE-EXISTING ANTIBODY Kapisi2, Jordan Tappero3, Mary Kakuru2, Emmanuel Arinaitwe2, LEVELS TO COMMON CHILDHOOD VACCINES Charlie Kim1, Grant Dorsey1, Moses Kamya2, Margaret Feeney1 Jill Coursen1, Silvia Portugal1, Aissata Ongoiba2, Boubacar Traore2, 1University of California San Francisco, San Francisco, CA, United States, John T. Harty3, Peter D. Crompton1 2Infectious Diseases Research Collaboration, Kampala, Uganda, 3Centers 1 2 for Disease Control and Prevention, Atlanta, GA, United States National Institutes of Health, Rockville, MD, United States, University of Sciences, Technique and Technology of Bamako, Bamako, Mali, 3University Plasmodium falciparum infection has been reported to induce of Iowa, Iowa City, IA, United States immunoregulatory cell populations such as FoxP3+ CD4 regulatory T Vaccine-induced protective immunity against many pathogens relies on cells (Tregs). Studies of malaria-naïve adults and low exposure regions long-lived plasma cells that maintain basal levels of antibody. Recent have indicated that Tregs expand during acute malaria infection and can be induced by malaria in vitro, however in children the relationship is evidence from animal models suggests that Plasmodium infection mediates a transient drop in antibody titers induced by prior influenza less clear. Treg expansion may be associated with the slow acquisition of immunity seen in children from malaria endemic areas. Indeed, other virus infection, raising the possible public health concern that malaria may be detrimental to previously generated vaccine responses. Prior studies in chronic diseases have been shown to induce Tregs which contributes to humans have assessed the impact of concurrent P. falciparum infection ongoing infection. We investigated the frequency and function of Tregs in well characterized cohorts of two-year-old (n=79) and four-year-old on responses to standard childhood vaccines, but to our knowledge (n=72) children in Tororo, Uganda. All prior malaria episodes from age 6 the impact of acute P. falciparum infection on previously induced months were documented and children were followed for 1 year following vaccine responses is unknown. To address this question we conducted a

astmh.org 480 longitudinal analysis of IgG titers specific for common viral (measles, polio, C-terminal region of merozoite surface protein 1 of P. vivax (PvMSP1-19) Hepatitis B) and bacterial (Haemophilus influenzae type b, meningococcus, is highly immunogenic conserved region that plays a major role in the tetanus) vaccine antigens in 54 children living in an area of Mali where the protective immunity against asexual blood stages of malaria parasites. 6-month malaria and dry seasons are sharply demarcated. Vaccine-specific Since this protective immunity has been shown to correlate with levels IgG titers were measured for each subject at five time points over an 18 of anti-MSP1-19 antibodies, this study aimed to evaluate the humoral month period: before and after the first dry season, during and 10 days immune response against PvMSP1-19 of individuals naturally exposed to after the first episode of febrile malaria of the ensuing malaria season, malaria, from endemic areas of Brazil, in order to assess the IgG and IgM and at the end of the second dry season. Preliminary analyses suggest profile, the avidity of IgG antibody (functional affinity) and their association that average IgG decay rates are not significantly accelerated by acute P. with different malaria clinical presentations. Serum samples from four falciparum infection; however, at the individual level a minority of children groups of individuals were studied: severe malaria (N=18), asymptomatic exhibited accelerated IgG decay rates following P. falciparum infection infection (N=17), non-severe symptomatic malaria undergoing their compared to decay rates over the dry season. In addition, preliminary first malaria episode (N=104) and non-severe symptomatic malaria with comparisons with studies of non-malaria exposed populations suggest previous malaria episodes (N=102). All were positive for P. vivax by thick that overall decay rates of vaccine-induced IgG responses may be higher blood smear and/or PCR and for IgG and/or IgM antibodies by ELISA- in the population of children who experienced febrile malaria. These PvMSP1-19. High avidity (>50%) IgG antibodies were observed in 92.9% data highlight the need for additional studies to understand the factors of patients who had previous malaria episodes (median reactivity index: underlying variability in vaccine-specific antibody decay rates in individuals IgG=8.7 and IgM=1.1) and in 88.0% of asymptomatic individuals (median residing in malaria endemic areas. reactivity index: IgG=2.4 and IgM=0.3). Low/moderate avidity (≤50%) was seen in 89.1% of patients undergoing their first malaria episode (median 1572 reactivity index: IgG=6.7 and IgM=3.0) and in 94.0% of severe malaria patients (median reactivity index: IgG=7.7 and IgM=6.7). Predominance THE IMPACT OF SCHISTOSOMA HAEMATOBIUM INFECTION of high-avidity antibody in individuals with non-severe malaria that had ON PLASMODIUM FALCIPARUM-INDUCED IMMUNE multiple episodes of malaria and in asymptomatic infections corroborates RESPONSES the protective role of humoral immunity. It likely reduces the risk to develop mild and severe malaria. Our results show that protective 1 1 2 Safiatou N. Doumbo , Jules Sangala , Jacqueline Moebius , immunity not only correlate with levels of anti-MSP1-19 antibodies but 2 2 1 Silvia Portugal , Tuan M. Tran , Aboudramane Bathily , Aissata also with the quality of these antibodies. Ongoiba1, Peter D. Crompton2, Boubacar Traore1, Ogobara K. 1 Doumbo 1574 1University of Sciences, Technique and Technology of Bamako, Bamako, Mali, 2National Institutes of Health, Rockville, MD, United States GENERATION OF “FULLY HUMAN” MONOCLONAL Plasmodium falciparum and Schistosoma haematobium often overlap ANTIBODIES AGAINST THE CIRCUMSPOROZOITE PROTEIN in tropical and subtropical countries and impose tremendous disease (CSP) OF PLASMODIUM FALCIPARUM USING HUMANIZED burdens. Evidence suggests that S. haematobium modifies the risk of HLA MICE febrile malaria; however, the nature of this putative immunomodulation 1 1 1 remains unclear. To investigate this question we analyzed S. haematobium- Sai P. Majji , Wathsala Wijayalath , Soumya Shashikumar , 1 1 1 infected individuals (n=15) and uninfected controls (n=24) within a Sandra Inoue , Andrea Akyeampong , Thomas L. Richie , Eileen 1 2 1 longitudinal cohort study in Mali. Before the malaria season, peripheral Villasante , Teodor-Doru Brumeanu , Sofia Casares blood mononuclear cells (PBMCs) were collected from S. haematobium- 1Naval Medical Research Center/Walter Reed Army Institute of Research, infected and uninfected individuals, all of whom were negative for Silver Spring, MD, United States, 2Uniformed Services University of Health P. falciparum by PCR. PBMCs were analyzed before and after in vitro Sciences, Bethesda, MD, United States stimulation with lysate of P. falciparum (3D7) infected red blood cells “Fully human” monoclonal antibodies (hmAb) are a novel approach (iRBCs) and analyzed by flow cytometry with intracellular cytokine staining; to treat infectious diseases. Fully human mAbs are devoid of the and supernatants of stimulated PBMCs were analyzed by a multiplex complications associated to the use of mAbs derived from mouse or cytokine assay. Compared to uninfected controls, S. haematobium-infected humanized antibodies. We have generated humanized HLA mice in NOD. individuals had a higher baseline percentage of dendritic cells, but no RagKO.IL2RgcKO background that develop a functional human immune differences were observed in the proportion of B cells, T cells, NK cells or system and respond to vaccination. Using human B cells from humanized their respective subsets. Stimulation with iRBCs showed a modest increase HLA mice immunized with irradiation-attenuated Plasmodium falciparum in IFN-γ producing CD4+ T cells in S. haematobium-infected individuals. sporozoites we have generated a panel of hmAbs against P. falciparum These preliminary data suggest that S. haematobium modulates the innate CSP. Herein we present data on anti-CSP hmAb immunocharacterization and adaptive immune response to P. falciparum infection. In an ongoing and in vitro and in vivo anti-parasitic activity. longitudinal study we are studying the impact of concurrent or recent S. haematobium infection on the host immune response to subsequent P. 1575 falciparum infection. MONITORING AND EVALUATION OF EFFECTIVENESS OF 1573 IFAKARA HEALTH AND DEMOGRAPHIC SURVEILLANCE SYSTEMS (HDSS) IN QUANTIFYING IMPACT OF MALARIA AVIDITY OF NATURALLY ACQUIRED ANTI-PLASMODIUM CONTROL STRATEGIES AND INTERVENTIONS VIVAX MSP1-19 ANTIBODIES IN INDIVIDUALS PRESENTING DIFFERENT CLINICAL EXPRESSIONS OF MALARIA Mwajabu M. Mwanaupanga Ifakara Health Institute, Dar es salaam, United Republic of Tanzania Maria Carmen A. Sanchez, Adeiana G. Lima, Arianni R. Sanchez, Eduardo M. Ramos-Sanchez, Silvia M. Di Santi, Marina P. Health and Demographic Surveillance Systems (HDSS) have been set up in Dornelas, Sandra L. Moraes, Hiro Goto various sites in Africa and Asia. In the absence of effective vital registration and information on mortality, these health and demographic surveillance São Paulo University, São Paulo, Brazil systems have well developed structures and standard operating procedures Plasmodium vivax is the most prevalent species in Brazil accounting for established to periodically monitor vital events like births, deaths and around 85% of clinical cases. P. vivax infections cause non-severe malaria migrations in well defined areas to produce population health information. in most cases, but may also be asymptomatic or cause severe disease. In Tanzania Ifakara DSS remain as an instrumental infrastructure of

astmh.org 481 producing key demographic indicators that are useful for planning and 1577 resource allocation to the community. The aim of my study is to determine the effectiveness of Ifakara HDSS in quantifying the impact of malaria QUALITY OF CARE FOR CHILDREN UNDER FIVE YEARS control strategies and interventions. A cross sectional study design will WITH UNCOMPLICATED MALARIA AT PRIVATE CLINICS IN be used to assess HDSS documentation mechanism on the impact of MAKINDYE DIVISION, KAMPALA DISTRICT different malaria control strategies and interventions performed in the 1 2 1 area, and to determine if the project effectively quantifies the impact of Ronald Mulebeke , Achillies Katamba , Aggrey Mukose , Pauline 2 1 1 2 those interventions and strategies. Systematic sampling will be used to Byakika , Ellizeus Rutebemberwa , Phyllis Awor , Jane Achan , 1 obtain number and types of documents that will be reviewed to fulfill the Adoke Yeka aim of our study. Collection and analysis of data will be both qualitative 1Makerere University School of Public Health, Kampala, Uganda, and quantities approach. Findings will be explained based on: 1) extent 2Makerere University College of Health Sciences, Kampala, Uganda the knowledge and skills of the project staff in content of the HDSS Over 60% of the population in developing countries seeks for care from malaria forms and data quality checking has increased from the pre to the private health facilities. In Uganda over 80% of the Population access post intervention periods of the project, 2) extent to which the malaria health care from small drug shops, private clinics and private-not-for profit information management has improved, 3) ways in which the project was providers because they are closer to the communities and are perceived to able to promote quality malaria data generation, 4) extents to which costs be affordable. However, health providers in private clinics more frequently were able to reach project goal of documenting impact of malaria control violate accepted medical standards and guideline because there primary strategies and interventions, and 4) how the HDSS understood and cared goal is making profit. Quality of care for children with uncomplicated about the importance of resources at work and if the managers supported malaria in such settings is generally sub-optimal with low adherence to them with training, supervision and needed resources. The conclusion will treatment guidelines. We are conducting a study to assess the quality of be based upon findings that will be obtained during the data collection. care provided to children under five years with uncomplicated malaria at private clinics in Makindye division, Kampala district, Uganda. The study 1576 is a cross-sectional cluster survey conducted in 30 private clinics. Data was collected using patient exit interviews, questionnaires for health BUILDING A CRITICAL MASS OF HEALTH WORKFORCE TO workers, observation of health workers during consultation, health facility FIGHT MALARIA IN NIGERIA audits and Key Informant Interviews. A total of 180 exit interviews were Muhammad K. Salihu1, Bolatito Aiyenigba1, Veronica Momoh2, conducted with caretakers of children and 30 healthcare workers were Abba Z. Umar1 observed while treating patients and were interviewed thereafter. A total of 8 Key informant interviews were conducted with heads of private 1FHI360, Abuja, Nigeria, 2Health Partners International, Abuja, Nigeria clinics. Data is being analyzed to determine predictors of quality of care for The Presidential Malaria Initiative (PMI) through the Malaria Action children with uncomplicated malaria at private clinics. A composite index Program for States (MAPS) in Nigeria currently supports seven states will be utilized to assess overall quality of care. to implement effective malaria control, prevention, diagnosis and treatment activities. Nigeria health workforces are diverse and cut across 1578 administrative, programmatic and clinical areas including community health services. Effective and sustainable malaria program will need FACTORS AFFECTING TREATMENT SEEKING FOR FEVER to build capacity in all these areas. This study documents the PMI BEFORE AND AFTER INTERVENTIONS TO IMPROVE achievement in Nigeria. In-service training using adult learning principles ACCESS AND TARGETING OF ARTEMISININ COMBINATION were conducted for all cadres of staff relevant to malaria prevention THERAPIES and control. Administrative and programmatic staffs were trained on malaria program management; clinical staff were trained on prevention Charles Festo1, Rebecca Thomson2, Boniface Johanes1, of malaria in pregnancy (MIP); malaria diagnosis using rapid test and Admirabilis Kalolella1, Happy Nchimbi1, Katia Bruxvoort2, Matthew microscopy; malaria case management including severe malaria. Other Cairns2, Mark Taylor2, Immo Kleinschmidt2, S. Patrick Kachur3, trainings conducted were focused on strengthening health information Catherine Goodman2 management; behavior change communication and community 1Ifakara Health Institute, Dar es Salaam, United Republic of Tanzania, acceptance of interventions. From October 2011 to September 2013, 2London School of Hygiene & Tropical Medicine, London, United Kingdom, 2770 health workers had been trained on MIP; over 9400 health workers 3Centers for Disease Control and Prevention, Atlanta, GA, United States and Community Care Givers had been trained on case management and Artemisinin combination therapies (ACTs) were introduced as the first line 3380 trained on malaria diagnosis. About 2498 health managers were antimalarial in Tanzania in 2006, but access and targeting were poor. The also trained on malaria program management. Over 4504 health workers main sources of treatment for fever included public and private health were trained on health information management; 3344 health educators facilities, drug stores, pharmacies and general shops, but the probability and journalists trained on malaria BCC. Programmatic results recorded of receiving both parasitological diagnosis and an ACT varied considerably include increase in fever cases tested from 45.9% in April 2013 to 70.2% across these outlet types. It is therefore essential to understand factors in March 2014; and referral of over 40,000 pregnant women and children that determine choice of provider. We assessed this using data from under 5 for ANC and treatment of fever. The critical mass of health large scale household surveys conducted before (2010) and after (2012) workers trained will contribute to the implementation of a sustainable, national rollout of rapid diagnostic tests in public facilities, and ACT efficient, integrated malaria program at state and local government subsidies under the Affordable Medicines Facility-malaria. We visited levels because many of these workers offer informal services in their a representative random sample of households in each of 3 regions communities. Pre-service training is needed to sustain malaria control (Mwanza, Mbeya, Mtwara) with varying malaria transmission and access program. to health care. 5,423 households at baseline and 5,511 at endline were sampled using a multi-stage design. All household members reporting fever were asked about treatment sought. There was no significant change in the percentage of people with fever seeking care between baseline and endline (69.5% and 73.6%, p=0.07). However, there were changes in treatment source, with an increase in the percentage using drug stores (41.3% to 54.1%, p<0.001), and a fall in use of public facilities (25.3% to 16.8% p<0.001). Overall, children <5 years old were more likely to be taken to a public facility at baseline and endline. We will present results

astmh.org 482 of multivariable analysis assessing the adjusted odds of seeking care, and 1580 of seeking care at specific provider types at baseline and endline. Relevant exposures include age, sex, region, urban/rural location, education of AUDITING VILLAGE HEALTH TEAMS’ CAPACITY FOR the household head, time to the nearest treatment source, enrolment in MANAGEMENT OF MALARIA: RESULTS OF THE 2013 health insurance scheme and local ACT stockout levels in public facilities. ACTWATCH UGANDA OUTLET SURVEY Key factors affecting treatment seeking decisions will be discussed, 1 2 3 4 including how these have changed following implementation of two key Julius Njogu , John H. Rogers , Henry Kaula , Vamsi Vasireddy , 5 interventions affecting malaria treatment provision, and the implications Megan Littrell for future policy to enhance access and targeting of ACTs. 1Population Services International, Nairobi, Kenya, 2Centers for Disease Control and Prevention, Dar es salaam, United Republic of Tanzania, 1579 3PACE, Kampala, Uganda, 4Independent Consultant, Chicago, IL, United States, 5Population Services International, Washington, DC, United States AN ASSESSMENT OF THE MALARIA-RELATED KNOWLEDGE Malaria is endemic across 95% of Uganda. The Ugandan government AND PRACTICES OF TANZANIA’S DRUG RETAILERS: aims at 85% of malaria cases receiving prompt and recommended EXPLORING THE IMPACT OF DRUG STORE ACCREDITATION treatment by the end of 2014. To achieve this goal, integrated Community Rebecca Thomson1, Boniface Johanes2, Charles Festo2, Case Management (iCCM) initiatives have been implemented in 34 Admirabilis Kalolella2, Mark Taylor1, Sarah Tougher1, Yazoume Ye3, districts by building capacity of Village Health Teams (VHTs) to manage Andrea Mann1, Ruilin Ren3, Katia Bruxvoort1, Barbara Willey1, Fred febrile children. Within these initiatives, all suspected malaria cases are Arnold3, Kara Hanson1, Catherine Goodman1 tested and confirmed cases are treated with Artemether-Lumefantrine (AL), and severe cases are referred for rectal Artesunate treatment. To 1 London School of Hygiene & Tropical Medicine, London, United Kingdom, enumerate VHT malaria capacity under iCCM, a census of VHTs was 2 3 Ifakara Health Institute, Dar es Salaam, United Republic of Tanzania, ICF conducted in 16 iCCM sub-counties using ACTwatch methodology. International, Calverton, MD, United States A questionnaire was administered to assess knowledge, availability of Since 2005 Tanzania has been upgrading its approximately 7,000 drug antimalarials and rapid diagnostic tests (RDTs). We present descriptive stores to Accredited Drug Dispensing Outlets (ADDOs), involving dispenser results on availability and knowledge. We surveyed a total of 1,862 VHTs. training, introduction of record keeping and enhanced regulation. Of these, 33.7% (628) stocked AL, 10.0% (187) rectal artesunate and ADDOs are permitted to stock 49 prescription-only medicines, including 21.6% (403) stocked malaria RDTs during the survey visit. Notably, when artemisinin combination therapies. Prior to accreditation drug stores AL was available, 56.5% (355/628) of VHTs stocked malaria RDTs. Of could officially stock over-the-counter medicines only, though many those with antimalarials, 91.6% (606/660) correctly stated recommended stocked prescription-only antimalarials. Oral artemisinin monotherapies first-line medicine for uncomplicated malaria (AL) and 83.3% knew and injectables were not allowed in any drug stores. By late 2011 ADDO its dosing regimen. Among RDT stockists, 93% (375/403) stated they conversion was complete in 14 of 21 regions. We explored variation in would never dispense antimalarial following a negative RDT result. The malaria-related knowledge and practices of drug retailers in ADDO and iCCM presents an important channel for increasing access to integrated non-ADDO regions. We excluded Dar es Salaam where market conditions case-management in Uganda. According to these results, VHTs are were not comparable to other regions. Data were collected as part of the highly knowledgeable but lack antimalarial and RDT stocks, which may Affordable Medicines Facility-malaria Independent Evaluation, involving undermine iCCM goals. Enhancing stable and reliable supply of first-line a nationally representative survey of antimalarial retailer in October- medicines and RDTs may sustain increased access to prompt and correct December 2011. We randomly selected 49 wards, and interviewed all drug malaria-case management. stores stocking antimalarials. Interviews were conducted in 148 drug stores in ADDO regions and 127 in non-ADDO regions. Drug stores in ADDO and 1581 non-ADDO regions were similar in terms of employing staff with health- related qualifications (96.1% and 96.2%, p=0.99); stocking the first ASSESSING THE COST OF PRIVATE SECTOR ACT SUBSIDIES line antimalarial (59.5% and 60.7%, p=0.89); and stocking artemisinin - THE FINANCIAL AND ECONOMIC COSTS OF THE monotherapy (0.9% and 0.0%, p=0.43). Drug stores in ADDO regions AFFORDABLE MEDICINES FACILITY - MALARIA (AMFM) IN performed better on knowledge of the first line antimalarial (99.5% and THREE AFRICAN COUNTRIES 91.5%, p=0.001). There was weak evidence of a lower price and higher market share of the first line antimalarial in ADDO regions. However, drug Catherine Goodman1, Fiona Cianci1, Daniel Cobos2, Sarah stores in non-ADDO regions were less likely to stock injectables (21.5% Tougher1, Kara Hanson1 and 3.6%, p=0.003). ADDO conversion is frequently cited as a model for 1London School of Hygiene & Tropical Medicine, London, United Kingdom, improving retail sector drug provision. Drug stores in ADDO performed 2Swiss Tropical and Public Health Institute, Basel, Switzerland better on some but not all indicators, possibly indicating weaknesses in AMFm was designed to improve uptake of quality-assured artemisinin ADDO regulation and high staff turnover. More evidence is needed on the combination therapies (ACTs). Hosted by the Global Fund to Fight HIV/ value-added and value for money of ADDO roll out to inform retail policy AIDS, Tuberculosis and Malaria, it operated in eight national scale pilots, in Tanzania and elsewhere. involving: (i) price negotiations with ACT manufacturers; (ii) a subsidy for ACTs at the top of the global supply chain; and (iii) supporting interventions, such as communications campaigns and provider training. An Independent Evaluation found that in most but not all settings AMFm improved the availability, affordability and market share of quality-assured ACTs. However, no data were available on AMFm costs. This study aimed to address this gap. We collected data on the costs of implementing AMFm in the private for-profit sector only in Nigeria, Kenya and Madagascar, representing a range of performance on AMFm indicators. Costs were included for all AMFm-related activities at both country level and the Global Fund headquarters. We adopted a “funders perspective”, covering all resources contributed by external funding agencies, NGOs and national governments, but excluding costs to commercial actors and households. Results are presented in terms of financial costs (actual expenditure) and economic costs (which include an annualised component

astmh.org 483 of start-up costs based on their expected useful life). All costs were an important source of treatment for many illnesses. In the case of malaria, converted to 2012 USD. The number of subsidised ACTs delivered for a large proportion of patients purchase antimalarial treatments from for- the private for-profit sector by the end of 2012 was 91,4 million (mn) profit providers. For example, data from 2009-2010 show that in Nigeria, in Nigeria and 28,3mn in Kenya, but only 2,1mn in Madagascar, where Kenya and Uganda, 97%, 67% and 40% of total antimalarial volumes imports were much lower reflecting the very limited communications were distributed by for-profit providers, respectively. However, medicines campaign, the predominance in the market of outlets not permitted to sell obtained are often inappropriate, because non-recommended treatments ACT, and political and economic disorder. Total financial costs for 2009-12 are widely and cheaply available. Subsidies for artemisinin combination were $110,2mn in Nigeria, $39,9mn in Kenya and $4,9mn in Madagascar. therapies (ACTs), the first-line treatment in most malaria-endemic Annual economic costs for 2011 were $57,2mn in Nigeria, $17,7mn in countries, have been implemented in a number of settings in order to Kenya and $2,2mn in Madagascar, implying respectively an economic improve coverage of ACTs and discourage use of other treatments. The cost per capita of $0.35, $0.42 and $0.10, and per ACT dose delivered largest initiative of this type was the Affordable Medicines Facility - malaria of $1.30, $1.51 and $1.91. The ACT subsidy itself accounted for 89% (AMFm), which was implemented at a national scale in eight pilots from of economic costs in Nigeria, 80% in Kenya, and 48% in Madagascar. 2010-2013. By the end of 2013, over 310 million treatments were ordered Sensitivity analysis will be presented to explore the impact of varying key for the private for-profit sector through the initiative, and an Independent assumptions and to estimate the costs of replication in other settings. The Evaluation reported large improvements in ACT availability, price and implications for the potential value for money of strategies to expand ACT market share in six of the eight pilots. However, little is known about the use through the private sector will be discussed. causes of inter- and intra- country variations in performance. The success of private sector ACT subsidy programmes is determined by providers’ 1582 decisions on whether to stock subsidized medicines and decisions on the pricing of subsidized medicines. This study used nationally-representative INVESTIGATING THE DETERMINANTS OF DEMAND FOR outlet-survey data from Kenya, Nigeria and Uganda to model provider ANTIMALARIAL MEDICINES IN BENIN, NIGERIA, UGANDA decisions to stock ACTs subsidized through AMFm, and set markups for AND ZAMBIA the drugs. These three countries were selected, because they have diverse contexts and AMFm had differing effects. For each country, multiple 1 1 1 Benjamin Palafox , Kara Hanson , Catherine Goodman , Aurélia regression analysis was used to examine the determinants of markups 1 1 2 Lépine , Virginia Wiseman , Marcos Vera Hernandez and stocking of subsidized ACTs. The determinants investigated were a 1London School of Hygiene & Tropical Medicine, London, United Kingdom, set of product, provider and market characteristics, including measures 2University College London, London, United Kingdom of competition. The analysis addressed the endogenous and hierarchical Artemisinin-based combination therapy (ACT) is an essential health nature of the data. The evidence presented will help identify settings systems input in malaria endemic countries. By 2008, all of these countries suitable for ACT subsidies and the types of supporting interventions and had adopted ACT as the recommended first-line treatment for falciparum their targeting that are most appropriate. malaria. Although ACT may be available free of charge through public facilities, use of non-ACTs is common. This is particularly true for children 1584 under 5 years of age who may be treated presumptively where diagnostic capacity is limited. Much of these non-ACTs are purchased easily from OPTIMAL PRICE SUBSIDIES FOR APPROPRIATE MALARIA private sector retailers who may also sell ACTs, but at much higher TESTING AND TREATMENT BEHAVIOR prices. Therefore, ensuring that children receive appropriate and quality Kristian S. Hansen1, Tine H. Lesner2, Lars P. Østerdal2 treatment for malaria requires an understanding of how responsive 1 household demand is to antimalarial prices and other determinants. We London School of Hygiene & Tropical Medicine, London, United Kingdom, 2 estimated econometric demand models for antimalarials to examine the University of Southern Denmark, Odense, Denmark determinants of antimalarial choice in Benin, Nigeria, Uganda and Zambia. Malaria continues to be a serious public health problem particularly In each country, data were collected through nationally representative in Africa. Limited access to antimalarials and over-treatment with surveys of households that experienced a recent paediatric febrile episode. antimalarials are two seemingly contradicting phenomena that co-exist. This was complemented by survey data from all possible public and private A global subsidy on selected antimalarial drugs has been suggested to sources of antimalarial medicines in the vicinity of these households. increase access to the most effective treatment both in the public and Treatment choices included ACTs, oral artemisinin monotherapies, non- private sectors. In order to also reduce over-treatment we propose a artemisinin therapies, and no treatment. The range of determinants combined price subsidy on malaria rapid diagnostic tests and antimalarial studied included various treatment price components (e.g. antimalarials, drugs. Focusing on the private sector, we analyse the optimal subsidy diagnostics, travel), and characteristics of the provider, household and combination that incentivises individuals suspecting themselves to have caregiver. Our findings will focus on the most significant determinants malaria to purchase a parasitological test before buying the recommended of which antimalarial households obtain and from where, and examine treatment using an expected utility model describing the health-seeking how responsive antimalarial demand is to changes in antimalarial prices behaviour of a representative individual. Solving our model numerically for and household income. Given the considerable resources directed toward individuals with a range of different health-seeking behaviours shows that improving access to appropriate malaria treatment, we will also discuss the optimal policy of the health planner is to redirect some of the subsidy how these findings may be applied to optimise the equitable impact of money from antimalarial drugs to parasitological tests. these investments in pluralistic health systems settings. 1583 DETERMINANTS OF STOCKING AND PRICING OF SUBSIDIZED ANTIMALARIAL TREATMENTS BY RETAILERS IN THE PRIVATE FOR-PROFIT SECTOR: EVIDENCE FROM NATIONAL SUBSIDY PROGRAMS IN KENYA, NIGERIA AND UGANDA Sarah Tougher, Catherine Goodman, Kara Hanson London School of Hygiene & Tropical Medicine, London, United Kingdom There is increased interest in improving quality of care in the private for- profit sector in low and middle income countries, as private providers are

astmh.org 484 1585 exhibited >90% transmission blocking activity, while 52-81% blocking was seen at 50 ug/ml. Our results suggest that CH-rPfs25 is strongly IDENTIFYING MINIMAL EPITOPES ON THE SURFACE OF immunogenic by different routes and as formulations with Alum and NE. PLASMODIUM VIVAX DUFFY BINDING PROTEIN REACTIVE We are continuing these studies to develop effective vaccine formulations WITH NEUTRALIZING MONOCLONAL ANTIBODIES for further evaluation and investigations into immune correlates of relative immunogenicity of CH-rPfs25 in various adjuvants. Miriam T. George1, Jesse L. Schloegel2, Francis B. Ntumngia1, Samantha J. Barnes1, Edwin Chen3, Christopher L. King4, Niraj H. 1587 Tolia3, Joanne L. Casey2, Michael Foley2, John H. Adams1 1University of South Florida, Tampa, FL, United States, 2La Trobe University, ENHANCING PRE-ERYTHROCYTIC STAGE VACCINE EFFICACY Victoria, Australia, 3Washington University School of Medicine, St. Louis, WITH THE DEVELOPMENT OF A HIGHLY IMMUNOGENIC MO, United States, 4Case Western Reserve University, Cleveland, OH, VIRUS-LIKE PARTICLE VACCINE AND A MULTI-COMPONENT United States VACCINE STRATEGY Plasmodium vivax Duffy Binding protein region II (PvDBPII) is an essential Katharine A. Collins, Matthew Cottingham, Rebecca Hillson, ligand for reticulocyte invasion, thereby making this molecule an attractive Sarah Gilbert, Adrian V. Hill vaccine candidate to protect against asexual blood-stage P. vivax. Similar to other blood-stage vaccine candidates, DBP allelic variation elicits a University of Oxford, Oxford, United Kingdom strain-specific immunity that may be a major challenge for development CSP-based subunit vaccines have been shown to protect against malaria of a broadly effective vaccine against vivax malaria. This study aims to in a range of models, but to date, none have been able to elicit high identify conserved epitopes of neutralizing anti-DBP monoclonal antibodies levels of durable sterilising efficacy in human field trials. Our aim is to (mAbs) using immunochemical and structural approaches to help design enhance pre-erythrocytic stage vaccine efficacy by two methods. Firstly a strain-transcending vaccine. The crystal structure of PvDBPII consists by increasing the immunogenicity of a CSP-based virus-like particle (VLP) of 2 α-helical bundles with an antiparallel β-hairpin near the N-terminus vaccine, and secondly by combining the VLP vaccine with a liver-stage viral and may be assigned into three subdomains delineated by six disulphide vector vaccine regimen. To enhance the immunogenicity of the CSP-based bonds. The various subdomains and combination of subdomains were particle we have developed an improved RTS,S like vaccine called R21. expressed in their correctly refolded and disulphide bonded conformation RTS,S is the leading CSP-based vaccine and consists of particles formed on the surface of the M13 filamentous phage. Additionally, a PvDBPII from a mixture of two proteins, with only ~20% of the total protein gene fragment library was used for biopanning to screen phage clones content being a CSP-HBsAg fusion protein. In the R21 particles, 100% of reactive with anti-DBP mAbs. Comparative analysis of specific targets of the total protein content is a CSP-HBsAg fusion protein and hence R21 will non-inhibitory anti-DBP mAbs with neutralizing anti-DBP mAbs will help contain a much higher percentage of CSP than RTS,S. This could result in determine essential regions of PvDBP for a subunit vaccine designed to enhanced immunogenicity and efficacy and is currently under evaluation. protect against blood-stage Plasmodium vivax malaria. The immunogenicity and efficacy of R21 + adjuvant was compared to non-particulate recombinant CSP + adjuvant in BALB/c mice. R21 was 1586 found to be more immunogenic and induced 10 fold greater levels of anti-CS antibodies as well as higher frequencies of CS specific T cells than EVALUATION OF FUNCTIONAL IMMUNOGENICITY OF CSP. These serum antibody titres were also durable and were maintained PLASMODIUM FALCIPARUM TRANSMISSION-BLOCKING at high titres when measured 3 months after vaccination. R21 was also ANTIGEN PFS25 PRODUCED IN E. COLI ADJUVANTED IN significantly more protective than non-particulate CSP in a BALB/c model VARIOUS NANOPARTICLES AND ADJUVANTS against P. berghei transgenic for P. falciparum CSP. Vaccination with R21 + Matrix M sterilely protected 82.5% of mice compared to only 42.5% with 1 1 2 2 Rajesh Kumar , Geetha Bansal , Grace Ledet , Richard Graves , CSP + Matrix M (p = 0.014). R21 was also assessed in combination with 2 1 1 1 Tarun Mandal , Dibyadyuti Datta , Evelina Angov , Nirbhay Kumar the ChAd63 ME.TRAP - MVA ME.TRAP vaccine regimen and there was 1Tulane University School of Public Health & Tropical Medicine, New no interference with the induction of vaccine specific immune responses Orleans, LA, United States, 2Center for and Drug Delivery, when the vaccines were mixed and administered together. In addition, Xavier University College of Pharmacy, New Orleans, LA, United States sterile efficacy against sporozoite challenge was significantly enhanced in Pfs25, expressed on the surface of gametes, zygotes and ookinetes, is the mixed vaccine group. R21 is now being taken forward for evaluation in an established target antigen for malaria transmission blocking vaccines. Phase I/IIa clinical trials. Previously, we have reported that codon harmonized recombinant Pfs25 (CH-rPfs25) produced in E. coli elicited highly potent transmission 1588 blocking antibodies using Montanide ISA51, Alum and CFA as adjuvants. In the current study, we have undertaken extensive evaluation of various PROJECTED COST-EFFECTIVENESS OF RTS,S VACCINATION IN nanoparticles/adjuvants via different routes of immunization to identify 43 SUB-SAHARAN AFRICAN COUNTRIES safer and effective adjuvants. Mice were immunized with CH-rPfs25 via Katya Galactionova, Fabrizio Tediosi, Thomas A. Smith, Melissa IM route (10 μg, three doses at 4 week intervals). Mice immunized with A. Penny CHrPfs25 in Alum via IP and IM routes induced comparable antibody Swiss Tropical and Public Health Institute, Basel, Switzerland titers (640,000). Since the protein was equally immunogenic by IP and IM routes, other adjuvant formulations were tested by IM route only. CH- Phase III trials of the RTS,S malaria vaccine are generating new estimates rPfs25 was adsorbed to nano-emulsions (4% & 8%NE) and PLGA particles of its efficacy and decay rate. To assess the appropriateness of introducing (2 different amounts). CH-rPfs25 formulated in 4% NE gave highest RTS,S into the EPI in endemic African countries cost-effectiveness estimates antibody response (ELISA titer 1,280,000) as compared to 320,000 in 8% need to be updated with the most recent clinical efficacy data. We NE. Antibody titers with PLGA (10 and 20 mg PLGA) were only 160,000. used methods for rapidly updating projections of country specific public We also evaluated NE formulations combined with MPLA and chitosan, health impact of the vaccine, and combined these with costing models and both demonstrated 640,000 antibody titers. Functional activity of for malaria case management and immunization to assess the cost- antibodies was evaluated by standard membrane feeding assay using effectiveness of RTS,S introduction. We consider deployment with a 3 dose purified IgG (50- 400 μg/ml) from immunized animals. 100 % transmission schedule targeting infants 6, 10 and 14 weeks of age, an older cohort blocking activity (no oocysts detected) was observed at 400 ug/ml of vaccinated at 6, 7.5 and 9 months, and a 4 dose schedule as above, IgG from Alum group (both routes IP and IM), NE (4%), and NE-MPLA. which includes a booster at 18 months after the third dose. Allowing for Purified IgG from various adjuvant groups at lower doses (100 μg/ml) still differences in epidemiological context and health systems we generate

astmh.org 485 predictions tailored to countries examined and apt to inform malaria 1590 control policy. An ensemble of individual-based stochastic simulation models of Plasmodium falciparum dynamics, with varied assumptions EFFICACY, SAFETY AND IMMUNOGENICITY OF about immune decay, transmission heterogeneity, and access to treatment HETEROLOGOUS PRIME-BOOST IMMUNIZATION WITH THE fit to an extensive library of field data were used to predict the impact of CANDIDATE MALARIA VACCINES CHAD63 ME-TRAP AND RTS,S. For each country average and incremental cost-effectiveness ratios MVA ME-TRAP IN 5-17 MONTHS OLD BURKINABE INFANTS were calculated relative to the routine case management and alternate AND CHILDREN vaccine deployment strategies. We show that RTS,S is likely to be a highly cost-effective intervention with a significant impact on malaria burden Alfred B. Tiono1, Issa N. Ouedraogo1, Jean-Baptist Yaro1, Katie in endemic settings and cost per DALY averted generally comparable to Ewer2, Edith C. Bougouma1, Issiaka Soulama1, Rachel Roberts2, routine malaria control interventions. Depending on vaccine properties, Amidou Diarra1, Alphonse Ouedraogo1, Alison Lawrie2, Sam coverage, and age-related disease burden vaccinating younger cohorts Coulibaly1, Thomas Rampling2, Carly Bliss2, Aïssata Barry1, Ines may avert more disease and at a lower cost compared to older age groups, Petersen3, Georgina Bowyer2, Nick Anagnostou2, Nicola Viebig3, even if initial efficacy is lower. As the vaccine targets only a small fraction Egeruan Babatunde Imoukhuede2, Philip Bejon4, Adrian Hill2, of the population susceptible to malaria and provides limited protection Sodiomon B. Sirima1 it does not eliminate the need for other control programs. Access to 1Centre National de Recherche et de Formation sur le Paludisme, effective treatment is particularly important to sustain health gains Ouagadougou, Burkina Faso, 2University of Oxford, Oxford, United achieved with the RTS,S. Kingdom, 3European Vaccine Initiative, Heidelberg, Germany, 4Kenya Medical Research Institute, Kilifi, Kenya 1589 The development of an effective vaccine against malaria is a high priority and of great importance in the context of coordinated efforts to reduce METHODOLOGY TO ESTIMATE THE COST OF INTRODUCING the burden of malaria. The protective efficacy of the candidate malaria RTS,S VACCINE INTO A NATIONAL IMMUNIZATION vaccines ChAd63 ME-TRAP and MVA ME-TRAP is being evaluated in an PROGRAM IN SUB-SAHARAN AFRICAN COUNTRIES ongoing phase I/IIb double-blind randomized trial. This immunization Katya Galactionova, Fabrizio Tediosi regime has shown significant partial efficacy in controlled human malaria infection trials and an initial time-to-infection trial in adults in Kenya. Swiss Tropical and Public Health Institute, Basel, Switzerland Initial efficacy results from this first efficacy trial in African infants will be Having demonstrated moderate levels of efficacy in Phase II and III trials available in May 2014. The primary objective is to evaluate the protective in Africa, the RTS,S vaccine against P. falciparum malaria is currently efficacy against clinical malaria, for a period of 6 months after the last considered for use within the Expanded Programme on Immunization vaccination. Clinical malaria is defined as fever (axillary temperature in a large number of endemic settings. Introduction of a new vaccines ≥37.5°C) together with Plasmodium falciparum count > 5,000 p/µl. Seven demands extensive resources from the health system. Aside from the hundred infants and children aged 5-17 months were randomized in 1:1 vaccine and related immunization supplies, resources are required for ratio to receive either ChAd63 ME-TRAP / MVA ME-TRAP in prime-boost storage and supply chain, training, development of educational materials, immunization or rabies vaccine. Immunization schedule was 0, 8 weeks. social mobilization, supervision, monitoring, vaccine delivery, and waste Clinical malaria episodes were captured through a health-facility based management. We propose a generalizable methodology to estimate passive case surveillance method. Venous blood samples for cellular and these costs related to vaccine introduction in African countries. Costs humoral immunogenicity were collected at various timepoints. Vaccine are evaluated from a broad provider perspective using the ingredients efficacy will be assessed using Cox regression models. For analysis of first approach; these reflect the economic value of resources, and take or only episodes of P. falciparum malaria, the incidence of episodes for into account overheads and cost of inputs shared with other health each group will be presented. Secondary analysis will examine multiple interventions. To address the uncertainty about the level of existing episodes, using the robust clustering method by individual. Analysis of capacity in the health system we consider several states of the EPI: no vaccination immunogenicity will describe the arithmetic and geometric spare capacity, estimated current capacity, and sufficient spare capacity means and median spots per million PBMC by vaccine group and to accommodate the new vaccine. At each capacity level we develop a timepoints. The safety analysis will include all solicited and unsolicited series of normative scenarios for service delivery and capacity scale-up in local and systemic adverse events including clinically significant laboratory accordance with the current operational guidelines. Scenarios are adapted abnormalities, and serious adverse events. These results will help define to a given country setting to take into account among other the structure the potential role of these recombinant viral vectors, either used alone or of the EPI program, distribution model, geography, and population as part of a multi-component vaccine, in malaria control in Africa. dynamics. Resource lists and quantity assumptions defined for each immunization scenario are matched with price and unit cost data via cost 1591 functions to assess the overall cost of the program. The methodology takes advantage of country data on prices of key inputs using routinely collected ANTIBODIES AGAINST A PLASMODIUM FALCIPARUM data from the cMYP, UNICEF, and WHO-CHOICE. The methodology is RHOPTRY NECK PROTEIN PFRON12 INHIBIT MEROZOITE applied to assess cost of RTS,S introduction in 6 endemic countries. We INVASION INTO ERYTHROCYTES test the robustness of estimates generated by varying core assumptions and prices of key inputs and validate against the literature on cost of EPI Daisuke Ito1, Eizo Takashima1, Shinya Hatano1, Tsutomu Yamasaki2, program and introduction of other new vaccines. Tomoyuki Hasegawa1, Kazutoyo Miura3, Amporn Thongkukiatkul4, Carole A. Long3, Jetsumon Sattabongkot5, Motomi Torii6, Takafumi Tsuboi1 1Ehime University, Matsuyama, Ehime, Japan, 2Okayama University of Science, Okayama, Japan, 3National Institute of Allergy and Infectious Diseases, Rockville, MD, United States, 4Burapha University, Chonburi, Thailand, 5Mahidol University, Bangkok, Thailand, 6Ehime University, Toon, Ehime, Japan Proteins coating Plasmodium merozoite surface and secreted from its apical organelles are considered as promising vaccine candidates for the blood-stage malaria. The rhoptry neck protein 12 of Plasmodium falciparum (PfRON12) was recently reported as a protein specifically

astmh.org 486 expressed in schizont and localized to the rhoptry neck of merozoite by and extremely well-tolerated. This is a 1st step toward establishing immunoelectron microscopy (IEM). The characteristics of the PfRON12 the conditions for operational implementation and logistics for mass suggest that it is a potential blood-stage vaccine candidate, and here we administration of PfSPZ Vaccine in Africa. assessed its potential in this regard. We expressed a recombinant PfRON12 protein by the wheat germ cell-free system to obtain anti-PfRON12 1593 antibody. Immunoblot analysis of schizont lysate detected a single band at approximately 40 kDa under reducing condition, consistent with the CHLOROQUINE NEITHER ELIMINATES NOR DELAYS predicted molecular weight. In contrast, the anti-PfRON12 antibody LIVER STAGE DEVELOPMENT OF PLASMODIUM DURING recognized a single band at approximately 80 kDa under non-reducing CHEMOPROPHYLAXIS VACCINATION condition, consistent with two-fold molecular weight of its reduced 1 2 2 form, suggesting the native PfRON12 forms a disulfide-bond-mediated Tejram Sahu , Lynn E. Lmbert , Jessica Hinderer , Solomon 1 2 1 homodimer. Immunofluorescence assay and IEM revealed that PfRON12 Conteh , Sachy Orr Gonzalez , Patrick E. Duffy localized to the rhoptry neck of merozoite in schizonts and to the surface 1National Institute of Allergy and Infectious Diseases, Rockville, MD, United of free merozoites. The biological activity of anti-PfRON12 antibody States, 2National Institute of Allergy and Infectious Diseases, Bethesda, was tested by an in vitro growth inhibition assay, and the antibody MD, United States significantly inhibits the merozoite invasion of erythrocytes. Since anti- Chloroquine (CQ) has been used in Chemoprophylaxis Vaccination (CVac) PfRON12 antibody inhibited the merozoite invasion in vitro, we decided against malaria, whereby sporozoite inoculations under the umbrella of to investigate whether PfRON12 is exposed to the human immune system CQ prophylaxis induce liver stage-specific immunity and long-lasting sterile in P. falciparum-infected individuals. The sera from P. falciparum infected protection against homologous parasite strain. CQ is known to kill blood individuals in Thailand and Mali reacted with the recombinant PfRON12, stage parasites, but its effect on liver stages is poorly studied, and the indicating PfRON12 is immunogenic in humans. Our findings suggest mechanism by which CVac-CQ induces strong protective immunity is not that PfRON12 plays an important role in the merozoite invasion process, understood. We used a luciferase expressing rodent parasite, Plasmodium and that it merits an additional evaluation as a P. falciparum blood-stage yoelii-Luc (Py-Luc), to monitor the effect of CQ on Plasmodium liver stage vaccine candidate. development. Balb/c mice with or without CQ prophylaxis were infected with Py-Luc sporozoites. Primaquine (PQ), a liver stage-specific anti- 1592 parasitic drug, was included as a positive control. We followed parasite development by intra-vital imaging at 44h, 54h and 60h post-infection. ADMINISTRATION OF PFSPZ VACCINE BY DIRECT VENOUS Parasite burden in liver was measured by quantifying bioluminescence INOCULATION IN AFRICA of whole body and isolated livers, as well as by quantifying liver stage parasite transcripts by qRT-PCR. Delay in appearance of parasites in the Freda Omaswa1, Sara Healy2, Mahamadou S. Sissoko3, blood was monitored by microscopic observation of Giemsa-stained thin Abdoulaye Katile3, Amagana Dolo3, Karamoko Niare3, Amadou blood smears. The parasite load in livers of CQ treated and untreated mice Niangaly3, Peter Billingsley4, Anita Manoj4, Eric James4, B. Kim Lee did not differ (p=0.714), and this was consistent at all three timepoints. PQ Sim4, Michael Walther2, Ogobara Doumb Doumbo3, Patrick Duffy2, treated mice had a significant reduction in parasite burden as compared to Stephen L. Hoffman4 both CQ treated and untreated groups (p=0.008), and were similar to the 1 2 University of British Columbia, Vancouver, BC, Canada, Laboratory of non-infected control mice. Parasites appeared in the blood stream of both Malaria Immunology and Vaccinology, National Institute of Allergy and CQ treated and untreated mice at the 54h time point. Taken together, our Infectious Diseases, National Institutes of Health, Rockville, MD, United findings indicate that CQ neither eliminates liver stage parasites nor delays 3 States, Malaria Research and Training Center, Mali-NIAID ICER, University their development. Further investigations into the mechanisms by which of Science, Techniques and Technologies of Bamako, Bamako, Mali, CVac-CQ induces protective immunity are required, and may give insights 4 Sanaria Inc., Rockville, MD, United States relevant to drug and vaccine development. Malaria is a major cause of morbidity and mortality despite considerable investment in existing anti-malarial measures, highlighting the need 1594 for a vaccine. The vaccine candidate, Sanaria® PfSPZ Vaccine, contains radiation-attenuated, aseptic, purified, cryopreserved Plasmodium DEVELOPMENT OF A METABOLICALLY ACTIVE, NON- falciparum (Pf) sporozoites (SPZ). Healthy 18-35 year Malians were REPLICATING, ASEPTIC, PURIFIED, CRYOPRESERVED, enrolled in a randomized, double-blind, placebo-controlled trial to assess GENETICALLY ATTENUATED PLASMODIUM FALCIPARUM safety, tolerability, immunogenicity, ease of administration, and protective SPOROZOITE VACCINE-PFSPZ (∆SLARP∆B9) VACCINE efficacy against naturally occurring malaria of PfSPZ Vaccine administered 1 1 2 by direct venous inoculation (DVI). PfSPZ Vaccine was thawed, diluted Adam M. Richman , B. Kim Lee Sim , Ben C. van Schaijk , 3 2 1 1 to a 0.5 mL volume in a 1 mL syringe with a 25-gauge needle, and Takeshi Annoura , Ivo H. Ploemen , Tao Li , Abraham Eappen , 1 1 1 then passed to local for administration. Local and systemic Patricia de la Vega , Sumana Chakravarty , Adam J. Ruben , 3 3 2 reactogenicity were solicited through 7 days after each vaccination. A Shahid M. Khan , Chris J. Janse , Robert W. Sauerwein , Stephen 1 survey on subject perception of vaccination procedure will be administered L. Hoffman at the end of the vaccination phase in July 2014. In total, 105 volunteers 1Sanaria Inc., Rockville, MD, United States, 2Department of Medical have been vaccinated, 12 were enrolled in a pilot safety group and 93 Microbiology, Nijmegen, Netherlands, 3The Leiden Malaria Research were randomized to receive 5 doses of 2.7x105 PfSPZ or normal saline Group, Parasitology, Leiden, Netherlands placebo. At this time, 207 DVIs with PfSPZ Vaccine or placebo have been Sanaria® PfSPZ Vaccine, composed of radiation attenuated, aseptic, administered. The time from vaccine request by the clinical team to thaw purified, cryopreserved Plasmodium falciparum (Pf) sporozoites (SPZ) and formulation in a syringe to completion of injection was on average 6 protected 6/6 (100%) of volunteers, who received the highest dose. minutes and injections on average took < 10 sec. Of the 207 vaccinations, By mid 2014 PfSPZ Vaccine will be in clinical trials in the U.S. and 5 only one has required a second injection attempt. This was for inability to countries in Africa and Europe. It is intended for elimination campaigns st locate a vein for the 1 dose in the pilot group. Vaccinations have been and prevention of malaria in travelers. There would be potential well tolerated with no local reactogenicity. Seven episodes of solicited manufacturing, potency and regulatory advantages if radiation-attenuated systemic reactogenicity, all grade 1, have been reported. Complete data parasites were replaced with genetically attenuated parasites (GAPs). on safety, tolerability, and administration after 5 doses of PfSPZ Vaccine We recently reported on a new target gene, b9 and its deletion in will be presented. DVI administration of PfSPZ has been rapid, efficient combination with the slarp gene. In rodent malarias ∆slarp∆b9 SPZs elicit excellent protective immunity and do not lead to blood stage infection.

astmh.org 487 Elimination of Pf slarp and b9 genes leads to attenuation similar to 1596 radiation. All prior Pf GAPs showed leaky attenuation and breakthrough liver stage development in vivo or in vitro. After characterization in GENETIC DIVERSITY OF THE PLACENTAL MALARIA VACCINE Nijmegen and Leiden, the parasites were transferred to Sanaria where CANDIDATE VAR2CSA IN TWO MALARIA ENDEMIC SETTINGS master and working cell banks (MCB and WCB) were made and an IN AFRICA USING PACBIO SEQUENCING engineering production run performed to demonstrate that Pf∆slarp∆b9 1 1 1 GAP was suitable for producing aseptic, purified, cryopreserved PfSPZ. Antoine Dara , Mark A. Travassos , Matthew Adams , Sarah 1 2 1 The parasites demonstrated all growth characteristics necessary for Schaffer DeRoo , Joana C. Silva , Christopher V. Plowe , Miriam K. 1 cGMP production. The Pf∆slarp∆b9 SPZ were assayed in Sanaria’s 6-day Laufer hepatocyte attenuation, 3-day hepatocyte potency, and sporozoite 1Howard Hughes Medical Institute/Center for Vaccine Development, membrane integrity (viability) assays. For the 6-day assay we used Pf University of Maryland, Baltimore, MD, United States, 2Institute of wild type and Pf∆p52∆p36 SPZ as positive controls; the wild type Pf, Genome Sciences and Department of Microbiology and immunology, Pf∆p52∆p36, and Pf∆slarp∆b9 SPZ produced 21+1, 1.5+1.25 and 0, University of Maryland, Baltimore, MD, United States 6-day liver stage schizonts respectively. Pf∆slarp∆b9 SPZ were potent and Pregnancy-associated malaria, a leading cause of maternal anemia and viable. We will next use this genetically attenuated double-mutant parasite low birth weight, is characterized by the sequestration of Plasmodium (Pf∆slarp∆b9) to manufacture, characterize and release a corresponding falciparum-infected erythrocytes in the placental microvasculature due to PfSPZ Vaccine, PfSPZ (∆slarp∆b9) Vaccine (also known as PfSPZ-GA1 expression of VAR2CSA. Epidemiological and serological studies indicate Vaccine) in compliance with cGMPs, conduct pre-clinical studies, submit to that VAR2CSA is a target of naturally acquired immunity, suggesting the appropriate U.S. and Dutch regulatory agencies, and conduct a clinical that VAR2CSA could be a potential target for a pregnancy-associated trial. malaria vaccine. However, based on limited data, there appears to be extensive genetic diversity within the var2csa gene that must be taken 1595 into account in the design of an effective vaccine. Genetic diversity has prevented the successful sequencing of var2csa from field isolates using FINE MAPPING OF ANTIBODY ISOTYPES AND standard sequencing platforms. Due to the low sequence complexity IMMUNODOMINANT B CELL EPITOPES INDUCED BY and high variant diversity of var2csa, applying a traditional Sanger MALARIA VACCINE, PFCELTOS/GLA-SE sequencing strategy on field samples is inefficient and costly. To overcome Casey K. Storme1, Sidhartha Chaudhury2, Sathit Pichyangkul3, this obstacle, we are characterizing var2csa genetic diversity in malaria Susan L. Baldwin4, Elizabeth Duncan1, Heather Hosie1, Darrick parasite isolates from two different endemic regions in Africa using a Carter4, Steven G. Reed4, Anders Wallqvist2, Christian F. combination of long range PCR amplification and the Pacific Bioscience Ockenhouse1, Jessica Cowden1, Elke Bergmann-Leitner1, Randall F. next generation sequencing platform. We performed a multiple alignment Howard4, Evelina Angov1 of the publicly available 20 coding sequences and 12 upstream promoter region sequences of var2csa, and designed primers based on three 1 Walter Reed Army Institute of Research, Silver Spring, MD, United States, of the 40 identified conserved regions with a minimum length of 25 2 Biotechnology HPC Software Applications Institute, Telemedicine and nucleotides. A first set of primers targets a 5 kilobase (kb) region spanning Advanced Technology Research Center, Fort Detrick, MD, United States, the upstream promoter region to the DBLepam4 domain, and a second 3 Armed Forces Research Institute of Medical Sciences, U.S. Army Medical primer set targets a 5 kb region spanning the DBLpam3 domain to the 4 Component, Bangkok, Thailand, Infectious Disease Research Institute, intracellular acidic terminal segment region. We have successfully amplified Seattle, WA, United States the two fragments for the reference strains 3D7 and NF54 and the first The secreted malarial protein Cell-Traversal protein for Ookinetes and fragment from the Dd2 and HB3 strains. We are employing this approach Sporozoites (CelTOS) is highly conserved among Plasmodium species, to determine the var2csa sequence of clinical isolates and characterize the essential to host hepatocyte invasion, and critical to malaria pathogenesis. extent of natural diversity in var2csa in Mali and Malawi. We previously reported that immunization of mice with full-length recombinant CelTOS from P. falciparum (PfCelTOS) adjuvanted in 1597 Montanide ISA-720 achieved 60% heterologous protection against a P. berghei sporozoite challenge. The immune mechanisms leading to this SIMULATION OF B CELL AFFINITY MATURATION EXPLAINS cross-species protection are based on both cellular and humoral effector ENHANCED ANTIBODY CROSS-REACTIVITY INDUCED BY THE mechanisms. Formulating PfCelTOS with the clinically relevant adjuvant POLYVALENT MALARIAL VACCINE AMA1 GLA-SE resulted in similar levels of protection in mice against challenge as seen with ISA-720, and these studies provided evidence to support its Sidhartha Chaudhury, Jaques Reifman, Anders Wallqvist evaluation in a Phase 1 safety, immunogenicity with Controlled Human BHSAI, U.S. Army Medical Research and Materiel Command, Fort Detrick, Malaria Infection (CHMI) clinical trial. Since protection is in part mediated MD, United States by antibodies, we sought to identify protective B-cell epitopes and, Polyvalent vaccines use a mixture of antigens representing distinct thereby, we determined the antibody fine specificity of preclinical and pathogen strains to induce an immune response that is cross-reactive clinical samples for PfCelTOS. To this end, various protein fragments were and protective. However, such approaches often have mixed results, and generated in E. coli or as synthesized peptides, and their reactivity with it is unclear how polyvalency alters the fine specificity of the antibody sera from PfCelTOS/GLA-SE immune mice, rats, non-human primates and response and what those consequences might be for protection. Here, human subjects was determined. We are currently implementing in vitro we present a coarse-grain theoretical model of B cell affinity maturation functional assays such as inhibition of sporozoite gliding motility and the during monovalent and polyvalent vaccinations that predicts the fine inhibition of sporozoite invasion and development within hepatocytes specificity and cross-reactivity of the antibody response. We stochastically (ILSDA) to identify whether this vaccine formulation induces responses to simulate affinity maturation using a population dynamics approach where functional B-cell epitopes within the identified immunodominant regions the host B cell repertoire is represented explicitly, and individual B cell sub- of CelTOS. Such characterizations will determine the role of these epitopes populations undergo rounds of stimulation, mutation, and differentiation. in mediating antibody-mediated protection from preclinical and clinical Antigens contain multiple epitopes and are present in subpopulations of studies of the CelTOS antigen. distinct pathogen strains, each with varying degrees of cross-reactivity at the epitope level. This epitope and strain-specific model of affinity maturation enables us to study the composition of the polyclonal response in granular detail and identify the mechanisms driving serum specificity and cross-reactivity. We applied this approach to predict the antibody

astmh.org 488 response to a polyvalent vaccine based on the highly polymorphic malarial 1599 antigen AMA1. Our simulations show that polyvalent AMA1 vaccination induces an enhanced cross-reactive antibody response primarily through PHASE 1 MALARIA VACCINE STUDIES IN AFRICA: WHAT a shift in affinity maturation that favors B cells specific to shared and DEFINES A NORMAL, HEALTHY VOLUNTEER? cross-reactive epitopes, and demonstrates how a polyvalent vaccine with 1 2 2 a small number of strains and only moderate allelic coverage may be Sara A. Healy , Issaka Sagara , Mahamadou S. Sissoko , 2 3 4 broadly neutralizing. These results present broad implications for general Merepen A. Guindo , Freda Omaswa , Joseph P. Shott , Stephen L. 3 2 1 polyvalent vaccine design. Hoffman , Ogobara Doumbo , Patrick E. Duffy 1Laboratory of Malaria Immunology and Vaccinology, National Institute 1598 of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, United States, 2Malaria Research and Training Center, Mali-NIAID HEPATITIS B, HEPATITIS C AND HIV INFECTION FREQUENCIES ICER, University of Science, Techniques and Technologies of Bamako, AMONG VOLUNTEERS SCREENED FOR MALARIA VACCINE Bamako, Mali, 3Sanaria, Inc., Rockville, MD, United States, 4Division of CLINICAL TRIALS IN MALI Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States Merepen A. Guindo1, Moussa L. Diakite1, Boucary Ouologuem1, As the malaria vaccine landscape continues to expand with new Souleymane Traore1, Amadou Konate1, Abdoulaye Katile1, candidates entering phase 1 trials in Africa, the concept of a normal, Yacouba Samake1, Bourama Kamate1, Sarro Yeya2, Mahamadoun healthy volunteer becomes difficult to consistently and accurately define H. Assadou1, Mahamadou S. Sissoko1, Issaka Sagara1, Boubacar in these settings. Inclusion/exclusion criteria and toxicity grading for these Traore1, Joseph P. Shott3, Sara A. Healy4, Stephen L. Hoffman5, studies can vary significantly between sites, impacting adverse event Patrick E. Duffy4, Ogobara Doumbo1 reporting, study stopping criteria, immunogenicity responses, known 1 Malaria Research and Training Center, Mali-NIAID ICER, University of susceptibility to malaria infection, and severity of disease. From May 2013 2 Science, Techniques and Technology, Bamako, Mali, SEREFO, University of to February 2014, 509 volunteers were screened for either a phase 1 3 Science, Techniques and Technology, Bamako, Mali, Division of Intramural malaria transmission blocking vaccine trial or a phase 1 whole sporozoite Research, National Institute of Allergy and Infectious Diseases, National malaria vaccine trial in Mali. All volunteers were screened by history, 4 Institutes of Health, Bethesda, MD, United States, Laboratory of Malaria physical examination, and standard laboratory testing (hematological Immunology and Vaccinology, National Institute of Allergy and Infectious and biochemical parameters, urinalysis, Hepatitis B/C virus and HIV 5 Diseases, Rockville, MD, United States, Sanaria Inc., Rockville, MD, United testing) with consistent screening ratios of 2 to 2.5 volunteers screened States to 1 volunteer enrolled. However, other exclusion criteria, such as Two clinical trials of malaria vaccine candidates (transmission blocking sickle cell disease/trait, electrocardiogram abnormalities, helminthiases, vaccine Pfs25-EPA/Alhydrogel® in collaboration with LMIV-NIAID) and schistosomiasis, and syphilis were not consistently evaluated nor whole organism vaccine PfSPZ (in collaboration with LMIV-NIAID and universally managed prior to enrollment, potentially creating variability Sanaria) have begun in Bancoumana and Doneguebougou, Mali. Both in adverse event reporting. The impact on immunogenicity responses, villages are located within 70km of Bamako, but are different in terms of malaria infection, and disease severity given the inconsistency in the ethnicity, surrounding terrain, and primary employment. We have screened definition of a healthy volunteer in this population prior to enrollment, 509 men and women volunteers aged 18-45 years in Bancoumana and is to be determined in these trials. Defining standard inclusion and 18-35 years in Doneguebougou. Screening has been conducted to exclude exclusion criteria increases the likelihood of producing reliable and those with human immunodeficiency virus (HIV) 1 and 2, hepatitis B virus reproducible results, but must be closely balanced with the research being (HBV; by hepatitis B surface antigen (HBsAg)) and hepatitis C (HCV; by representative of the general healthy population under study. The creation anti-HCV core antibodies)). For HCV and HBV testing, AccuDiag ELISA of extremely narrow inclusion exclusion criteria can have significant was used in Doneguebougou and standard rapid diagnostic tests (RDT) in implications on the ability of the study to be representative of the Bancoumana. Determine HIV1/2 Alere® RDT with positivity confirmation by population under study and generalizability of the research results. ELISA GENSCREEN ULTRA Ag-Ab were used at both sites for HIV 1/2. Ten of 204 (4.9%; 95%CI [2.37-8.82]), and ten of 218 (4.6%; 95% CI [2.22- 1600 8.27]) volunteers were positive by HIV RDT respectively in Doneguebougou and Bancoumana. Four (2.0%; 95% CI [0.54-4.94])and five (2.3%; 95% A GENOMIC EPIDEMIOLOGY APPROACH TO ASSESSING AND CI [0.75-5.27]) were confirmed positive by HIV ELISA GENSCREEN ULTRA IMPROVING STRAIN-SPECIFIC WHOLE ORGANISM VACCINE Ag-Ab. HIV prevalence rates were comparable (p> 0.05) at the two sites. EFFICACY Forty-six volunteers (22.5%; 95% CI [17.01-28.90]) and 21 volunteers 1 2 2 1 (9.6%; 95% CI [6.06-14.35]) were HBsAg positive respectively in Kara Moser , Mark Travassos , Amed Ouattara , Elliot Drábek , 1 1 3 3 Doneguebougou and Bancoumana, while six (2.9%; 95% CI [1.08-6.29]) Cesar Arze , Claire M. Fraser , Kim Lee Sim , Stephen L. Hoffman , 4 4 and four (1.8%; 95%CI [0.5-4.63]) were anti-HCV positive at the sites Ogobara K. Doumbo , Mahamadou A. Thera , Shannon Takala- 2 1 2 respectively. The HBsAg positive frequency in Doneguebougou is higher Harrison , Joana C. Silva , Christopher V. Plowe than in Bancoumana, and higher than in previous studies, indicating that 1Institute for Genome Sciences, University of Maryland, Baltimore, MD, the prevalence of HBV infections may be increasing in this area. Together United States, 2Howard Hughes Medical Institute/Center for Vaccine these viral diseases are an important consideration in the screening process Development, University of Maryland School of Medicine, Baltimore, for malaria vaccine trials. MD, United States, 3Sanaria Inc., Rockville, MD, United States, 4Malaria Research and Training Center, University of Sciences, Techniques and Technology of Bamako, Bamako, Mali Genetic diversity in Plasmodium falciparum is an obstacle for broadly efficacious malaria vaccines. A subunit blood stage malaria vaccine provides allele-specific efficacy, preventing only clinical malaria caused by parasites identical to the vaccine antigen at key polymorphic loci. A similar phenomenon may apply to whole-organism vaccines, albeit potentially at many, currently unknown, loci throughout the parasite genome. The attenuated whole-organism malaria vaccine PfSPZ Vaccine, which protects against homologous challenge, is based on the African strain NF54, the parent stock of the reference P. falciparum strain, 3D7. Controlled human

astmh.org 489 malaria infection trials to assess heterologous protection will initially use 1602 the South American strain 7G8. To determine how NF54 and 7G8 relate to genetic variation in natural populations, whole genome sequence data INJECTION OF PURIFIED, ASEPTIC CRYOPRESERVED from NF54, 7G8 and from twelve mono- or polyclonal clinical samples of PLASMODIUM FALCIPARUM SPOROZOITES (PFSPZ P. falciparum from a Malian village were analyzed. SNPs for all 12 Malian CHALLENGE) IS AN ALTERNATIVE TO MOSQUITO BITE strains, NF54 and 7G8 were called against the 3D7 genome using GATK. ADMINISTRATION FOR CONTROLLED HUMAN MALARIA Using the most reliable of the SNP filters tested, an average of 37,000 INFECTION (CHMI) SNPs were called for Malian strains. In contrast, 383 and 17,667 SNPs were called for NF54 and 7G8, respectively. As expected, NF54 was nearly Thomas L. Richie1, Peter F. Billingsley1, B. Kim Lee Sim1, Eric identical to 3D7, and the Malian strains were on average considerably R. James1, Meta Roestenberg2, Else M. Bijker3, Kirsten E. Lyke4, more dissimilar genetically from 3D7, and by proxy from NF54, than is Matthew B. Laurens4, Susanne H. Hodgson5, Adrian V. Hill5, Seif 7G8. Principal coordinate analysis was used to compare genetic diversity Shekalaghe6, Salim Abdulla6, Benjamin Mordmüller7, Peter G. between strains, both at the genome-wide level and in a subset of 26 Kremsner7, Gloria P. Gómez-Pérez8, Almudena Legarda8, Pedro L. single-copy antigenic genes. This analysis placed NF54 centrally among the Alonso8, Bernhards Ogutu9, Kevin Marsh10, Robert W. Sauerwein3, 12 Malian samples, suggesting a fairly representative genetic composition. Stephen L. Hoffman1 NF54 and 7G8 clustered tightly with one another and with a subset of 1Sanaria, Inc., Rockville, MD, United States, 2Leiden University Medical the monoclonal strains, possibly representing an artifact of polyclonality. Center, Leiden, Netherlands, 3Radboud University Medical Center, Principal coordinate analysis of single-copy antigens showed some of the Nijmegen, Netherlands, 4Center for Vaccine Development, University of field isolates to be more distant from NF54 than 7G8 is, suggesting that Maryland School of Medicine, Baltimore, MD, United States, 5The Jenner additional suitable challenge strains can be easily identified. Institute, University of Oxford, Oxford, United Kingdom, 6Ifakara Health Institute, Bagamoyo, United Republic of Tanzania, 7Tropical Medicine 1601 Institute, University of Tübingen, Tübingen, Germany, 8Barcelona Centre for International Health Research (CRESIB), Hospital Clínic - University of HUMANIZED DRAG MICE SUSTAIN THE VERTEBRATE Barcelona, Barcelona, Spain, 9Centre for Clinical Research, Kenya Medical LIFE CYCLE OF PLASMODIUM FALCIPARUM AND ELICIT Research Institute, Nairobi, Kenya, 10KEMRI Wellcome Trust Research PARASITE-SPECIFIC IMMUNE RESPONSES Programme, Kilifi, Kenya Wathsala Wijayalath1, Sai Majji1, Eileen F. Villasante1, Teodor D. Plasmodium falciparum (Pf) sporozoite (SPZ)-infected mosquitoes for Brumeanu2, Thomas L. Richie1, Sofia Casares1 controlled human malaria infections (CHMI) are produced routinely only in the USA and the Netherlands, by a small number of restricted-access 1Naval Medical Research Center/Walter Reed Army Institute of Research, insectaries. In the 28-day CHMI procedure, healthy adults are infected with Silver Spring, MD, United States, 2Uniformed Services University of Health malaria by the bites of 5 PfSPZ-infected mosquitoes and followed to assess Sciences, Bethesda, MD, United States efficacy of antimalarial drugs or vaccines. Although CHMI permits critical Plasmodium falciparum is one of the deadliest protozoan parasites decisions regarding product advancement without the need for expensive among the five species of human malaria, which accounts for the and lengthy field evaluations, its use is limited by the requirement for a highest morbidity and mortality in tropical and sub-tropical countries. secure insectary capable of producing infected mosquitoes. This restricts In addition of having multiple life cycle stages in the human host, P. performance of CHMI to local sites or requires transport of PfSPZ-infected falciparum parasites shows high antigenic diversity and cytoadherence mosquitoes to sites with a mosquito-secure facility. Costs may exceed properties, which increases the severity and complexity of the disease. $100K per CHMI. These limitations have been largely bypassed by Numerous efforts have been conducted over decades to address disease Sanaria® PfSPZ Challenge, a cGMP product consisting of highly purified, pathogenesis, immunity, and vaccine development, mostly in in-vitro aseptic, cryopreserved PfSPZ for parenteral use that is easily stored and or in rodent or non-human primate models. However, none of these transported to distant sites. Seven trials enrolling 178 volunteers have been models completely represent the disease, as it is in the natural human conducted to test the safety, tolerability and infectivity of PfSPZ Challenge host, thus demanding the necessity of developing an accurate animal given by intradermal (ID), intramuscular (IM), intravenous (IV), or direct model. We generated a HLA- class II expressing humanized DRAG mice, venous inoculation (DVI) routes, first in the Netherlands and subsequently which develop a functional human immune system, following human in the UK, Tanzania, USA, Germany, Spain and Kenya. PfSPZ Challenge has hematopoietic stem cell infusion. DRAG mice develop human hepatocytes, been uniformly safe, and has infected 100% of volunteers by each route kupffer cells, liver endothelial cells and erythrocytes and sustain the in 5 of the 7 trials using well-tolerated doses. IV, DVI and IM injection have complete life cycle of P. falciparum malaria parasite. Our data also achieved pre-patent periods of 11.0-11.5 days, matching those following demonstrate that the infected DRAG mice self-cure blood-stage infection mosquito bites, and IV and IM demonstrated a dose response. PfSPZ following intravenous inoculation of live P. falciparum sporozoites and Challenge has enabled CHMI in Africa, emphasizing the potential for this elicit humoral responses characterized by IgM and IgG antibodies against “challenge in a bottle” to accelerate development of novel antimalarial ring, trophozoite and schizont stage parasites. The infected DRAG mice drugs and vaccines and to promote understanding of innate and acquired also elicit cellular responses mediated by TNF-alpha against P. falciparum immunity to malaria. blood-stage parasites. Thus the DRAG mice represent the first small animal model, which has the ability to sustain the complete P. falciparum life cycle and to elicit parasite-specific immune responses.

astmh.org 490 1603 Ad vectors; the first four antigens are designed to induce T cell responses targeting sporozoite and liver stages, while those encoding AMA1 and CD8 T CELLS MEDIATE STERILE PROTECTION OF OUTBRED MSP1 are designed to induce antibody responses targeting asexual blood MICE FROM PLASMODIUM YOELII CHALLENGE FOLLOWING stages. We have previously demonstrated that Ad and DNA/Ad prime/ RECOMBINANT DNA-PRIME/AD5-BOOST IMMUNIZATION boost vaccines are immunogenic in murine, swine, and nonhuman primate EXPRESSING TWO CANDIDATE VACCINE ANTIGENS models, and that a PfCA DNA/Ad vaccine is protective in humans. Here, BALB/c mice were immunized i.m. with either the individual components, Emily C. Smith1, Rebecca Danner1, Jessica S. Bolton1, Andrea the 5 antigen PfCSLAM mixture, or a 4 antigen PfCLAM mixture, K. Strein1, Nonenipha Rangel1, Esteban Abot1, Arnel Belmonte1, administered as a single Ad dose (1x108 pu) on study day (SD) 28, two Noelle Patterson1, Keith Limbach1, Martha Sedegah2, Eileen F. Ad doses on SD1 and SD28, or pDNA (50 µg) on SD1 and Ad (1x108 pu) Villasante2, Thomas L. Richie2, Joao C. Aguiar3 on SD28. Animals were bled pre-, 2 and 6 weeks post-boost for antibody 1Henry M. Jackson Foundation/Naval Medical Research Center, Silver assays, and spleens were harvested 2 and 6 weeks post-boost for T cell Spring, MD, United States, 2Naval Medical Research Center, Silver Spring, assays. Results established that the 5- and 4-antigen mixtures, CSLAM MD, United States, 3CAMRIS/Naval Medical Research Center, Silver Spring, mixture ± SSP2, induced antigen-specific T cell and antibody responses to MD, United States each antigen comparable to those induced by the individual components, Previously, we identified two antigens, UIS3 and Falstatin, which conferred as assessed by IFN-γ ELISpot or ELISA. These data support that the addition sterile protection of outbred mice from P. yoelii sporozoite challenge of antigens to the PfCA mixture which is protective in humans can following recombinant DNA-prime/Adenovirus serotype 5 (Ad5)-boost broaden the vaccine specificity. Future plans include GMP manufacture and immunization. In the present study, we aimed to identify the immune clinical testing of the CSLAM or CLAM DNA prime /Ad boost vaccine. component(s) mediating this protection. CD1 mice were primed with Py UIS3 and Py Falstatin DNA vectors followed by a boost with recombinant 1605 Ad5 vectors. Protection ranged from 7% to 57% in multiple experiments, with the highest level of protection observed for Ad5 boost of 1010 PLASMODIUM FALCIPARUM AMA1-BASED SUBUNIT pu. Sporozoite IFA titers ranged from 1:40 to 1:640, while blood-stage VACCINE FMP2.1/AS02A ELICITS A DIVERSE AND STRONG titers ranged from 1:2560 to 1:20480. A wide range of ELISA titers YET UNPROTECTIVE IMMUNE RESPONSE IN A PEDIATRIC was observed to UIS3 protein (range 1:25 to 1:78125) while those to COHORT IN BANDIAGARA, MALI Falstatin were more uniformly high (range 1:34576 to 1:144317). High Jason A. Bailey1, Mark A. Travassos1, Christopher G. Jacob1, frequencies of CD8 T cells producing IFN-g following Falstatin stimulation Amed Oattara1, Drissa Coulibaly2, Matthew B. Laurens1, Shannon were observed in the spleens of immunized mice (range 9.6% to 18.9% Takala-Harrison1, Carter L. Diggs3, Lorraine Soisson3, Tyler S. of CD8 T cells). A fraction of the responding T cells also produced TNF Brown1, Kirsten E. Lyke1, David E. Lanar4, Sheetj Dutta4, D. Gray and/or IL-2 in addition to IFN-g. Interestingly, the frequencies of Falstatin- Heppner4, Jozelyn Pablo5, Rie Nakajima5, Algis Jasinskas5, Amadou specific CD8 T cells producing IFN-g were significantly increased among Niangaly2, Andrea A. Berry1, Bourema Kouriba2, Mahamadou A. mice immunized with both UIS3 and Falstatin (above) compared to those Thera2, Ogobara K. Doumbo2, Philip L. Felgner5, Christopher V. immunized with Falstatin alone (range 4.7% to 7.4% of CD8 T cells, Plowe1 p<0.01). Similarly, endpoint serum ELISA titers targeting Falstatin were also 1 increased among mice immunized against both antigens (above) compared Howard Hughes Medical Institutes, Center for Vaccine Development, 2 to those immunized against Falstatin alone (range 1:15663.4 to 1:108539, Baltimore, MD, United States, Malaria Research and Training Center, 3 p<0.05). In vivo depletion of CD8 T cells prior to challenge resulted in Bamako, Mali, United States Agency for International Development, 4 complete loss of the protection. These data indicate that UIS3 and Falstatin Washington, DC, United States, Walter Reed Army Institute of Research, 5 are promising candidate malaria vaccine antigens. Further study is required Silver Spring, MD, United States, Department of Medicine, University of to fully understand the individual antigen contribution to protection and California Irvine, Irvine, CA, United States its duration, as well as to identify an optimal platform eliciting high level FMP2.1/AS02A is a blood stage malaria subunit vaccine candidate based durable T cell immunity. on the ectodomain of apical membrane antigen 1 (AMA1) of the 3D7 strain of Plasmodium falciparum. In a Phase 2 vaccine trial in 400 children 1604 aged 1-6 years in Bandiagara, Mali, West Africa, the vaccine had only a statistically insignificant 17% efficacy against all clinical malaria episodes D/HUAD5-PFCSLAM, A PLASMODIUM FALCIPARUM MULTI- when compared to the rabies control vaccine. However, it had 64% ANTIGEN MULTI-STAGE ADENOVIRUS VECTORED VACCINE efficacy against clinical malaria caused by homologous strains with respect CANDIDATE, IS IMMUNOGENIC IN BALB/C MICE to eight pre-specified polymorphic amino acid positions. To assess anti- AMA1 antibody specificity in AMA1 vaccine trials, we developed a protein 1 1 1 Noelle B. Patterson , Maureen E. Stefaniak , Joseph J. Campo , microarray for measuring seroreactivity to 263 unique AMA1 ectodomain 1 1 2 Kalpana Gowda , Keith Limbach , Joseph T. Bruder , C. Richter variants detected by sequencing the ama1 gene in field samples. We 2 3 3 3 King , Pamela Resch , Marilyn Ferrari , Larry R. Smith , Thomas L. evaluated AMA1 seroreactivity in a random sample of 40 children (aged 1 1 1 Richie , Denise L. Doolan , Eileen Villasante 1-6 years) and 20 adults (aged 18-55 years) pre- and post-vaccination 1 2 Naval Medical Research Center, Silver Spring, MD, United States, GenVec, with FMP2.1/AS02A or rabies control vaccine. Both children and adults Inc., Gaithersburg, MD, United States, 3Vical, Inc., San Diego, CA, United immunized with the AMA1 vaccine had broad and strong immune States responses to diverse AMA1 variants compared to controls 90 days after We have demonstrated that a malaria DNA/human adenovirus serotype the first immunization. Due to the broad cross-reactivity of antibodies 5-vectored vaccine platform (Ad) prime/boost vaccine expressing PfCSP generated by the vaccine, we were unable to pinpoint specific AMA1 and PfAMA1 antigens (PfCA) is well tolerated, immunogenic, and variants or polymorphic epitopes associated with protection from clinical efficacious in a Phase 1 clinical study (27%, 4/15 volunteers sterilely malaria. Further analysis using multivariable logistic regression, as well protected against controlled human malaria infection administered by as principle components analysis, Random Forest, and receiver-operating mosquito bite) (Chuang et al., 2013). We are now evaluating strategies characteristic curves, suggest that antibodies stimulated by the FMP2.1/ to increase efficacy. Here, we report the addition of antigens to broaden AS02A vaccine may be biased, binding preferentially to immunodominant vaccine immunogenicity, with a focus on inducing a multi-valent, multi- but unprotective AMA1 epitopes. This form of deceptive imprinting has stage, and multi-immune (cellular and humoral) response against both been described in HIV and influenza as a tool for immune escape and pre-erythrocytic and erythrocytic parasite life cycle stages. PfCSLAM is a also has been characterized in malaria as the ‘smokescreen effect’. These cocktail of PfCSP, PfSSP2/TRAP, PfLSA1, PfAMA1, and PfMSP1 DNA and

astmh.org 491 results suggest that a priori knowledge of a functional epitope map could long hypothesized that antibodies raised against epitopes outside the inform the selection of malaria subunit vaccine antigens that would repeat domain can also confer significant protection against sporozoite generate protective antibody populations. invasion. Using a newly developed chimeric Plasmodium berghei strain bearing the N-terminal region of the P. falciparum CSP, we report the 1606 characterization of a monoclonal antibody (MAb) recognizing the P. falciparum CSP. We mapped the fine epitope specificity of this MAb (5D5) A FULL-LENGTH PLASMODIUM FALCIPARUM RECOMBINANT and established that it recognizes an amino acid sequence immediately CIRCUMSPOROZOITE PROTEIN EXPRESSED BY adjacent to Region I of the P. falciparum CSP. Using both the novel P. PSEUDOMONAS FLUORESCENS PLATFORM AS A MALARIA berghei- P. falciparum chimera and P. falciparum parasites, we further VACCINE CANDIDATE characterized the 5D5 MAb epitope specificity and show that it can bind both live and air-dried fixed sporozoites. Most importantly, we 1 1 1 Gabriel Gutierrez , Amy R. Noe , Steve Giardina , Diego demonstrate that 5D5 can strongly inhibit parasite infection in vivo and 2 3 3 3 Espinosa , Ryan Haverstock , Jeff Allen , Hongfan Jin , Xiangming can inhibit cleavage of CSP, and so provide additional evidence that 4 4 4 5 Li , Jordana Coelho-dos-Reis , Ryota Funakoshi , John Sacci , antibodies targeting epitopes other than those at CSP’s repeat region can 3 6 6 1 Diane Retallack , Tom Vedvick Vedvick , Chris Fox , Ramses Ayala , be highly protective. Furthermore, we propose that this MAb could be 6 1 4 Steven G. Reed , Fidel Zavala , Moriya Tsuji utilized as an antibody-based, therapeutic prophylaxis, which may be a 1Leidos Life Sciences, Frederick, MD, United States, 2Johns Hopkins critical tool in the face of growing malaria drug resistance. Bloomberg School of Public Health, Baltimore, MD, United States, 3Pfenex Inc., San Diego, CA, United States, 4Aaron Diamond AIDS Research Center, 1608 New York, NY, United States, 5University of Maryland School of Medicine, Baltimore, MD, United States, 6Infectious Disease Research Institute, SAFETY AND PROTECTIVE EFFICACY OF INTRAVENOUS Seattle, WA, United States IMMUNIZATION WITH CRYOPRESERVED PLASMODIUM The circumsporozoite protein (CSP) of Plasmodium falciparum is a major FALCIPARUM SPOROZOITES UNDER CHEMOPROPHYLAXIS - surface protein, which forms a dense coat on the sporozoite’s surface. TUECHMI-002 Preclinical research on CSP and clinical evaluation of a CSP fragment-based Benjamin Mordmueller1, B. Kim Lee Sim2, Peter F. Billingsley2, RTS,S/AS01 vaccine have demonstrated a modest degree of protection Eric R. James2, Meral Esen1, Markus Gmeiner1, Guezin Surat1, against P. falciparum, mediated in part by humoral immunity and in part Heimo Lagler1, Thomas L. Richie2, Stephen L. Hoffman2, Peter G. by cell-mediated immunity. Given the partial protective efficacy of the Kremsner1 RTS,S/AS01 vaccine in a recent Phase 3 trial, further improvement of CSP- based vaccines is crucial. Here we describe the preclinical evaluation of 1University of Tuebingen, Tuebingen, Germany, 2Sanaria Inc., Rockville, a full-length, recombinant CSP (rCSP)-based vaccine candidate against MD, United States P. falciparum malaria suitable for current Good Manufacturing Practice Exposure to 12-15 Plasmodium falciparum (Pf)-infected mosquitoes, (cGMP) production. Utilizing a novel high-throughput Pseudomonas administered three times at monthly intervals under continuous fluorescens expression platform, we demonstrated greater efficacy of chemoprophylaxis with chloroquine is highly efficacious in preventing full-length rCSP as compared to N-terminally truncated versions, rapidly asexual blood stage infection following subsequent controlled human down-selected a promising lead vaccine candidate, and developed a malaria infection (CHMI) in healthy, adult, malaria-naïve individuals. To high-yield purification process to express immunologically active, intact translate and expand this immunization strategy, mosquito bites need to antigen for clinical trial material production. The rCSP, when formulated be replaced by a pharmaceutical product that can be easily administered with various adjuvants, induced antigen-specific antibody responses as and exactly dosed. In addition, the chemoprophylactic regimen needs to be measured by ELISA and immunofluorescence assay (IFA), as well as CD4+ adjusted to the specific requirements of this approach. Our previous study T-cell responses as determined by ELISpot. The adjuvanted rCSP vaccine (TUECHMI-001) showed that one mosquito bite corresponds to ~620 conferred protection in mice when challenged with transgenic P. berghei intravenously (IV) injected, cryopreserved Pf sporozoites (PfSPZ) produced sporozoites containing the P. falciparum repeat region of CSP. Furthermore, by Sanaria Inc. Here, we report first results on the safety, tolerability, heterologous prime/boost regimens with adjuvanted rCSP and an immunogenicity and preliminary protective efficacy of escalating doses of adenovirus type 35-vectored CSP (Ad35CS) showed modest improvements Sanaria’s PfSPZ (PfSPZ Challenge) under chemoprophylaxis (PfSPZ-CVac in eliciting CSP-specific T-cell responses and anti-malarial protection, approach) with chloroquine. During immunization, PfSPZ were injected by depending on the order of vaccine delivery. Collectively, these data support direct venous inoculation (DVI) three times at 4-week intervals. Volunteers the importance of a further clinical development of adjuvanted rCSP, either receive staggered doses of 3,200 (Group A), 12,800 (Group B) or 51,200 as a stand-alone product or as one of the components in a heterologous (Group C) PfSPZ per injection, corresponding to approximately 5, 20 or prime/boost strategy, ultimately acting as an effective vaccine candidate 80 mosquito bites. In every group, 9 volunteers received PfSPZ and 5 for the mitigation of P. falciparum-induced malaria. placebo, while all received 10 mg/kg chloroquine 2 days before the first injection, followed by 5 mg/kg every week for a total of 10 doses. To 1607 assess efficacy of the immunization regimen, CHMI will be done 8 weeks after completion of chemoprophylaxis by DVI of 3,200 PfSPZ Challenge. A MONOCLONAL ANTIBODY AGAINST THE N-TERMINAL Subsequently, a PfSPZ-CVac dose that shows at least 75% efficacy, good REGION OF THE PLASMODIUM FALCIPARUM safety and tolerability will be tested using an experimental ultra-short CIRCUMSPOROZOITE PROTEIN STRONGLY INHIBITS chemoprophylaxis with azithromycin and chloroquine. Here, 2 g extended- SPOROZOITE INVASION OF HEPATOCYTES release azithromycin will be given on the day of each PfSPZ injection followed by 10 mg/kg chloroquine 5 days later. Complete results for the 1 2 1 2 Diego Espinosa , Amy Noe , Photini Sinnis , Gabriel Gutierrez , first two dose groups will be presented together with safety, tolerability 1 Fidel Zavala and preliminary immunogenicity data of the highest dose (51,200 PfSPZ). 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States, 2Leidos Life Sciences, Frederick, MD, United States Studies in animals and human volunteers have demonstrated that antibodies against the Circumsporozoite Protein (CSP) can protect against infection by Plasmodium sporozoites. Due to its repetitive nature and tandem disposition, the epitopes at CSP’s repeat region are the most prominent target of protective antibody responses. However, it has been

astmh.org 492 1609 homologous and heterologous Pf parasites? 4) Can the number of doses required to provide short term and long term homologous protection LONGITUDINAL ANALYSIS OF HUMORAL AND CELLULAR be reduced (to 3 doses of 4.5x10^5 PfSPZ)? We will present safety and IMMUNITY FOLLOWING VACCINATION WITH AN immunogenicity results for both the 3 and 5 dose regimens. Additionally, ATTENUATED PLASMODIUM FALCIPARUM SPOROZOITE we will present our plans to conduct late Phase 2 and Phase 3 trials VACCINE supporting a BLA for licensure in adults. The potential for worldwide benefit results from the fact that DoD requirements - excellent safety Andrew Ishizuka1, Kailan-Sierra Davidson1, Kim Lee Sim2, Stephen and tolerability, efficient administration, and sterile protection lasting for Hoffman2, Robert A. Seder1 at least six months - are characteristics that equally support deployment 1Vaccine Research Center, National Institutes of Health, National Institute to malaria endemic areas to prevent disease and death and to promote of Allergy and Infectious Diseases, Bethesda, MD, United States, 2Sanaria campaigns aiming to halt malaria transmission and eliminate the disease Inc., Rockville, MD, United States from defined geographical areas. A highly effective and durable vaccine for preventing Plasmodium falciparum (Pf) malaria infection is a critical need for preventing 1611 the substantial morbidity and mortality incurred by this infection. Pf sporozoites (PfSPZ) administered by mosquito bites are the only MAN VERSUS MOSQUITO: HOW VECTOR-BORNE immunogens shown to induce high-level sterilizing protection (>80%) PATHOGENS MOVE AROUND THE WORLD in humans. We previously reported that attenuated, aseptic, purified, Michael Johansson1, Andrew Tatum2 cryopreserved PfSPZ (PfSPZ Vaccine) administered 4 or 5 times 1Centers for Disease Control and Prevention, San Juan, PR, United States, intravenously (IV) conferred high-level protection in humans in a dose- 2University of Southampton, Highfield, United Kingdom dependent manner. Moreover, initial analysis of humoral and cellular immunity showed that there was a dose-dependent increase in CSP Vector-borne pathogens, such as malaria and dengue, are major causes antibody titer, functional inhibition of sporozoite invasion in vitro and the of morbidity and mortality, worldwide. These pathogens often move frequency of sporozoite specific IFN-g producing CD4 and CD8+ T cell between endemic areas and from endemic to non-endemic areas via responses. Here, we substantially expanded this analysis and performed airplanes, which can rapidly transport infected humans and vectors over a longitudinal assessment of antibody and cellular responses through the long distances. Upon introduction, either the human or vector may initiate course of vaccination and after controlled human malaria ~ 3 weeks after new transmission cycles in other locations under the right ecological the final immunization and ~ 5 months later. For T cell assessment, multi- conditions. While numerous countries have established regulations for parameter flow cytometry with two recently developed 16-color panels as disinsection of airplanes arriving from certain locales, it has not been used to assess the magnitude, phenotype and quality of sporozoite specific clear whether this significantly reduces transmission risk. We developed T cell responses. Together, these data provide insights into the durability of branching process models to assess the probability of traveling humans immunity and protection after vaccination with the PfSPZ Vaccine and will and vectors initiating transmission resulting in at least one local human guide ongoing and future studies to define the mechanistic correlates of infection in a new location. For humans, this depends on the probability protection. of an infected person traveling, infecting a mosquito, and that mosquito infecting a human, as well as several constituent processes. For mosquitos, 1610 it is the probability of an infected mosquito traveling and infecting a human, again with constituent processes. We assessed these models CLINICAL DEVELOPMENT OF THE PFSPZ VACCINE TO for Plasmodium falciparum, a causative agent of malaria. For a plane PROTECT THE WARFIGHTER FROM MALARIA moving from a highly endemic area to another area highly suitable for P. falciparum transmission, the probability of introduction of P. falciparum by Judith E. Epstein1, Kristopher Paolino2, Sharina Reyes1, Robin a human is approximately 100%. However, for a mosquito it is less than Nielsen2, Alexandra Singer1, Martha Sedegah1, Richard Ruck2, 0.1%. Analysis of the sequence of events leading to introduction makes Carlos Vasquez1, Elizabeth Saverino3, Silas Davidson2, Peter F. it clear that mosquitoes have lower probabilities of travel, infection, and Billingsley3, Anita Manoj3, Sumana Chakravarty3, Kavita Tewari3, further transmission compared to humans for whom the risk of pathogen Eric James3, Anusha Gunasekera3, Adam Richman3, Matthew introduction is many times larger. While controlling the transportation Stewart3, Deborah Padilla3, Tooba Murshedkar3, Thomas L. Richie3, of mosquitoes may be critical for avoiding the introduction of vector B. Kim Lee Sim3, Eileen Villasante1, Stephen L. Hoffman3 or pest species, our model indicates that it has little benefit for vector- 1Naval Medical Research Center, Silver Spring, MD, United States, 2Walter borne pathogens. Given the ever-increasing volume of travel, it is critical Reed Army Institute of Research, Silver Spring, MD, United States, 3Sanaria to develop new ways to reduce the risk of pathogen spread by infected Inc., Rockville, MD, United States humans. A vaccine to prevent malaria for use by the Department of Defense (DoD) must provide sustained sterile protection against infection. The Sanaria® 1612 PfSPZ Vaccine has been developed to address this need. It is composed of aseptic, purified, cryopreserved, vialed PfSPZ manufactured in compliance DURABILITY OF POLYESTER-BASED LONG-LASTING with all regulatory standards. In a recent clinical trial, PfSPZ Vaccine INSECTICIDAL NETS IN THREE GEOGRAPHICAL ZONES OF protected 6/6 (100%) subjects against controlled human malaria infection NIGERIA - A THREE YEAR FOLLOW-UP OF NETS DISTRIBUTED (CHMI) at the highest dosage regimen administered (5 intravenous THROUGH CAMPAIGNS doses of 1.35x10^5 PfSPZ), and there was a dose response in regard to Albert Kilian1, Emmanuel Obi2, Richmond A. Selby3, Hannah antibody and T cell responses. The protective regimen was safe and well Koenker4, Matt Lynch4 tolerated, indicating that further dose escalation could be undertaken. 1 2 Since completion of that study, 6 additional PfSPZ Vaccine trials have been Tropical Health LLP, Montagut, Spain, Malaria Consortium, Abuja, 3 4 or will be initiated, designed to optimize dose size, dose interval, number Nigeria, Malaria Consortium, Kampala, Uganda, Centers for of doses and duration of protection. This study, conducted by the DoD, Communication Programmes, Baltimore, MD, United States addresses the following questions: (1) Is PfSPZ Vaccine safe and tolerable The average survival of Long-lasting Insecticidal Nets (LLIN) has increasingly administered by direct venous inoculation? (2) Can 5 doses of 2.7x10^5 become of interest to malaria program managers and the international PfSPZ provide protection 3 weeks (short term) after immunization against community as this measure of durability will determine the frequency of CHMI carrying a heterologous Pf strain? (3) Can 5 doses provide protection LLIN replacements at household level and the cost of sustaining universal when subjects undergo a second CHMI at 24 weeks (long term) with coverage. With the recent publication of a WHO-recommended method

astmh.org 493 to estimate net survival, comparative analyses from different areas or 1614 for different brands have now become possible. Following the mass distribution campaigns of 2010/11 in Nigeria annual follow-up surveys ANALYSIS OF DETERMINANTS OF INSECTICIDE TREATED to measure attrition and physical integrity of campaign LLIN (polyester, NETS (ITNS) USE AMONG CHILDREN UNDER FIVE YEARS 100 Denier) were undertaken in three locations of Nigeria representing USING LOGISTIC REGRESSION different eco-geographic and climatic zones: Shinkafi district in Zamfara 1 2 1 State in the northern dry-savannah, Toto district in Nasarawa State in the Wahjib Mohammed , Anthony J. Turkson , David Y. Mensah central guinea-savannah and Abi district in the rain-forest area of Cross 1National Malaria Control Programme, Accra, Ghana, 2Takoradi River State. In each district a population representative sample was drawn Polytechnic, Accra, Ghana using a 20 cluster sampling design and in each selected community 15 Malaria causes between three hundred and fifty (350) and five hundred households that had received nets from the campaign were included in (500) million clinical episodes and over one million deaths annually. the interview and net assessment. In the questionnaire reasons for any loss Children and pregnant women are the most vulnerable group and of campaign nets were explored as well as attitudes and practices towards most endangered by the disease. Insecticide-Treated bednets (ITNs) net care and repair. The assessment of physical integrity of the nets was range among the most effective measures of malaria prophylaxis, yet its done according to WHO recommendations and the proportionate Hole implementation level in sub-Saharan Africa is still low. The goal of this Index used to evaluate the outcome for each net. In the first round of study was to assess factors influencing the use of ITNs by children under surveys a total of 900 households were sampled and 1,571 campaign five years in Ghana. A cross-sectional study was conducted using pre- nets assessed while for the second round the figures were 896 and tested, interviewer-administered questionnaires. Possible factors were 1,367 respectively. Two years after the campaign the survival of LLIN measured using five hundred (500) mothers or guardians of children varied considerably between locations with 69.6% (95% CI 62.7-75.7) under five years, in twenty three (23) communities in the Asamankese still in serviceable condition in Nasarawa State, 81.4% (75.0-86.5) in sub-municipality in the West Akim District in the Eastern Region of Ghana. Zamfara and 89.4% (84.2-93.5) in Cross River State. The final round of Logistic regression was used to assess the influence of five possible factors data collections is currently ongoing and will be presented together with on ITNs use. In order of importance in determining one’s use of LLIN, estimates of median LLIN survival. Reasons for differences by location will ‘Sleeping area allowing for the use of Long Lasting Insecticides Net (LLIN)’ be explored and discussed. was the most important factor influencing the use of ITNs by children under five years in Ghana, followed by ‘Household’s expected monthly 1613 income’, then ‘Mother’s SHS level of education’ compared to ‘none’, while the least was ‘Number of children under five years’. The study also MODELING THE EFFECTS OF TRANSNATIONAL MIGRATION revealed that 86.2% of the participants owned ITNs out of which 92.81% ON PUBLIC HEALTH POLICIES IN SOUTHEAST ASIA: AN used it the night before the study. The study revealed that there is a INVESTIGATION INTO THE IMPACT ON ELIMINATION relationship between influencing factors and use of LLINs. To improve ITNs STRATEGIES usage, there should be continuous distribution of LLINs and Insecticide Treated Materials (ITMs), such as insecticide treated curtains for doors and Amruta Dixit1, Ming-Chieh Lee1, Edward Wenger2, Guiyun Yan1 windows. This should be heavily supported by education and Behavior 1 2 University of California Irvine, Irvine, CA, United States, Institute for Change Communication (BCC) via radio, TV, and other media especially on Disease Modeling, Seattle, WA, United States the hanging techniques and need to provide adequate space for sleeping Human migration plays an important role in the spread of infectious area. diseases. In order to determine the impact of human movement between areas of unequal malaria endemicities on malaria transmission and the 1615 resulting control and elimination strategies, we need to improve our understanding of: the migration patterns present in the study areas of EVALUATING DIFFERENCES IN HOUSEHOLD INSECTICIDE- interest, the malaria acquisition risk borne by the migrants, and the impact TREATED BEDNETS (ITN) OWNERSHIP BETWEEN UGANDA of migration on malaria transmission. We hypothesized that persistent and AND ZIMBABWE DURING 2005-2011: AN ECONOMETRIC unmonitored flow of people between porous borders between areas of APPROACH unequal transmission, presents a significant challenge to the elimination 1 2 of malaria in the area of low disease transmission. The present study aims Yesim Tozan , Selver Derya Uysal to provide a quantitative assessment of malaria risks due to transnational 1New York University, New York, NY, United States, 2Institut für Höhere migration and to evaluate subsequent intervention strategies for malaria Studien, Vienna, Austria control and elimination. Within the EMOD framework developed by the The rapid scale-up of insecticide-treated bednets (ITNs) in African countries Institute for Disease Modeling, we have constructed simulations with have received heightened attention with the availability of significantly two geographically-connected populations in order to model the impact greater resources for national malaria control efforts, particularly after of varying degrees of human migration rates and intervention methods 2005. In this study, we investigate the mean differences in household ITN on malaria transmission. Our preliminary simulation results suggest that ownership in Uganda and Zimbabwe for the years 2005 and 2011, using the proportion of humans infected was markedly different between the DHS and GPS data and MARA malaria endemicity maps and econometric various rates of human migration. We will continue to develop increasingly methods. The probability of owning an ITN in Uganda was 12.8% higher complex simulations that explore both human and vector-based than in Zimbabwe in 2005. Although ITN ownership increased steadily in interventions such as the administration of primaquine and the application both countries, the difference in the probability of owning an ITN widened of long-lasting microbial larvicides. The simulations results can be used significantly and became 33.5% over this period. The Blinder-Oaxaca to consider various feasible pathways to sustained local elimination as technique can be used to study the mean outcome differences between a function of cross-border migration rates. This approach has the ability two groups (in our case two countries). Using this technique, we divide to make a timely and significant contribution to public health by filling a the ITN ownership differential between two countries into a part that is critical gap in our knowledge of human migration patterns, especially in “explained part” by group differences in ITN ownership determinants, Southeast Asia. The utility of these data and this novel modeling tool is such as household characteristics and malaria risk, and a residual part multidisciplinary and has the potential to inform sophisticated models in that cannot be accounted for by such differences in ITN ownership other research fields. determinants. This “unexplained” part captures the effects of group differences in unobserved predictors of ITN ownership. Our preliminary results showed that the larger fraction of the increase in the ITN ownership differential in this period was due to the unexplained part. To understand

astmh.org 494 this result we investigated what happened in these countries in terms of assessed change in disparity in ITN ownership and use among different malaria control efforts between 2005 and 2011. Published literature points socioeconomic groups in Uganda between 2006 and 2011. The authors to a significant difference in malaria financing in these countries, yet the used Lorenz Concentration Curve and Index (C-Index) to assess equity in earmarked proportion of funding for increasing the ownership and use household ITN ownership and use among children under five between of ITNs is usually high in all countries. Next we will study to what extent wealth quintiles separately in 2006 (Demographic and Health Survey malaria control efforts in these two countries affected ITN usage and all- data) and 2011(Malaria Indicator Survey data). C-Index values range cause child mortality rates and potentially extend our analysis to include between -1 to 1, a value of 0 suggests no difference in ownership and other African countries. use among different socioeconomic groups. Household ownership of at least one ITN rose significantly from 16% (2006) to 60% (2011). Similarly, 1616 ITN use among children under five was very low (10%) in 2006 and increased substantially to 47% in 2011. The increase in ITN ownership was UNDERSTANDING INTRA-HOUSEHOLD DECISION MAKING associated with significant reduction in inequity among wealth quintiles FOR INSECTICIDE-TREATED MOSQUITO NET ALLOCATION IN (C-Index 0.11, 95% CI: 0.08;0.34) in 2006 versus 0.02, 95% CI: 0.01;0.04 UGANDA in 2011). Similarly the disparity in use of ITN use among children under five from different wealth quintiles greatly reduced from 2006 (C-Index: Emily E. Ricotta, Steve A. Harvey, Hannah Koenker, Matthew 0.04, 95% CI:-0.10;0.19) to 2011 (C-Index: 0.01, 95% CI:-0.04;0.06). Lynch This achievement is probably due to the shift to universal coverage in 2009 Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United which led to free mass distribution campaigns of Long-lasting Insecticidal States Nets (LLINs), with 7.2 million LLINs distributed by 2010. This achievement Intra-household allocation of insecticide-treated mosquito nets (ITNs) is in parity between wealth quintiles should be sustained; however, efforts an important concern for malaria prevention programs. Behavior change are needed to further increase overall ITN ownership coverage and use communication campaigns stress the need to prioritize children under five in Uganda. This is achievable through additional free mass campaign and pregnant women for net distribution and use, as these two groups distribution combined with traditional distribution channels. are the most vulnerable to malaria infection. This study utilized a pile sort activity to help understand net allocation decision making in Luwero and 1618 Nebbi, two Ugandan districts. Sixty-four respondents, half men and half women, were asked to assign 10 individuals ranging in age from infant ASSESSING THE CONTRIBUTION OF MALARIA CONTROL to older adult and including adults and children of both sexes as well as INTERVENTIONS ON REDUCTIONS IN ALL-CAUSE UNDER-5 one visibly pregnant woman to one of four beds. One bed had a new net, MORTALITY IN ZAMBIA one a slightly used net with a few holes, one an old net with large holes, Katherine E. Colson1, Laura Dwyer-Lindgren1, Nancy Fullman1, and the last with no net. After sorting, respondents were asked why they Tom Achoki2, Matthew Schneider3, Peter Hangoma4, Marie Ng1, allocated each individual as they had. The number of times a household Felix Masiye5, Emmanuela Gakidou1 member was placed under each net was evaluated, and a hierarchical cluster analysis was completed to determine which household members 1Institute for Health Metrics and Evaluation, Seattle, WA, United States, were typically grouped together. Responses were compered between male 2Ministry of Health of Botswana, Gaborone, Botswana, 3United States and female, urban and rural, and district of residence. Results demonstrate Agency for International Development, Washington, DC, United States, that net allocation differs by gender, rural/urban, and district, however the 4Department of Economics, University of Bergen, Bergen, Norway, pregnant woman and baby were always clustered together and were given 5Department of Economics, University of Zambia, Lusaka, Zambia the best net greater than 50 and 60 percent of the time, respectively. Between 2000 and 2010, Zambia rapidly scaled-up malaria control Children ages 5-14 were also typically clustered together and given the interventions such as insecticide-treated nets and indoor residual best or second-best net. Young adults (around age 25), older adults, and spraying. At the same time, Zambia recorded substantial declines in elders were given the worst net or no net a majority of the time. Reasons under-five mortality. Zambia’s experience is heralded as a malaria-control given for these placements varied, but common themes include the baby success story, but it is unclear whether the expansion of malaria control and pregnant women being the most vulnerable to malaria, the older contributed to improved childhood survival beyond the impacts of other children being able to help repair nets, and the heads of household and child health interventions. To quantify the impact of malaria control efforts young adults being robust enough to not succumb to malaria infection in Zambia, we estimated trends for all-cause under-five mortality and a and having sufficient resources to purchase additional nets. The general range of child and maternal health interventions at the subnational level. consensus was that this household did not have enough nets, and that We quantified the reduction in child mortality associated with malaria the family should strive to provide nets for all of their household members. control while taking into account trends in other key interventions as well This exercise supports current behavior change campaign messaging, as socio-demographic, health system, and environmental factors across demonstrating that, at least in their reported behavior, people are likely to districts. Our estimation methods included generalized linear models and prioritize vulnerable populations when it comes to net use. functional data analysis and were validated with cross-validation and simulation techniques. We found that the bivariate relationship between 1617 malaria control and child mortality was strong and significant, but this relationship was attenuated when other factors were considered. Several REDUCTION IN DISPARITY OF INSECTICIDE-TREATED NETS other child health interventions also scaled up dramatically during the OWNERSHIP AND USE AMONG SOCIOECONOMIC GROUPS same time period, including pentavalent immunization, prevention of AFTER SCALE UP IN UGANDA mother-to-child-transmission of HIV/AIDS, exclusive breastfeeding, and nutrition programs. Because of this simultaneous expansion, it was Cameron Taylor, Ana Claudia Franca-Koh, Samantha Herrera, Jui statistically infeasible to isolate the effects of malaria control efforts. In Shah, Lia Florey, Yazoume Ye the absence of the combined scale-up of these interventions between ICF International, Rockville, MD, United States 2000 and 2010, we estimated that child mortality would have been 11% The increase in funding for malaria control in the past decade resulted higher in 2010. The scale up of these interventions accelerated declines in an increase in Insecticide-Treated Nets (ITNs) ownership and use in in mortality by 1% each year. Our findings emphasize the importance of many countries in sub-Saharan Africa, particularly in Uganda. However, constructing a comprehensive landscape of the drivers of progress in child with the shift in programmatic focus from target groups to universal mortality. A greater quantity of high-quality and localized data is critical coverage there is a need to ensure equal access and use of ITNs for all for evaluating the independent impact of each intervention on childhood sub-populations regardless of their socioeconomic status. This study survival.

astmh.org 495 1619 PfSPZ-based products without the use of mosquitoes, developed technology for the in vitro production of Pf oocysts. We have optimized INCREASING ROLE OF ANOPHELES FUNESTUS AND gametocyte culture conditions that are optimal for the in vitro production AN. ARABIENSIS IN MALARIA TRANSMISSION IN THE of ookinetes and oocysts, established culture conditions to reproducibly KILOMBERO VALLEY, TANZANIA produce and quantify 3 and 7 to 8 day oocysts, and demonstrated that in vitro produced oocysts are similar in size and morphology to mosquito 1 2 1 Dickson W. Lwetoijera , Caroline Harris , Samson Kiware , produced oocysts. These developments have made it possible to assess 3 4 5 1 Stefan Dongus , Gregor Devine , Philip McCall , Silas Majambere the transmission blocking activities of candidate vaccines in an in vitro 1Ifakara Health Institute, Ifakara, Morogoro, United Republic of Tanzania, transmission blocking (iTB) assay without the need for mosquitoes. 2Lancaster University, Lancaster, United Kingdom, 3Swiss Tropical and Preliminary data indicate correspondence between the results of SMFA and Public Health Institute, Basel, Switzerland, 4QIMR Berghofer Medical iTB assays. The transmission blocking activity of antibodies against Pfs25 Research Institute, Brisbane, Queensland, Australia, 5Liverpool School of and Pfs48/45 in SMFA and iTB assay will be presented. Tropical Medicine, Liverpool, United Kingdom This longitudinal study demonstrates the trends in malaria vector dynamics 1621 and their relative contribution to malaria transmission in hyper-endemic transmission settings in Tanzania. The study was conducted in two IDENTIFICATION OF A NOVEL PLASMODIUM FALCIPARUM villages within the Kilombero valley, in rural Tanzania for five consecutive HEAT SHOCK PROTEIN 70 (HSP70Z), ALSO IDENTIFIED AS years (2008 - 2012). 72 houses were selected per village and each house CG4, AS A TRANSMISSION BLOCKING VACCINE CANDIDATE was sampled for mosquitoes monthly using a CDC light trap. Collected Mark DeCotiis, Joan Aebig, Buddhadeb Mallik, Nikiah Dawson, mosquitoes were assessed for species identity and sporozoite infection Olga Muratova, Karine Reiter, Martin Burkhardt, Vu Nguyen, status using PCR and ELISA respectively. Anopheles funestus susceptibility Patrick E. Duffy, Nicholas J. MacDonald, David L. Narum to insecticides was assessed using WHO guidelines. A total of 100,810 malaria vectors were collected, of which 76% were An. gambiae s. l. and National Institutes of Health, Rockville, MD, United States 24% were An. funestus. Of all An. funestus samples that amplified with A malaria transmission blocking vaccine (TBV) is critical to achieve the PCR (n = 2,737), 97% were An. funestus s.s., 2% were An. rivorulum and goal of malaria elimination in some areas. To date, only one sexual stage 1% An. leesoni. Whereas for An. gambiae s.l. (n = 8,117), 93% were An. protein, Pfs25, has been evaluated in humans and a second sexual stage arabiensis and 7% were An. gambiae s.s. The proportion of An. gambiae protein, Pfs230, will begin human testing soon. Other sexual stage s.s. identified by PCR (2,924) declined from 0.2% in the year 2008 to proteins have failed to reach the clinic due to an inability to produce undetectable levels in 2012. An. arabiensis dominated the wet season them at pilot-scale following cGMP. In order to expand the number of whereas An. funestus dominated the dry season. Malaria transmission TBV candidates, a panel of monoclonal antibodies (mAbs) was produced intensity significantly decreased from an EIR of 78.14 infectious bites/ against Plasmodium falciparum macrogametes to identify novel surface person/year in 2008 to 35ib/p/yr in 2011 but rebounded to 226 ib/p/ proteins that, when targeted by antibodies could interfere with parasite yr in 2012 coinciding with an increased role of An. funestus in malaria development in the mosquito midgut. Here we report on the evaluation of transmission. Insecticide susceptibility tests indicated full susceptibility one mAb, identified as 1C7, which recognized a heat shock protein (HSP), of An. funestus to deltamethrin (100% mortality), reduced susceptibility identified as HSP70z or Cg4, by Western blot of a sexual stage parasite to dieldrin (95%), permethrin (93%), and confirmed resistance to lysate and by pull-down studies using LC/MS/MS techniques. HSP70z is DDT (86%). Similar findings were also recorded for An. arabiensis, in expressed in asexual and sexual stage parasites with a molecular mass of separate study in same area. The results indicate the continuing role of approximately 100 kDa. In macrogametes, HSP70z may be localized on An. arabiensis and the increasing importance of An. funestus in malaria the macrogamete cell surface by a live immunofluorescence assay. Most transmission. These findings call for complementary vector control and importantly, 1C7 blocked P. falciparum transmission in mosquitoes, with surveillance tools that target these specific vector species, their behaviour similar activity to that of a Pfs230 domain 1 specific mAb using an ex vivo and their ecology and an insecticide resistance management plan to membrane feeding assay. A recombinant form of HSP70z (named rHSP70z) preserve the efficacy of LLINs. was produced in Pichia pastoris that was comprised of approximately 20% of the native protein which was recognized by 1C7 in Western blots. 1620 rHSP70z specific IgG purified from sera of immunized rabbits failed to block parasite transmission. In a competition ELISA, rHSP70z specific rabbit DEVELOPMENT OF AN IN VITRO TRANSMISSION BLOCKING antibodies failed to compete for 1C7 binding to rHSP70z, likely explaining (ITB) ASSAY AGAINST PLASMODIUM FALCIPARUM WITHOUT the lack of transmission blocking activity. Currently, the 1C7 epitope is THE USE OF MOSQUITOES being mapped to evaluate whether a synthetic peptide mimicking the 1C7 epitope will induce transmission blocking antibodies. Abraham G. Eappen, Tao Li, Meghan Marquette, Peter F. Billingsley, Adam Richman, B. K. Sim, Stephen L. Hoffman Sanaria Inc., Rockville, MD, United States Major efforts are underway aimed at developing malaria vaccines that can be used for elimination of Plasmodium falciparum by preventing the development of the parasite within the mosquito and thereby preventing transmission of the parasite to humans. To assess candidate vaccines, antibodies are induced by immunization of animals and humans, and assessed for their transmission blocking activity against sexual erythrocytic and mosquito stage parasites in a standard membrane-feeding assay (SMFA). These SMFAs are biologically relevant, but highly consumptive of personnel time and resources, require a mosquito colony and are not easily adaptable to high through-put. Thus, it is difficult, time consuming and expensive to assess large numbers of candidate anti-sera in SMFAs. Sanaria’s technology platform generates live, aseptic, purified, cryopreserved P. falciparum sporozoites (PfSPZ) that can be administered as a highly protective malaria vaccine. PfSPZ are produced using Anopheles stephensi mosquitoes as bioreactors. Sanaria, in its quest for developing

astmh.org 496 1622 particularly groups from different socio-economic status. This analysis used Lorenz concentration curves and indices to assess the equity of ITN IMPACT OF INDOOR RESIDUAL SPRAYING ON household ownership, access and use in Mozambique, using data from ENTOMOLOGICAL INDICES OF MALARIA TRANSMISSION IN the 2007 Malaria Indicator Survey and 2011 Demographic Health Survey. THE BUNKPURUGU-YUNYOO DISTRICT IN THE NORTHERN Concentration Index (C-Index) values range between -1 and 1, with a SAVANNAH ZONE OF GHANA value of 0 representing perfect equality. From 2007 to 2011, equity in ITN household ownership (C-Index: 0.06 in 2007 and 0.04 in 2011), ITN access Samuel Dadzie1, Sylvester Coleman2, Peter Mumba2, Aklilu (C-Index: 0.12 in 2007 and 0.09 in 2011), and ITN use among children Seyoum2, Philip Ricks3, Daniel Szumlas4, Paul Psychas5, Jacob under five (C-Index: -0.04 in 2007 and 0.03 in 2011) showed slight Williams6, Maxwell Appawu1, Daniel Boakye1 improvements, while equity in ITN use among pregnant women remained 1Noguchi Memorial Institute for Medical Research, Accra, Ghana, 2Ghana the same (C-Index: .06 for both years). It is important to highlight that Africa Indoor Residual Spraying Project, Abt Associates, Accra, Ghana, while improvements where shown, equity in ITN household ownership, 3Malaria Branch, Division of Parasitic Diseases and Malaria, Atlanta, GA, access and ITN use among children under five, all were fairly equitable United States, 4Centers for Disease Control and Prevention, Atlanta, in 2007. The results demonstrate that ITN scale-up efforts have been GA, United States, 5University of Florida, Gainesville, FL, United States, successful as well as equitable across the population, however further 6Research Triangle International, Washington, DC, United States improvements in access and ITN coverage are still needed to reach targets. Malaria remains a major public health problem in Ghana, especially in the northern savannah zone. This study was conducted to evaluate the 1624 impact of U.S President’s Malaria Initiative (PMI) and the Ghana Health Services Indoor Residual Spraying (IRS) program on malaria transmission DIFFERENCES IN DEET AND PICARIDIN SENSITIVITY in the Bunkpurugu-Yunyoo District in northern Ghana. In 2011 and 2012, BETWEEN SOUTHEAST ASIAN VECTORS OF MALARIA the district was sprayed with a pyrethroid, alphacypermethrin, at an AND ARBOVIRUSES, RESULTS OF A FIELD EVALUATION OF application rate of 25mg/m2. In 2013, an organophosphate, pirimiphos- TOPICAL REPELLENTS 2 methyl at a rate of 1g/m ), was used based on declining susceptibility Lies Durnez1, Mao Sokny2, Karel Van Roey1, Vincent Sluydts1, Siv of local vectors to pyrethroids. Indoor resting densities (IRD), parity, Sovannaroath2, Tho Sochantha2, Marc Coosemans1 sporozoite rate and entomological inoculation rates (EIR) of the local 1 2 vector species were monitored through pre- and post-IRS monthly human Institute of Tropical Medicine, Antwerp, Belgium, National Center for landing and pyrethrum spray collections. The IRD of Anopheles gambiae Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia s.l. (the predominant vector species, 99.2% of all Anopheles collected) Scaling up of insecticide treated nets has contributed significantly to was reduced from a mean of 2.91 mosquitoes/room recorded from the a substantial malaria decline. However, some malaria vectors, and baseline surveys to 2.10 mosquitoes/room (27.7% reduction) in 2012 after most vectors of arboviruses, bite outdoors and in the early evening. spraying with alphacypermethrin. In 2013, the mean IRD of An. gambiae Therefore, topically applied insect repellents may provide a crucial s.l. was further reduced to 0.22 mosquitoes/room, representing 89.2% additional protection against mosquito-borne pathogens. Among topical decline compared to 2012. In 2012 there was a non-significant reduction repellents, DEET is the most commonly used, followed by others such (p=0.289) in the mean parity rate for An. gambiae s.l. from 75% to 43% as PMD or picaridin. A study was carried out in Cambodia to determine (57% reduction). Spraying with Actellic in 2013, resulted in 67% reduction the entomological efficacy of DEET and picaridin repellents on wild in parity rate from 43% to 32% (p = 0.130). A comparison of the pre populations of several mosquito genera, including vectors of arboviruses and post-IRS EIRs also revealed a significant (p<0.05) reduction, from 0.35 (Aedes aegypti and Ae. albopictus) and malaria (Anopheles dirus, An. infective bites/man/night (ib/m/n) in 2011 to 0.021 ib/m/n in 2012. In minimus, An. maculatus and An. barbirostris). During 230 survey days 2013, there was slight decrease in EIR to 0.018 ib/m/n after spraying with in two consecutive years, the lower limbs of 5 persons were treated Actellic 300CS that year. The results show that the IRS operations resulted with repellents (‘DEET 20%’, ‘picaridin 20%’, or ‘picaridin 10%’) or in reduction in key entomologic indicators. IRS with pirimiphos-methyl ethanol (2 negative controls), followed by mosquito collections on the had the greatest impact on indoor resting densities and parity rates of An. treated limbs during 5 consecutive hours. The treatments were grouped gambiae s.l. but not EIRs. The PMI funded IRS program after three years following a 5x5x5 Graeco-latin square to equalize the effects of treatment contributed to 94.8% reduction in malaria transmission in the study area. days, collection sites, and test persons. Protection rates were high (91- 99.2%), with significant differences between treatments, genera, and 1623 species. For malaria vectors, ‘DEET 20%’ performed better than ‘picaridin 20%’ or ‘picaridin 10%’. The protection rate against An. barbirostris SCALING UP OF INSECTICIDE-TREATED NETS (ITN) was significantly lower as compared to the other vectors, especially for OWNERSHIP AND USE IN MOZAMBIQUE: HAS THE SCALE-UP the picaridin repellents. As malaria endemic areas often differ in their BEEN EQUITABLE? vector species composition, this heterogeneity in repellent sensitivity between vector species might result in a geographically heterogeneous Samantha Herrera, Ana Claudia Franca-Koh, Jui Shah, Cameron epidemiological impact of repellent use for malaria control. Taylor, Lia Florey, Yazoume Ye ICF International, Rockville, MD, United States 1625 Insecticide-treated nets (ITNs) are effective tools for malaria prevention and can significantly reduce morbidity and mortality due to malaria, MOLECULAR AND ENVIRONMENTAL INTERACTIONS IN especially among children under five in endemic areas. ITN ownership ANOPHELES GAMBIAE INFLUENCE BOTH REPRODUCTIVE and use have rapidly increased in Mozambique, due to scaling up of ITN CAPACITY AND PLASMODIUM DEVELOPMENT distribution efforts within the country over the past seven years, which Perrine Marcenac1, Francesco Baldini2, Nicola Segata3, Abdoulaye have aimed to achieve universal coverage. The percent of households that Diabate4, Flaminia Catteruccia1 owned at least one ITN rose by more than three-fold from 15.7% in 2007 1 2 to 51.4% in 2011. ITN access, defined as one ITN for every two people Harvard School of Public Health, Boston, MA, United States, Università 3 4 in the household also rose significantly, from 9.4% in 2007 to 37.0% di Perugia, Terni, Italy, University of Trento, Trento, Italy, Institut de in 2011. Similarly during this time, ITN use among children under five Recherche en Sciences de la Sante/Centre Muraz, Bobo-Dioulasso, Burkina and pregnant women increased significantly, from 6.7% to 35.7% and Faso from 7.3% to 34.3%, respectively. With the aim of achieving universal Reproduction in the major malaria vector Anopheles gambiae is influenced coverage, it is important to also assess distribution across subgroups, by a number of molecular and environmental factors. Male-female

astmh.org 497 molecular interactions following mating are important determinants maintains malaria hotspots/pops of mainly asymptomatic carriers. of fertility and fecundity. At the same time, an increasing amount of Targeting malaria transmission in low transmission settings requires a evidence points to reproductive processes playing an important role in better understanding of this heterogeneity. Plasmodium parasite development. Male transfer of the steroid hormone 20-hydroxy-ecdysone (20E) during mating activates the transcription of 1627 a Mating-Induced Stimulator of Oogenesis (MISO) gene that transduces the mating signal into an increase in egg development. Silencing MISO NO PRODUCT? NO PROGRAM: TREATMENT UPTAKE by RNA interference in laboratory colonies reduces egg development to AND AVAILABILITY OF ANTIMALARIAL DRUGS FOR levels observed in virgin females. This phenotype is caused by reduced INTERMITTENT PREVENTIVE TREATMENT OF MALARIA IN expression of yolk protein precursors (YPPs) after MISO silencing, impairing MALAWI lipid accumulation in the oocyte. Previous research shows that the same 1 1 2 YPPs essential for lipid accumulation in the developing mosquito egg Katherine Wolf , Loren Bausell , Evance Moyo help parasites escape the immune system. We have found evidence that 1United States Agency for International Development | DELIVER PROJECT, MISO depletion decreases Plasmodium infection in A. gambiae, further Arlington, VA, United States, 2United States Agency for International reconstructing the molecular pathways linking egg development and Development | DELIVER PROJECT, Lilongwe, Malawi Plasmodium infection. Functional studies performed in field A. gambiae Throughout Africa, 30 million pregnant women are exposed to malaria show that in natural populations MISO is essential for egg development, each year. Several interventions are central to malaria control efforts, as its silencing abolishes oogenesis after blood feeding. These results including dispensing sulfadoxine-pyrimethamine (SP) for intermittent suggest that in natural mosquito populations MISO is a key switch that preventive treatment in pregnancy (IPTp). While preventing malaria in directs resources derived from the blood meal towards oogenesis only pregnancy is a key focus of global interventions, trends in using SP to in mated females. Besides molecular factors, environmental factors prevent malaria during pregnancy have largely stagnated. Twenty years also affect A. gambiae reproductive biology in natural populations. after adopting IPTp as an official policy, the rates of pregnant women in Sequencing analysis of the microbiota from reproductive tissues of wild Malawi who receive at least two doses of SP is only 55 percent. Research A. gambiae demonstrates enrichment of bacteria in specific villages and has identified factors impeding adherence to IPTp policies: antenatal mating swarms, which may impact reproductive success and isolation. clinic (ANC) client behavior/attitudes, provider actions/attitudes, and SP In addition we have identified bacteria in reproductive tissues that have availability at facilities. Qualitative data indicate that stockouts significantly been previously shown to impact Plasmodium survival. We are currently impact IPTp coverage. However, only a quantitative analysis will show if determining the effects of these bacteria on reproductive biology and there is a relationship between IPTp use and stockouts. For four years, mosquito fitness, two factors relevant for malaria transmission. By the USAID | DELIVER PROJECT examined trends for three complementary elucidating the different molecular and environmental interactions data sources: SP availability at heath facilities using the country’s logistics regulating reproduction and parasite development in A. gambiae, we hope management information system; SP uptake at health facilities using to develop novel targets for vector control. national ANC service statistics; and IPTp coverage in households reported in the Malawi Demographic Health Surveys. Preliminary results show a 1626 general decline in SP stockouts between 2010 and 2013. SP stockouts peaked in late 2011 and the first quarter of 2012 (80 percent); low HETEROGENEITY OF HUMAN AND MOSQUITO BEHAVIOR stockout rates were reported during the last half of 2012 and all of 2013 IN RELATION TO OUTDOOR AND EARLY MALARIA (6 percent). High stockout rates in early 2012 correlate with a sharp drop TRANSMISSION IN SOUTHEAST ASIA in SP uptake during the same time (62 to 31 percent). As SP availability improved, beginning in May 2012, the IPT coverage rates also improved. Charlotte Gryseels1, Lies Durnez1, Sambunny Uk2, Vincent Women attending their first ANC visit after 12 weeks, who received any Sluydts1, Tho Sochantha2, Somony Heng2, Karel Van Roey1, SP, increased to almost 100 percent by mid-2013. These results show a Lydie Canier3, Didier Menard3, Sokny Mao2, Saorin Kim3, Marc close relationship between SP availability and the uptake of preventive Coosemans1, Koen Peeters Grietens1 treatment during pregnancy. Using a mixed-method case study approach, 1 2 Institute of Tropical Medicine Antwerp, Antwerp, Belgium, National this analysis will explore the impact of SP stockouts on MIP programming Center for Parasitology, Entomology and Malaria Control, Phnom Penh, efforts. Cambodia, 3Institut Pasteur du Cambodge, Phnom Penh, Cambodia Outdoor transmission is one of the key factors for malaria control and 1628 elimination in the Greater Mekong Region, including Cambodia, where malaria is now mainly reduced to forested regions inhabited by ethnic AN EXTENDED MOLECULAR BARCODE FOR TRACKING minorities and mobile migrants. Tackling outdoor transmission requires a PLASMODIUM FALCIPARUM PARASITE POPULATIONS better understanding of the heterogeneity in human and vector behavior 1 1 2 1 during times when people are still active outdoors. Results from a mixed- Nicholas K. Baro , Rachel Daniels , Eli Moss , Wesley Wong , 3 2 1 methods social science study in Ratanakiri, Cambodia, indicate that local Daniel L. Hartl , Daniel E. Neafsey , Sarah K. Volkman , Dyann F. 1 ethnic minorities have different socio-cultural characteristics than the Wirth majority society targeted by malaria control programs. (i) Mobility caused 1Harvard School of Public Health, Boston, MA, United States, 2Broad by a multiple residence system increases exposure to the sylvatic vector Institute, Cambridge, MA, United States, 3Harvard University, Cambridge, An. dirus, as during the malaria peak season people usually reside on MA, United States their farms in the forest. (ii) Open housing blurs the boundary between The lack of a generalizable approach to malaria control requires a indoor and outdoor biting. (iii) Differences in sleeping times between real-time assessment of both epidemiological and parasite population villages, farms and during forest activities creates diverse evening biting genetics at the local level. The major epidemiological shifts associated opportunities, if assumed that people sleep under insecticide-treated nets with the transition into a malaria-eliminating country are mirrored by at night. However, (iv) evening resting is frequently done without nets, changes in the genetic diversity profile of malaria parasite populations. and (vi) even night sleeping often occurs under non-impregnated bought These genetic signatures can be monitored using population genetic - nets, which are often torn, making it hard to establish to what extent based tools to determine efficacy of malaria intervention efforts prior transmission actually occurs early or outdoors. Results seem to suggest to measurable changes in disease prevalence. For these reasons, and to that the evolving interplay of vector and human behavioral heterogeneity further understand the basic biological processes of parasite transmission, we have improved upon an interim panel of neutral, unlinked, single nucleotide polymorphisms (SNPs) that is specifically designed for resolving

astmh.org 498 individual parasites in highly related parasite populations and allows for 1630 quantification of allele balance at polymorphic loci to infer changes in transmission dynamics. The extended molecular barcode includes SNPs OPTIMAL COVERAGE AT MINIMAL COST: A DYNAMIC exhibiting high minor allele frequency and low divergence (e.g FST) filtered MODELING APPROACH TO SIMULTANEOUS ALLOCATION OF from screening over 500 whole genome sequenced samples representing MULTIPLE ANTI-MALARIA INTERVENTIONS populations from Africa, South East Asia and South America. The accuracy 1 2 3 and sensitivity of Sequenom’s Matrix-assisted laser desorption/ionization Beatrix Balogh , David W. Hutton , Ravi M. Anupindi , Leslie 1 4 4 1 time-of-flight (MALDI-TOF) mass spectrometry genotyping technologies Arney , Meiting Liu , Haipeng Wu , Peter S. Larson , Prashant 1 4 allow for quantitative output that reflects proportions of each allele in Yadav , Mark L. Wilson multiplexed reactions. The increased number of loci and low dynamic 1William Davidson Institute, University of Michigan, Ann Arbor, MI, United range of the minor allele improve our ability to predict the number of States, 2Department of Health Management and Policy, School of Public distinct parasite genomes in samples with higher complexity of infection Health, University of Michigan, Ann Arbor, MI, United States, 3Stephen M. (COI). In silico testing of the extended molecular barcode demonstrates its Ross School of Business, University of Michigan, Ann Arbor, MI, United utility for detecting recent common ancestry among parasites in a sample States, 4Department of Epidemiology, School of Public Health, University of on a much more cost-effective basis than using variants called with whole Michigan, Ann Arbor, MI, United States genome sequence data. We are exploring the use of these approaches to Malaria control efforts often involve a multipronged approach, including identify shared regions of the genome that are identical by descent among insecticide-treated nets (ITNs), indoor residual spraying (IRS), and highly related Senegalese populations and assess COI in high transmission pharmaceutical-based treatment and prevention. Public health agencies settings. (government, private, NGOs) are constrained by conflicting demands on limited budgets. Determining the appropriate combination and magnitude 1629 of interventions is both complicated and context specific. Using a dynamic modeling approach, we analyzed the system dynamics of multiple malaria RECEPTIVITY OF ANOPHELINE MOSQUITOES IN SOUTHERN interventions functioning in concert, evaluated the cost-effectiveness of ZAMBIA: TWO YEARS OF BIONOMIC DATA FROM AN AREA treatment and prevention strategies, and developed a resource allocation TARGETED FOR MALARIA ELIMINATION model to apportion resources across intervention types to maximize Jennifer C. Stevenson1, Limonty Simubali2, Harry Hamapumbu2, overall cost effectiveness. This compartmental mathematical model Sungano Mharakurwa1, Douglas E. Norris1 (using MATLAB) includes age, human immunity, and seasonality in the Plasmodium transmission cycle, with interventions of IRS, ITNs, intermittent 1 Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United preventive treatment for pregnant women (IPTp), and mass screening 2 States, Macha Research Trust, Choma, Zambia and treatment (MSAT). We assessed individual and combined effects on Malaria prevalence in Macha, southern Zambia has reduced significantly various transmission parameters such as biting rate, force of infection, over the past decade, with community parasite prevalence currently and mosquito death. For a set of parameters considered typical of high below 1%. The province is now targeted for elimination strategies. It transmission malaria endemic settings, results show that after five years is important, however, to establish the potential of the local mosquito into the model run and compared to a baseline with no interventions, population to transmit malaria to estimate the risk of resurgence if the with a combination of 65% coverage of IRS and ITNs, infection prevalence parasite were re-introduced. To address this, light trap collections of in children under five years of age and pregnant women would decrease mosquitoes were carried out monthly in randomly selected geo-referenced by about 18%. Those above five years of age exhibited decrease in households in the catchment area of Macha Hospital from February 2012 prevalence by about 16% with 70% combined IRS and ITNs. The addition to February 2014. Peak times of host-seeking were studied using collection of IPTp and MSAT decreased long-term community prevalence in all age bottle rotators. Approximately 1200 female anophelines were collected groups by an overall additional 12%. This modeling approach allows a from 856 traps across 336 different households. 60% were identified careful assessment and optimization of costs and benefits to particular as the vector An. arabiensis to give a mean catch of 0.88 An. arabiensis combinations of malaria interventions and should assist public health per trap-night over the study period. Catches ranged from 0 to 148 groups in maximizing benefit under constrained budgets. An. arabiensis in a single trap, with highest catch recorded in February 2013 (monthly mean 7.2 per trap-night). Households with anophelines 1631 appeared to be clustered. Spatial analyses are ongoing to overlay mosquito distribution on malaria risk maps. 10.8% of An. arabiensis were HOUSEHOLD EXPENDITURES FOR MALARIA TREATMENT blood fed and the human blood index was calculated as 0.93. At least FROM A POPULATION-BASED SURVEY 9 other anopheline species were identified, some of which were highly 1 2 anthropophagic. One specimen was found to be positive for Plasmodium Donald S. Shepard , Peter Mangesho , Robert Mpagala 1 1 2 2 falciparum sporozoites by ELISA. Preliminary analysis of host-seeking Kihomo , Aggrey Kihombo , Joseph Mugasa , Amos Malongo , 2 2 1 times indicated peaks between 22:30 and 02:30, but vector activity was Ruth Mnzava , Filemoni Tenu , Angelique K. Rwiyereka , Yara 1 3 4 5 recorded as early as 20:30. Despite substantial reduction in malaria cases A. Halasa , Aaron M. Samuels , Mark Rowland , Frank Mosha , 2 in Macha, large numbers of the vector An. arabiensis exist at certain times William Kisinza of the year and in certain localities. Here An. arabiensis demonstrates high 1Brandeis University, Waltham, MA, United States, 2National Institute for endophily and anthropophagy. There is the potential for human exposure Medical Research, Tanga, United Republic of Tanzania, 3Centers for Disease outside the times of net use. Whilst drug-based elimination strategies Control and Prevention, Atlanta, GA, United States, 4London School of are encouraged, vector control methods should be maintained and Hygiene & Tropical Medicine, London, United Kingdom, 5Kilimanjaro entomological surveillance continued to monitor any increase in dominant Christian Medical Centre, Moshi, United Republic of Tanzania or secondary vectors. Effective malaria control technologies not only improve health, but also reduce health expenditures by government and households. In Muheza, Tanzania, a randomized trial is assessing the effectiveness and cost- effectiveness of a non-pyrethroid insecticide-treated wall liner (ITWL) and indoor residual spraying (IRS). To project savings, the investigators are determining current expenditures for malaria treatment in conjunction with a cross-sectional epidemiological household survey. Following mapping and household enumeration, from December 2013 through February 2014 epidemiological teams visited 4200 randomly chosen

astmh.org 499 residents aged>6 months across 60 village clusters. Respondents who 1633 reported a malarial episode within the past 30 days were selected for a follow-up economic interview. Its questions included household income, FIGHT AGAINST MALARIA BY STUDENTS AND treatment received, time required for travel and treatment, and expenses SCHOOLCHILDREN IN KINSHASA, DR CONGO incurred. Overall, 18% of sampled residents reported a malaria illness episode within the last 30 days. Preliminary data are currently available Sophia M. Mangala, Junior Musenga, Ornella L. Mampuya, for 467 representative malarial cases. Of these, 6% received inpatient Viviane Mitshiabu, Steve Tulantched, Thierry L. Bobanga, Thierry L. hospital treatment, 91% received ambulatory treatment outside the home, Bobanga and 3% received only in-home or no treatment. Overall combined travel Protestant University of Congo, Kinshasa, Democratic Republic of the and treatment time averaged 5 hours. Household expenditures per case Congo averaged US$5.24 (TZS 8,545), but the median value of US$2.33 (TZS Malaria is a real public health problem in Africa. Despite the efforts 3,800) was below the mean. This arose because expenditures on malaria made by countries, 655,000 deaths were recorded worldwide in 2010. treatment were highly skewed with a standard deviation equal to 213% And the Democratic Republic of the Congo accounts for nearly 10 % of of the mean. Analysis of household expenditures found 54% was for the mortality And the fight against this disease should be of interest all direct medical expense (consultations, beds, tests, and medications), 27% groups in the community. And new and innovative approaches should be for transportation, and 19% for other expenses. Average expenditures of considered to reduce disease Thus, students of the Protestant University hospitalized patients US$32.82 (TZS53,491) were an order of magnitude of Congo have been trained in the fight against malaria with the aim of above those of non-hospitalized patients of US$2.15 (TZS5,787). training schoolchildren Students’ knowledge was initially evaluated. And By comparison, household monthly revenue was under US$46.01 training was organized to provide knowledge about the transmission, (TZS75,000) for 71% of respondents. Average inpatient expenses prevention, particularly on ITN and finally what to do in case of fever. represent about a month’s median income. Given low rural incomes, Method: 823 students in 9 schools in the city of Kinshasa sensitized by even routine medical expenses can strain household budgets. Adjustment 20 university students who were trained by the managers of the National of censored expenditures for episodes in progress and valuing time lost Program of Academic Medicine and University of Kinshasa Parasitology would increase estimated costs. If ITWL and IRS prove efficacious in this Department. The age of children was between 9 to 11 years old This district, households will save on treatment expenses. awareness was marked by a strong interaction between children and sensitizers. At the end of awareness, the children were evalued to assess 1632 their understanding of the subject. And the best schoolchildren were awarded. It was noted that over 70 % of children could answer basic IMPACT OF MALARIA INTERVENTIONS ON REDUCTIONS questions on malaria. In conclusion, at the end of this work, over 800 IN NEONATAL MORTALITY IN MALAWI, RWANDA AND schoolchildren in the city of Kinshasa and 20 students have contributed to MAINLAND TANZANIA the fight against malaria, which is a tiny proportion of the population of said city. It is therefore important to continue these actions as awareness Rebecca Winter1, Lia Florey1, Anne Langston2, Tom Pullum1 through children, future actor and relay in the transmission of knowledge 1 The Demographic and Health Surveys Program, ICF International, from their family. This, could be, an innovative approach to malaria 2 Rockville, MD, United States, ICF International, Rockville, MD, United control. States Between 2000 and 2010 sub-Saharan Africa’s impressive gains in under- 1634 five mortality have been accompanied by a more modest reduction in neonatal mortality. As a result, neonatal mortality now accounts for about INSECTICIDE-TREATED NET (ITN) UTILIZATION AND a third of all under-five deaths in the region. This study selected three MAINTENANCE IN KINSHASA, DR CONGO countries in SSA with significant reductions in NMR during this period_ Nono B. Mangaza1, Fabrice B. Ekomisa2, Solange E. Umesumbu3, Malawi, Rwanda, and mainland Tanzania_ in order to identify factors that 4 2 contributed to the observed reductions, with special interest in the relative Billy S. Kunyu , Thierry L. Bobanga importance of scale-up of interventions to protect women against malaria 1University of Technology Belcampus, Kinshasa, Democratic Republic of the during pregnancy. In Malawi, the neonatal mortality rate among women’s Congo, 2Protestant University of Congo, Kinshasa, Democratic Republic most recent children born fell from 26 deaths per 1,000 live births in the of the Congo, 3National Malaria Control Program, Kinshasa, Democratic five years preceding the 2000 DHS to 20 deaths per 1,000 live births in Republic of the Congo, 4National Institute of Biomedical Research, the five years preceding the 2010 DHS. In Rwanda, the NMR declined Kinshasa, Democratic Republic of the Congo from 29 to 14 deaths per 1,000 live births, and in mainland Tanzania Malaria remains at the beginning of the first century killer in DRC with from 32 to 18 deaths per 1,000 live births between the 1999 and 2010 more than 60,000 deaths, especially among children under 5 and surveys. Multivariate decomposition procedures were used to examine the pregnant women Among the strategies, the LLIN is a tool of choice but extent to which the scale-up of malaria interventions contributed to these its availability and hanging are operational problems. Furthermore its observed reductions. Results show that in all three countries the rapid efficiency and sustainability are elements to evaluate. The present study increase in mosquito net ownership was associated with the observed was conducted with the aim to determine the proportion of households reductions in neonatal mortality, even after adjusting for changes in the with at least one LLIN ; the proportion screens installed ( used); the distribution and effects of sociodemographic characteristics and key proportion of pregnant women who slept under LLINs ; the proportion of maternal and delivery services. In Malawi - where information on mothers’ children who slept under LLINs ; determine the average age of the net and use of IPTp was available - the study did not find evidence that the scale- the number of washing on average per month. A cross-sectional study up of IPTp was associated with the reduction in NMR. In conclusion, the on the use and sustainability of ITN Long (LLINs) in the city of Kinshasa. findings reinforce the importance of consistent and universal mosquito net We formed a cluster sampling and multistage. The sample size was 104 use in areas with high prevalence of malaria. While malaria interventions households per municipality. And 24 selected communes we got 2,512 are most often geared towards saving the lives of children at older ages households. The use of the net in Kinshasa was 59.4 %. Pregnant women (6 months to 5 years), the study findings contribute to a growing body of and children under 5 were using respectively 70% and 60%. Through the evidence pointing to the importance of malaria interventions to neonatal city’s most important use is in the center where the mosquito nuisance survival. with the Culex is the greatest with 72.7 %. Whereas the periphery of the city use is low (44 to 55%) where anopheles populations are most abundant. The proportion of pregnant women and children under 5 years under nets was 70 % and 60 % respectively. In 2,512 households visited

astmh.org 500 nearly 4,812 LLINs were counted. The average duration of the net in the may travel hours to reach targeted communities, and they may not be household was in the range 19 to 24 months with 1.5 washes by month. able to return to a centralized location for clean-up at the end of the day. More than 50% of household use detergent for washing ITN. The majority AIRS has developed a mobile soak pit (MSP) that can be transported from of nets found in the households have probably lost their effectiveness site to site with the spray team, can be installed in less than one half hour, before 18 months of utilization. In conclusion, the use of LLINs was and can remove pesticide contamination from wash waters. Advantages still low in Kinshasa. The inhabitants do not respect manufacturer’s of this mobile soak pit include a marked reduction in the time that spray recommendations in term of washing. Study on bio efficacy and durability operators spend traveling from site to wash area, better adsorption of the in field use condition must be conducted to make evidence of efficacy pesticide due to the characteristics of the filter material, better control of soak pit materials, and better protection of the community because the 1635 pesticide contamination is taken away in the filter. EFFECT OF MASS DRUG ADMINISTRATION OF IVERMECTIN 1637 TO HUMANS ON MALARIA TRANSMISSION AND EPIDEMIOLOGY IN WEST AFRICA BASELINE STUDIES ON ANGLOGOLD ASHANTIS’ INDOOR RESIDUAL SPRAYING PROGRAM (IRS) FOR MALARIA Haoues Alout CONTROL IN GHANA Colorado State University, Fort Collins, CO, United States Kwaku P. Asante1, Dominic B. Dery1, David K. Dosoo1, Kinsley Mass drug administration (MDA) of ivermectin to control filariasis and Kayan1, Sylvester Segbaya2, Victor Asoala3, Kofi Tchum1, Frank onchocerciasis has been shown to reduce malaria vector survivorship Amoyaw2, Nicholas Amoako1, Abraham Oduro4, Seth Owusu- and the proportion of mosquitoes that are able to transmit Plasmodium Agyei1 falciparum in the same communities. Therefore, we have proposed that ivermectin MDA be considered in malaria control and elimination 1Kintampo Health Research Centre, Brong Ahafo Region, Ghana, strategies. In our ongoing efforts to fully characterize the effect of 2AngloGold Ashanti (Ghana) Malaria Control Limited, Ashanti Region, ivermectin MDA on malaria transmission, we compared its effect on Ghana, 3Navrong Health Research Centre, Upper East Region, Ghana, Anopheles gambiae and Plasmodium falciparum natural populations in 4Navrongo Health Research Centre, Upper East Region, Ghana three different West African countries (Senegal, Liberia and Burkina Faso) Malaria is still high in many parts of Africa including Ghana. Private -public across different seasons. Blood fed mosquitoes were collected indoors partnership are essential in significantly reducing the burden of malaria before and after MDA in treated villages by health authorities for either in high risk countries. In Ghana, there is rapid scale up of indoor residual lymphatic filariasis (ivermectin+albendazole) or for onchocerciasis control spraying in various parts of the country with funding from the Global (ivermectin alone), and concomitant mosquito sampling was performed Fund and the United States President’s Malaria Initiative (PMI). AngloGold in untreated control villages. We compared the blood fed mosquito Ashanti (a mining company) is implementing an IRS programme in 25 survivorship, sporozoite rate and parasite genetic diversity in mosquitoes, districts in Ghana. The results of baseline studies (sensitivity of insecticides, taking account for temperature, humidity, mosquito species and type of prevalence of childhood malaria parasitemia and anemia) conducted prior treatment. The mosquitocidal effect was consistent in all field sites and to the IRS programme are presented here. Malathion was most effective seasons, and did not vary with the addition of albendazole. The reduction with 100% An. gambiae mortalities in seven districts. Fenitrothion was in sporozoite rates were significant when compared to control villages effective in three districts whiles Propoxur worked in one district. Few but the observed reductions vary across field sites. In Burkina Faso in (14) kdr susceptible strains were detected in samples analyzed with the treated village, sporozoite rate was significantly reduced by 79% majority being homozygous kdrRR(120) resistant species compared to 32 following MDA (from 8% to 1.7%) and increased to pre-treatment levels Heterozygous kdrRS. Preliminary data shows high prevalence of malaria after two weeks. Additionally, MDA completely eliminated sporozoite parasitemia (range: 30 - 50%) and anemia (range 40% - 60%). An transmission from outdoor host-seeking mosquitoes for a period of organophosphate class of insecticide is considered most appropriate for two weeks. The potential impact of ivermectin on Plasmodium genetic IRS in eleven districts currently earmarked in Ghana. Rotation of different diversity in mosquitoes and its consequence on malaria epidemiology and classes of insecticides over time is however recommended as it offers a transmission will be discussed. practical solution for resistance management in light of rapid resistance development. Monitoring of malaria parasitemia and anemia during the 1636 IRS programme is required. IMPROVED ENVIRONMENTAL COMPLIANCE AND 1638 OPERATIONAL EFFICIENCY USING MOBILE SOAK PITS TWO VARIANTS OF MULTIDRUG RESISTANT VIBRIO Peter J. Chandonait CHOLERAE O1 BIOTYPE EL TOR INVOLVED IN TWO Abt Associates, Inc., Cambridge, MA, United States CONSECUTIVES OUTBREAKS OF CHOLERA IN CAMEROON The President’s Malaria Initiative (PMI) employs thousands of spray (2004 - 2005 AND 2010 - 2012) operators and annually sprays millions of homes in Africa with a residual 1 1 1 pesticide to kill malaria-bearing mosquitos. At the end of the work day, the Ariane A. Nzouankeu , Antoinette Ngandjio , Vanessa Nkamga , 1 2 1 spray operators must clean their spray tank and their personal protective Pierrette Simo , Marie-Laure Quillici , Marie-Christine Fonkoua equipment (helmets, face shields, gloves, and boots), resulting in wash 1Centre pasteur du Cameroun, Yaounde, Cameroon, 2Institut Pasteur de water that is contaminated with pesticide residue. This contaminated Paris, Paris, France water is treated before release in soak pits, which are pits 1 meter wide x Outbreaks of cholera due to toxigenic Vibrio cholerae are always 2 meters length x 1 meter deep, filled with a bed of commonly available dangerous and need a quick taking care of patient. From may 2010 to cooking charcoal and other materials to filter out and break down the February 2012, an outbreak of cholera raged in Cameroon. The epidemic pesticide. These soak pits are centrally located within targeted spray areas started in the North and reached the south of Cameroon 4 months later, so that many operators can travel to and use these facilities for clean-up at in September 2010. The isolated strains, V. cholerae O1 serotype Ogawa, the end of the day. PMI has built and/or refurbished hundreds of soak pits were multidrug resistant with additional resistance to nalidixic acid. over the past several years of IRS project implementation. Ensuring that isolated from 2 subsequent outbreaks of cholera in Cameroon, 2004 - these soak pits are built properly so as to minimize negative environmental 2005 and 2010 - 2011. Geographically, the 2004-2005 outbreak was impacts from IRS is a major environmental compliance responsibility of the localized in the south of Cameroon, while the current outbreak (2010- implementing partner. However, in sparsely populated spray areas, teams 2011) covered the whole area of Cameroon. A total of 200 V. Cholerae O1

astmh.org 501 isolated during the last outbreak of cholera in Cameroon were used in this 1640 study; the strains belonged to biotype El Tor, serotype Ogawa. The strains were resistant to multiple antimicrobials especially nalidixic acid, which SENDING PEACEKEEPING TROOPS INTO AN ONGOING was the newest character. Molecular detection of their virulence factors CHOLERA OUTBREAK - THE CHILEAN EXPERIENCE DURING revealed that tcpa gene which encodes the toxin coregulated pilus was MINUSTAH (MISSION DES NATIONS UNIES POUR LA characteristic to El Tor biotype, while nuleotide sequence of ctxB which STABILISATION EN HAÏTI) encode the sub-unit B of the cholera toxin, was closer to the classical biotype. A total of 3 mutations was observed on the ctxB nucleotide Reinaldo Rosas1, Alberto Fica1, Dannette Guiñez1, Myriam sequence of which 2 were PFGE fingerprinting types showed different Polanco2, Alejandra Saldivia1, Wilson Wigant1, Vjera Triantafilo1, patterns, This study reveals that V. cholerae strains isolated in 2010-2011 Thomas Weitzel3 were different from strains isolated in 2004-2005, using antimicrobial 1Hospital Militar de Santiago, Santiago, Chile, 2Comando de Salud del susceptibility phenotypes, characterization of antimicrobial resistance, Ejército, Santiago, Chile, 3Universidad del Desarrollo Clinica Alemana, cholera toxin genotyping, PFGE. PFGE analysis revealed two different Santiago, Chile unrelated profiles All the strains harboured tcpa El Tor allele, which was a Since 2004, Chile participates in MINUSTAH with a permanent contingent supplemental argument ranging these strains in El Tor biotype. of 500 soldiers, who change every 6 months. The Preventive Medicine Service prepares troops and medical units for potential threats. After the 1639 2010 earthquake, a cholera outbreak rapidly spread through Haiti, which was probably related to soldiers participating in the same UN mission. This PREVALENCE OF ENTEROAGGREGATIVE ESCHERICHIA work describes the experience with pre- and post-deployment measures of COLI VIRULENCE GENES IN YOUNG CHILDREN FROM the Chilean Forces to reduce the cholera risk in Haiti as well as importation RURAL SOUTH AFRICA AND PATHOGENESIS OF POTENTIAL into Chile by asymptomatic carriers. Those actions included sanitary and VIRULENCE MARKERS IN A MOUSE MODEL: THE MAL-ED hygiene education of troops, training for medical officers, pre-deployment COHORT immunizations with whole-cell oral cholera vaccine (WC/rBS), and post- deployment microbiological surveillance. In 2011, vaccine side effects Nicoline Tanih1, David T. Bolick2, Amidou Samie1, Emanuel were monitored using a standardized questionnaire. After deployment, Nyathi1, Leah Barrett2, Relana Pinkerton2, James P. Nataro2, Araceli all troops were screened for Vibrio cholerae carriership. Data on WC/ Santiago2, Richard L. Guerrant2, Pascal O. Bessong1 rBS vaccination were available for 569 soldiers of whom 9.7% reported 1 2 University of Venda, Thohoyandou, South Africa, University of Virginia, systemic and 10.5% gastrointestinal (GI) side effects. A shorter interdose Charlottesville, VA, United States interval (7 vs 30 days) was associated with more GI disturbances (17.4% Enteroaggregative Escherichia coli (EAEC) is recognized as a cause of vs 7.7%, p<0.001). In the subgroup receiving WC/rBS 30 days apart, growth shortfalls with or without diarrhea worldwide. A number of EAEC systemic side effects were more common if peacekeepers simultaneously virulence-related genes (VRGs) have been described but their role in the received other pre-deployment vaccines (11.9% vs 4.3%, p<0.05). All clinical outcome of infection is not completely defined and may vary from surveillance stool cultures were negative, except for 2 (in 2012) which both one geographical location to another. Recently, we have characterized Aar grew strains of non-pathogenic V. cholerae. Cholera is still a threat for (AggR activated regulator), a new negative regulator in EAEC. Deletion underdeveloped countries and military operations within these countries of Aar upregulated the expression of the EAEC master regulator AggR. need to take measures to prevent infection and further spread the disease Accordingly, we have found that an EAEC strain lacking Aar causes growth by deployed troops. Our experience showed an acceptable safety profile of failure without diarrhea in our zinc deficient mouse model. We extracted WC/rBS, especially if doses are separated by 30 days. This vaccine provides DNA from 207 strains of EAEC isolated from the stool of 109 children a fairly high protection rate for the first 6 months, but its influence on followed from birth to 12 months of age from the Dzimuali community the rate of asymptomatic carriers is uncertain. Therefore, additional in the Limpopo Province of South Africa. We investigated the prevalence post-deployment stool cultures seem an appropriate surveillance tool for of EAEC VRGs using multiplex polymerase chain reaction. Samples were returnees to non-industrialized countries such as Chile. analyzed for identification of 18 VRGs. Plasmid encoded haemolysin (aar) was the most frequently detected (86.5%), followed by aggregative 1641 adherence regulator (aggR, 53.6%) and EAEC HilA homologue (eilA, 47.3%). Secreted autotransporter toxin gene (sat) was observed at lowest NOVEL IMMUNOLOGICAL SIGNALS FOR DIAGNOSING frequency (0%). Although only (5%) of participants had diarrhea in their ACUTE TYPHOID FEVER first 12 months, children with EAEC had greater growth shortfalls (P= Nga Tran Vu Thieu1, Elizabeth Klemm2, Gordon Dougan2, Sabina 0.03) and one child had an aar(-) EAEC with striking growth failure. These 3 3 1 data confirm a high prevalence, endemicity and heterogeneity of EAEC Dongol , Buddha Basnyat , Stephen Baker strains in the Limpopo Province of South Africa and their association with 1Oxford University Clinical Research Unit, Vietnam, Ho Chi Minh, Vietnam, growth failure. However, investigations are on-going to determine the 2Sanger Institute, Cambridge, United Kingdom, 3Oxford University Clinical impact of potential virulence determinants of the EAEC strains (aar (-) and Research Unit, Nepal, Kathmandu, Nepal aggR +) in a murine model and their impact on the host on diarrhea and Salmonella Typhi is the causative agent of typhoid. There are a variety growth impairment. of methods for diagnosing typhoid; all lack sensitivity and specificity. Serological assays targeting LPS and/or flagella are commonly used settings to diagnose typhoid. However, cross-reactivity makes these antigens unsuitable for assessing disease burden and diagnosing acute illness. There have been few studies focusing on studying the immunological response to specific Salmonella antigens. Using an S.Typhi antigen array we identified a number of antigens that elicited a significant IgM or IgG response greater than that of uninfected controls. We selected the best twelve (6 IgM and 6 IgG) antigens that gave a differential response between the groups for further investigation. Firstly, we expressed and purified these antigens and immunized mice to study the ability of the mouse serum to stimulate bacterial killing. All of the serum samples from immunized mice were able to stimulate a bactericidal response against S. Typhi, inhibiting >80% of the bacterial growth over three hours.

astmh.org 502 Additionally, eight of the serum samples had a bactericidal effect on S. 1643 Paratyphi A. None of the serum from immunized mice demonstrated any bactericidal activity against S. Typhimurium. We further investigated this INCIDENCE AND ETIOLOGY OF INFECTIOUS DIARRHEA FROM bactericidal response by repeating the experiments with gene knockout A MULTIYEAR FACILITY-BASED SURVEILLANCE SYSTEM IN strains. There was a marked reduction in bactericidal activity with the GUATEMALA, 2008-2012 immunized mouse serum on the strains of S.Typhi strains harboring the 1 1 1 2 respective specific antigen encoding gene knockout. Specifically, serum Wences Arvelo , Aron Hall , Olga Henao , Beatriz Lopez , Chris 2 3 3 1 from mice immunized with CdtB (subunit of typhoid toxin) demonstrated Bernart , Juan C. Moir , Lissette Reyes , Susan P. Montgomery , 1 2 2 a significant reduction bactericidal activity against a cdtB S.Typhi mutant Oliver Morgan , Alejandra Estevez , Maria R. Lopez , Michele B. 1 1 1 1 in comparison to wild-type S.Typhi. These preliminary data show that Parsons , Jan Vinje , Nicole Gregoricus , Jon Gentsch , Umesh 1 1 1 1 S.Typhi 12 antigens appear to stimulate specific and strong immunological Parashar , Eric Mintz , Joe Bryan , Kim A. Lindblade responses in patients with acute typhoid. Furthermore, we suggest these 1Centers for Disease Control and Prevention, Atlanta, GA, United States, antigens may be candidate diagnostics or subunit vaccines, and provide a 2Universidad del Valle de Guatemala, Guatemala City, Guatemala, 3Ministry novel insight for further understanding of host immune responses induced of Health and Welfare, Guatemala City, Guatemala during acute typhoid. Diarrheal diseases are a major cause of morbidity and mortality worldwide, yet data on etiology and population-level incidence in developing countries 1642 are limited. Diarrhea surveillance was conducted at two hospitals and 10 ambulatory clinics in the departments of Santa Rosa and Quetzaltenango THE SPATIOTEMPORAL DYNAMICS AND PHYLOGENETICS OF in Guatemala. A case was defined as a person of any age having≥ 3 SALMONELLA PARATYPHI A IN KATHMANDU, NEPAL loose stools in a 24-hour period and was admitted or presented to the Duy Pham, Stephen Baker surveillance facilities. Epidemiologic and clinical data were collected. Stool specimens were tested for bacterial, parasitic, and viral enteric pathogens. Oxford University Clinical Research Unit, Ho Chi Minh, Vietnam Estimated incidence rates were calculated by adjusting for healthcare Enteric fever is a life threatening systemic disease caused by the bacteria seeking behaviors, based on results from a survey in the surveillance Salmonella Typhi and Salmonella Paratyphi A, B, and C. Typhi is the major area assessing the proportion of those with reported diarrhea who agent of enteric fever, yet Paratyphi A is emerging at an unprecedented visited a surveillance facility during their illness. From November 2008 to rate. In our location in Patan Hospital, Kathmandu, standardised blood December 2012 there were 5,331 diarrhea cases. The weighted estimated culture surveillance over the last ten years has shown annual increases community incidence averaged 659 diarrhea cases per 10,000 persons in the proportions of individuals with Paratyphi A. More than two per year during the four year period. The estimated incidence was highest thirds of the culture confirmed enteric fever cases are now caused by among children aged <5 years, averaging 1,584 cases per 10,000 children this serovar. Both Typhi and Paratyphi A are systemic pathogens that per year, while among those aged ≥5 years the estimated incidence induce indistinguishable syndromes. However, they exhibit contrary averaged 311 cases per 10,000 persons. From 2008-2009 samples from epidemiologies, different geographical distributions, and different 1,401 (26%) cases were tested for all the pathogens of interest. Among propensities to develop resistance to antimicrobials. Additionally, they these, 846 (60%) specimens were from children aged <5 years in whom are genetically and phenotypically distinct, having gone through a a virus was identified in 211 (25%) patients; of which, 178 (84%) tested lengthy process of convergent evolution to cause an identical disease. To positive for norovirus and 101 (48%) for rotavirus, including co-infections. understand the emergence and the molecular epidemiology of Paratyphi Among the 555 patients aged ≥5 years the most frequently identified A in our setting we genome sequenced 182 organisms isolated from etiology was bacterial with 134 (24%) cases. Diarrheagenic Escherichia coli patients with acute or relapsed enteric fever, and a number isolated from was detected in 94 (70%) cases, Shigella spp in 31 (23%), Campylobacter the gallbladder of asymptomatic carriers. Performing phylogenetic analysis spp in 5 (4%), and Salmonella spp in 4 (3%) cases. Identification of and evolutionary reconstruction we find that Paratyphi A is isolated and parasites was low (24 cases, 9%), and most cases were among those genetically distinct from Typhi. Our data show that Paratyphi A has been aged 5-19 years. These data demonstrate a substantial burden of viral and through a major clonal expansion in Kathmandu, apparently driven by bacterial diarrheal diseases in Guatemala, which may help guide public resistance to fluoroquinlones and increased virulence through multi- health policies aimed at reducing the burden of illness and death due to copy effector proteins. Contemporary isolates of Paratyphi A have been diarrhea. introduced from other parts of Asia and induced a clonal replacement of the native strain(s). We surmise that Typhi and Paratyphi A have a dissimilar epidemiology in Nepal with Paratyphi A associated with spatiotemporal 1644 outbreaks and person-to-person transmission. Our study is the first to tackle the local phylogenetics and spatiotempral dynamics of Paratyphi A. SALMONELLA ENTERICA SEROVAR ENTERITIDIS OUTBREAK Our work outlines new perspectives on enteric fever and will pave the way AT A LODGE IN MOKOPANE, LIMPOPO PROVINCE, JANUARY for future genomic epidemiology investigations of this important emergent 2014 pathogen. Akhona N. Tshangela National Institute for Communicable Diseases, Johannesburg, South Africa Salmonella enterica serovar Enteritidis is a leading cause of foodborne disease worldwide, but there is little data available in South Africa. Asymptomatic food-handlers have also been associated as a source of infection. We investigated the aetiology of an acute gastroenteritis outbreak among persons staying at a lodge in Limpopo Province, South Africa in January 2014. A retrospective cohort analysis was used to determine the risks of illness associated with consuming foods and/or beverages at the lodge. The at-risk population were contacted to complete a standard questionnaire related to food and beverages consumed at the lodge, symptoms of illness, visits to healthcare facilities and specimen submission for pathogen testing. Food and water samples were tested, as well as completion of an environmental assessment questionnaire by staff and external caterers. The data was categorised and STATA version 12 was used for multivariate analyses. A total of 73 ill persons, including astmh.org 503 3 laboratory-confirmed infections, were identified: 69/109 (63%) of the 1646 selected cohort were seen at health facilities. Of the at-risk population 87% (109/124) completed the standard questionnaire: 66 cases of INTESTINAL INFLAMMATION AND ALTERED BONE gastrointestinal illness and 43 healthy individuals were identified, with a METABOLISM IN PERUVIAN INFANTS corresponding attack rate of 61%. Most of the cases were females (86%, n=57) with a mean age of 33 years (S.D=7.1), and 36% (n=24) of the Gwenyth Lee, Pablo Peñataro Yori, Margaret Kosek cases were hospitalised. Epidemiological data suggested a point source Johns Hopkins School of Public Health, Baltimore, MD, United States outbreak with no further transmission. Statistical analysis of survey data Low-grade inflammation resulting from frequent enteric episodes and indicated consumption of diluted fruit juice (from concentrate) adjusted by resulting enteropathy is believed to be a causes of growth faltering among other food and beverage items, presented a risk ratio of 1.5 (95% CI, 1.1- children in the developing world. In order to clarify these associations, 1.8, p=0.032). Environmental analysis indicated increased risks for cross- markers of bone metabolism may be beneficial, as they are more dynamic contamination. The outbreak was possibly due to cross-contamination of over short intervals than anthropometry. In order to test this hypothesis, food/ beverages prepared in the lodge kitchen, and fruit juice consumption urine samples from 139 6-month olds and 64 15-month old Peruvian was the main exposure associated with ill cases. Feedback on food safety Amazonian infants, and serum samples from the same children at 7 and hygiene practices to prevent cross-contamination at the kitchen lodge and 15 months, were tested for markers of bone collagen formation were provided. (plasma osteocalcin (OC)) and resorption (urinary deoxypyridinoline (DPD)/ creatinine (Cr)), as well as the acute phase protein plasma alpha-1-acid 1645 glycoprotein (AGP) and the plasma cytokines TNF-α, interleukin-6 (IL-6), and IL-1β , and fecal alpha-1-antitrypsin (AAT), a marker of intestinal FECAL SHEDDING OF ROTAVIRUS FOLLOWING ROTARIX inflammation. The mean plasma osteocalcin at seven months was VACCINATION IN A COHORT OF BOLIVIAN INFANTS 41.2µg/L in boys and 36.4µg/L in girls, by 15 month this had fallen to Rachel M. Burke1, Paulina A. Rebolledo2, Anna Fabiszewski and 34.1 µg/L and 25.2 µg/L, respectively. The mean DPD/Cr at 6 months de Aceituno1, Lucia Inchauste3, Volga Iñiguez3, Rita Revollo4, was 58.9 nmol/mmol Cr and 65.1 nmol/mmol Cr for boys and girls, Parminder Suchdev1, Juan Leon1 respectively, and at 15 months, 68.3 nmol/mmol Cr and 52.1 nmol/mmol Cr for boys and girls, respectively. The mean length-for-age Z score (LAZ) 1 Rollins School of Public Health, Emory University, Atlanta, GA, United at 6 months was -1.3 and 19.4% were stunted (LAZ< -2). By15 months of 2 States, Emory University School of Medicine, Atlanta, GA, United States, age the mean LAZ was -2.0 and 53.1% were stunted. The mean weight- 3 Universidad Mayor de San Andrés, La Paz, Plurinational State of Bolivia, for-length Z-score (WLZ) was 0.9 at 6 months , 0.8 at 7 months, and 0.4 4 Hospital Municipal Los Andes, El Alto, Plurinational State of Bolivia at 15 months. 76.4% of children at 6mo, and 69.1% at 15 month were Worldwide children under the age of five suffer from an estimated 138 classified as having subclinical inflammation, defined by AGP> 1g/L. Bone million cases of rotavirus (RV) diarrhea per year. Currently available oral collagen metabolism was altered by nutritional status, as OC was inversely RV vaccines, such as the human attenuated monovalent vaccine Rotarix, associated with both length-for-age and weight-for-length, and DPD/Cr increase RV-specific immunity and prevent severe diarrhea. Following was positively associated with length-for-age. Correspondingly, the ratio of vaccination, RV vaccine strains are expected to replicate in the intestinal OC/DPD was highest among shorter and leaner children. After adjusting tract and be excreted in feces. Fecal shedding of vaccine strains could for anthropometric status, age, and gender, OC and the ratio of OC/DPD lead to horizontal transmission of such strains and theoretically afford were both inversely associated with fecal AAT, but not with plasma AGP, protection to unvaccinated contacts, which would be of substantial benefit tnf-alpha, IL-6, or IL-1beta. Our findings suggest that bone metabolism is in impoverished countries such as Bolivia where rates of immunization are suppressed among children with chronic intestinal inflammation. suboptimal or RV associated morbidity is high. With the ongoing rollout of RV vaccines in low-middle income countries, it is imperative to evaluate 1647 and describe RV vaccine-virus shedding in vaccine recipients. Between June 2013 and April 2014 a birth cohort of 462 Bolivian infants were enrolled EPIDEMIOLOGY OF SELF-REPORTED HEALTH EVENTS and followed through receipt of Rotarix to quantify the prevalence of AMONG DEPLOYED U.S. MILITARY PERSONNEL fecal shedding within 7 days of the initial vaccine dose (at approximately 2 1 2 2 months of age). Shedding was assessed using enzyme immunoassay (EIA) Ruvani M. Chandrasekera , Mark S. Riddle , Chad K. Porter and in a subset of infants real-time reverse transcription-polymerase chain 1Cherokee Nation Technology Solutions, Silver Spring, MD, United States, reaction (RT-PCR) was used for confirmation. Bivariate logistic regression 2Naval Medical Research Center, Silver Spring, MD, United States was used to identify potential predictors of shedding. The mean age at Deployed military personnel are at risk of experiencing numerousadverse the initial (pre-vaccine) visit was 35 days (SD 8 days), and 55% of the health events to include diarrhea, vomiting, fever and muscle aches infants were male. Baseline prevalence of stunting was 20%, prevalence while on humanitarian and war-fighting operations. These symptoms of preterm birth was 19%, and prevalence of low birth weight was 7%. can negatively impact servicemember effectiveness and epidemiologic The mean maternal age was 26 years (SD 6 years), and 61% of mothers studies are needed to describe their frequency and associated risk factors; had completed secondary school. Shedding was identified in 6 infants out however, extensive prospective epidemiological studies in operational of 305 tested (2%). All before-mentioned predictors were tested, but no environments are challenging. We utilized self-reported data collected significant associations with shedding were detected (likely due to lack through the Post-Deployment Health Assessment from individuals of power). Initial hypotheses for the low shedding rate include the high following operational deployments and compared prevalence estimates prevalence of exclusive breastfeeding, role of maternal antibodies, and across region/country of deployment, multiple demographic characteristics circulating RV in the community; these and other potential explanations as well as pre-existing medical conditions. Univariate and multivariate will be addressed in future investigation. logistic regression methods were also used to identify unique risk factors while controlling for important covariates. Of 21,982 subjects, the top five self-reported symptoms included back pain (15.9%), feeling tired/problems sleeping (15.7%), swollen, stiff or painful joints (13.6%), diarrhea (12.7%) and muscle aches (11.1%). Among those reporting diarrhea/vomiting, a high proportion were assigned to limited duty/bed rest (36% and 54%, respectively). Further data will be presented on the estimated level of care required for reported adverse health events, and potential risk factors for increased self-report of diarrhea and vomiting. While these results are limited to self-report, the data support prior studies highlighting diarrhea

astmh.org 504 and vomiting as significant causes of morbidity and troop down-time that past estimates of the costs of vaccination per life year saved have during operational deployment. Furthermore, recent studies highlighting been overestimated when compared to an analysis that takes into account the link between acute gastroenteritis and long-term adverse health accurate estimates of indirect vaccine protection at different pricing levels. outcomes raise the importance of these common, deployment-related health events. Continued evaluation of primary prevention strategies is 1650 needed. CHARACTERIZING THE REGIONAL AND GLOBAL 1648 DISTRIBUTION AND BURDEN OF CHOLERA REGULATION OF SMALL RHO GTPASES IN REDUCING Sean M. Moore, Andrew A. Azman, Carla E. Zelaya, Justin Lessler INTESTINAL CELLS MIGRATION INDUCED BY STRAINS OF Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United ENTEROPATHOGENIC ESCHERICHIA COLI States There are an estimated 2.8 million cholera cases per year globally, but 1 1 1 Paloma A. Cavalcante , Mara M. Prata , Antônio V. Silva , Josiane the majority of these cases are not detected or reported. Because of 1 1 1 1 S. Quetz , Alexandre B. Havt , Ila F. Lima , Eunice B. Carvalho , inadequate surveillance and reporting the global distribution of cholera 1 1 1 Pedro H. Quintela , Tamara S. Rodrigues , Rafael H. Gonzaléz , risk and its public health burden are poorly known. Previous attempts to 1 2 3 Rosemeyre S. Freire , Sean R. Moore , James Nataro , Richard L. determine the global burden of cholera have relied on a limited number 3 1 Guerrant , Aldo A. Lima of case studies that do not capture the broad range of settings and 1Federal University of Ceara, Fortaleza, Brazil, 2University of Cincinnati, environmental conditions where cholera occurs. Here we assemble a Cincinnati, OH, United States, 3University of Virginia, Charlottesville, VA, database of cholera surveillance and incidence reports from a variety of United States government, scientific, and non-governmental agency sources, with a Intestinal epithelial migration provides an important early response particular focus on sub-Saharan Africa where a majority of cholera cases when intestinal pathogens damage the host intestinal barrier. Small Rho have been reported in the past several decades but where the distribution GTPases Rac1, RhoA and Cdc42 are important regulators of this migratory of risk and burden is still poorly understood. We will then use a formal response. We sought to test the hypothesis that EPEC impairs intestinal modelling framework to associate cholera transmission with environmental epithelial migration in vitro via small Rho GTPase-dependent mechanisms. and socioeconomic variables and map the global distribution of cholera Methods: We investigated the effects of EPEC strain E2348/69, EPEC strain risk and incidence. As a preliminary analysis we developed cholera LDI001 (isolated from a malnourished child), and commensal E. coli HS on incidence maps for the West African country of Guinea-Bissau using IEC-6 cell migration; as well as on the regulation of transcription and gene a hierarchical Bayesian framework with cholera data at spatial scales expression of small Rho GTPases by qPCR and confocal immunofluorescent ranging from neighborhood-level incidence in the capital city of Bissau microscopy, respectively. Results: We observed a significant reduction in to country-level reports. Cholera incidence from 1986-2009 was highest IEC-6 cell migration for all E. coli strains tested. However, pathogenic EPEC in the island and coastal districts (including Bissau city). Incidence did not strains reduced migration to a greater degree than E. coli HS. Only EPEC change significantly between the 1990s (which included outbreaks in E2348/69 induced significant cellular necrosis. Gene analyses of small Rho 1994 and 1996-1997) and the 2000s (which included outbreaks in 2005 GTPases revealed an increase in rac1 transcription in EPEC LDI001 infected and 2008), except for an increase of 26-280% in the islands of the Bijagos cells and upregulation of rhoA transcription following infection with all Archipelago. Improving our understanding of the spatial distribution strains. Confocal imaging showed an increase in Rac1 expression and of cholera in Guinea-Bissau and associating incidence with climate, decrease in RhoA in response EPEC LDI001 infection. We further observed environmental and socioeconomic factors will provide a basis for planning increased expression of Cdc42 in all infected groups. Conclusions: The public health preventions to reduce cholera transmission in this region. results suggest differential suppression of migration and co-regulation of small Rho GTPases in response to infection with enteropathogenic vs. 1651 commensal E. coli strains. These in vitro data corroborate an emerging in vivo and clinical understanding of the pathobiology of this infection and its BIOMARKERS OF ENVIRONMENTAL ENTEROPATHY FOR associations with malnutrition and intestinal barrier injury. POSITIVELY ASSOCIATED WITH TOXIN-SPECIFIC B AND T CELL RESPONSES TO AN ORAL CHOLERA VACCINE IN 1649 BANGLADESHI CHILDREN INCORPORATING HERD PROTECTION INTO A COST-BENEFIT Daniel T. Leung1, Taufiqur R. Rahman2, Muhammad Ikhtear 2 2 2 2 ANALYSIS OF TYPHOID FEVER VACCINE INTERVENTIONS Uddin , Naoshin S. Nishat , Mohammad Rubel Hoq , Amit Saha , Jason B. Harris1, Stephen B. Calderwood1, Firdausi Qadri2, Edward Marina Antillon, Virginia E. Pitzer T. Ryan1 Yale University, New Haven, CT, United States 1Division of Infectious Diseases, Massachusetts General Hospital, Boston, Recent studies have shed light on the costs and savings of typhoid fever MA, United States, 2International Centre for Diarrhoeal Disease Research, vaccination campaigns, but no studies been able to quantify the potential Bangladesh, Dhaka, Bangladesh economic savings attributable to realistic estimates of herd protection, the Environmental enteropathy (EE) is a poorly understood condition that protection conferred to individuals who did not receive the vaccine. In light refers to chronic alterations in intestinal permeability, absorption, and of numerous budget constraints in resource-poor settings, it is necessary inflammation that affects young children in resource limited settings. While to more accurately estimate the indirect effects of vaccine campaigns as EE has been linked to suboptimal oral vaccine performance in children, the well as the additional potential financing mechanisms that may lower causative immunological mechanisms are poorly defined. The objective of the costs of vaccination per DALY saved. Field studies are now available this study was to determine how markers of enteropathy are associated to assess the burden of typhoid fever and the possible impact of Vi- with immune responses to an oral cholera vaccine (OCV). We collected polysaccharide and Vi-conjugate vaccines. Using mathematical models for blood and stool from 40 Bangladeshi children who received two doses of typhoid transmission, we can quantify the indirect protection of vaccines an OCV given 14 days apart. We measured five EE markers, including stool under different vaccine strategies. Moreover, surveys on the private myeloperoxide (MPO), a marker of intestinal inflammation, stool alpha demand for these vaccines in South Asian contexts, where the disease is anti-trypsin (AAT), a marker of intestinal absorption, as well as plasma endemic, inform calculations on the optimal vaccine subsidies necessary to endotoxin core antibody (EndoCab), plasma intestinal fatty acid binding achieve a desired level of vaccine coverage while recuperating some of the protein (iFABP), and plasma soluble CD14 (sCD14), all markers of microbial programmatic costs through user fees. With that in mind, we will show translocation. We measured cholera toxin (CT)- and lipopolysaccharide

astmh.org 505 (LPS)-specific antibody responses by ELISA, toxin-specific 1653 responses by flow cytometry following whole blood culture, and T cell culture cytokines by Luminex array. Using a multiple linear regression OPTIMIZING THE CHEMOTHERAPEUTIC APPROACH FOR THE model, we assessed each vaccine-associated immune response outcome TREATMENT OF BURULI ULCER: POSSIBLE OPTIONS AND as separate dependent variables, and used log-transformed EE marker RESEARCH NEEDS measurements, along with gender, blood group, and age, as independent 1 2 2 3 variables. We found stool MPO to be a positive predictor of antibody Charitha Gowda , Martina Casenghi , Julien Potet , Eric Comte , 2 responses to CT, plasma iFABP a positive predictor of gut-homing memory Jennifer Cohn T cell responses, and stool AAT a positive predictor of interferon-gamma 1University of Pennsylvania, Philadelphia, PA, United States, 2Medecins responses. No marker predicted antibody responses to LPS. Variance Sans Frontieres Access Campaign, Geneva, Switzerland, 3Medecins Sans inflation factor for all independent variables were < 1.6, suggesting Frontieres, Geneva, Switzerland no multi-collinearity. In summary, we demonstrate that biomarkers of Buruli ulcer (BU) is a serious necrotising skin infection caused by the environmental enteropathy are positive predictors of toxin-specific immune environmental pathogen Mycobacterium ulcerans, and represents the responses to an OCV. third most common mycobacterial infection. Current WHO treatment has simplified the delivery of care for BU by recommending that 1652 early, limited lesions be treated with antibiotics alone. However, these recommendations still present significant disadvantages including the LEPROSY IN NIGERIA (2008-2012): AN EVALUATION OF THE use of injectable agents. There is an urgent need to identify alternative NATIONAL SURVEILLANCE SYSTEM oral regimens that are effective, short-course, have few drug-drug Aishatu Abubakar Sadiq1, Joshua A. Obasanya2, Patrick Nguku1, interactions with antiretrovirals and can be used in children. We Gabrielle Poggensse1, Kabir Sabitu3 performed a systematic literature review for publications focused on chemotherapy for M. ulcerans infection. We searched PubMed, EMBASE, 1Nigeria Field Epidemiology and Laboratory Training Programme, Abuja, Scopus, WHO Global Index Medicus, CAB Abstracts and Cochrane Nigeria, 2National Tuberculosis and Leprosy Control Programme, Abuja, Library with standardized search terms, screened abstracts submitted Nigeria, 3Department of Community Medicine, Ahmadu Bello University, to international conferences and assessed the ClinicalTrials.gov registry. Zaria, Nigeria While there were no restrictions by publication date or type, only articles Leprosy is a chronic infectious disease caused by Mycobacterium leprae in English, French and Italian as of December 31, 2012 were included. an acid fast, rod shaped bacillus. It was historically associated with social We included in vitro and clinical studies, with the primary outcomes isolation and psychological consequences. Although eradicated in most of clinical resolution of the ulcer without (clinical studies) and countries, ongoing transmission in African and Asian countries requires assessment of in vitro activity (pre-clinical data). We excluded all studies enhanced surveillance and monitoring. In 2012 Nigeria reported 3805 without microbiological confirmation of M. ulcerans infection. 49 clinical cases and no deaths. We evaluated the national Leprosy surveillance studies including 6 RCTs, 14 observational cohorts, 11 case series and 18 system to assess its usefulness and attributes. Centers for Disease case reports were identified. Various drugs and drug combinations were Control(CDC) guidelines for evaluating surveillance systems were used. identified as having clinical efficacy against BU disease in resource-poor Ten Stakeholders from partner agencies, national and state level were settings. In particular, the combinations of clarithromycin+rifampin and interviewed using structured questionnaires. Secondary data (2008- clarithromycin+fluoroquinolones demonstrated good efficacy and safety. 2012) was abstracted from surveillance data submitted to the National In vitro data reveal a number of promising compounds. Although recent Tuberculosis and Leprosy Control Program. Laboratory diagnostic studies indicate that a fully oral regimen for BU may be as equally effective capacity was also assessed.Out of 20,623 cases reported, 3805(18.45%) as regimens containing aminoglycosides, further research is needed to were received in 2012, 3623(17.57%) in 2011,3913(18.97%) in identify and evaluate new treatments. The anti-tuberculosis research & 2010,4383(21.25%) in 2009 and 4899 (23.76%) in 2008.The prevalence development (R&D) pipeline represents a potentially rich source of novel reduced from 4.3x10-3 /10,000 population in 2008 to 1.2x10-4/10,000 compounds for BU treatment. We propose an R&D agenda aimed at population in 2011.Child proportion was 10.7% in 2008 and 8.0% in delivering new, more efficacious and readily implementable treatments 2011.Grade 2 disability rate ranged between 11.7% and 13.4%.Being an against Buruli ulcer in resource-limited settings. adult (OR=1.97; 95% CI =1.37-2.82) and male (OR=1.25; 95% CI=1.05- 1.48) was found associated with Multibacillary Leprosy. Residents of the 1654 Northwest region (OR=0.73; 95%CI=0.59-0.90) were less likely to have Multibacillary Leprosy. The system is active and rated simple by 17(85%) PERSISTENT GUT MICROBIOTA IMMATURITY IN of respondents. Review of weekly,quarterly and monthly reporting forms MALNOURISHED BANGLADESHI CHILDREN and records at the national level showed timeliness. All suspected cases 1 2 3 (100%) were laboratory tested within 24hours of presentation.There is a Sathish Subramanian , Sayeeda Huq , Tanya Yatsunenko , 2 2 2 high laboratory turnover of staff and low numbers of personnel trained Rashidul Haque , Mustafa Mahfuz , Mohammed A. Alam , Amber 4 3 3 3 in laboratory diagnosis.The system is fully integrated with surveillance Benezra , Joseph DeStefano , Martin F. Meier , Brian D. Muegge , 3 3 5 of Tuberculosis at all levels. Leprosy transmission is still ongoing and Michael J. Barratt , Laura G. VanArendonk , Qunyuan Zhang , 5 6 2 the WHO elimination target (<1 case/10,000 population) has been Michael A. Province , William A. Petri , Tahmeed Ahmed , Jeffrey 3 achieved at the national level. Pockets of leprosy exist in the northwest I. Gordon region.We recommend intensification of surveillance activities in all 1Center for Genome Sciences and Systems Biology, Washington University, zones,improvement of Laboratory diagnostic capacity and recruitment of St. Louis, MO, United States, 2International Centre for Diarrhoeal Disease additional personnel. The system is acceptable,flexible, simple and timely. Research, Bangladesh, Dhaka, Bangladesh, 3Center for Genome Sciences The system is meeting its purpose and promotes the achievement of the and Systems Biology, Washington University, St. Louis, MO, United States, global elimination target for leprosy. 4Department of Anthropology, New School for Social Research, New York, NY, United States, 5Division of Statistical Genomics, Washington University, St. Louis, MO, United States, 6Departments of Medicine, Microbiology and , University of Virginia School of Medicine, Charlottesville, VA, United States Therapeutic food interventions have reduced mortality in children with severe acute malnutrition (SAM) but incomplete restoration of healthy growth remains a major problem. The relationships between the type of

astmh.org 506 nutritional intervention, the gut microbiota, and therapeutic responses seen anymore. The Polymerase Chain Reaction (PCR) for IS2404 positivity are unclear. In the current study, bacterial species whose proportional rate among patients referred by the community health volunteers was not representation define a healthy gut microbiota as it assembles during the systematically lower than in patients referred by other systems. This study first two postnatal years were identified by applying a machine-learning- clarifies the role played by community health volunteers. It highlights that based approach to 16S rRNA datasets generated from monthly fecal in Bénin, the community health volunteers are an important link in the samples obtained from a birth-cohort of children, living in an urban slum control of BU. of Dhaka, Bangladesh, who exhibited consistently healthy growth. These age-discriminatory bacterial species were incorporated into a model that 1656 computes a ‘relative microbiota maturity index’ and ‘microbiota-for-age Z-score’ that compare development (defined here as maturation) of a GREEN SYNTHESIS OF SILVER NANOPARTICLES USING child’s fecal microbiota relative to healthy children of similar chronologic FLORAL EXTRACT OF GOMPHRENA GLOBOSA AND ITS age. The model was applied to twins and triplets (to test for associations ANTIMICROBIAL ACTIVITY AGAINST MULTI DRUG RESISTANT of these indices with genetic and environmental factors including BACTERIA diarrhea), children with SAM enrolled in a randomized trial of two food 1 2 interventions, and children with moderate acute malnutrition. Our Kovendan Kalimuthu , Arokiyaraj Selvaraj , Murugan 1 3 results indicate that SAM is associated with significant relative microbiota Kadarkarai , Vincent Savariar immaturity that is only partially ameliorated following two widely used 1Bharathiar University, Coimbatore, India, 2National Institute of Animal nutritional interventions. Immaturity is also evident in less severe forms Science, Suwon, Republic of Korea, 3Loyola College, Chennai, India of malnutrition and correlates with anthropometric measurements. Antibiotic resistance has become a major clinical and public health Microbiota maturity indices provide a microbial measure of human problem within the lifetime of most people living today. Silver postnatal development, a way of classifying malnourished states, and a nanoparticles (AgNPs) are well known biocidal substances that can parameter for judging therapeutic efficacy. More prolonged interventions be incorporated as antimicrobial agents in phar¬macology, veterinary with existing or new therapeutic foods and/or addition of gut microbes medicine, implants, wound dressings, and topical ointments. AgNPs were may be needed to achieve enduring repair of gut microbiota immaturity in also found to exhibit antimicrobial activities. The present work reports one childhood malnutrition and improve clinical outcomes. step ecofriendly method for the synthesis of AgNPs using Gomphrena globosa and its antibacterial effects against drug resistant bacteria. In the 1655 present results, AgNPs was characterized by ultraviolet-visible spectroscopy, X-ray diffraction spectroscopy, Transmission electron microscopy and CONTRIBUTION OF THE COMMUNITY HEALTH VOLUNTEERS particle size analyzer. The synthesized particles were found to be spherical IN THE CONTROL OF BURULI ULCER IN BENIN in shape and sizes ranged between 55-60 nm. Further energy-dispersive Yves T. Barogui1, Ghislain E. Sopoh2, Roch C. Johnson3, Janine de X-ray spectroscopy confirmed the presence of silver. Furthermore these Zeeuw4, Ange D. Dossou2, Jean G. Houezo2, Annick Chauty5, Julia green synthesized AgNPs were found to show significant antimicrobial Aguiar6, Didier Agossadou7, Patrick A. Edorh3, Kingsley Asiedu8, effect against the drug resistant Methicillin resistant Staphylococcus Tjip S. van der Werf4, Ymkje Stienstra4 aureus, cipro flaxin resistant Escherichia coli, and carbapenem resistant Acetobacter baumanii. This outcome may pave a way for using floral 1 Centre de Dépistage et de Traitement de l’Ulcère de Buruli de Lalo, extract of the AgNPs a drug carrier system to cure bacterial diseases. Lalo, Benin, 2Centre de Dépistage et de Traitement de l’Ulcère de Buruli d’Allada, Allada, Benin, 3Université d’Abomey-Calavi, Abomey-Calavi, Benin, 4University of Groningen, University Medical Center Groningen, 1657 Department of /Infectious Diseases, Groningen, FREQUENCY OF VIRULENCE GENOTYPES IN ESCHERICHIA 5 Netherlands, Centre de Dépistage et de Traitement de l’Ulcère de Buruli COLI STRAINS ISOLATED FROM URINARY TRACT INFECTIONS 6 de Pobè, Pobè, Benin, Centre de Dépistage et de Traitement de l’Ulcère OF MEXICAN PATIENTS de Buruli de Zangnanado, Zangnanado, Benin, 7Programme National de Lutte Contre la Lèpre et l’Ulcère de Buruli, Ministère de la Santé, Cotonou, Gloria Luz Paniagua Contreras1, Bianca Hidalgo Hernández1, Benin, 8Department of Control of Neglected Tropical Diseases, World Selene Acebo Falcón1, Eric Monroy Pérez1, Raymundo Rodríguez Health Organization, Geneva, Switzerland Moctezuma2, Pablo Domínguez Trejo2, Sergio Vaca1 Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium 1Universidad Nacional Autónoma de México, Tlalnepantla, Edo. de México, ulcerans. Usually BU begins as a painless nodule, plaque or edema, Mexico, 2Instituto Mexicano del Seguro Social, Tlalnepantla, Edo. de ultimately developing into an ulcer. Striking is the high number of patients México, Mexico presenting with ulcers in an advanced stage. Such late presentation will Uropathogenic Escherichia coli (UPEC) is responsible for a high percentage complicate treatment and have long term disabilities as a consequence. of urinary tract infections (UTIs) worldwide, among them cystitis and The disease is mainly endemic in West Africa. Strategy for control this pyelonephritis. The purpose of this work was to determine the frequency disease is early detection using community village volunteers. This study of the genotypes: pap (pilus associated with pyelonephritis), papGI and aims to understand the contribution of different actors in the current papGII (pilus associated with pyelonephritis GI and GII), hlyA (haemolysin), referral pattern in Benin, the role of the different referral systems on afa (afimbrial adhesin), sfa (S fimbriae), iron (iron), iuc (aerobactin), cnf the stage of disease at presentation in the hospital and the diagnostic (cytotoxic necrotizing factor), astA (enteroaggregative toxin), sap (she precision of Buruli ulcer. Patient information of Buruli ulcer patients pathogenicity island marker) and set (Shigella enterotoxin 1) in a group of that reported to one of the four BU centers in Benin between January E. coli strains isolated from Mexican patients suffering UTIs. E. coli strains 2008 and December 2010 was collected using the WHO/BU01 forms. were identified by biochemical tests and by PCR amplification of 16S rRNA. Information traced from these forms were general characteristics of Genes pap, papGI, papGII, hlyA, afa, sfa, iron, iuc, cnf, astA, sap and set the patient, the results of diagnostic tests, the presence of functional were detected by multiplex PCR and by end-point PCR. Urine samples of limitations at start of treatment, lesion size, patient delay and the referral 100 urinary tract infected patients were microbiologically analyzed. E. coli system. The role of the different referral systems on the stage of disease at was identified in 60% of the samples (n=60). Of the E. coli strains, 48.3% presentation in the hospital was analyzed by a logistic regression analysis. (n=29) carried the set gene; 41.6% (n= 25) carried papGI; 26.6% (n=16) About a quarter of the patients (26.5%) were referred to the hospital carried hlyA; 23.3% (n=14) carried afa; 21.6% (n=13) carried pap; 20% by the community health volunteers. In our data, community health (n=12) carried papGII; 18.3% (n=11) carried sfa; 16.6% (n=10) carried volunteers seemed to refer patients more frequently in an earlier stage of iron; 13.3% (n=8) carried iuc; 6.6% (n=4) carried cnf1; 10% (n= 6) carried disease but after adjustment for the health center, this effect could not be

astmh.org 507 astA and 5% (n=3) carried sap. The high frequency of the identified genes from ICU health care workers’ cell phones twice monthly. Microbiological in the UPEC strains suggests that they are virulent and able to produce identification and resistance patterns were determined by standard cystitis and/or pyelonephritis. methods. Suspicious ESBL-producing bacteria in the antibiogram were confirmed by the phenotypic CLSI ESBL confirmatory test. We then 1658 performed PCR for detection of blaTEM, blaSHV and blaCTX-M genes to characterize the ESBL. A total of 114 employees were enrolled, 114 COMMUNITY-ACQUIRED METHICILLIN-RESISTANT devices were tested, resulting in 491 samples. Twenty-two percent STAPHYLOCOCCUS AUREUS INTRODUCED INTO A (25/114) of providers phones were colonized with nosocomial pathogens. BRAZILIAN PUBLIC PEDIATRIC CLINIC Among 105 Enterobacteriaceae isolated, 33.3% (35/105) produced ESBLs, including 18.8% (9/48) of Enterobacter spp., 55.9% (19/34) of Escherichia 1 2 Mariel A. Marlow , Claudete Araújo Cardoso , Marcos Gabriel coli, 26.1% (6/15) of Klebsiella pneumoniae and 12.5% (1/8) of Klebsiella 3 2 2 Pinheiro , Ivete Martins Gomes , Cláudia Aguiar Barros , Lee W. oxytoca. blaCTX-M was the most prevalent ESBL. ESBLs resulted in a 1 3 Riley , Fábio Aguiar-Alves phenotype of Multidrug resistance: Tobramycin resistance represented 1University of California Berkeley, Berkeley, CA, United States, 2School 74.3% of isolates, both Ciprofloxacin and Sulfamethoxazole/Trimethropim of Medicine, Fluminense Federal University, Niterói, Brazil, 3Pathology 68.6%, Gentamicin 62.9%, Amikacin 17.1%, and Cefoxitin 5.7%. Program, Laboratory of Molecular Epidemiology and Biotechnology, No carbapenem resistance was detected and Metallo-beta-lactamases Rodolfo Albino University Laboratory, Fluminense Federal University, (MBLs), Carbapenemases and AmpC beta-lactamases were not identified Niterói, Brazil in isolated Enterobacteriaceae. Our data suggest cell phones can be an Low-income communities, health care professionals, and adults in important source of ESBL spread in developing world ICUs. Methods to frequent contact with children are all populations known to be at higher prevent outbreaks and transmission of these bacteria from commonly used risk for methicillin-resistant Staphylococcus aureus (MRSA) colonization. fomites, such as cell phones, are needed. The objective of our investigation was to determine risk factors for colonization with MRSA as well as staphylococcal cassette chromosome 1660 mec (SCCmec) genotypes among pediatric health care workers from a public hospital in Rio de Janeiro, Brazil. We collected nasal swabs and data THE ROLE OF YERSINIA PESTIS PRESENSIBILIZATION AND on potential risk factors from 178 health care workers from all pediatric GENETIC BACKGROUND IN RESISTANCE OF BLACK RATS sectors from January to December 2012. Swab cultures were evaluated for AGAINST PLAGUE antimicrobial resistance against Cefoxitin and Oxacillin, and resistance was Voahangy Andrianaivoarimanana1, Guillaume Soubigou2, confirmed by identifying the mecA gene by PCR. MRSA colonization was Minoarisoa Rajerison1, Michel Ranjalahy1, Ronan Jambou2 5.1% (n=9/178). Logistic regression analysis showed being a nurse and working in an inpatient unit as potential risk factors for MRSA colonization 1Institut Pasteur de Madagascar, Antananarivo, Madagascar, 2Institut (adjusted OR= 11.6, 95% CI 1.2 - 110.7 and 2.7, 95% CI 0.3 - 23.7, Pasteur, Paris, France respectively). No non-work related risk factors were identified. Four of In Madagascar, plague remained endemic in the rural areas of the central the nine isolates were found to be SCCmec type IV, the genotype most highlands with the black rat, Rattus rattus, as the main reservoir. The commonly associated with community-acquired MRSA (CA-MRSA). Five occurrence of plague cases from one season to another in the same isolates were observed to be SCCmec type III, with four collected from villages is questioning. One hypothesis is that part of black rats can acquire nurses of various pediatric sectors in August/September alone. We then resistance to plague allowing survival of rat populations and maintenance compared these isolates with nasal MRSA isolates sampled from children of infected fleas and thus of the disease. We previously described within 48 hours of being admitted to the pediatric ward from December resistance of field black rats living in endemic area against plague whereas 2011 to July 2012. Of 11 CA-MRSA isolates from 92 children sampled those from non endemic ones remained sensitive. This study investigates (12.0%), ten were found to be SCCmec IV, including two which preceded whether this resistance is genetically driven or if pre immunization of rats those collected from nurses in the same sector. Given SCCmec type III with Yersinia pestis could increase survival during subsequent infections. F1 isolates appear to have circulated among various pediatric sectors, the generation of black rats, originating either from plague endemic or plague introduction of the more virulent CA-MRSA genotype to the pediatric free zones were obtained and challenged once or twice with Y. pestis. inpatient ward is of high concern. Prevention in the hospital setting may Rat survival, antibody production and gene expression were compared also depend on interventions at the community level in low-income during the acute phase of the disease. First inoculation of a low dose settings. of Y. pestis greatly increases survival of rats against a lethal dose of the bacteria. This protection of primed rats can likely be related to anti-F1 IgG. 1659 Transcriptome analysis of leukocytes five days after infection revealed that genes related to inflammation but also to apoptosis were more expressed EXTENDED-SPECTRUM BETA-LACTAMASE (ESBL)- in rats from non endemic than in those from endemic ones. In the other PRODUCING ENTEROBACTERIACEAE ISOLATED FROM hand, anti-apoptotic BcL2 pathway was highly expressed in resistant HEALTH CARE WORKERS’ CELL PHONES IN FIVE PERUVIAN rats. This suggested that rat susceptibility to infection could be driven by INTENSIVE CARE UNITS: ANTIBIOTIC RESISTANCE PATTERNS apoptosis of activated leukocytes. Transmission of a resistance phenotype AND MOLECULAR CHARACTERIZATION to the F1 generation for R. rattus from endemic plague foci is highlited. These findings highlight the role of low transmission of bacteria in a 1 2 2 2 Steev Loyola , Luz Gutierrez , Gertrudis Horna , Jorge Osada , resistance phenotype of R. rattus to plague. A genetic component of this 1 1 2 Andres G. Lescano , Kyle Petersen , Jesus Tamariz resistance is also supported This study provides critical insights on the role 1U.S. Naval Medical Research Unit - 6, Lima, Peru, 2Universidad Peruana of R. rattus in plague persistence in Madagascar Cayetano Heredia, Lima, Peru Technological advances allow rapid and efficient communication through cell phones; their unsupervised use in hospital environments however, is common. Outbreaks associated with ESBL-producing (Extended-spectrum beta-lactamase) Enterobacteriaceae have been widely described worldwide in intensive care units (ICU), we therefore hypothesized that cell phones might represent a source of these infections in Peru. We conducted a 5-month passive surveillance in 3 Pediatric ICUs and 2 ICUs from 3 hospitals during February to June 2012. Swabs were collected

astmh.org 508 1661 hospitals. Also, the VIM-2 positive isolate demonstrate great than 95% homology with a reference to wild type strain reflecting the capacity of CHARACTERIZATION OF ACINETOBACTER ISOLATES carbapenem-sensitive isolates to acquire to carbapenem resistant. POSITIVE FOR IMP CARBAPENAMASE FROM PERUVIAN HOSPITALS 1663 Nathanael D. Reynolds GENETIC CHARACTERIZATION OF RECOVERED BACILLUS U.S. Naval Medical Research Unit - 6, Lima, Peru ISOLATES FROM THE ENVIRONMENTAL SURVEILLANCE Carbapenem antibiotics constitute the last resort in treating multi-drug SWABS BY SEQUENCING OF GYRB GENE: A PUBLIC HEALTH resistant Acinetobacterinfections typically acquired during hospitalization. APPROACH The effectiveness of carbapenems against Acinetobacter has been compromised in the last decade largely due to the emergence of Irshad M. Sulaiman, Emily Jacobs, Steven Simpson, Khalil carbapenem-hydrolyzing metallo-beta-lactamases (MBL). Metallo-beta- Kerdahi lactamases include the enzymes IMP, VIM, NDM, GIM, and SIM and Food and Drug Administration, Atlanta, GA, United States account for the majority of carbapenem resistance in Gram negative The primary mission of FDA is to enforce the Food, Drug and Cosmetic bacteria. While extensively characterized in East Asia and Europe, currently Act and regulate food, drug and cosmetic products. To assess adulteration there exist limited reports on MBL resistance in Acinetobacter infections in these commodities, FDA uses presence of pathogenic microorganisms from South America. Surveillance of hospital acquired Acinetobacter in manufacturing and distribution areas as one of the regulatory action infections in Lima and Iquitos, Peru between March 2011 and February criteria and to ensure that the firm is following good manufacturing 2013 identified 32 suspected nosocomial isolates (11 from Lima and 21 practices. Further, FDA provides guidance to achieve the said goal by from Iquitos) and 20 clinical and environmental isolates associated within establishing an environmental monitoring program in these facilities. This ICU outbreaks in Lima. Four of the 52 Acinetobacter spp. isolates were study was conducted to verify the effectiveness of pathogen control in positive for blaIMP, consisting of one A. baumannii, one A. haemolyticus, a pharmaceutical compounding facility located in Southeast region of and two A. junii. Phenotypic carbapenem resistance as defined by United States. A total of 28 environmental swabs were collected from minimum inhibition concentrations to imipenem indicates resistance in one several locations of a compounding company premises. The swab samples A. junii and A. haemolyticus. Whole genome sequencing of the blaIMP- were initially examined by conventional microbiologic protocols. Of these, positive isolates identified multiple resistance genes and characterized the several swabs were found positive for the presence of rod-shaped, gram- IMP-16 variant in the A. baumannii and A. junii isolates and IMP-18 in positive bacteria, Bacillus. It is a diverse group of bacteria, and some of its the A. haemolyticus isolate. Phylogenetic analysis indicates no relativeness species are human-pathogenic that can cause range of infections including between the A. junii isolates or any IMP-positive isolates with previously ear infections, meningitis, urinary tract infections and septicemia. Species- identified the IMP-positive Acinetobacter isolates referenced by the NCBI identification of recovered Bacillus isolates were completed by our recently Whole Genome Shotgun Database. developed protocol based on nucleotide sequencing of PCR amplified gyrB gene products. Analysis of data confirmed four species of Bacillus (B. 1662 cereus, B. pumilus, B. subtilis, and B. thuringiensis) in the swabs examined. This newly developed gyrB-based molecular diagnostic protocol can be CHARACTERIZATION OF CARBAPENEMASE-POSITIVE used as a suitable genetic marker for rapid detection of Bacillus in the PSEUDOMONAS AERUGINOSA ISOLATES IN LIMA environmental monitoring program of public health importance. HOSPITALS 1664 Drake H. Tilley U.S. Naval Medical Research Unit - 6, Lima, Peru PRODUCTION AND EVALUATION OF A32 KDA FRAGMENT OF Pseudomonas aeruginosa is an opportunistic pathogen that accounts THE IMMUNOGLOBULIN-LIKE B PROTEIN FROM LEPTOSPIRA for ten percent of all hospital-acquired infections. Therapeutic options Hua-Wei Chen, Giulia Weissenberger, Chien-Chung Chao, Wei- for treatment of P. aeruginosa infections are increasingly limited due to Mei Ching inherent antimicrobial resistance an ability to acquire new mechanisms of resistance. Recently, carbapenemases have emerged as one of Naval Medical Research Center, Silver Spring, MD, United States the major mechanisms of acquired resistance in P. aeruginosa and Leptospirosis is caused by spirochaetes of the genus Leptospira. It is represent a significant clinical concern due to its ability to hydrolyze considered to be the most widespread zoonotic disease in the world. the majority of beta-lactam antibiotics. The most clinically relevant and Symptoms of leptospirosis are easily confused with a variety of other widely disseminated carbapenemases include KPC, IMP, VIM and NDM. febrile illnesses (e.g., dengue and malaria) that require different treatment Currently, little information has been reported on the prevalence of regimens. Currently, the Microscopic Agglutination Test (MAT) is the carbapenemase genes present in P. aeruginosa isolates in Peru. To begin standard method for the diagnosis of leptospirosis. It is not only technically to define carbapenemase resistance in Peru, 124 carbapenem-resistant complex but also time-consuming. With the publication of the whole Pseudomonas aeruginosa isolates were collected from nosocomial and genome sequences of several pathogenic species of Leptospira, hundreds outbreak infections from three hospitals in Lima. Antibiotic resistance to of genes encoding surface-exposed lipoproteins and outer membrane beta-lactam was identified in 18 percent (23/124) of the P. aeruginosa, proteins have been identified as candidates for the development of rapid as defined by disk diffusion assay according to the CLSI guidelines. PCR diagnostics of leptospirosis. An ELISA using recombinant antigens (rLipL32, was performed on all 124 isolates to detect the carbapenemase genes rLipL41, and rLigA-Rep) for the detection of Leptospira-specific antibodies blaKPC, blaIMP, blaVIM and blaNDM. From the 124 isolates, 22 (18%) P. has been developed in our laboratory with sensitivity close to 90%. Here, aeruginosa isolates were identified as the IMP-16 variant, one (1%) isolate we prepared a recombinant protein containing the coding region of amino positive for the VIM-2 varient, and none positive for KPC or NDM, being acids 630-931 of LigB (rLigB-Rep). The over-expressed rLigB-Rep, which this 23 isolates extensively drug-resistant. Finally, in order to determine contains a six-histidine tag at the N-terminus, was primarily found in the genomic-relatedness, the 23 isolates with carbapenemase genes were inclusion body. The solubilized rLigB-Rep in 8 M urea was purified with a analyzed using rep-PCR on the Diversilab system. The 23 isolates clustered nickel column under denatured conditions. We achieved greater than 90% into four genomically distinct groups. Interestingly, several of IMP positive purity as demonstrated by SDS-PAGE. The purified rLigB-Rep was refolded isolates clones were dispersed throughout different hospitals, suggesting by dialysis in buffer (20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA) possible clonal spread of IMP-16 positive P. aeruginosa between Lima containing 6M, 4M, 2M, 1M, and no urea at 4oC. The refolded rLigB-Rep

astmh.org 509 has been shown that it was recognized by confirmed leptospirosis patient ampicilin + sulbactam (10/10), cephalothin (30),ciprofloxacin (5), sera in western blot. These data suggest that rLigB-Rep antigen can be norfloxacin (10) e nitrofurantoin (300) (standard disc diffusion method used to further improve the ELISA assay’s sensitivity. as per CLSI guidelines using discs of standard potency. Furthermore the costs of different classes of antibiotics were compared. Statistical analysis 1665 was performed using the program “Prism” from Graphpad. The results showed that all antibiotics tested here are effective (p <0.05). However EPIDEMIOLOGICAL PROFILE OF LEPTOSPIROSIS CASES, there was an increase in the sensitivity of nitrofurantoin in relation to GUATEMALA: 2008-2013 other antibiotics with a decrease of the resistance of the same antibiotic in compared with others (p <0.05), and a variation in the sensitivity and Maria R. Lopez1, John P. McCracken1, Danilo Alvarez1, David resistance among the fluoquinolonas with the ampicillin more inhibited Moran1, Rafael Ciraiz2, Joe Bryan3, Celia Cordon-Rosales1 beta-lactamase. There was also a significant difference in cost, and 1Universidad del Valle de Guatemala, Guatemala, Guatemala, 2Ministry of showed are more affordable the nitrofurantoin (p <0.05), followed by Health Guatemala, Guatemala, Guatemala, 3Centers for Disease Control fluoroquinolones, beta-lactamase inhibitor and cefalotinas. In conclusion, and Prevention, Atlanta, GA, United States this work shows that should be considered an alternation of treatments Leptospirosis is a widespread zoonosis with a higher incidence in countries for infection by E. coli, thus favoring the control of bacterial resistance and with humid subtropical or tropical climates. Leptospira are usually the cost effective to the population. transmitted to humans through soil or water contaminated with infected urine from mammal hosts (mainly rodents). Outbreaks are associated 1667 with flooding, agriculture, as well as recreational, sporting and military activities. In Guatemala, there are reports of cases in humans and animals ANTIBIOTIC RESISTANCE PATTERNS OF ENTEROBACTER but the epidemiology of leptospirosis is unknown. Active surveillance for SPECIES ISOLATED FROM CHILDREN WITH CYSTITIS IN IRAQ suspected leptospirosis cases was conducted during 2008-2013 among Maitham G. Yousif patients with acute febrile illness (AFI) attending the Cuilapa National Hospital (932 M) or Nueva Santa Rosa ambulatory facilities (1005 M) in Al-Qadysia University, Dewania, Iraq the Department of Santa Rosa, in southern Guatemala. Leptospirosis is Awareness of antibiotic sensitivity patterns among gram negative bacterial typically an undifferentiated AFI but suspected cases are seldom confirmed pathogens is clinically important not only to empirically guide therapy but with laboratory diagnosis. In this study, AFI was defined as self-reported also to monitor the emergence of new drug resistant strains. We report fever or measured temperature ≥38ºC that began <7 days before here a survey of antibiotic resistance in Enterobacter species cultured presentation with no other diagnosis (e.g. pneumonia or diarrhea). Blood from children with cystitis in Iraq. We collected 1,474 urine cultures from samples were taken and tested for IgM anti-Leptospira by enzyme-linked children aged 1-7 years treated at Al-Hakeem Hospital in Najaf governate, immunosorbent assay. Of 553 patients studied (396 hospitalized cases Iraq between January and September of 2010. Enterobacter species were and 157 ambulatory cases), 25 (6%) hospitalized patients were positive identified in 3.9% of cultures (n=57), and were found more commonly in while 8 (5%) ambulatory patients were positive. The median of age (IQR) females (68.4%) than males (13.6%). Cultures positive for Enterobacter of cases was older (22 years (16-34)) than the leptospirosis negative cases occurred most frequently in February followed by July. The following seven (16 (7-29)). Most cases (79%) were between 10-39 years and 53% were antibiotics were tested on isolates: cephalexin, cefotaxime, ceftriaxone, male. The majority of the patients presented with nonspecific signs, such gentamicin, nalidixic acid, ciprofloxacin and amikacin. Antibiotic resistance as headache (91%), nausea (85%), myalgia (81%), vomiting (79%), variations were measured monthly and appeared to have a seasonal arthralgia (67%), and hemorrhages (12%). During 2008-2013, 22 cases dependence. In January Enterobacter isolates were strongly resistant (67%) were detected in the rainy season (May-Oct). Cases were higher to cephalexin, in February to cefotaxime, in March to ceftriaxone, in 2010, the year with the most rain in a decade due to tropical storm cephalexin and gentamycin, and in April to cefotaxime and nalidixic acid. Agatha, with 18 (55%) cases detected and 14 (78%) in the rainy season. In July isolates showed no resistance to amikacin and low resistance Given endemic nature of leptosporosis in Guatemala and Central America, to ciprofloxacin, while in August and September strong resistance to efforts on prevention and control should focus on these events and the cephalexin. Identification of factors which lead to an apparent seasonal greater risk among adults. variations in antibiotic resistance patterns among Enterobacter will require further study that includes careful evaluation of demographic 1666 and therapeutic histories of patients from whom Enterobacter strains are isolated, speciation of these isolates and collection of larger numbers EXTENDED EMPIRICAL TREATMENT CONTRIBUTES TO of isolates over longer periods of time in order to control for sampling CHANGES IN BACTERIAL RESISTANCE PROFILE variability. Wellington F. Rodrigues1, Camila B. Miguel1, Ana Paula O. Nogueira2, Carlo J. Oliveira1, Marcelo C. Araújo1, Javier E. Lazo- 1668 1 Chica SIGNIFICANT DIFFERENCES IN ULTRASOUND FINDINGS 1Universidade Federal do Triângulo Mineiro, Uberaba, Brazil, 2Universidade BETWEEN MALNOURISHED AND NON-MALNOURISHED Federal de Uberlândia, Uberlândia, Brazil SCHOOLCHILDREN IN MADAGASCAR Extra intestinal infections acquired in the community with Escherichia coli 1 2 3 (E. coli) showed in the United States, a rate of frequency of six million Maria Teresa Giordani , Daniela Sardei , Indi Trehan , Tom 4 5 5 to 8 million cases of uncomplicated cystitis and 127,500 cases of sepsis Heller , Jacopo Nicoletti , Enrico Brunetti per year. Empirical treatment has been associated with the emergence 1San Bortolo Hospital, Vicenza, Italy, 2Pediatrician, Carrè, Vicenza, of bacterial resistance. although some countries are concerned to switch Italy, 3Washington University, St. Louis, MO, United States, 4Klinikum drugs used to treatment infectious, others keep the same treatment ( Muenchen-Perlach, Muenchen, Germany, 5University of Pavia, IRCCS S. for treatment empirical ) over a long period This study aims to assess the Matteo Hospital Foundation, Pavia, Italy sensitivity and resistance profile of E. coli across the different drugs used Childhood malnutrition contributes to high mortality rates and decreased for treatment. A cross-sectional and retrospective study (January 2008 to educational and adult capacity among survivors. Not just closely linked December 20013) at a public hospital, university, belonging at System of to an increasing burden of infectious diseases, it is also increasingly Health of Minas Gerais/Brazil (empirical treatment = fluoroquinolones), recognized as a cause of chronic morbidity in later life. We explored the was considered only in patients ambulatory that presented symptoms utility of bedside ultrasound in identifying specific pathologic findings (dysuria). The antibiotics tested were as follows (potency in µg/disc): in malnourished children in Madagascar, with the aim of improving the

astmh.org 510 management of co-morbidities. 83 children with stunting and/or severe and animals. Once considered an esoteric tropical disease, research on acute malnutrition underwent bedside abdominal sonography and B. pseudomallei has gained prominence due to its potential for epidemic compared with 76 non-malnourished children. Liver and spleen size, liver spread, increasing numbers of case reports from non-endemic regions, and echogenicity, intraluminal bowel evidence of massive helminthic infections, classification by the United States as a potential bioterrorism agent. Lack enlargement of abdominal lymph nodes, thickening of the gallbladder of awareness among physicians, along with a wide variability in disease wall, and other pathologic findings were assessed. Malnourished children manifestations, contributes to underdiagnosis and delayed treatment, had hepatosplenomegaly (36% vs 18%) and fatty liver (41% vs 18%) and also confounds accurate assessment of global prevalence. Although more frequently than non-malnourished children. Hepatosplenomegaly melioidosis is endemic in Northern Australia and parts of Southeast Asia, was more common in the Antaimoro area, where malaria and sickle-cell there are no published reports from the Thai-Myanmar border. Here we anemia are more prevalent. Evidence of intestinal helminth infections report the first two documented cases of fatal melioidosis in this region. were common in both groups, but in non-malnourished children were The discussion of cases in as-yet-unrecognized foci of disease is of great mostly associated with fever and acute diarrhea. Other pathologic findings public health importance and may help to better elucidate environmental were present In 17 malnourished children (20%) compared with 8% of and host determinants of infection. Our study highlights the need to both non-malnourished children. Preliminary results of this study suggest that increase clinical awareness of melioidosis on the Thai-Myanmar border, ultrasound evaluation of malnourished children is feasible and can be and to better assess the true burden of disease in the region through aid in identifying co-morbidities. The high rates of fatty liver infiltration improved case detection and rigorous B. pseudomallei prevalence studies. particularly deserves more attention, as a possible marker for the development of metabolic diseases and liver fibrosis in adulthood. 1671 1669 HISTOPLASMOSIS IN OREGON EX ECUADOR 1 1 2 LEPROSY, A MIMICKER OF OTHER DISEASES IN A Jon E. Lutz , Jonathan Brewer , John Blizzard DEVELOPED COUNTRY 1Bend Memorial Clinic and St. Charles Medical Center, Bend, OR, United States, 2Department of , Bend, OR, United States 1 2 2 2 Rone Lin , Winnie Ooi , Samuel Moschella , Stephanie Burns A 41 year old river raft guide returned from a 2 week long rafting trip 1Tufts Medical Center, Boston, MA, United States, 2Lahey Hospital and in a jungle region of Ecuador on March 30, 2012. He had received Medical Center, Burlington, MA, United States appropriate travel vaccinations and had taken mefloquine for malaria Leprosy (Hansen’s disease) is an uncommon disease in the U.S. and prophylaxis. Shortly after his return he developed fever to 103 F range, therefore unfamiliar to many health-care providers. This disease primarily chills, sweats, headache, myalgias, arthralgias and fatigue. He self- occurs in immigrants from countries with higher endemicity such as medicated with amoxicillin and subsequently ciprofloxacin was prescribed India and Brazil. Initial misdiagnosis can lead to significant delay in by a physician. On examination on April 9th he was afebrile, had some therapy, morbidity and loss of function. Here we reviewed our database shotty cervical and inguinal but otherwise had a of 172 patients with leprosy, who were evaluated and treated in our normal physical examination including that of the lungs. Initial laboratory Hansen’s disease public health satellite clinic from 1998 to 2014. There investigations including malaria smears, blood cultures and stool studies were 18 patients (10%) with prior misdiagnoses; eleven of them initially were unrevealing. A chest x-ray had evidence of a right sided infilatrate. presented with skin rash, six with neuropathy and one with rheumatologic Ceftriaxone and doxycycline were given but fevers persisted. A CT symptoms. Skin rash from Hansen’s disease can be difficult to distinguish scan of the chest was carried out and revealed extensive mediastinal from more common dermatologic conditions such as fungal skin and hilar lymphadenopathy and too numerous to count lung nodules. infection, allergic dermatitis and cutaneous sarcoidosis. Idiopathic mono- Fiberoptic bronchoscopy was nondiagnostic. On April 20th therapy with neuropathies with wrist or foot drops, as well as mononeuritis multiplex, ketoconazole was initiated to treat possible paracoccidioidomycosis. were among the common misdiagnoses for patients with leprous neuritis. On April 24th, histoplasmosis serologies were reported positive. He Leprosy can also mimic rheumatoid arthritis with solely joint symptoms was started on liposomal amphotericin B. On April 25th he underwent without initial involvement of skin or nerve. In our review, the diagnosis a minithoracotomy for definite diagnosis as there was concern about could be delayed for a significant amount of time, for even up to 10 years possible coexisting malignancy. Pathology and intraoperative cultures were in 2 cases. This led to irreversible loss of neurologic function (foot/wrist consistent with histoplasmosis. He subsequently completed a 3 month drops), neuropathic ulcers and osteomyelitis. We will present 3 illustrative course of itraconazole and did well. We herein discuss briefly travel- cases: one of leprous neuritis misdiagnosed as mononeuritis multiplex from associated histoplasmosis and the current recommendations for drug sarcoidosis, a second of polyarthritis from leprosy mimicking rheumatoid therapy of severe primary histoplasmosis infection. arthritis, and lastly, a case of borderline tuberculoid leprosy with skin rash thought to be from cutaneous sarcoidosis based on initial evaluation of 1672 a skin biopsy. To summarize, it is of crucial importance to include leprosy in the differential diagnoses for patients with chronic rash, neuropathy or DESCRIPTION OF DENGUE-RELATED HOSPITALIZATION joint symptoms in the appropriate epidemiologic setting, because early AND DISEASE SEVERITY FROM AN ENHANCED DENGUE recognition and treatment can prevent progression of the disease and its SURVEILLANCE SYSTEM IN PUERTO RICO morbidities. Janice Pérez-Padilla1, Eric González2, Aidsa Rivera-Sánchez1, Brenda Torres-Velásquez1, Juan P. Torres2, Luisa Alvarado2, Kay M. 1670 Tomashek1 TWO FATAL CASES OF MELIOIDOSIS ON THE THAI- 1Centers for Disease Control and Prevention, San Juan, PR, United States, MYANMAR BORDER 2San Lucas Episcopal Hospital, Ponce, PR, United States Dengue is an acute febrile illness (AFI) that is endemic in Puerto Rico. The Miriam B. Torchinsky1, Cindy S. Chu2, Clare Ling2, Francois clinical spectrum of dengue ranges from mild AFI to a life-threatening Nosten2 illness. Although timely identification of dengue patients can reduce 1University of British Columbia, Vancouver, BC, Canada, 2Shoklo Malaria medical complications and mortality, this complicated by clinical Research Unit, Mae Sot, Thailand manifestations that overlap with other AFI. To identify early clinical Melioidosis, caused by the gram-negative environmental bacterium features that can be used as predictors for severe dengue, we evaluated Burkholderia pseudomallei, is an infectious disease of clinical importance in the clinical course of laboratory-positive (i.e., DENV nucleic acid detected endemic areas, and is associated with a high case-rate fatality in humans by RT-PCR or anti-DENV IgM antibody detected by ELISA) dengue patients

astmh.org 511 enrolled in the Sentinel Enhanced Dengue Surveillance System (SEDSS) 1674 site located in Ponce, Puerto Rico. Patients were those presenting with AFI during May 7, 2012 to May 6, 2013 that were hospitalized (n = 262) or SURVEILLANCE FOR ARTEMISININ, QUININE, AND were out-patients that returned for follow-up evaluation (n = 120). Of all MALARONE RESISTANCE AMONG IMPORTED PLASMODIUM 382 patients, there were no significant differences in age or sex between FALCIPARUM MALARIA - CALGARY, CANADA (2013-14) hospitalized and non-hospitalized patients. Admitted patients sought care later than non-hospitalized patients (mean day of presentation = 4 vs.2 Asongna Folefoc, Abu N. Mohon, Lydia Lee, Dylan R. Pillai days), and had a mean hospital stay of 4 days. Clinical findings associated University of Calgary, Calgary, AB, Canada with hospitalization were anorexia (p = 0.002), diarrhea (p = 0.021) and Calgary has seen an increase in imported malaria cases in recent years with dengue warning signs of persistent vomiting (p < 0.001), abdominal a large proportion coming from sub-Saharan Africa where Plasmodium pain (p < 0.001) and bleeding (p = 0.013). Laboratory findings at falciparum is hyper-endemic. Surveillance for emerging resistance to presentation associated with hospitalization were leukopenia (p = 0.021) antimalarial drugs such as Artemisinin, Quinine, and Malarone is essential and thrombocytopenia (p < 0.001). Mean platelet count was significantly in returning travelers. Artemisinin resistance has been reported in different between hospitalized and non-hospitalized patients (mean = Cambodia and mutations within the K13-propeller gene associated with 81,000 vs. 151,000) (p < 0.001). Patients that presented 4-7 days after resistance to this drug. Mutations in the cytochrome B (cytB) are linked illness onset had greater odds of having thrombocytopenia (OR = 2.18; to Malarone resistance. In this study, we prospectively determined the CI: 1.24-3.83) or elevated liver transaminases (OR = 4.74; CI: 1.53-19.45). susceptibility and resistance genotype of imported P. falciparum malaria Enhanced dengue surveillance revealed that hospitalized dengue patients to Calgary from April, 2013 to April, 2014. Travelers were mostly male presenting late for clinical care were more likely to present with dengue (n=12, 67%) VFR (n=17, 94%) with mean age of 35.3 years destined warning signs, and were hospitalized more frequently. Further analysis will for sub-Saharan Africa (n=17, 94%) predominantly West Africa (n=11, assess correlation between early presentation and ultimate disease severity. 61%). Malarone (oral) was the commonest treatment option (n=11, 61%) followed by artesunate (n=3, 17%) and quinine (n=3, 17%). Positive 1673 malaria samples from patients (n=18) were tested with a standardized panel of antimalarials using the ELISA-based HRP2 ex vivo drug sensitivity ACCESS MATTERS: AFRICAN-BORN U.S. MILITARY protocol developed by WWARN. DNA was extracted from patient EDTA TRAVELERS UTILIZE TRAVEL CARE AT HIGH RATES blood samples (n=18) and primers flanking the K-13 propeller gene Kristina St. Clair1, Anthony R. Artino2, Patrick W. Hickey2 and cyt B were used for PCR amplification of this gene. PCR products were bidrectionally sequenced and analyzed for mutations. Our ex-vivo 1Naval Medical Center Portsmouth, Portsmouth, VA, United States, results showed IC values of 17.36 ± 11.92nM, 4.34 ± 2.34 nM, 4.06 ± 2Uniformed Services University, Bethesda, MD, United States 50 1.66 nM, 4.00 ± 1.39 nM for Artemisinin, Artesunate, Artemether and Immigrant travelers that visit friends and relatives (VFRs) are less likely to Dihydroartemisinin, respectively; mean IC50s of 39.04 ± 15.73 nM, 16.33 have a pre-travel health encounter and more likely to experience serious ± 4.36 nM, 80.44 ± 25.75 nM, 17.23 ± 3.65 nM and 127.38 ± 36.97nM illness. Immigrants comprise more than 5% of active duty service members for Chloroquine, Mefloquine, Quinine, Amodiaquine and Piperaquine in the United States Department of Defense and have free access to travel respectively; and mean IC50 = 27.2 ± 22.26 for Atovaquone. Analysis of medicine care, yet their travel health seeking behaviors are unknown. 351 the K-13 propeller and CytB gene showed that all imported malaria were African-born service members and 470 U.S.-born comparators completed wild type to date. Our results confirm that imported P. falciparum malaria an Internet-based survey to assess pre-travel health care utilization, to Calgary from sub-Saharan Africa remains wild-type at key resistant loci perceived potential barriers, and health outcomes. While overall use (K13 and cyt B) and susceptible to Artemisinin, Malarone and quinine the of services was equivalent if official duty was included commonest treatment options when tested ex vivo. (p=0.94), when traveling on leave status (not on official duty) African-born service members were more likely than U.S.-born to see a physician, 65% 1675 vs. 35% (p<0.001) prior to their most recent travel to a low, low-middle or upper-middle income country. This persisted when stratified by malaria risk A SURVEY ON KNOWLEDGE, ATTITUDES AND PRACTICES at destination with African-born service member VFRs reporting pre-travel AMONG INTERNATIONAL TRAVELERS IN UGANDA health care more than American-born comparators, 65% vs. 45% (p < 1 2 2 0.001) Both African-born and American-born service members reported Zain Chagla , Palmyra Immordino , Claire Webb , Ronald M. 2 3 easy access to medical care. African-born military service members perceive Mbwasi , Christopher Sanford less risk of illness when traveling to Africa compared to American born 1McMaster University, Hamilton, ON, Canada, 2London School of Hygiene travelers (p<0.001) yet, somewhat paradoxically, place more importance & Tropical Medicine, London, United Kingdom, 3University of Washington, on pre-travel medical services (p = 0.007). African-born service members Seattle, WA, United States are more willing to self-diagnose and treat illnesses such as malaria In an increasingly international environment, global travel rates are (p<0.001) and rely on locally purchased medications (p<0.001). There was increasing. However, significant risk exists amongst travelers. Counseling no difference in reported adherence to malaria chemoprophylaxis. This against preventable morbidity and mortality, such as transit related United States Military Health System study revealed data that opposes injuries, vaccine preventable diseases, violent crimes, and tropical illnesses, previous civilian studies: African-born VFRs in the military sought pre- is essential for international travelers. The majority of data around travel health care more often than their U.S.-born counterparts. Access traveler attitudes and counseling is based on pre-travel assessments, or to care and positive beliefs about the benefits of travel medicine services in evaluation of returned ill travelers, but is biased towards those with contribute to this finding. These findings have implications for the role health seeking behaviors. This study was designed to look at attitudes of national health-care reform and community engagement programs. and practices amongst international travelers while actively travelling. The Disclaimer: The Views expressed are those of the author(s) and do not study took place Jinja, Uganda, a popular tourist destination near Kampala necessarily reflect the official views of the Uniformed Services University of known for white-water rafting and the Nile River. Participants were the Health Sciences, the U.S. Army, or the Department of Defense. recruited at tourist locales in Jinja, and a semi-structured questionnaire was administered to tourists after obtaining voluntary informed consent. A total of 153 travelers were surveyed. The majority was female, with average age 31 years, and predominantly from the USA, Australia, and the Netherlands. Most participants had received pre-travel advice through a travel clinic (64.0%) or a general practitioner (25.4%), and others cited the Internet, friends, and relatives as sources of information. Participants

astmh.org 512 endorsed malaria (94.7%), vaccines (92.1%), and diarrhea (59.5%) were therapy with two more standards RA medications, methotrexate (MTX) the most important pre-travel counseling items, but few mentioned and etanercept (EPT). The study group included Iraqi patients with traffic accidents (15.0%) or sexually transmitted infections (13.7%). moderate to highly active RA. A double blind, randomized, placebo - Malaria prophylaxis was prescribed to the majority (80.1%), but only controlled clinical trial was conducted in which 100 RA male and female three quarters of those people took prophylaxis, with adherence issues patients were enrolled from a group who were already on MTX or EPT attributed to side effects or a lengthy duration of stay. Finally, when asked for at least 1 month. This pool of subjects was divided into two groups, about health and safety issues experienced during the trip, nearly a third one to receive 20 mg atorvastatin tablet and the other to receive placebo had encountered an issue, most of which either related to gastroenteritis, capsules for three consecutive months. This study revealed first that only malaria, or traffic accidents. Although many travelers seek medical care 49 patients completed the 3 months trial, 25 patients in atorvastatin prior to departure, counseling regarding non-infectious issues such as road and 24 patients in placebo group. All patients were clinically evaluated traffic accidents, personal safety, and risk behaviors are lacking, despite by measuring swollen joint count (SJC), tender joint count (TJC), visual these being a major cause of morbidity and mortality. Integration of analogue scale (VAS) and disease activity score (DAS28). Blood samples this information into Internet resources and clinical practice may help to of all subjects patients were evaluated for erythrocyte sedimentation rate decrease mortality amongst global travelers. (ESR), C reactive protein (CRP) at baseline, monthly and at the end of the study. RA patients undergoing 20 mg atorvastatin treatment showed a 1676 significant (P < 0.05) reduction in CRP, SJC and TJC compared to those who received placebo. In addition, atorvastatin treatment groups trended FACTORS ASSOCIATED WITH MORTALITY BY DENGUE toward reduced ESR, VAS, and DAS28, but these differences did not achieve statistical significance (p > 0.05). In conclusion we believe that 20 Elizabeth Borrero mg atorvastatin is a safe and well-tolerated drug that has modest anti- Fundación Santa Fe de Bogotá, Bogotá, Colombia inflammatory effect in patients with moderate to severe active RA. Dengue is a public health priority in Colombia due to the significant increase in the number of cases from 26817 in 2008 to 57589 in 2013. 1678 In 2010 a 183% increase was noted in respect to 2009 and 9393 cases of severe dengue was reported and 217 confirmed deaths. A case- GLUCOSE-6-PHOSPHATE DEHYDROGENASE DEFICIENCY control study was conducted in the departments of Cundinamarca, Norte AND DENGUE IN SINGAPOREAN MALES: A CASE-CONTROL Santander, Santander, Cauca Valley and Meta to identify the clinical factors STUDY associated with mortality in patients with severe dengue in hospitals of level II and III attention between the periods 2009-2013. A case was Tun-Linn Thein, Linda K. Lee, Joshua G. Wong, David C. Lye, considered to be death by dengue and a control pertained to survivors, Yee-Sin Leo all were confirmed through IgM or RT-PCR. The sample size was 50 cases Tan Tock Seng Hospital, Singapore, Singapore and 150 controls. For the analysis logistic regression was utilized. 42% (63) Clinical presentations of dengue infection range from asymptomatic, of the patients originated from Cauca Valley, 24% (36) Meta, 18% (27) non-severe to severe disease. We aim to test the hypothesis that patients North of Santander, 10% (15) Santander and 6% (9) Cundinamarca. 60% with glucose-6-phosphate dehydrogenase (G6PD) enzyme deficiency (90) were less than 16 years and 52% were women. The factors associated may present with severe disease and hemolysis. We analyzed a cohort of independently with mortality were: comorbidities OR 3,18 (IC 95%: 1,33; adult dengue patients treated at Tan Tock Seng Hospital, Singapore from 7,60), social risk OR 3,33 (IC 95%: 1,21; 9,17), tachycardia OR 8,94 (IC January 2005 to December 2008. Dengue infection was confirmed by 95%: 2,57; 31,05), tachypnea OR 2,71 (IC 95%: 1,16; 6,28), altered state positive polymerase chain reaction or dengue serology with World Health of consciousness OR 12,09 (IC 95%: 2,72; 53.70), respiratory difficulty Organization (WHO) probable dengue definition. Singaporean males with OR 5,61 (IC 95%: 2,24; 14,06), pleural effusion OR 2,85 (IC 95%: 1,33; documented G6PD status were defined as cases. For each case, three 6,11), main organ damage OR 3,14 (IC 95%: 1,29; 7,64), severe bleeding controls were selected by matching citizenship and year of infection. OR 2,66 (IC 95%: 1,19; 5,95), and previous consultation OR 2,68 (IC Hemolysis was defined as low hemoglobin concurrent with low serum 95%: 1,16; 6,19). In the multivariate analysis the factors associated with haptoglobin, or high reticulocyte or lactate dehydrogenase or bilirubin. increased death were: social risk social OR 9.88 (IC 95%: 1,26; 77,11), Dengue hemorrhagic fever and severe dengue were classified according altered state of consciousness OR 11.48 (IC 95%: 1,34, 97,93), respiratory to WHO 1997 and 2009 dengue guidelines. Compared with cases (n=30), difficulty OR 9.84 (IC (95%: 1,96; 49,36) main organ damage OR 9.55 (IC controls (n=120) were significantly younger (median 26 vs. 35 years, 95%: 1,77; 51,41) and severe bleeding OR 8,08 (IC 95%: 1,95; 33,66). p0.05). During their clinical course, cases had significantly higher rates of Patients with these clinical characteristics should be hospitalized for jaundice (10% vs. 1%, P<0.05), serum bilirubin (median 27 vs. 10 mmol/L, extended observation and opportune treatment to avoid death. p<0.001), aspartate transaminase (median 148 vs. 91 U/L, p<0.05), and lower hematocrit (45% vs. 46%, p<0.001), haemoglobin level (13 vs. 14 1677 mg/dL, p<0.001). There was no difference in rates of dengue hemorrhagic fever (23% vs. 22%, p>0.05). However, cases had higher tendency to ATORVASTATIN FOR THE TREATMENT OF RHEUMATOID develop severe dengue and hemolysis than controls ([23% vs. 12%] ARTHRITIS IN IRAQ and [14.29% vs. 2.7%] respectively) although the difference was not Sami Salman1, Samer Shuker2 significant (p>0.05). The two groups had similar rates of blood and platelet transfusions, intravenous fluid and length of hospitalization (p>0.05). The 1 Unit, College of Medicine, Baghdad University, Baghdad, observed differences should be prospectively validated in larger cohorts 2 Iraq, Pharmacology Unit, College of Medicine, Baghdad University, and in different populations. Baghdad, Iraq Rheumatoid arthritis (RA) is a common chronic inflammatory and disabling disorder that is characterized by synovitis, articular destruction, and many systemic extra articular features. In addition, RA is associated with both morbidity and mortality due to accelerated atherosclerosis and risk of increased cardiovascular disease. Atorvastatin is well known anti dyslipidemic agent that is considered to have potential anti-inflammatory and immune modulatory functions in patients with RA. To explore the possible anti-inflammatory effects of Atorvastain in patients with RA, we designed a study to evaluate the effect of atorvastatin compared to

astmh.org 513 1679 semi-structured questionnaire, vaccination cards were also reviewed. We defined a completely immunized child as a child who had received one A RANDOMIZED, DOUBLE BLIND, CLINICAL TRIAL OF dose of BCG, three doses of oral polio vaccine, three doses of Diptheria- TWO DOSE REGIMENS OF VINS POLYVALENT ANTIVENOM Pertusis-Tetanus vaccine and one dose of measles vaccine by 12 months of FOR THE TREATMENT OF SNAKEBITE WITH NEUROTOXIC age. Bivariate and multivariate data analysis was performed using Epi-info ENVENOMATION IN NEPAL software. Of the 750 mothers interviewed, (36.6%) were fully immunized. Children of mothers with poor knowledge on immunization were less Emilie Alirol1, Sanjib K. Sharma2, Anup Ghimire2, Antoine Poncet1, likely to be fully immunized (Odds ratio (OR) =0.55, 95% CI=0.23-0.51). Christophe Combescure1, Khaled Mostaguir1, Chabilal Thapa3, Children whose mothers possessed primary or no formal education were Vijaya Paudel4, Robert W. Taylor5, Deb Pandey6, David A. Warrell7, less likely to be fully immunized compared to children of mothers with at Ulrich Kuch6, François Chappuis1 least a secondary level education (OR=0.50, 95%CI=0.34-0.73). Children 1Geneva University Hospital, Geneva, Switzerland, 2B.P. Koirala Institute of delivered at health facilities were more likely to be fully immunized Health Sciences, Dharan, Nepal, 3Dumkauli Primary Health Care Centre, (OR=1.81, 1.21-2.69). The major determinants of complete immunization Nawalparasi, Nepal, 4Bharatpur Hospital, Chitwan, Nepal, 5Mahidol Oxford status were knowledge level, maternal educational status and place Research Unit, Bangkok, Thailand, 6Forschungszentrum Biodiversität und of birth of the children. Raising the level of knowledge and increasing Klima (Bio+C), Frankfurt am Main, Germany, 7University of Oxford, Oxford, maternal literacy level as well as encouraging health facility births are United Kingdom essential to improve immunization coverage in these rural communities. Snakebite is an important medical emergency in rural Nepal. Although accurate figures are lacking, over 20 000 cases are recorded annually, 1681 with a case fatality rate close to 15%. Important variations exist between hospitals in the management and outcome of snakebite envenomation. VARIATIONS IN PRESENTATION OF ERYTHEMA NODOSUM In particular, striking disparities are observed in the dosage of antivenom, LEPROSUM: REPORT OF THREE CASES SEEN AT A U.S. reflecting poor adherence to the complex national protocol. Although HANSEN’S DISEASE CLINIC clinical studies have been conducted on viperid snake envenomation, Kristoffer E. Leon1, Robert W. McDonald2, Anandi N. Sheth2, well-designed dose-finding studies are almost non-existent for elapid Jessica K. Fairley2 envenomation. The purpose of this trial was to compare the efficacy and 1Emory University, Atlanta, GA, United States, 2Emory University School of safety of two antivenom dosing schemes in the treatment of neurotoxic Medicine, Atlanta, GA, United States envenomation. The trial was conducted between May 2011 and March 2013 in 3 health facilities of southern Nepal. 157 patients presenting Hansen’s disease (leprosy) remains the leading infectious cause of disability, with signs of neurotoxic envenomation were randomized either to a high with 250,000 new cases reported globally yearly. Erythema nodosum initial dose regimen (intervention) or to the low initial dose regimen as leprosum (ENL) reactions, or Type 2 reactions, are humoral responses of recommended by the Nepalese national protocol (control). The primary the immune system that can cause systemic illness, including fever, skin composite outcome included death, requirement for manual ventilation lesions, joint pains and neuritis. ENL reactions occur in patients on the and worsening of neurotoxicity. Secondary outcomes included time lepromatous end of the spectrum, classified by the Ridley-Jopling system. to recovery, occurrence of adverse reactions, and cost. There was no In non-endemic countries, Hansen’s disease (HD) remains rare and often statistically significant difference between arms in the proportion of underrecognized with the literature lacking in clinical descriptions of patients reaching the primary endpoint (control 48.7% vs intervention leprosy complications in the United States. To fill this gap, we report three 38.5%, p=0.264). No differences were observed in the analysis of safety patients with lepromatous leprosy who were seen at a HD clinic in Atlanta, outcomes. In 51 patients the snake species could be identified. 29 had GA with complicated ENL reactions within the last three years. The first been bitten by cobras (Naja spp) and 22 by kraits (Bungarus spp.). Those patient was a 33-year-old Bangladeshi woman who presented with high bitten by kraits experienced more primary outcomes (68.2% versus fever, abdominal pain, and arthralgias. She lacked the distinctive skin 27.1%, p=0.004), and recovered less often (40.9% vs 96.5%, p<0.001) or lesions usually seen in ENL, but was incidentally found to have splenic more slowly (mean time 18 hours vs 5 hours, p<0.001) than did patients lesions. She responded well to prednisone and was able to be weaned off bitten by cobras. These findings suggest that there is no difference in after 6 months. Second, a 42-year-old Vietnamese man initially presented efficacy and safety between low and high initial dose of antivenom for with classic ENL lesions, fevers, and lymphadenopathy that progressed neurotoxic snakebite, and that envenomation due to krait bites is less in severity despite increasing doses of corticosteroids. He eventually was responsive to antivenom than that following cobra bites. admitted to the intensive care unit with a severe systemic inflammatory response syndrome. He was subsequently started on thalidomide without 1680 recurrences. The last patient was a 68-year-old U.S.-born man, who displayed symptoms representative of both Type 1 and Type 2 reactions as FACTORS ASSOCIATED WITH COMPLETE ROUTINE his initial presentation of HD. These included joint pain, severe extremity IMMUNIZATION STATUS OF CHILDREN 12-23 MONTHS IN swelling, skin nodules and a progressive neuropathy. He had been RURAL AREAS OF OSUN STATE - SOUTHWESTERN NIGERIA misdiagnosed with a seronegative arthritis prior to this presentation. While all three cases reported are ENL, the differences of clinical courses and Elizabeth B. Adedire1, Ikeoluwapo O. Ajayi2, Olufemi Ajumobi1 presentations highlight the complexity of the disease and the need for 1Nigerian Field Epidemiology and Laboratory Training Program, FCT Abuja, increased awareness of unique manifestations of lepromatous leprosy. Nigeria, 2University of Ibadan, Ibadan, Oyo State, Nigeria Immunization is a cost-effective public health intervention to reduce morbidity and mortality associated with infectious disease. Incomplete immunization status especially in rural areas has led to a high burden of VPD in children. The Nigerian Demographic and Health Survey, 2008 showed that only 57.8% of children had received all recommended vaccines in Osun State far below WHO target of 80%. We conducted this study to identify the factors associated with complete immunization status of children in rural areas of Osun State. A total of 750 mothers of children aged 12-23 months were interviewed, using the WHO 30 cluster sampling technique. We collected data on socio-demographic characteristics, history of vaccination and factors associated with immunization status using

astmh.org 514 1682 1684 SEROLOGICAL STUDY OF ANTIBODIES ANTI-TOXOCARA THE EPIDEMIOLOGY OF ORAL HUMAN PAPILLOMAVIRUS CANIS EVALUATED BY ELISA AND WESTERN BLOT IN INFECTION AMONG HEALTHY MEN AND WOMEN IN LIMA, PEDIATRIC PATIENTS WITH CRYPTOGENIC EPILEPSY PERU Maria de Lourdes Caballero-Garcia1, Jocelyn Simon-Salvador1, Brian J. Rosen1, Leora Leora Walter2, Robert H. Gilman1, Lilia Juan Hernandez-Aguilar1, Benjamin Nogueda-Torres2, Enedina Cabrerra3, Patti E. Gravitt1, Morgan A. Marks1 1 Jiménez-Cardoso 1Johns Hopkins University Bloomberg School of Public Health, Baltimore, 1Hospital Infantil de Mexico Federico Gomez, Mexico City, Mexico, 2Escuela MD, United States, 2New York Hospital Queens, New York, NY, United Nacional de Ciencias Biologicas, Mexico City, Mexico States, 3Asociacion Benefica Prisma, Lima, Peru It has been suggested a correlation between infection by Toxocara canis The incidence of head and neck cancers associated with human and epilepsy and it is thought that epileptic seizures can be derived papillomavirus infection has been increasing in Peru. However, the from the immune response generated by the presence of the parasite or burden of oral HPV infection in Peru has not been assessed in a healthy direct lesions that can cause in the brain. The objective was to determine population. The objective of the study was to estimate the prevalence antibody anti-T. canis against antigen of the parasite through the and correlates of oral HPV infection in healthy male and female residents technique of ELISA and Western blot in pediatric patients with cryptogenic from Las Pampas, a shantytown in Lima, Peru. A population-based sample epilepsy attending outpatient consultations in the area of of the of 1,500 healthy men and women between the ages of 8-85 in low Children Hospital of México Federico Gomez (HIMFG). We analyzed 111 to middle income areas of Lima, Peru was identified through random patients from 6 to 16 years of age with confirmed diagnosis of epilepsy household sampling between January and August 2010. Adjusted with clinical and epidemiological background who attend the external odds Ratios (aOR with 95% CI) were used to assess the association of consultation of Neurology of the HIMFG. We analyzed the presence demographic factors, sexual practices, and oral hygiene on the prevalence of antibodies anti-T. canis by ELISA using excretion-secretion antigens of oral HPV infection. The prevalence of any HPV and any high-risk HPV obtained from larvae of L2 T. canis cultured in vitro and the children who (HR-HPV) was 6.8% and 2.0%, respectively. The three most common types tested positive by this technique were evaluated by the technique of were HPV 55 (3.4%), HPV 6 (1.46%), and HPV 16 (1.09%). Male sex (aOR, Western blot to determine the molecular weight of excretion-secretion 2.32; 95% CI: 1.29, 4.18), age 19-27 (aOR, 2.77; 95% CI: 1.02, 7.56) and proteins recognized by sera from patients with antibodies anti-T. canis. It 46-55 years (aOR, 3.52; 95% CI: 1.07, 11.5) were significantly associated was found that 12.5%. sera had antibodies against antigens of excretion- with prevalent HPV infection after adjustment. The prevalence of oral HPV secretion for T. canis. Nine children were evaluated by the technique of in this population-based sample of healthy men and women from Peru Western blot and only 5 were positive for this technique, recognized was similar to estimates observed in the United States. Higher prevalence two main antigens of 24 and 35 kDa. The analysis in sera from pediatric of oral infections in men were consistent with a male predominance of patients with epilepsy, showed a rate of 6.9% to antigens from T. canis, HPV-associated HNC and may signal a sex-specific etiology in the natural after analysis by ELISA and Western blot. history of infection. 1683 1685 MAPPING THE POTENTIAL RISK OF MYCETOMA IN DIAGNOSIS OF LIVER TUMORS USING IMAGE-BASED SUDAN USING MAXIMUM ENTROPY ECOLOGICAL NICHE STATISTICAL FEATURES MODELLING Alyaa H. Ali, Intithar Malk, Sabah N. Mazhar Abdallah M. Samy, A. Townsend Peterson Department of Physics, College of Science for Women, Baghdad University, Kansas University, Lawrence, KS, United States Baghdad, Iraq WHO recognized mycetoma as one of 17 neglected tropical diseases Tumors located in the liver can be detected using CT imaging and (NTDs) worldwide. Studies revealed a soil-borne mediated or thorn comparison of intensity histograms with the associated statistical prick-mediated origin of mycetoma, but no studies are available to parameters for characterizing different regions of the liver including investigate the effect of soil type and Acacia distribution on mycetoma normal and cancerous. Identifying whether the tumor is benign or in Sudan. Here, we report efforts to investigate risk factors associated malignant is an important step in image-based liver cancer diagnosis. with mycetoma risk in Sudan using ecological niche modeling. Records In this paper we describe an automated system for image-based liver of mycetoma cases were obtained from the scientific literature, PubMed, segmentation of CT imagery using a multi-stage process. Each CT liver and GIDEON. Acacia records were obtained from the Global Biodiversity image is pre-processed to remove noise and enhance image quality Information Facility. We developed ecological niche models (ENMs) based to recognize structures within the liver. A key challenge is related to on digital GIS data layers summarizing soil, land-surface temperature, separating the liver from the rest of the abdominal cavity in CT imagery. and greenness, summarizing environmental variation across Sudan. ENMs We used a statistical feature descriptor to characterize healthy tissue calibrated in endemic districts were transferred across all of Sudan, and versus cancerous regions and then applied a modified K-means classifier to suggested that greatest risk was in a belt across central and southern improve the accuracy of the tumor segmentation process. Sudan. We visualized mycetoma in environmental dimensions, and the results revealed that mycetoma in ecologically diverse landscapes under 1686 wide ranges of environmental conditions. We tested niche similarity between Acacia and mycetoma, and found significant niche similarity. ASSOCIATION OF HEMOGLOBIN LEVELS AND SELECTED These results revealed contributions of different environmental factors to BIOCHEMICAL MARKERS WITH DIABETIC NEPHROPATHY mycetoma risk, identify suitable environments for disease emergence, raise DISEASE the concerns for mycetoma-acacia association, and provide steps towards Khalid G. Al-Fartosi, Manal B. Al-Tamimi, Saad H. Al-Badry a robust, predictive risk map for the disease. Department of Biology, College of Science, University of Thi-Qar, Thi-Qar, Iraq The present study was designed to investigate the relationship between hemoglobin levels (Hb), serum creatinine, lipid profile and urine albumin excretion in diabetic patients in Thi-Qar province, Iraq with various degrees

astmh.org 515 of nephropathy. The study cohort included 60 patients and 30 healthy regimen we used will be described. Pediatric meningitis needs special subjects (control group) presented at the Al-Nasiriyah Endocrine Centre attention and a high rate of clinical suspicion as the yield of microbial in Thi-Qar province, Iraq. The diabetic subjects were divided into three isolation is low primarily due to the use of antibiotics prior to hospital groups each with 20 subjects presenting diabetes mellitus (DM) disease for admission. The choice of empirical antimicrobial usage might need to be 1-5 years, 6-10 years and more than 10 years. The clinical results showed reviewed from a clinical and public health perspective. a significant decrease in the levels of Hb (p<0.01) in patients with DM compared to the control group. Also, there was a significant increase in 1689 blood sugar and urine albumin excretion in patients with DM compared with the control group (p<0.01). Serum creatinin increased significantly in UTILIZING THE COMMUNITY HEALTH WORKER NETWORK patients with more than 10 years of DM compared with the control group. FOR LYMPHATIC FILARIASIS (LF) MORBIDITY MONITORING: These results indicate a dyslipidemia in patients with DM compared with THE DEVELOPMENT OF AN SMS-BASED SURVEILLANCE the control group. TOOL 1687 Michelle C. Stanton1, Square Z. Mkwanda2, Alexander Y. Debrah3, Linda B. Debrah3, Achim Hoerauf4, Nana-Kwadwo THE EFFECT OF METFORMIN ON GHRELIN SERUM LEVEL IN Biritwum5, Abigail Best1, Matthew Cliffe1, Louise A. Kelly-Hope1 TYPE 2 DIABETES MELLITUS 1Liverpool School of Tropical Medicine, Liverpool, United Kingdom, 2Ministry of Health, Lilongwe, Malawi, 3Kwame Nkrumah University of Mohammed N. Mohammed, Nahla O. Tawfik Science and Technology, Kumasi, Ghana, 4University of Bonn, Bonn, Department of Dental Basic Sciences, College of Dentistry, University of Germany, 5Ghana Health Service, Accra, Ghana Mosul, Mosul, Iraq Lymphatic filariasis (LF) is a parasitic infection that is responsible for over Ghrelin is an orexigenic peptide hormone. A great deal of evidence 15 million lymphoedema and 25 million hydrocele cases globally, resulting suggests that ghrelin is involved in the development of Type 2 diabetes in LF morbidity being one of the leading causes of long-term disability. mellitus (DM). The aim of this study was to investigate the effect of Currently, there are no standardised methods available for quantifying and metformin on ghrelin serum level in Type 2 DM patients. This clinical mapping LF morbidity burden. The purpose of this study was therefore to control study was carried out at the Al-Wafaa Medical Center for Diabetic pilot a novel method for collecting LF morbidity data in endemic areas of and Endocrine Disorder patients in Mosul from October 2011 to March Malawi and Ghana. The premise of this method is that community health 2012. Fifty-five Type 2 diabetic patients and 20 control healthy subjects workers are able to quickly identify lymphoedema or hydrocele cases in were enrolled. Patients and subjects were divided into 4 groups. Blood their villages, but do not currently have a standardised method of collating samples were collected from all subjects and the body mass index (BMI) this information. We have therefore developed an SMS-based tool which was calculated for each person. Fasting blood sugar (FBS) level and ghrelin enables health workers to submit information on each identified case in serum level were estimated for each patient. This study demonstrated their communities using a basic mobile phone. This tool was trialled under a non-significant lower mean ghrelin serum level in the diabetic group two scenarios: in March 2014 the tool was trialled by qualified, salaried compared to healthy controls. There were, however, significant differences health workers in southern Malawi; in May 2014 the study was repeated in ghrelin serum levels between the diabetic group without metformin and in Ghana using volunteer community health workers. In both scenarios, the diabetic group treated with 1,000 mg metformin daily (p < 0.05). In each health worker was asked to submit each identified case’s village of this study we found that ghrelin serum levels had a negative correlation residence, age, sex, condition and severity of condition (if lymphoedema) with age of patients over 30 years and BMI in both healthy and diabetic via SMS to a smartphone housed in-country. This information was then individuals. instantly compiled into a single database. A random sample of cases was visited by a medically qualified person to confirm the health workers’ 1688 diagnoses, and GPS coordinates of their villages were recorded. The feasibility of the method was assessed in terms of the ease in which health PEDIATRIC MENINGITIS IN THE AL-ABBASEYA FEVER workers were able to correctly identify cases (true positive rate), and the HOSPITAL OF CAIRO ease of use of the SMS-based tool (data entry error rates). A comparison A. M. Al-Abbasi, Amro Shaee, Haifa I. Tawfeek between the performance of salaried health workers and volunteer health workers was also undertaken. Preliminary results for Malawi indicate that Infectious Diseases and Clinical Immunology, College of Medicine, the true positive rate for reported lymphoedema and hydrocele cases using Baghdad University, Baghdad, Iraq this method were 90% (95% CI [80%, 97%]), and 92% (95% CI [77%, Meningitis in children can be a fatal disease. Early discovery and prompt 97%]) respectively. treatment using effective antimicrobials greatly reduces the mortality and resulting complications of the disease. Since meningitis is caused by a number of different microorganisms, detection of the causative agent is essential for proper treatment and to establish any preferential pattern of prevalence in different age groups or geographical regions. We conducted our clinical study on 61 patients admitted to Al-Abbasseya Fever Hospital in Cairo during the period of December 2010 to June 2011. The patients ranged in age from one month up to 17 years. Diagnosis of these cases was based on history, clinical data, laboratory tests, cerebrospinal fluid (CSF) examination and other visual diagnostic tests. There were 61 cases with 35 males and 26 females. Based on the CSF culture, gram stain, cell count and cell type diagnostic information, the patients were grouped into two classes: Acute bacterial meningitis (Group 1) and acute non-bacterial meningitis (Group 2). Group 1 consisted of 31 cases (20 males and 11 females) while Group 2 had 29 cases (15 males and 14 females), and one case of recurrent meningitis. The yield of microbial isolation from Group 1 was only 32.2% with four cases of N. meningitides, two cases of S. pneumonia, three cases of H. influenza and one case of Gram negative rods. The clinical and laboratory information and antimicrobial treatment

astmh.org 516 1690 GPELF with a goal to eliminate LF as public health problem by 2020 A.D. Two key strategies are: Interruption of transmission of LF infection in EPIDEMIOLOGY OF PODOCONIOSIS IN ETHIOPIA: RESULTS endemic countries by reducing microfilariae prevalence levels (below 1%) FROM A FIRST NATIONWIDE MAPPING through Mass Drug Administration (MDA); Prevention and alleviation of disabilities and sufferings in individuals already affected by LF. Nepal: A 1 2 2 Kebede Deribe , Simon J. Brooker , Rachel L. Pullan , Heven total of 61 districts are considered LF endemic. Some districts with high 3 3 3 3 Sime , Abeba Gebretsadik , Ashenafi Assefa , Amha Kebede , prevalence are as high as 40% . The population at risk in Nepal are 25 4 5 6 Asrat Hailu , Maria Rebollo Polo , Oumer Shafi , Moses millions. The causative agent are Wuchereria bancrofti, and transmission 5 7 8 J. Bockarie , Abraham Aseffa , Richard Reithinger , Fikre vector is Culex quinquefasciatus. The reported chronic conditions in 9 10 10 Enquselassie , Melanie J. Newport , Gail Davey 2012 are 28,835, majority were hydrocele. The 10 most morbid districts 1Brighton and Sussex Medical School and School of Public Health, Addis with hydrocele were Morang, Jhapa, Bardia, Banke, Saralahi, Dhading, Ababa University, Addis Ababa, Ethiopia, Falmer, Brighton, United Nuwakot,Kapilbastu, Bara, and Mahottari.LF Elimination Strategies: Kingdom, 2Faculty of Infectious and Tropical Diseases, London School of Interruption of transmission by Mass Drug Administration (MDA) using Hygiene & Tropical Medicine, London, United Kingdom, 3Ethiopian Public two drugs regimen, Diethylcarbamazine (DEC) and Albendazole, once Health Institute, Addis Ababa, Ethiopia, 4School of Medicine, Addis Ababa yearly for six years. Morbidity management by self care and with support University, Addis Ababa, Ethiopia, 5Center for Neglected Tropical Diseases, using intensive but simple, effective and local hygiene technique.MDA Liverpool School of Tropical Medicine, Liverpool, United Kingdom, 6Federal 2013: The number of MDA districts were 56. The total population in MDA Ministry of Health, Addis Ababa, Ethiopia, 7Armauer Hansen Research districts was 25087450. The estimated eligible population for MDA: Phase Institute/ALERT, Addis Ababa, Ethiopia, 8International Development Group, I : 37 districts of eastern, central and western regions and Phase II : 19 Research Triangle Institute, Washington, DC, United States, 9School of districts of mid western and far western regions. Public Health, Addis Ababa University, Addis Ababa, Ethiopia, 10Brighton and Sussex Medical School, Falmer, Brighton, United Kingdom 1692 Podoconiosis (endemic non-filarial elephantiasis) is a major cause of tropical lymphoedema and is endemic in Ethiopia. To guide targeting REAL-TIME PCR AND MELT-CURVE ANALYSIS (QPCR-MCA) and implementation of the National Neglected Topical Diseases control AS A REFERENCE LABORATORY TOOL FOR THE DETECTION strategy, an integrated nationwide survey of lymphatic filariasis (LF) OF ONCHOCERCA VOLVULUS AND ITS IMPORTANCE FOR and podoconiosis was conducted between June and September MONITORING AND EVALUATION ACTIVITIES 2013. Here we present a description of podoconiosis epidemiology in Elizabeth A. Thiele1, Vitaliano A. Cama1, Thomson Lakwo2, Ethiopia resulting from this survey. District health offices’ reports of 3 1 podoconiosis and LF were used to guide selection of survey sites. Data Sindew Mekasha , Paul T. Cantey and cluster level GPS coordinates were collected via smartphones by 1Centers for Disease Control and Prevention, Atlanta, GA, United States, trained local health workers. Individual level data were available for 2Ministry of Health, Kampala, Uganda, 3Ethiopian Public Health Institute, 129,959 randomly-sampled individuals from 1,315 communities in 659 Addis Ababa, Ethiopia districts. Blood samples were tested for Wuchereria bancrofti antigen The efforts to eliminate onchocerciasis in most of Africa by 2025 using immunochromatographic card tests (ICT). A clinical algorithm was necessitate re-examination of current monitoring and evaluation used to diagnose podoconiosis by excluding other potential causes of tools. In particular, assessment of hypoendemic zones, stop-treatment lymphoedema of the lower limb. Mixed-effects logistic regression was determinations, and post-intervention surveillance will require sufficiently used to identify individual-level correlates, adjusting for dependence within sensitive tools to detect low-intensity infections. Mass drug administration district and municipality. Overall, 8,110 of 129,959 (6.2%, 95%CI; 6.1 with ivermectin (IVM) is expected to decrease microfiladermia and thereby to 6.4%) surveyed individuals were identified with lymphoedema with decrease the usefulness of skin snip microscopy, currently the standard 5253 (4.0%, 95% CI; 3.9 to 4.1%) confirmed as podoconiosis cases. assessment tool. Using a pan-filarial qPCR-MCA, we assessed 1) the utility Prevalence among men and women was 3.4% (95%CI; 3.3 to 3.5%) of this single-step assay for detecting evidence of microfilaria (MF) in and 4.7% (95%CI; 4.5 to 4.8%) (p<0.001), respectively. During the residual skin snips and 2) the sensitivity of skin snip microscopy relative to survey 85.2% (95%CI: 84.9 to 85.3%) of respondents were wearing our PCR-based assay. Specificity of the qPCR-MCA to Onchocerca volvulus shoes, but only 57.9% (95%CI: 57.6 to 58.2%) of them were wearing was verified using DNA from O. volvulus macrofilariae; MF of B. malayi protective shoes. Female sex, older age, wearing shoes after 12 years of and pahangi, L. loa, M. ozzardi, and W. bancrofti; and uninfected human age, washing feet less frequently than daily were significantly associated controls. Utility of the qPCR-MCA assay and the relative sensitivity of with increased odds of having podoconiosis. Attending formal education, microscopy were evaluated with residual skin biopsies (i.e., after 24-hour living in a house with a covered floor were associated with decreased odds incubation in saline) collected from hyperendemic regions of Uganda and of having podoconiosis. The survey confirmed that podoconiosis remains Ethiopia (n=500 each) which had received limited rounds of IVM. qPCR- a significant public health problem and is widely distributed in Ethiopia; MCA detected over 94% of known positive skin snips (139/147 total it is endemic throughout 30% of the country’s landmass, where more microscopy positive), identifiable by consistent, well-defined dissociation than 40% of the population live. Results provide a current benchmark curves at 79.35°C (S.D. 0.22) with a minimum 1°C difference from other of the burden of the disease, against which future podoconiosis control filarial species. Using qPCR-MCA as the reference test, the sensitivity of programmes can be measured. the skin snip microscopy was only 74.7% (121/162) and 28.1% (18/64) in Uganda and Ethiopia, respectively. Combined across countries, qPCR-MCA 1691 detected an additional 87 positive samples (38.5%), indicating a combined microscopy sensitivity of 61.5% (139/226). When evaluating low-intensity EFFECTS OF ELIMINATION CAMPAIGN OF LYMPHATIC infections (≤2 MF/snip), the sensitivity of microscopy was only 46% FILARIASIS SEEN IN NEPAL (74/154). Thus, skin snip microscopy does not appear to be sufficiently sensitive to assess transmission in areas with low microfiladermia or to Harishchandra Shah make stop-treatment decisions in the absence of other transmission Ministry of Health and Population, Eastern Health Development assessments (e.g., vector data). qPCR-MCA can augment sensitivity Region,Dhankuta, Nepal, Dhankuta, Nepal and provide diagnostic confirmation of skin biopsies and will be useful Effects of Elimination Campaign of lymphatic Filariasis seen in Nepal, for validating new monitoring tools that may be developed to support 2014* Lymphatic filariasis (LF) is one of the five infectious diseases elimination efforts. targeted by WHO for elimination as public health problem in Nepal. They are LF, Kala-azar, Leprosy, Yaws and Chagas disease. WHO launched

astmh.org 517 1693 1694 FIFTY-EIGHT YEARS OF MAN AGAINST THE WORM IN EVALUATION OF LYMPHATIC FILARIASIS AND BUDONGO ONCHOCERCIASIS FOCUS OF UGANDA- ONCHOCERCIASIS IN THREE SENEGALESE DISTRICTS INTERRUPTION OF TRANSMISSION IS FINALLY IN SIGHT TREATED FOR ONCHOCERCIASIS WITH IVERMECTIN FOR MORE THAN 15 YEARS Moses N. Katabarwa1, Tom Lakwo2, Peace Habomugisha3, Ephraim Tukesiga4, Edson Byamukama3, Edson Byamukama3, Nana O. Wilson1, Alioune Badara Ly2, Ngayo Sy3, Lamine Galex Orukan Ochieng5, William Mugayo6, Thomson Isingoma6, Diawara4, Abdel Direny5, Mawo Fall6, Vitaliano A. Cama1, Paul T. Moris Okumu7, Martini Mborekere8, David Oguttu9, Annet Cantey1, Mark L. Eberhard1, LeAnne M. Fox1, Daouda Ndiaye7, Khainza3, Edridah Tukahebwa9, Rolf Garms10, Tom Unnasch11, Christine Dubray1 12 Frank O. Richards 1Division of Parasitic Diseases and Malaria, Center for Global Health, 1The Carter Center/Emory University, Atlanta, GA, United States, 2Ministry Centers for Disease Control and Prevention, Atlanta, GA, United States, of Health, Vector Contol Division, Kampala, Uganda, 3The Carter Center, 2Ministère de la Santé et de l’Action Sociale, Dakar, Senegal, 3Service de Kampala, Uganda, 4Kabarole District Health Services, FortPortal, Uganda, Lutte Antiparasitaire, MSAS, Dakar, Senegal, 4World Health Organization 5Mbale District Health Services, Mbale, Uganda, 6Masindi Health Services, and African Programme for Onchocerciasis Control Representative, Masind, Masindi, Uganda, 7Bulisa District Health Services, Bulisa, Uganda, Dakar, Senegal, 5IMA World Health, Washington, DC, United States, 6RTI/ 8Hoima District Health Services, Hoima, Uganda, 9Ministry of Health, ENVISION, Dakar, Senegal, 7Cheikh Anta Diop University, Dakar, Senegal 10 Vector Control Division, Kampala, Uganda, Bernhard Nocht Institute for In Africa, there is significant overlap between onchocerciasis and lymphatic 11 Tropical Medicine, Hamburg, Germany, Department of Global Health, filariasis (LF). Current efforts to eliminate these two diseases are through 12 University of South Florida, Tampa, FL, United States, The Carter Center, mass drug administration (MDA) with ivermectin alone for onchocerciasis Atlanta, GA, United States or with ivermectin and albendazole for LF. Years of ivermectin distribution The battle against onchocerciasis in hyperendemic Budongo focus (where for onchocerciasis may have decreased or interrupted LF transmission the baseline microfilaria (mf) rate was 80% and fly infection rate 20%) in certain areas. The Kedougou region, Senegal, has been historically has lasted over a half a century. Vector control of Simulium neavei using known to be co-endemic with LF (immunochromatographic test [ICT] DDT commenced in1955, and larviciding was halted when no larval/ prevalence ≥1%) and onchocerciasis (microfilaria prevalence 9.0-68.4%). pupal stages of S. neavei were observed on crabs, and no adults flies were MDA for onchocerciasis started in 1988 and as of 2014, albendazole had found in human landing captures. Six months after stopping larviciding, not been added to target LF. The objective was to assess in an integrated however, rapid repopulation of S. neavei was observed. In August 1956, manner the status of LF and onchocerciasis in three districts of Kedougou a new series of DDT larviciding was initiated. In 1958, the early stages after ≥15 years of ivermectin MDA. Sixteen villages close to rivers and of the vector could not be found in the rivers and the adult fly had breeding sites for onchocerciasis vector, Simulium spp, were selected. disappeared by 1962 indicating elimination of S. neavei from the focus. LF antigenemia testing (ICT) was added to skin snip microscopy for Repeated courses of DEC were provided to individuals in the communities onchocerciasis evaluation. Convenience sampling of residents ≥5 years who were infected. To avoid vector reinfestation, maintenance larviciding was performed. Dried blood spots were collected to test for antibodies continued until about 1971. Political unrest in Uganda led to the against Wb123 (LF) and Ov16 (onchocerciasis) antigens (results pending). collapse of this work, and by 1989 S. neavei had again repopulated One village refused to participate and one was excluded because it the area and onchocerciasis recrudescence had occurred. Annual mass was treated with ivermectin too recently. Forty percent (1154/2925) of drug administration (MDA) with ivermectin was provided from 1989 to residents participated; 50% were males and the median age was 15 years. 2007 to all the 184 affected communities and a population of 150,195 In two districts, no participants were ICT or skin snip positive. In the third people. However, a 2008 serosurvey of 3159 children showed an OV16 district, 3.4% (6/176) were ICT-positive (village range 1.9-6.4%) and 0.7% antibody rate of 9.5%, indicating continued transmission. After Uganda (1/150) were skin snip-positive. The mean age of ICT-positive participants established a policy for onchocerciasis elimination in 2007, biannual was 49 years (range 25-79); the participant with a positive skin snip was treatment was launched in Budongo in 2008, and continues to date. 79 years old. After ≥15 years of ivermectin distribution, LF prevalence was However, in assessments done in 2011 vector infectivity rates still ranged still above treatment threshold in one of the three districts included in the up to 8.7%. In June 2012, temephos (Abate®) larviciding was added to evaluation. The integrated evaluation of LF and onchocerciasis provided compliment twice per year ivermectin treatments. By February, 2014, only important information on both diseases that should help program 2 (0.7%) crabs out of 300 were infested; no adult fly has been collected managers make decisions about treatment interventions. since September, 2013. Budongo focus is an example of a difficult onchocerciasis ‘hot spot’ requiring both twice yearly ivermectin MDA and vector control to break transmission. 1695 THE USE OF HUMAN SWEAT METABOLITES AS BAIT FOR MONITORING VECTORS OF ONCHOCERCIASIS IN WEST AFRICA AND LATIN AMERICA Ryan M. Young1, Nathan D. Burkett-Cadena1, Tommy W. McGaha2, Mario A. Rodríguez-Pérez3, Laurent Toé4, Bill J. Baker1, Raymond Noblet2, Eddie W. Cupp5, Andrew J. Shilling1, Javier Garza-Hernández3, Kristel Rodríguez-Morales3, José Torres- Estrada6, Thomas R. Unnasch1 1University of South Florida, Tampa, FL, United States, 2University of Georgia, Athens, GA, United States, 3Instituto Politécnico Nacional, Reynosa, Mexico, 4Multidisease Surveillance Centre, World Health Organization, Ouagadougou, Burkina Faso, 5Auburn University, Auburn, AL, United States, 6Instituto Nacional de Salud Pública, Tapachula, Mexico Onchocerciasis, a.k.a. river blindness, is a parasitic disease caused by infection from the nematode Onchocerca volvulus. The parasite is transmitted to humans by the bite of infected black flies (genus

astmh.org 518 Simulium). The World Health Organization (WHO) estimates 18 million 1697 people suffer from onchocerciasis, the vast majority occurring in central Africa, and isolated foci in the American Tropics. Treatment of the POTENTIAL RE-EMERGENCE OF WUCHERERIA BANCROFTI disease relies on accurate epidemiological data, which is best achieved TRANSMISSION IN A PREVIOUSLY CONTROLLED though real-time data of infection prevalence in the vectors. The need HYPERENDEMIC REGION (SIKASSO) IN SOUTHERN MALI for a new monitoring method is crucial. To this end we identified key 1 1 1 primary metabolites in human sweat, which putatively attract black flies Siaka Yamoussa Coulibaly , Yaya Ibrahim Coulibaly , Ilo Dicko , 1 1 1 to humans. Laboratory studies were then conducted in Southern Mexico Siaka Konate , Housseini Dolo , Moussa Brema Sangare , Michel 1 1 and West Africa to test which compounds attracted these vectors of Emmanuel Coulibaly , Lamine Soumaoro , Abdallah Amadou 1 1 1 2 onchocerciasis, using electroantennography and y-tube olfactometry. The Diallo , Modibo Sangare , Benoit Dembele , Massitan Dembele , Pj 3 3 3 1 attractive compounds will be developed into baits to lure black flies to a Hooper , Eric Ottesen , Dominique Kyelem , Sekou F. Traore , Amy 4 4 novel trap for monitoring vector abundance and infection prevalence in D. Klion , Thomas B. Nutman both Latin America and Africa. In this study we describe the identification 1ICER Mali, Bamako, Mali, 2National Lymphatic Filariasis Elimination of key human sweat components via GC-MS, the identification of Program, Mali, Bamako, Mali, 3Neglected Tropical Diseases Support attractive metabolites to the two major species of Simulium as well as trap Center, Task Force for Global Health, Atlanta, GA, United States, 4National development and bait formulation for the continued monitoring of these Institute of Allergy and Infectious Diseases/National Institutes of Health, important disease vector. Bethesda, MD, United States Mass drug administration (MDA) for the elimination of lymphatic filariasis 1696 (LF) has led to potential transmission interruption in several endemic regions of Mali based on transmission assessment surveys (TAS). To assess LYMPHATIC FILARIASIS ELIMINATION: ASSESSMENT OF the utility of TAS approach, we utilized standard TAS methodology (ICT TWO VILLAGES WITH DIFFERENT ENDEMICITY LEVELS IN A positive prevalence in 6-7 year olds) and compared it to xenomonitoring, PREVIOUSLY HIGHLY ENDEMIC REGION (SIKASSO) OF MALI night blood microfilariae counts and IgG4 antibody to Wb123 (for the Yaya Ibrahim Coulibaly1, Salif Seriba Doumbia1, Ilo Dicko1, last 3 years) over a five year period (2009-2014) following the cessation Lamine Soumaoro1, Massitan Dembele2, Sekou Fantamady Traore1, of MDA in 6 villages in the region of Sikasso in southern Mali. In 2009 Joseph Kubofick3, Amy Klion3, Louise Kelly-Hope4, Thomas B. (at the start of the surveillance period) all 289 children aged 6-7 years Nutman3, Moses J. Bockarie4 were negative for circulating filarial antigen (CFA) by ICT, by calibrated thick smears of blood collected at night, and by IgG4 antibody to Wb123. 1 2 ICER Mali, Bamako, Mali, National Lymphatic Filariasis Elimination Despite this, 2/4391 (0.11%) dissected mosquitoes were positive for larvae 3 Program, Mali, Bamako, Mali, National Institutes of Health, Bethesda, of Wuchereria bancrofti (Wb). In 2011, there was a CFA prevalence by 4 MD, United States, CNTD, Liverpool School of Tropical Medicine, United ICT of 2.6% (8/301) in the 6-7 year olds, a prevalence of 1.09% (1/92) Kingdom, United Kingdom for antibody responses to Wb123, but negative xenomonitoring. In the Lymphatic filariasis (LF) is a disfiguring neglected tropical disease that is subsequent 2 years (2012 and 2013), there were consistent and significant targeted for elimination by 2020 using annual mass drug administration increases in the prevalence of CFA (Trend Chi2= 11.49, p=0.0007) to 3.9% (MDA) in endemic areas. We previously reported the Anopheles gambiae (11/285) in 2012, and 4.1% (13/316) and in the prevalence of anti-Wb123 complex frequencies in 2 neighboring villages of Kolondièba, one of IgG4 to 3.2% (10/316) in 2013. Despite this increase in both ICT and the highly LF endemic district of Sikasso, Mali. To assess transmission Wb123 IgG4 antibody prevalence, no infected anopheline mosquitoes interruption after 6 annual MDA in the district of Kolondièba and were found in 2011, 2012 and 2013. These data suggest that despite determine the potential impact of vector density on MDA-induced having met the criteria for cessation at the beginning of the surveillance, transmission interruption. A cross sectional study with quantitative and that there appears to be low level emergence of Wb transmission and that qualitative data collection methods in 2 villages of Sikasso region. The antibody monitoring may provide a better early warning tool than more village of Boundioba had a lower Anopheles density as compared to standard TAS tools. Bougoula (1,494 versus 251 specimen from July to December 2011). A total of 481 volunteers in Bougoula including 340 female (70.5%) and 1698 332 in Boundioba including 221 female (66.6%) were included in this study. The 6-7 years/15 years and above composition was 113/368 and SYSTEMIC NON-COMPLIANCE: A POTENTIAL FACTOR INTHE 127/205 respectively in Bougoula and Boundioba. Microfilaremia was RE-EMERGENCE OF LYMPHATIC FILARIASIS TRANSMISSION significantly more frequent in the15 years and above in Boundioba(1.95%, IN SIKASSO, MALI 4/205) as compared toBougoula (0%, 0/368), (p=0.02, Fisher exact test). 1 1 1 Additionally, the 2 villages showed comparable low prevalences in 6-7 Ilo Dicko , Yaya Ibrahim Coulibaly , Siaka Yamoussa Coulibaly , 1 1 1 years olds with respectively 1/127 and 0/113 for Boundioba and Bougoula. Housseini Dolo , Moussa Brema Sangare , Siaka Konate , Michel 1 1 Anopheles vector density may be misleading because it is not necessarily Emmanuel Coulibaly , Lamine Soumaoro , Abdallah Amadou 1 1 1 2 associated with a higher endemicity in a village under MDA. Diallo , Modibo Sangare , Benoit Dembele , Massitan Dembele , Pj Hooper3, Eric Ottesen3, Dominique Kyelem3, Thomas B. Nutman4, Sekou Fantamady Traore1, Amy D. Klion4 1ICER Mali, Bamako, Mali, 2National Lymphatic Filariasis Elimination Program, Mali, Bamako, Mali, 3Neglected Tropical Diseases Support Center, Task Force for Global Health, Atlanta, GA, United States, 4National Institute of Allergy and Infectious Diseases/National Institutes of Health, Bethesda, MD, United States Recent transmission assessment surveys (TAS) suggest low level re- emergence of Wuchereria bancrofti transmission after cessation of mass drug administration (MDA) in 6 previously hyperendemic villages in Sikasso, Mali. Coverage rates in the villages ranged from 67% to 89.6% over the 7 years of MDA and all stopping criteria were met at the beginning of the surveillance period. To begin to identify potential causes for this re-emergence, a questionnaire was administered to randomly selected adult residents of the six villages to assess the prevalence of and

astmh.org 519 reasons for systematic non-compliance with MDA. A total of 486 subjects 1700 (170 men and 316 women) were questioned, of whom 16.1% (79/486) reported never swallowing MDA drugs. The most common reasons given INTEGRATED FILARIAL MICRO-MAPPING TO DETERMINE were being unaware of MDA (24/486; 4.9%), being pregnant or breast- IMPLEMENTATION STRATEGIES IN LOA LOA CO-ENDEMIC feeding (8/486; 1.6%) and not willing to take the drugs (6/486; 1.2%). AREAS: THE ANGOLAN EXPERIENCE Although systematic non-compliers were more likely to be younger [OR = 1 1 2 2 1.7 (1.006-2.921) for individuals 15-30 vs. >30 years of age], compliant Miguel Brito , Rossely Pauloi , Tom Unnasch , Robert Novak , 2 3 3 and systematically non-compliant subjects were similar with respect to Benjamin Jacob , Michelle Stanton , David Molyneux , Louise 3 participants’ instruction level [OR = 1.2 (0.59-2.51)] and the presence of Kelly-Hope lymphoedema / hydrocele [OR = 0.5 (0.11-2.63)]. These data suggest that 1CISA - Health Research Centre in Angola, Caxito, Angola, 2College of significant rates of systematic non-compliance can be present despite Public Health, Department of Global Health, University of South Florida, adequate overall coverage rates. Whether persistent infection in systematic Tampa, FL, United States, 3Liverpool School of Tropical Medicine, Liverpool, non-compliers provided the reservoir for re-emergence of transmission in United Kingdom the 6 study villages requires further study. The co-distribution of Loa loa (loiasis or tropical eye worm) is a significant impediment to the lymphatic filariasis (LF) and onchocerciasis elimination 1699 programmes in Angola, due to the potential risk of severe adverse events (SAEs) associated with the drug ivermectin when given to individuals with NON-MANSONELLA OZZARDI ATYPICAL MICROFILARIASIS high L. loa microfilarial loads in the blood. This has significant implications IN THE PERUVIAN AMAZON BASIN for mass drug administration campaigns and alternative strategies may Salomon Durand1, Dolores Rimarachin2, Amy C. Morrison2, be required in selected areas. One of the highest L. loa risk areas is Dande Karen Campos2, Jorge Nunez1, Moises Sihuincha3, Geral C. Municipality, Bengo Province in Northern Angola, an area which historically Baldeviano1, Andres G. Lescano1 has reported cases of LF and onchocerciasis. To better determine the safest treatment strategy in this area, this study conducted an integrated filarial 1U.S. Naval Medical Research Unit - 6, Callao, Peru, 2U.S. Naval Medical micro-mapping survey to understand the geographical overlap of the Research Unit - 6, Iquitos, Peru, 3Hospital Apoyo Iquitos, Iquitos, Peru three diseases. GIS-Remote Rapid Eye satellite data was also employed to Filariasis is a neglected tropical disease around the world. This disease is a provide the foundation for empirical information on vector and parasite vector-borne infection caused by nematodes (roundworms) of the family populations. In total 23 villages, distanced approximately 10-15km apart, Onchocercidae. Since 1950s, filariasis caused by Mansonella ozzardi has across peri-urban and rural areas were surveyed during January-February been regularly reported in the Peruvian Amazon basin. Isolated cases 2014. In each village, up to 100 individuals were assessed using the rapid have been diagnosed as M. perstans, Bruguia spp, Onchocerca spp. and assessment procedure for loiasis (RAPLOA) and rapid epidemiological Dirofilaria spp. Since 2010, by examining de-identified blood samples mapping of onchocerciasis (REMO), and two questions on LF morbidity (thick smear and Knott’s concentration techniques) we have characterized (presence of lymphedema, hydrocele). The study found low levels of the prevalence of filarial infections and associated clinical symptoms in endemicity of all three diseases (<20%), with different overlapping both rural (n=383) and urban (n=755) human populations located on distributions, with most villages reporting at least one filariasis case. tributaries of the Amazon River near and in Iquitos, Peru. In the rural To confirm the hypo-endemic levels of LF and onchocerciasis, a further communities, prevalence of microfilariae was 28.5% (109/383) overall; but seroprevalence survey using rapid diagnostic tests in the same villages is the majority 99% (108/109) were M. ozzardi. Prevalence rates were not planned for June 2014. This will provide additional micro-epidemiological heterogeneous ranging from 0 to 72.5% in 11 communities. In contrast, information to help determine if the recommended alternative strategy of 755 samples from residents of Iquitos, 2% and 4% were infested of albendazole twice yearly and long-lasting/insecticide treated bednets with M. ozzardi and an atypical microfilaria, respectively. Interestingly, (LLINs/ITNs) should be used for LF elimination, and if an alternative to those infected with M. ozzardi tended to be febrile adult males with ivermectin for hypoendemic onchocerciasis elimination, such as the drug occupations associated with rural areas, whereas those infested with the doxycycline or vector control, needs to be considered. atypical parasite were rarely febrile and were often children or housewives. Surveillance in local hospitals identified at least one morphologically 1701 distinct atypical microfilariae and one co-infection with M. ozzardi in seven symptomatic patients; one had subacute skin lesions and the others fever. SERO-PREVALENCE AND RISK FACTOR SURVEY FOR The atypical microfilariae were macroscopically distinct from M. ozzardi, LYMPHATIC FILARIASIS IN PAPUA NEW GUINEA measuring 600 x 8 µm, with no sheath and no nuclei in the tail. In a 1 2 2 subset of samples tested by PCR. All M. ozzardi were confirmed, but five Melinda Susapu , Moses Bockarie , Lisa Reimer , Louise Kelly- 2 of the atypical microfilariae tested negative the internal transcribed spacer Hope rDNA sequence of M. perstans and Onchocerca volvulus. In conclusion, 1Department of Health, Port Moresby, Papua New Guinea, 2Liverpool atypical microfilariae with a distinct epidemiology from M. ozzardi, and not School of Tropical Medicine, Liverpool, United Kingdom related to M. perstans, or O. volvulus, are sufficiently prevalent to warrant Papua New Guinea (PNG) has an estimated population of 7 million investigation of their health impact in the Peruvian Amazon. inhabitants; of which 4 million are predicted to be at risk of lymphatic filariasis (LF). LF is a debilitating disease caused by lymphatic-dwelling nematodes Wuchereria bancrofti, which is transmitted by Anopheles mosquitoes in PNG, similar to malaria. Some national and published data exists, however, the geographical distribution, burden of disease and associated risk factors are currently not well defined. This study investigated the prevalence and potential risk factors of LF in one endemic province of PNG through the administration of a prevalence survey and household questionnaire related to the LF transmission and the national LF programme. In April 2013, four villages in a southern remote area of Madang Province were selected. Approximately 100 individuals in each village were interviewed and examined for LF infection, using ICT rapid diagnostic kits, and evidence of clinical disease. This study found that 32 individuals out of 389 surveyed (8.2%) were LF antigen positive, and 3 individuals had lymphodema (elephantiasis) of the leg (0.8%). All of those

astmh.org 520 interviewed did not know about the disease, what caused it, how it was A systematic review identified 9 published randomized controlled trials transmitted or were aware of the national programme to eliminate LF. A evaluating the effects of various Doxycycline regimens on Wuchereria follow-up microfilaremia (MF) survey was conducted in the study site with or Brugia microfilaria levels at 12 months post-treatment in 5 countries, the most ICT positive individuals, and included the majority of community with or without interim single-dose microfilaricide (Ivermectin or DEC). members (n=300). Preliminary results indicate the average Mf prevalence For elimination of microfilariae at 12 months (compared to placebo), was 41.5%, and ranged from 10.5% in children under 10 years, to 54.6% the pooled Risk Ratio was 3.22 (1.95, 5.32) , with high heterogeneity in adults over 50 years of age. Mf prevalence was found to be higher in (I2=68%). Subgroup analysis showed: Doxycycline 6-8 week regimens, males (46.4%) than females (34.9%), and among those living in houses RR= 3.96 (2.07, 7.59); Doxycycline 3-4 week regimens, RR= 2.14 (1.08, made of bush material (45.8%) compared with other semi-permanent 4.26); Ivermectin or DEC 4 months after Doxycycline, RR= 2.29 (1.73, materials (23.1%). The field work is still in progress and expected to be 3.04); no interim microfilaricide, RR= 5.80 (2.36, 14.24). Multi-day finish with final results by November 2014. This research highlights that LF Doxycycline regimens effectively eliminate LF microfilariae at 1 year after is endemic in remote areas of the country and the national LF programme treatment. Applicability of such multi-day regimens to population-wide has to scale up its efforts to control and eliminate the spread of infection control programs is limited. Further studies should evaluate shorter-term with particular emphasis on LF advocacy and education to those most at treatment. risk. 1704 1702 SIMULTANEOUS DETECTION OF ONCHOCERCA VOLVULUS METHODS FOR ASSESSING LYMPHATIC FILARIASIS AND O. OCHENGI IN INFECTED SIMULIUM FLIES USING TRANSMISSION IN LOW ENDEMIC AREAS OF BANGLADESH: ANEW MULTIPLEX REAL-TIME PCR ONE STEP CLOSER TO THE ELIMINATION GOAL Anna Albers1, Christina Weier1, Sandra Arriens1, Samuel Wanji2, A. S. M. Sultan Mahmood1, Rouseli Haq1, Mujibur Rahman1, Kenneth Pfarr1, Achim Hoerauf1 1 2 Shamsur Rahman , Louise Kelly-Hope 1University Hospital of Bonn, Bonn, Germany, 2Research Foundation for 1Directorate General of Health Services, Dhaka, Bangladesh, 2Liverpool Tropical Diseases and Environment, Buea, Cameroon School of Tropical Medicine, Liverpool, United Kingdom Onchocerca volvulus parasitic worms infect ~37 million people in sub- Bangladesh had one of the highest burdens of lymphatic filariasis (LF) at Saharan Africa and parts of Latin America, causing dermatitis, skin atrophy the beginning of the Global Programme to Eliminate LF (GPELF), with an and visual impairment. In 27 endemic countries in Africa, 130 million estimated 80 million people at risk of infection. Baseline mapping in 2000 people at risk of the disease. The January, 2014 report of the International using the rapid diagnostic ICT cards, found that 34 of the 64 districts in Task Force for Disease Eradication has stated the continued need for the country were endemic, however, only 19 districts required mass drug improved diagnostics to assess when mass drug administration efforts administration (MDA) using the drugs albendazole and DEC to interrupt can be halted and to monitor for recrudescence. Currently transmission is transmission. The Bangladesh LF Programme has successfully scaled up monitored by identifying larvae in dissected Simulium damnosum species, MDA in these districts, and is moving into the elimination phase and using the vector of O. volvulus, which are also able to transmit O. ochengi, a the WHO recommended Transmission Assessment Survey (TAS) to assess parasitic worm of cattle that does not infect humans. We developed a their success in interrupting transmission - so far with excellent results. The multiplex real-time PCR based on the ND5 gene of the Onchocercidae outstanding important question for the LF Programme was how to assess genus with specific TaqMan probes to differentiate O. volvulus and O. the 15 endemic districts that were found to have low prevalence levels ochengi from other Onchocercidae. A blinded study with 217 flies from O. (<1%) and not be eligible for MDA. Follow-up night blood microfilaria volvulus and O. ochengi endemic and O. volvulus/O. ochengi co-endemic (Mf) and community clinical surveys undertaken in 2008 -2010 in selected areas in Cameroon (n=23) showed 100% specificity in all analyzed areas of these districts found little or no evidence of infection and disease Simulium flies. Vector monitoring to assess transmission potential in such as lymphedema and hydrocele. Currently, there is no recommended endemic areas is reliable. Our multi-plex real-time PCR offers time and cost strategy for assessing low endemic districts, therefore, in order to address savings over species identification via microscopy. this issue and provide more rigorous evidence that LF is not a public health problem, the TAS method is being used as an assessment tool 1705 with additional systematic patient searching at household level. The assessments are planned for each month of 2014 and being carried out MODELING THE EFFECTS OF MASS DRUG TREATMENTS AND by trained field teams visiting schools for TAS (targeting children) and VECTOR CONTROL ON CO-INFECTION WITH MALARIA AND using local community clinic workers and volunteers to visit households LYMPHATIC FILARIASIS (targeting individuals with clinical manifestations). To date five districts have been assessed with good results, and if the remaining districts are Sarah Lukens, Brajendra K. Singh, Edwin Michael also found to have little or no LF infection or disease, the national LF University of Notre Dame, Notre Dame, IN, United States programmes can ‘shrink the LF map’ by approximately 30 million people Malaria and lymphatic filariasis (LF) are both transmitted by anopheles and move one step closer to their elimination goal, with an increased mosquitoes and are co-endemic in many regions of the tropics. For focus on the new priorities of surveillance and morbidity management. ongoing global campaigns to eliminate LF and malaria, it is important to understand interactions between both parasites and hosts where they 1703 co-exist. The use of mass drug administrations (MDA) to reduce prevalence and intensity of microfilariae may increase the lifespan of anopheles SYSTEMATIC REVIEW AND META-ANALYSIS OF mosquitoes and thereby potentially increasing the transmission of malaria, DOXYCYCLINE IN CONTROL OF LYMPHATIC FILARIASIS while interactions at the host level may affect susceptibility, disease severity, and co-transmission of both diseases. Each parasite system alone Patricia A. Woodall exhibits complex dynamics where factors such as vector biting rates and Emory University, Atlanta, GA, United States threshold prevalence of human infection contributes to either extinction or Lymphatic Filariasis control programs rely on periodic population-wide stabilization to an endemic level. Knowledge of how interactions between administration of microfilaricidal agents repeated over several years to both systems may affect co-infection endemicity and extinction dynamics decrease transmission. Effective macrofilaricidal agents could decrease is important for designing effective disease management programs such the years necessary to achieve elimination. Doxycycline has emerged as MDA and vector control (VC). We extend a mathematical model of as a possible agent due to its effects on the Wolbachia endosymbiont.

astmh.org 521 malaria-LF co-infection to describe how the interplay between these two evaluation of the impact MDA on LF. More than 3,900 people were infections influence threshold behaviors in the system, and how MDA and tested for filarial antigenemia in 40 villages in 6 districts in the central and VC interventions can influence elimination or resurgence of both diseases. Southeastern part of the country. Antigen rates ranged from 4-22% in Lakota, 4-21% in Tiebissou, 15-41% in Akoupe, 21-25% in Agboville, 1706 9-14% in Bettie, and 6-35% in Abengourou districts. Microfilaremia rates ranged from 1% in Lakota to 11% in Abengourou and Agboville. IMPACT OF MEROWE DAM ON ONCHOCERCIASIS VECTORS Prior ivermectin distribution in areas with coendemic onchocerciasis may OF ABU HAMED, NORTHERN SUDAN partially explain the highly variable Mf rates in these areas. This study has helped to establish the current LF situation in Côte d’Ivoire, and this Isam Zarroug1, Arwa Elaagib2, Sara Abuelmaali3, Hanan information will be used to plan and implement the national LF elimination Mohamed1, Wigdan Elmubarak1, Kamal Hashim4, Tong Chor M program based on MDA. Deran1, Nabil Aziz5, Tarig Higazi6 1National Onchocerciasis Control Program, Khartoum, Sudan, 2University 1708 of Khartoum, Khartoum, Sudan, 3National Health Laboratories, Khartoum, Sudan, 4National Programme for Prevention of Blindness, Khartoum, PROGRAMMATIC IMPLICATIONS OF EXTENSIVE VECTOR Sudan, 5The Carter Center, Khartoum, Sudan, 6Ohio University, Zanesville, CONTROL ON THE ELIMINATION OF LYMPHATIC FILARIASIS OH, United States IN ZAMBIA Abu Hamed, the northernmost onchocerciasis focus in the world, is 1 2 3 located along the River Nile banks in the Nubian Desert. Hydroelectric Mutale Nsakashalo , Michelle Stanton , Enala Mwase , Elizabeth 1 4 5 2 dams can alter activity of black flies and may provide breeding sites for Chizema , Victor Mukonka , Peter Songolo , Brent Thomas , Maria 2 2 2 2 black fly. Merowe Dam, the largest hydropower project in Africa, was Rebollo , Moses Bockarie , J. Russell Stothard , Louise Kelly-Hope built west of Abu Hamed focus in 2009. The impact of the Dam on 1Ministry of Health, Lusaka, Zambia, 2Liverpool School of Tropical onchocerciasis and its black fly vectors in Abu Hamed focus was measured Medicine, Liverpool, United Kingdom, 3University of Zambia, Lusaka, in this study. Entomological surveys for aquatic stages and adult Simulium Zambia, 4Copperbelt University of Zambia, Ndola, Zambia, 5World Health hamedense were conducted before and after the inception of Merowe Organization Country Office, Lusaka, Zambia Dam in 2007/2008 and 2010/2011. There was no black fly breeding or Lymphatic filariasis (LF) is widely endemic in Zambia, and the National adult activity in the previously known breeding sites upstream of the LF Programme is planning the first nationwide mass drug administration Merowe Dam with the western most breeding site found in Al Sarsaf (MDA) to interrupt transmission in 2014. Overall the LF prevalence is low village near the center of the focus. No adult or aquatic stages of black (<10%) in most regions of the country and it is possible that the well flies were found downstream of the Dam. The artificial lake of the Dam establish vector control programme for malaria has already impacted LF flooded all the breeding sites in the western region of the focus and transmission over the past decade. To better understand the distribution no aquatic stages and/or adult black fly activity were established in the of vector control that has occurred across the country and the potential study area upstream of the Dam. The Dam seems to have positive impact implications for the LF Programme as it moves to scale up MDA, this on onchocerciasis and its black fly vectors in Abu Hamed focus. These study developed a spatially modelled vector control map and examined outcomes of the Merowe Dam might have contributed to the recently it in relation to the baseline LF prevalence data collected across 108 geo- declared interruption of onchocerciasis transmission in Abu Hamed focus. referenced sites in 2003 and 2011, and the sentinel site data collected Continuous entomological surveys are needed to monitor presence of across 32 geo-referenced sites in 2014. Information on bed nets, including black fly vectors and its impact on the disease. long-lasting/insecticide treated bed nets (LLIN/ITNs) and indoor residual spraying (IRS) distributions was obtained from the Ministry of Health, and 1707 public data sources such as the Demographic Health Survey (DHS) data, President’s Malaria Initiative (PMI) reports, and combined in a weighted THE CURRENT STATUS OF LYMPHATIC FILARIASIS IN COTE sum to form a multiple vector intervention score, which was then used D’IVOIRE PRIOR TO IMPLEMENTATION OF A NATIONAL to produce district-level maps of vector control intensity. Each district was PROGRAM OF MASS DRUG ADMINISTRATION classified according to LF prevalence and the vector intervention score which included the following combinations i) low LF /high vector control, Aboulaye Meite1, Andrew C. Majewski2, Gary J. Weil2, Peter U. ii) low LF / low vector control iii) high LF/ high vector control and iv) high Fischer2 LF/low vector control. These groups will help the LF Programme as it 1Programme National de lutte contre la Schistosomiase, les scales up MDA to determine if a district has potential for elimination, in Géohelminthiases et la Filariose Lymphatique, Ministry of Health, Abidjan, need of very high MDA coverage and will require standard or enhanced Côte D’Ivoire, 2Washington University School of Medicine, St. Louis, MO, surveillance. United States Côte d’Ivoire is planning to implement a coordinated national program 1709 of mass drug administration (MDA) for elimination of onchocerciasis (Oncho) and lymphatic filariasis (LF) in the near future. The country has MODELING IMPACTS OF INTEGRATED VECTOR CONTROL complex patterns of endemicity for these infections with extensive areas ON LYMPHATIC FILARIASIS TRANSMISSION DYNAMICS AND of coendemicity, areas that have received mass drug administration with ELIMINATION PROCESS ivermectin for variable periods, and extensive migration to and from neighboring countries (Liberia, Ghana, and Burkina Faso). For example, Brajendra K. Singh, Edwin Michael some areas in the northern part of the country have received many rounds The University of Notre Dame, Notre Dame, IN, United States of ivermectin for onchocerciasis control during OCP and APOC, and LF Lymphatic filariasis (LF) is a target for global eradication by 2020. Launched rates tend to be low in the North. In contrast, Oncho is uncommon in in 2000, the Global Programme to Eliminate Lymphatic Filariasis relies coastal areas (with some exceptions), and little ivermectin has been used in mainly on large-scale preventative chemotherapy programs via mass the South. Some 47 of the country’s 82 health districts (mainly in Central drug administration (MDA) to eliminate this major vector-borne parasitic and Southern districts) are considered to be co-endemic for LF and Oncho. disease from all endemic settings. Modeling studies of LF transmission LF mapping circa 2001 was based on antigen testing (Binax Now Filariasis, and control have been crucial for designing national control programs by card test) of 50-100 people in two villages per district. The current study establishing and quantifying the presence of infection and vector biting was performed to obtain more current information on the distribution thresholds below which the transmission is interrupted. Renewed malaria of LF in the country and to identify sentinel sites for monitoring and control efforts have witnessed large-scale applications of vector control

astmh.org 522 (VC) through the use of long lasting insecticidal nets (LLIN) alone or in 1711 some combination with indoor residual spray (IRS) across the majority of LF endemic regions. LLIN/IRS affects the transmission of mosquito-borne DIFFERENCES IN OV-16 ELISA IMMUNE RESPONSES AMONG infections either by directly killing, or by preventing mosquitoes from CHIMPANZEES INOCULATED WITH ONCHOCERCA VOLVULUS coming into contact with infected hosts through several mechanisms. Recent community trials have shown the substantial impact that VC may Circe E. McDonald, Paul N. Cantey, Vitaliano Cama, Mark L. have in enhancing LF transmission interruption particularly when infection Eberhard prevalence has been depressed to low levels using MDA. Despite these Centers for Disease Control and Prevention, Center for Global Health, observations, theoretical and quantitative modeling of the impact of VC on Atlanta, GA, United States LF transmission dynamics that takes explicit account of the various effects Onchocerciasis, also known as river blindness, is a neglected tropical the different VC options may have on mosquito populations is scarce. Such disease caused by the filarial parasite Onchocerca volvulus. Detection of analysis is vital when chemical insecticides require repeated applications in immunoglobulin G4 (IgG4) antibodies to Ov16 recombinant antigen is the affected communities due to their variable durations of effectiveness. the basis for serological tests for onchocerciasis. However, the dynamics This need for frequent insecticide applications introduces a number of of this immune response have yet to be thoroughly characterized. A factors such as the effects of adherence to the recommended timeframe non-human primate model was used to evaluate the temporal evolution for the replenishment of LLIN/IRS and the maintenance of the required of antibody responses under controlled infection conditions. Nine community coverage, which may contribute to different outcomes from chimpanzees were inoculated with third-stage larvae of O. volvulus. Three VC between communities. We aim to extend our present Bayesian chimpanzees were inoculated with approximately 100 stage three larva Melding LF modeling framework by incorporating the specific effects that (L3) of Guatemalan origin, either one, three, or five times. Six chimpanzees the application of LLIN and IRS, used separately or in combination, may each were inoculated once with 200, 300, or 400 L3 of Liberian origin. have on the effective mosquito biting rate to quantify and gain better On a monthly basis, serum was collected and the presence of microfilariae insights on the role of VC in the MDA-based LF control programs. (Mf) was determined via skin snip microscopy. Seven of nine chimpanzees developed patent infections, and six were used to evaluate the temporal 1710 responses over a median number of 1,660 days post-inoculation (PI). The seventh chimpanzee with patent infection was not evaluated due to AXENICALLY-DERIVED CAENORHABDITIS ELEGANS ANTIGEN health complications and was withdrawn at 535 days PI. Infections were FOR THE TREATMENT OF AUTOIMMUNITY categorized based on average microfiladermia of three consecutive dates Belinda M. Jackson1, Marina N. Torrero2, Edward Mitre1 as: weak (<10Mf/snip), mild (≥10 and <20) and strong (≥20 Mf/snip). One chimpanzee had a weak infection, two developed mild infections, and 1 2 Uniformed Services University, Bethesda, MD, United States, National three had strong infections. No positive IgG4 responses to Ov16 were Institutes of Health, Bethesda, MD, United States detected in the two inoculated but uninfected chimpanzees. The mean The potential use of helminth infections as a protective measure against time to develop IgG4 responses and detection of Mf were 414 and 485 inflammatory disease has been validated in both animal and clinical days PI. Four chimpanzees showed decreases in IgG4 values towards studies. However, several obstacles impede the use of parasitic helminths the end of the study. In three of these chimpanzees, decreased IgG4 in clinical practice. These include difficulties in obtaining large quantities responses were detected with decreasing microfiladermia. These findings due to complex lifecycles, challenges in purification from lifecycle hosts, indicate that positive serology to Ov16 occurs only among chimpanzees inter-batch variability in product, and the potential of live infections to that developed patent infections, and suggest that anti-Ov16 antibody cause clinical symptoms. To overcome these hurdles, we tested whether responses may decrease over time after reductions in detectable Mf loads a homogenate of soluble antigens prepared from axenically grown in skin snips. Caenorhabditis elegans (aCeAg) would protect against autoimmunity. Using a TLR4 reporter cell line, we demonstrated that, in contrast to 1712 soluble antigen prepared from C. elegans grown on Escherichia coli lawns, aCeAg lacks LPS and does not activate TLR4. Twice weekly intraperitoneal CHARACTERIZING REACTIVITY TO ONCHOCERCA VOLVULUS injections of 100 mcg of aCeAg protected against the development ANTIGENS IN MULTIPLEX BEAD ASSAYS of type 1 diabetes in non-obese Diabetic (NOD) mice (80% T1DM in 1 1 1 PBS-injected controls, vs 10% in aCeAg group). Histological analysis Karla Feeser , Vitaliano Cama , Elizabeth A. Thiele , Thomson 2 3 1 1 demonstrated twice as many pancreatic islets in aCeAg-treated mice Lakwo , Sindew Mekasha , Jeffrey Priest , Paul T. Cantey (p<0.001) as well as greater numbers of uninfiltrated islets. As observed 1Centers for Disease Control and Prevention, Atlanta, GA, United States, in studies using antigens from parasitic helminths, aCeAg treatments 2Ministry of Health, Vector Control Division, Kampala, Uganda, 3Ethiopian increased the levels of basophils, eosinophils, and polyclonal and Public Health Institute, Addis Ababa, Ethiopia helminth-specific IgE immunoglobulins. Further, we observed increased Onchocerciasis is a neglected tropical disease targeted for elimination production of the suppressive cytokine IL-10 (p<0.05), but not of the pro- in Africa by 2025. Multiplex assays can provide a powerful platform inflammatory cytokine IFN-gamma, from splenocytes of aCeAg-treated for monitoring and evaluation, as well as integrated surveillance of animals. This study demonstrates proof-of-concept that antigens obtained onchocerciasis and other co-endemic diseases; however, the specificity from the non-parasitic nematode C. elegans can be used to obtain the and sensitivity of O. volvulus antigens have not been well-characterized same immune responses, and same immunoprotective effects, as parasitic within this context. A multiplex immunoassay was developed and used helminths. Given that C. elegans can be grown axenically in controlled to evaluate three antigens (Ov16, Ov17 and Ov33) for onchocerciasis. conditions without the need of any intermediate hosts, aCeAg may be The performance of each antigen was characterized using a panel of 499 able to overcome many of the current obstacles facing helmintic therapies specimens. One hundred ten samples were positive for onchocerciasis by for inflammatory diseases. skin snip microscopy and PCR, while the 389 controls were from people living in areas where onchocerciasis was not endemic and had infection with Wuchereria bancrofti, Brugia malayi, Loa loa, Mansonella spp, Strongyloides stercoralis, Hymenolepis nana, cysticercosis, schistosomiasis, or other human pathogenic parasites. All samples were analyzed in duplicate for IgG and IgG4 reactivity. Receiver Operator Characteristics (ROC) analyses were used to determine optimal cutoffs for all antigens. High sensitivity and specificity were detected for Ov16 and Ov33, while the Ov17 assays had specificities below 80%, identifying 75 false positives

astmh.org 523 among controls with lymphatic filariasis (LF). The Ov16 cutoff values for 1714 IgG or IgG4 were 379 and 32 fluorescent units (MFI), with sensitivities of 96.3 and 96.3% and specificities of 98.7 and 99.7%, respectively. HOOKWORM INFECTION IN SCHOOL-AGED KENYAN For Ov33, a cutoff of 5,216 MFI in IgG reactivity resulted in 90.8% CHILDREN IS ASSOCIATED WITH LOWER PHYSICAL FITNESS sensitivity and 97.2% specificity. The IgG4 cutoff was 67 MFI with a 1 2 3 higher sensitivity of 96.3% and specificity of 98.5%.The IgG4 assay for Monica Nayakwadi Singer , Indu Malhotra , Francis Mutuku , 4 4 4 both Ov16 and Ov33 detected few false positives, although the Ov33 Jackson Muinde , Peter Mungai , Dunstan Mukoko , Ginny 1 2 1 assay detected 5 additional false positives among onchocerciasis-negative Gildengorin , Charles King , A. Desiree LaBeaud samples that were positive for either LF (3) or schistosomiasis (2). While 1University of California San Francisco Benioff Children’s Hospital Oakland, no statistical difference was detected between the IgG and IgG4 assays Oakland, CA, United States, 2Case Western Reserve University, Cleveland, for Ov16 and Ov33 (p>0.3), assays with the highest specificity and lowest OH, United States, 3Technical University of Mombasa, Mombasa, Kenya, cutoff values will help to ensure the ability of programs to monitor their 4Division of Vector Borne and Neglected Tropical Diseases, Nairobi, Kenya work towards reaching desired elimination endpoints. Overall, Ov16 and Children living in parasite-endemic resource-limited areas are heavily Ov33 are highly sensitive and specific antigens in the multiplex platform. burdened by infection and the comorbidities associated with these Further analysis of these antigens, either alone or in combination, may be diseases. Reduced physical fitness can result from anemia, undernutrition useful for monitoring and evaluating progress towards the elimination of and chronic parasitic infections including soil transmitted helminths (STH), onchocerciasis. Entamoeba histolytica, Giardia lamblia, malaria and schistosomiasis. Our goal was to determine the prevalence of parasitic infections and their 1713 association with physical fitness as measured by the validated multi- stage 20 meter shuttle run (20mSRT) method. From January to March EVALUATION OF HLA IMMUNOINFORMATICS FOR THE 2014, a cohort of 101children aged 4-7 years in coastal Kenya was IDENTIFICATION OF BRUGIA MALAYI PUTATIVE T CELL evaluated. At the visit, blood, stool, and urine were collected and tested EPITOPES CONSERVED WITH WUCHERERIA BANCROFTI AND for presence of infection as follows: blood smear for malaria, Ritchie stool LOA LOA examination for STH, E. histolytica, and G. lamblia, and urine filtration for S. haematobium. 20mSRT were scored based on level achieved. Melissa Torres1, Nils Pilotte1, Tomabu Adjobimey2, Sabine 2 3 4 5 Descriptive statistics were used to estimate infection rates. Wilcoxon scores Specht , Frances Terry , Alexander Debrah , Linda Batsa , determined the association between each type of infection and 20mSRT Alexander Kwarteng2, Anne De Groot3, Steven Williams1, Achim 2 level achieved. The cohort included 101 children with a mean age of 5.8 Hoerauf years, 53% male. 43% reached 20mSRT level 1, 48% reached level 2, 1Smith College, Northampton, MA, United States, 2Institute of Medical 5% reached level 3 and 4% reached level 4. Age, sex, and hemoglobin Microbiology, Immunology and Parasitology, University Hospital Bonn, level (mean 10g/dL) were not significantly associated with the shuttle run Bonn, Germany, 3EpiVax, Providence, RI, United States, 4Faculty of Allied level achieved. Trichuris was the most prevalent parasitic infection (13%), Health Sciences, Kwame Nkrumah University of Science and Technology, followed by hookworm (7%), E. histolytica (7%), giardia (5%), malaria Kumasi, Ghana, 5Kumasi Centre for Collaborative Research in Tropical (4%), Ascaris (1%), and schistosomiasis (1%). Hookworm infection was Medicine, Kumasi, Ghana associated with a lower level achieved in the 20mSRT (p=0.002, 95% CI The availability of filarial genome sequences and improved 0.014-0.113). Parasitic infections are common in school-aged children in immunoinformatics tools promises to accelerate the identification of coastal Kenya and may impair physical fitness. Hookworm infection, in highly conserved and immunogenic filarial vaccine components. The particular, is associated with decreased physical fitness as measured by the work done in this study has taken advantage of the functional similarities, 20mSRT. rather than genetic diversity, of HLA binding residues in efforts to further identify putative T cell epitopes as potential vaccine antigens to combat 1715 lymphatic filariasis (LF). Predictions were previously made using the iVAX websuite containing both the EpiMatrix and ClustiMER immunoinformatics INFERENCE OF PARASITE BURDEN FROM INDIRECT tools. 20-mer peptide sequences were selected for peptide synthesis from INTENSITY DATA AND IMPLICATIONS FOR STUDY DESIGN proteins within the Brugia malayi secretome. The 20 sequences were James E. Truscott, Arminder Deol, Roy Anderson selected based on predictions to bind up to 8 of the most common HLA alleles represented within the software toolkit: DRB1*0101, DRB1*0301, Imperial College London, London, United Kingdom DRB1*0401, DRB1*0701, DRB1*0801, DRB1*1101, and DRB1*1501. In A substantial proportion of data collected in monitoring and evaluation silico filarial conservation analyses were done using basic local alignment (M&E) of macro-parasites records secondary indicators of the infection, tools within BROAD Institute’s Filarial Worms Database. These analyses such as egg counts or microfilaria, as a measure of parasite burden. identified cross conservation for 6 out of the 20 sequences that shared Arguably, however, the most useful information for diagnosis, estimation sequence identity between B. malayi and Wuchereria bancrofti and/or Loa of morbidity and for understanding the transmission dynamics of loa. Proof-of-principle assays for all 20 putative epitopes were performed disease are quantities such as mean adult parasite burden and levels using rHLA binding assays tested with 4 out of the 8 HLA alleles predicted of aggregation of this burden within the population. Since adult worm by the software: DRB1*0101, DRB1*0401, DRB1*1101, and DRB1*1501. counts are often very difficult to obtain, and we have only a few Results from these competitive binding assays demonstrated allele-specific studies from which to estimate the relationship between adult worms binding biases. The 6 putative sequences sharing conservation with W. and transmission stages, we need robust statistical descriptions of the bancrofti and/or L. loa were tested on PBMCs from patients living in LF distribution of measured quantities, such as egg counts, in terms of basic endemic areas that had been exposed to W. bancrofti. Upon peptide parameters of parasite biology. These will enable reliable inference of stimulation, subset CD4+ and CD8+ T cell populations from patients quantities such as mean worm burden, parasite aggregation and density- infected with W. bancrofti were selected for determination of cytokine- dependent processes from standard M&E data. We have developed a specific responses by ELISpot and flow cytometry. Results demonstrated method for doing this calculation using extensive egg count and worm the predicted peptides derived from the B. malayi secretome were capable count data to identify and parameterize models of the distribution of of inducing T cell responses, which differed dependent on infection and Ascaris lumbricoides egg production as a function of worm burden. disease status. These results suggested that the cross- conserved peptides Results show that egg counts are best described by a negative binomial were capable of binding to HLA from patients exposed to W. bancrofti. distribution with mean egg production from individual subject to an exponentially decreasing fecundity. Models of this kind will allow the optimal use of available data sources to extract reliable estimates of

astmh.org 524 basic biological parameters and their associated confidence intervals. 1717 This has strong implications for the tailoring of study design and choice of diagnostic techniques to optimize information gained against cost EPIDEMIOLOGY OF ANTHELMINTHIC TREATMENT FAILURE incurred. As an example, we discuss the reappraisal of M&E as elimination IN HOOKWORM (NECATOR AMERICANUS) INFECTIONS is approached and egg intensities fall. IN CHILDREN IN THE KINTAMPO NORTH MUNICIPALITY, GHANA. II. BASELINE PARASITOLOGY DATA 1716 Michael D. Wilson1, Josephine Quagraine1, Gertrude Ecklu- EFFECT OF A SINGLE DOSE OF 8 MG MOXIDECTIN OR Mensah1, Joseph Otchere1, Yvonne Ashong1, Millicent Opoku1, 150 µG/KG IVERMECTIN ON INTESTINAL HELMINTHS Lisa Harrison2, Debbie Humphries3, Kwadwo A. Koram1, Michael IN PARTICIPANTS OF A CLINICAL TRIAL CONDUCTED IN Cappello2 NORTHEAST DRC, LIBERIA AND GHANA 1Noguchi Memorial Institute for Medical Research, Legon-Accra, Ghana, 2Yale School of Medicine, New Haven, CT, United States, 3Yale School of 1 2 3 Simon K. Attah , Kambale Kataliko , Maurice M. Nigo , Mawolo Public Health, New Haven, CT, United States Kpawor4, Nicholas O. Opoku1, Eric Kanza2, Didier Bakajika3, Hayford Howard5, Germain Mambandu3, Kambale Kasonia2, Previous studies in multiple communities in the Kintampo North Michel Vaillant6, Christine Halleux7, Piero L. Olliaro7, Annette C. Municipality (KNM), Ghana in 2007 revealed a high prevalence of Kuesel7 hookworm infection (45%) and albendazole (single dose, 400mg) treatment failure (39%) school age children. Subsequent study in 2010 1 2 Onchocerciasis Chemotherapy Research Center, Hohoe, Ghana, Centre confirmed the high prevalence of hookworm (39%) and an even higher de Recherche Clinique de Butembo, Université Catholique du Graben, rate of albendazole treatment failure (56%) in this population. The 3 Butembo, Democratic Republic of the Congo, Centre de Recherche overall goal of an NIH-funded longitudinal study is to fully characterize en Maladies Tropicales de l’Ituri, C.R.M.T- ITURI, Hôpital Général de the epidemiology and molecular basis of benzimidazole treatment failure Référence de Rethy, Rethy, Province Orientale, Democratic Republic of the in KNM. Parasitological, anthropometric, socioeconomic, nutritional and 4 Congo, The Liberian Institute for Biomedical Research, Clinical Research immune status data were obtained from a cohort of consented 273 5 Center Bolahun, Monrovia, Liberia, The Liberian Institute for Biomedical school-aged children (48.1% females and 51.9% males). Fecal samples, Research, Clinical Research Center Bolahun, Lofa County, Liberia, demographic data were also obtained from three consented household 6 7 Monrovia, Liberia, CCMS CRP-Santé, Luxembourg, Luxembourg, World members of participants in two communities. We report here, the baseline Health Organization, Geneva, Switzerland parasitology results. Stool and blood samples were analyzed for intestinal In the Phase 3 study comparing the effects of a single dose of 8 mg helminths (Kato-Katz) and malaria (RDT and microscopy) infections, and moxidectin vs. ivermectin standard dose (150µg/kg) on O. volvulus skin PCR used to identify the hookworm species. All the participants gave fecal microfilariae levels and participant well-being in 941 males and 531 samples and 260 donated blood, and those found infected treated with females, including 79 children 12-17 years, , participants were randomized albendazole. Fifty-eight (21.2%) were infected and 16 of 46 (34.7%) in a 2:1 ratio to moxidectin : ivermectin. 1465 participants underwent failed treatment and the overall cure rate was 61.7%. The geometric a single sample Kato-Katz test at screening. 876/1465 (60%) were mean of intensity infection was 376.1epg (± 890.5) at pre-treatment and infected with ≥1 type of intestinal helminth: Ascaris lumbricoides 44 was 57epg (± 46.3) post treatment. The fecal egg count reduction rate (3%), Trichuris trichuria 22 (1.5%), hookworm 783 (53.4%), Schistosoma was 81.2%. Sixty nine hookworm specimens were all identified as Necator mansoni 233 (15.9%), Strongyloides 4 (0.3%). 835 with ≥1 type of americanus. 76.54% (199/260) and 67.7% (176/260) were positive intestinal helminth were treated with either moxidectin or ivermectin and by RDT and microscopy respectively. Geometric mean of Plasmodium tested again 1 month later with a single sample Kato-Katz test. Results falciparum intensity was 1377.3 (± 3623.9) parasites/ml of blood. 16.6% obtained are expressed as cure rate (CR) and egg reduction rate (ERRam) (45/173) participants were co-infected with both parasites. Hookworm and using arithmetic mean egg counts post treatment relative to arithmetic malaria co-infection rate was 17.3% (45/260). The cross sectional survey mean egg counts pretreatment (EPG) for each species. Results for revealed hookworm prevalence of 33.3% (46/138) among household ivermectin treated subjects: A. lumbricoides n=10, EPG=408, CR=100%, members, and 56.9% (29/51) of households, and the percent positive ERRam=100%; T. trichuria n=6, EPG=2856, CR=83%, ERRam=76%; child with at least one household member positive was 73.3% (11/15). hookworm n=259, EPG=842, CR=29%, ERRam=52%; S. mansoni n=67, EPG=236, CR=54%, ERRam=73%. Results for moxidectin treated subjects: 1718 Ascaris lumbricoides n=34, EPG=386, CR=97%, ERRam=97%; T. trichuria n=11, EPG=1409, CR=91%, ERRam=99%; hookworm n=491, EPG=601, EPIDEMIOLOGY OF HOOKWORM (NECATOR AMERICANUS) CR=48%, ERRam=82%; S. mansoni n=143, EPG=168, CR=64%, INFECTIONS IN CHILDREN AND ANTHELMINTHIC ERRam=66%. Co-administration of either ivermectin or moxidectin with TREATMENT FAILURE IN THE KINTAMPO NORTH drugs like benzimidazoles and/or praziquantel may help achieve high MUNICIPALITY, GHANA. III. EVALUATION OF NUTRITIONAL efficacy in preventive chemotherapy programmes for soil-transmitted RISK FACTORS AT BASELINE nematodes and schistosomiasis. Sena Apeanyo Noguchi Memorial Institute for Medical Research, Legon, Accra, Ghana An ongoing NIH-funded longitudinal study of school-age children aims to fully characterize the epidemiology of hookworm in the Kintampo North Municipality (KNM) where previous studies have found high albendazole treatment (400mg) failures in children. This arm of the study is to investigate the influence of modifiable host factors including overall nutritional status,food security and dietary diversity as risks factors of hookworm infestationat baseline. Structured questionnaires and food frequency questionnaires were used to collect data from 271 study participants. Haemoglobin concentration, weights and height z- scores were used to determine nutritional status of the study participants, WHO criteria was used to determine food security and food diversity. Fifty-eight participants (21.2%) and sixteen out of 46 studied participants (34.8%) were hookworm infected pre- and post-treatment respectively. astmh.org 525 The overall mean Hb levels(anemia defined as <12.0g/dl) was 11.47g/ 79.1%, 77.9%, and 23.6% for Sj, Sh and Sm; for the 10th centile it was dl (±1.28), 11.50g/dl (±1.25)and 11.35g/dl (±1.38) for negative and 100%, 88.2%, and 70.3%, and for the 25th centile 100%, 97.8%, and positive cases respectively, which were not significantly different between 100%, respectively. them. The mean weight z-scorefor the study participants was -0.02 (±0.97), 0.65 (±1.19) and -0.39 (±0.89) for negative and positive cases 1720 respectively, which were significantly between the two groups (P=0.02). The mean height z-score overall, was -0.01 (±1.0), -0.01 (±0.97)and EPIDEMIOLOGY OF HOOKWORM (NECATOR AMERICANUS) -0.12 (±1.07) for negative and positive cases respectively, which were INFECTIONS IN CHILDREN AND ANTHELMINTHIC not significantly between the two groups (P=0.087). 74.2% of the study TREATMENT FAILURE IN THE KINTAMPO NORTH participants experienced food insecurity and so were 72.4% and 74.6% MUNICIPALITY, GHANA. I. BASELINE INDICATORS OF of the negatives and positives cases respectively. Above-average dietary EPIDEMIOLOGY AND SOCIOECONOMIC RISK FACTORS diversity was observed in 46.5% of the study participants, and was 48.8% 1 1 2 and 37.9% for negative and positive cases respectively. In conclusion, Hector L. Samani , Sena Apeanyo , Karen Zhang , Debbie 2 1 3 most of the study participants were anemic, underweight, stunted and Humphries , Kwadwo A. Koram , Michael Cappello , Michael D. 1 consumed less diverse food. Moreover, infected children were significantly Wilson underweight than their non-infected counterparts. 1Noguchi Memorial Institute for Medical Research, Legon-Accra, Ghana, 2Yale School of Public Health, New Haven, CT, United States, 3Yale School 1719 of Medicine, New Haven, CT, United States In 2007 and 2010, high prevalence rates of hookworm infections COMPARING METHODS TO ASSESS SCHISTOSOMA (45%) and (39%) respectively in school age children was observed in the RESPONSE TO PRAZIQUANTEL TREATMENT Kintampo North Municipality (KNM), Ghana. Also observed were high Piero L. Olliaro1, Michel Vaillant2, Aïssatou Diawara3, Jean T. rates of albendazole (single dose 400mg) treatment failure; (39%) and Coulibaly4, Amadou Garba5, Jennifer Keiser5, Charles H. King6, (54%) respectively. As part of an NIH-funded study which aims to fully Stefanie Knopp5, Aly Landouré7, Eliézer K. N’Goran4, Giovanna characterize the epidemiology and molecular basis of benzimidazole Raso5, Alexandra U. Scherrer8, José Carlos Sousa-Figueiredo9, treatment failure in KNM, a census/enumeration survey was conducted Katarina Stete5, Xiao-Nong Zhou10, Jürg Utzinger5 in eight communities in KNM, then 271 children aged between 8 and 12 years were randomly selected for a longitudinal study. To define the 1World Health Organization, Geneva, Switzerland, 2Methodology and epidemiology of hookworm infection at baseline and the specific host Statistical Unit, Centre for Health Studies, CRP-Santé, Luxembourg, factors associated with albendazole treatment failure, demographic, Luxembourg, 3New York University Abu Dhabi, Abu Dhabi, United Arab socioeconomic and environmental information was collected using Emirates, 4Unité de Formation et de Recherche Biosciences, Université Félix structured questionnaires. The cohort comprised of 138 males and 133 Houphouët-Boigny, Abidjan, Côte D’Ivoire, 5Department of Epidemiology females. The cohort’s mean age was 9.49 (± 1.69). The overall hookworm and Public Health, Swiss Tropical and Public Health Institute, Basel, prevalence was 21.4% (58/271), of which 67.2% of infected children Switzerland, 6Center for Global Health and Diseases, Case Western Reserve were the group 10 years and above. Significant associations were found University, Cleveland, OH, United States, 7Institut National de Recherche between hookworm infections and possession of cattle and dogs (P=0.049 en Santé Publique, Bamako, Mali, 8Division of Infectious Diseases and and P=0.029), ownership of shoes (P=0.015) and wearing shoes daily Hospital Epidemiology, University Hospital Zürich, University of Zürich, (P=0.020). No significant associations were found with gender, age, Zürich, Switzerland, 9Department of Infectious and Tropical Diseases, access to agricultural land, and types of water sources and toilet facilities London School of Hygiene & Tropical Medicine, London, United Kingdom, (P>0.05). No associations were also found between treatment response 10National Institute of Parasitic Diseases, Chinese Center for Disease and any of the socio-economic parameters (P>0.05). Control and Prevention, Shanghai, China To diagnose Schistosoma infection, stool or urine samples are examined for 1721 worm eggs, but there is no definitive agreement as to how best express treatment efficacy. We analyzed data from 24 trials conducted in Africa, RICE FORTIFIED WITH IRON AND OTHER MICRONUTRIENTS Asia and Latin America enrolling overall 4740 individuals infected with S. IMPACTS HOOKWORM INFECTION RISK IN mansoni (Sm, n=1804; 38.1%), S. haematobium (Sh, n=2633; 55.5%), or SCHOOLCHILDREN S. japonicum (Sj, n=303; 55.5%), and treated with praziquantel at doses of 40 (n=3713), 60 (n=690) and 80 (n=337) mg/kg. Efficacy was measured Brechje de Gier1, Maiza Campos Ponce1, Marlene Perignon2, using cure rate (CR) and egg reduction rate (ERR) calculated using either Marion Fiorentino2, Khov Kuong3, Chhoun Chamnan3, Megan geometric or arithmetic means (ERRgm, ERRam). We compared efficacy Parker4, Kurt Burja5, Jacques Berger2, Katja Polman6, Frank outcome measures: 1) ERRam vs. ERRgm, 2) ERR vs CR, and 3) ERR and CR Wieringa2 based on quadruplicate vs single Kato-Katz thick smear examination for 1VU University Amsterdam, Amsterdam, Netherlands, 2IRD, Montpellier, Sm. We found that: 1) ERRam and ERRgm can be used interchangeably France, 3DFPTQ, Fisheries Administration, Phnom Penh, Cambodia, 4PATH, only if treatment efficacy is very high (>95%); as efficacy falls, estimates Seattle, WA, United States, 5WFP, Phnom Penh, Cambodia, 6ITG, Antwerp, are higher with ERRgm than ERRam. Modeling data shows that Belgium consistency between means is better for Sh and Sj than for Sm; 2) poor Soil-transmitted helminth (STH) infections often co-exist with micronutrient correlation between ERRgm/am and CR except when ERRs are very high deficiencies in the developing world. There is evidence that these major (>97%). 3) using a single rather than quadruplicate Kato-Katz thick smear health issues can influence and exacerbate each other. Fortification of excluded 19% of Sm-infected individuals; the effect on estimating ERR staple foods such as rice can be an effective tool to improve micronutrient was negligible by individual studies; however, on aggregate ERRam and status of vulnerable groups, but its impact on STH infection is currently CR were 8-9% higher (no effect on ERRgm.) A valid complement for drug unclear.In a cluster randomized, placebo-controlled, double-blinded trial, efficacy monitoring is to study the distribution of individual responses to 3 different types of fortified rice were introduced through the World Food identify suboptimal responders. Of the 2358 Sh-infected individuals with Program (WFP) School Meal program in Cambodia. The 3 types of fortified complete data records 61.3% were negative post-treatment (cure rate, rice differed in micronutrient compositions and production method. CR), 32.4% had reduced egg counts (rEC), 6.3% had no change/increased Children (6-15 y) received 1 type of rice, unfortified rice (placebo), or no egg counts (nEC). For Sm (n=1699) individuals CR was 75.4%, rEC school meal (control) for 7 months. Stool samples were collected and 20.5%, nEC 4.1%. For Sj (n=300) CR was 90%, eEC 8.3%, nEC 1.7%. analyzed by Kato Katz method at baseline, 3 months and 7 months. The response achieved by the 5th centile (the 5% worse responders) was After baseline, all children received a single dose of 400mg albendazole.

astmh.org 526 The effects of consumption of fortified rice on hookworm infection were and low egg mortality at high temperatures, but also a drop in proportion analyzed by multiple logistic regression. Baseline prevalence of STH was of eggs reaching maturity above 30°C. To demonstrate implications for 17.0%, which were mainly hookworm infections (16.6%) of light intensity. the optimal timing of mass treatment campaigns, models were applied A risk factor for hookworm infection was being a boy (P=0.011). After 7 to districts with differing levels of A. lumbricoides transmission across months (n= 1236 children), hookworm infection prevalence was between Kenya. District-level prevalence estimates were generated using predictive 17-24% in control, placebo and in the NutriRice fortified rice groups. risk maps developed by the Global Atlas of Helminth Infection and In the children receiving 2 types of UltraRice fortified rice, hookworm monthly temperature patterns were derived from MODIS. Results suggest prevalence was 33-34% (P<0.001). The new infection rate was highest in timing of treatment could have important consequences for programme the UltraRice group with the highest iron content (24.6%), intermediate in impact. Depending on region, changing the treatment date resulted in an the UltraRice group with less iron (21.8%), and lowest in the control and estimated 18-55% comparative decrease in prevalence after four yearly placebo and groups (12.5% and 11.9%, P=0.001 for difference among treatment rounds. This highlights the potential importance of appropriate groups). Fortifying rice with micronutrients, especially iron, can increase timing of the established Kenya National Deworming Programme. risk of hookworm infection. Type of fortificant appears to be a major More generally, this approach provides insight into the epidemiology effect modifier. These findings have big implications for policies aiming to of A. lumbricoides infection, methods for testing and validating these improve child health and nutritional status in tropical regions. predictions, and can help guide optimal long-term helminth control strategies in diverse settings. 1722 1724 HOW DOES THE SCALE OF DEWORMING PROGRAMS AFFECT THEIR COST-EFFECTIVENESS? KEY RESIDUES OF CRY5B STRUCTURE AND FUNCTION: MUTAGENESIS BY ALANINE SCANNING Hugo C. Turner, James M. Truscott, Roy M. Anderson Imperial College London, London, United Kingdom Jillian Sesar, Yan Hu, Hui Fan, Partho Ghosh, Raffi Aroian The predominant control strategy for the soil-transmitted helminths (STHs) University of California San Diego, La Jolla, CA, United States is regular periodic mass drug administration (MDA), targeting school-aged Soil-transmitted helminthes infect more than 2 billion people worldwide children. The World Health Organization (WHO) and London declaration and only one drug (albendazole) is able to show a high enough efficacy on Neglected Tropical Diseases (NTDs) has set goals to scale up MDA, against parasite worms under conditions for mass drug administration. so that by 2020, 75% of the pre-school and school-aged children in However, recent studies have shown an increase in resistance to this drug, need, will be treated regularly. It has been observed that increasing the stressing the importance of finding a new treatment option. Crystal (Cry) number treated can reduce the per capita costs of MDA programmes proteins produced from the soil bacterium Bacillus thuringiensis have been (economies of scale). This is because a number of the costs associated with used for decades as a means to control insects that destroy crops and MDA delivery are fixed (i.e. do not depend on the number treated), and transmit human diseases, and studies have shown these proteins to be therefore increasing the number treated reduces the average fixed cost safe to humans. Our lab has shown that crystal proteins, specifically Cry5B, per treatment. However, the implications this has on the cost-effectiveness are able to kill both the free-living nematode Caenorhabditis elegans, as of scaling up control for STH infections, and the optimum treatment well as parasitic roundworms (eg. Ancylostoma ceylanicum, hookworm). strategy have not been explored. We developed costing functions which Cry proteins intoxicate invertebrates by acting as pore-forming toxins. account for the changes in the per capita costs of treatment with scale, Several defined steps in their mechanism of action have been suggested and incorporated them into STH dynamic transmission models. We found from insect studies, but there is still great uncertainty as to the importance that, due to these economies of scale, the cost effectiveness of STH of these various steps. We believe that the nematode - Cry5B system has control programmes markedly increased with the number treated. This has great potential to unlock mysteries surrounding Cry proteins and to be a notable implications for programmes considering scaling up MDA, in line potential therapeutic agent. Here, I have mutated all of the 698 amino with the current goals set by the London declaration. acids in the toxin domain of Cry5B, and subsequently tested these mutants on C. elegans to assess for changes in toxicity levels, screening for variants 1723 with an increase in activity as compared to the wild type. From this screen and subsequent quantitative LC50 killing assays to confirm the screen THE EFFECT OF SEASONALITY ON THE PREVALENCE OF results, I have identified several key variants of interest that are additionally ASCARIS LUMBRICOIDES AND IMPLICATIONS FOR THE more active against A. ceylanicum both in vitro and in vivo. Additionally, OPTIMAL TIMING OF MASS TREATMENT PROGRAMS these residues most likely play a key role in Cry5B protein function, with the eventual goal being to correlate these changes in activity with specific 1 2 3 Emma L. Davis , Rachel Pullan , Leon Danon , Deirdre changes in protein functionality. These improved Cry protein variant 1 Hollingsworth candidates have the potential to be used in therapeutics for treating one 1University of Warwick, Coventry, United Kingdom, 2London School of of the most neglected diseases of our time, parasitic worms. Hygiene & Tropical Medicine, London, United Kingdom, 3Queen Mary University of London, London, United Kingdom 1725 Previous studies have shown that changes in temperature affect maturation times, development proportions, and mortality rates of Ascaris A NOVEL NEXT-GENERATION SEQUENCING APPROACH TO lumbricoides and suum eggs, which suggests that climate could impact DEVELOP IMPROVED MOLECULAR DIAGNOSTICS FOR THE both overall prevalence and reinfection rates after treatment. Depending DETECTION OF SOIL TRANSMITTED HELMINTH INFECTIONS on the size of this effect, this could have implications for the optimal time Nils Pilotte, Marina Papaiakovou, Lou Ann Bierwert, Steven A. of year for mass treatment in order to get the maximum impact out of the Williams large number of donated drugs. However, there has been little quantitative investigation of the influence of seasonal timing of mass chemotherapy on Smith College, Northampton, MA, United States treatment programmes. Using historical data from experimental studies on The soil transmitted helminths (STHs) are a group of parasitic worms A. suum eggs – which have been shown to display equivalent behaviour to responsible for causing extensive morbidity in many of the world’s A. lumbricoides eggs – to characterise relationships between temperature most economically depressed locations. With an estimated 880 million and egg maturation and survival, models were developed which children infected with one or more species of STH parasite, accurate investigated the effect of temperature dependent development of eggs on and cost-effective diagnostic measures are of premier importance to mean worm burden in the human population. These reveal fast maturation global control and elimination efforts. Accordingly, the use of molecular

astmh.org 527 diagnostic measures, such as real-time PCR, has shown great promise for 1727 the improved detection of STH infections. To date, such molecular assays have utilized ribosomal and mitochondrial target sequences, as previously HELMINTH INFECTIONS DURING PREGNANCY MAY reported. While effective, such sequences are frequently not the highest DECREASE NUTRITIONAL FITNESS OF THE OFFSPRING copy number target and will not yield the most sensitive assay possible. 1 1 2 The most sensitive assay will utilize the most highly repetitive, unique, Emily A. McDonald , Sunthorn Pond-Tor , Blanca Jarilla , 2 2 2 non-coding DNA sequences found within the genome of each species. Marianne J. Sagliba , Analisa Gonzal , Remigio Olveda , Luz 2 1 1 Consequently, we have coupled next-generation sequencing technology Acosta , Jonathan D. Kurtis , Jennifer F. Friedman with the Galaxy-based software RepeatExplorer, to identify the most 1Rhode Island Hospital, Providence, RI, United States, 2Research Institute of numerous, non-coding DNA sequences within multiple species of STH Tropical Medicine, Manila, Philippines parasites including Trichuris trichiura, Necator americanus, Ancylostoma Helminth infections represent a significant disease burden in endemic duodenale, and Ascaris lumbricoides. Following the application of this regions of the world, and polyparasitism may have a bigger impact on approach to each STH species, we designed TaqMan-based primer-probe overall health than any individual infection. We have previously shown that combinations for each candidate sequence. Species-specificity was verified Schistosoma japonicum results in a profound pro-inflammatory response for each assay and repeatable detection of genomic DNA isolated from at the maternal-fetal interface during pregnancy and decreased invasion each parasite was demonstrated at concentrations ranging from 1.0ng to characteristics of placental trophoblast cells in vitro. Another critical 1.0fg. Through this novel approach to the identification of species-specific, function of the placenta is to regulate nutrient exchange between mother high copy-number target sequences, we have developed a new strategy and fetus. Herein, we have shown that treatment in vitro of primary for the design of a PCR-based diagnostic assay with improved sensitivity. trophoblasts with schistosome soluble egg antigens (SEA), resulted in a significant drop in gene expression of specific amino acid transporters. 1726 These include the sodium-coupled neutral amino acid transporter 1 (SNAT1; 80% reduction) and large neutral acid transporter (LAT1; 70% THE ROAD TOWARDS EFFICIENT CONTROL OF reduction). To investigate the metabolic impact of helminth infections SCHISTOSOMIASIS IN THE DEMOCRATIC REPUBLIC OF during pregnancy, we utilized samples from a cohort of pregnant women CONGO: PRE-ASSESSMENT OF STAFF PERFORMANCE AND from Leyte, the Philippines. Most subjects had polyparasitic infections, MATERIAL RESOURCES IN ENDEMIC REGIONS including schistosomiasis and geohelminth infections, with prevalence rates of 70%, 79%, and 40% for Ascaris lumbricoides, Trichuris trichiura, Sylvie Linsuke1, Liliane Mpabanzi2, Joules Madinga3, Sabin 1 4 1 2 and hookworm, respectively. Given the relatively low intensity of Nundu , Faustin Mukunda , Pascal Lutumba , Katja Polman schistosome infection and the high prevalence rates of geohelminths, we 1Institut National de Recherche Biomédicale, Kinshasa, Democratic assessed the relationship between the number of helminth infections and Republic of the Congo, 2Institute of Tropical Medicine, Antwerp, Belgium, metabolic parameters in utero. After controlling for SES and gestational 3University of Kinshasa, Kinshasa, Democratic Republic of the Congo, age, leptin levels were found to be lower in the cord blood of infants 4Programme National de Lutte contre la Bilharziose et Parasitoses born to mothers with one or more helminth infections. In addition, cord Intestinales, Kinshasa, Democratic Republic of the Congo blood leptin levels were positively associated with birth weight (107g Schistosomiasis is a disease affecting approximately 200 million people heavier on average in those infants in the highest tertile of leptin levels), worldwide. Since a long time, schistosomiasis has been known to be and increased leptin levels were associated with a reduced risk of fetal endemic in certain provinces of the Democratic Republic of Congo. growth restriction. These data suggest that helminth infections can impact However, the most recent national data available on schistosomiasis the transport of nutrients across the maternal-fetal interface, providing a prevalence and distribution were published in the early sixties. . Recently, possible link between fetal metabolic hormones and growth in utero. the Ministry of Health adopted a national plan against schistosomiasis aiming at to distribute Praziquantel (PZQ), the treatment of choice for 1728 schistosomiasis, to all individuals infected. For effective introduction of control strategies, data on national prevalence and distribution of GUT INSTINCTS: EVALUATING PARENTAL ATTITUDES schistosomiasis in the DRC urgently need to be updated. The present TOWARD INTESTINAL WORM TREATMENT IN RURAL CHINA study assessed the knowledge of health workers on schistosomiasis as Louise Lu1, Chengfang Liu2, Alexis Medina1, Darvin Scott Smith1, well as the availability of the facilities needed for adequate diagnosis Scott Rozelle1 and management of the disease in the endemic provinces of Kinshasa 1 2 and Bas-Congo in the DRC. This study was conducted in 9 health zones Stanford University, Stanford, CA, United States, Center for Chinese (HZ) of Kinshasa and 2 HZ in Bas-Congo. Health workers could name all Agricultural Policy, Beijing, China symptoms of schistosomiasis. Kato-Katz, urine filtration or sedimentation Nearly forty percent of elementary schoolchildren in rural provinces of were not available as diagnostic methods in any health facilities. Southwest China are infected with soil-transmitted helminths. However, Diagnosis therefore almost solely relied anamnesis. The knowledge on parasitic worm infection is neglected as a major public health problem schistosomiasis did not differ between the rural Bas-Congo and urban in these villages and deworming treatment is rarely sought, despite its Kinshasa. The fees for consultation, diagnostics and treatment were three efficacy and low cost (one treatment dosage costs less than 3 cents USD). times higher in Kinshasa than Bas-Congo. Health workers in Kinshasa and Surveys and interviews were conducted in six rural villages in Guizhou, Bas-Congo are able to name the symptoms related to schistosomiasis. China to evaluate what factors influence parental decisions to seek or not However there is a lack of availability of adequate diagnostic tools and seek deworming among rural Chinese schoolchildren. It was found that treatment. The fees of diagnostics and treatment are high for a population knowledge about helminth infection and prevention was severely lacking often living in extreme poverty. and often influenced by deep-rooted myths, such as the local belief that deworming medicine can harm a child’s future fertility. The majority of household interviewees were highly skeptical of high worm prevalence in their children, despite the nearly universal practice of regularly deworming their pigs. A comprehensive deworming program involving biannual administration of deworming treatment, household health education, and village health system strengthening is necessary to effectively mitigate the disease burden of helminth infection in rural China.

astmh.org 528 1729 triatomine vectors combined with high incidence of feeding on humans underscore the importance of Chagas disease surveillance in Texas and RANDOMIZED CONTROLLED TRIAL OF TWO IVERMECTIN prompt for urgent measures for vaccine development, vector control, and REGIMENS FOR STRONGYLOIDES STERCORALIS: EARLY increasing public awareness. FINDINGS 1731 Guadalupe Jimenez1, David Goodman-Meza1, Herbert B. Tanowitz2, Christina M. Coyle2 POPULATION GENETIC STRUCTURE OF THE TSETSE FLY: 1Jacobi Medical Center, Bronx, NY, United States, 2Albert Einstein College TARGETING REPRODUCTIVE REFRACTORY INTERVENTIONS of Medicine, Bronx, NY, United States FOR WILDLIFE AND LIVESTOCK TRYPANOSOMIASIS IN Humans can be chronically infected with Strongyloides stercoralis for KENYA decades because of the mechanism of autoinfection. Although most Benard Kulohoma infected individuals are asymptomatic, S. stercoralis is capable of transforming into a fatal illness in setting of HTLV-1 and steroids. Chronic International Centre for Insect Physiology and Ecology, Nairobi, Kenya infection with S. stercoralis presents both diagnostic and therapeutic Tsetse flies (Glossina species) are major vectors of both human and challenges. Parasitologic diagnosis of chronic infection is difficult, livestock trypanosomiasis, and efficiently transmit the causative parasites because the larval output can be low and irregular. The diagnostic trypanosomes. Tsetse flies infest up to 10 million km2 of land stretching accuracy of serologic testing has a sensitivity and specificity of 93% across 40 countries in sub-Saharan Africa. Multiple drugs that treat African and 95%, respectively. Ivermectin is the treatment of choice for this animal trypanosomiasis exist, and have substantially improved veterinary parasitic infection, although optimal dosing is yet to be determined. This management of livestock with both susceptible and resistant trypanosome study evaluated the serologic response to S. stercoralis infection after strains. Although, multi-drug resistant trypanosome strains have lower treatment with ivermectin 200 mcg/kg given 2 weeks apart based on the fitness and are therefore thought to be less persistent, the effect of auto-infective cycle (Group A) vs. given on two consecutive days (Group increased communal use of multiple antibiotics on transmission rates of B). Patients were referred from outpatient clinics or identified on the in- these pathogenic species is still not fully clear. Moreover, widespread multi- patient services and invited to participate in the study. Participants were drug resistance due to prolonged usage or under-dosing could also have randomized to either treatment arm and repeat serologies were performed adverse repercussions on public health, further complicating management at 3 months intervals for 9 months after treatment. Forty-seven cases were disease management. Our study, will exploit genomic approaches to enrolled, mean age was 54.1 (SD 15.2), 61.7% male, 55.3% Hispanic, understand the population structure of tsetse flies in circulation in disease mean eosinophil count 0.55/nl (SD 0.53), IgE 608.0 mg/dl (SD 680.4) and endemic regions of Kenya so as to determine the impact of drug use on HTLV-1 was negative in all cases. There were no significant differences in the prevalence of multi-drug resistance. It’s effect on tsetse infectivity baseline demographic or clinical variables between the two groups. Of and transmissibility of multi-drug resistant trypanosome strains. This the 47, 51.0% had completed the 9 months follow-up. Mean eosinophil study also aims to identify genes essential for successful reproduction as count (p=0.002) and IgE value (p=0.045) both decreased after treatment potential long-term vector control targets. This study holds the promise of of cases in both treatment groups. Of the 47 patients, 9 cases remained identifying socio-demographic independent vector control strategies, and sero-positive on follow-up; five (22.7%) in Group A and four (16.0%) will enable the judicious use of appropriate drugs to which trypanosome in Group B (p=0.751). In this randomized controlled trial, there was no strains are not resistant. Thereby enabling prolonged trypanosome and difference in serologic outcome in the two treatment arms, but treatment tsetse control while avoiding widespread multiple-drug resistance. resulted in decreased eosinophil and IgE values. 1732 1730 EMERGENCE OF A TROPICAL DISEASE IN U.S. DOGS: A TRYPANOSOMA CRUZI INFECTION PREVALENCE AND BLOOD PROSPECTIVE STUDY OF LEISHMANIA IN U.S. FOXHOUNDS MEAL ANALYSIS IN VECTORS OF CHAGAS DISEASE IN Angela J. Schneider, Robert Schaut, Christine Petersen SOUTHWEST TEXAS, 2013-2014 University of Iowa, Iowa City, IA, United States 1 2 3 Rodion Gorchakov , Edward J. Wozniak , Teresa P. Feria , Melissa Leishmania is the causative agent of leishmaniasis, a deadly protozoan 1 1 1 N. Garcia , Peter J. Hotez , Kristy O. Murray disease which affects approximately 12 million people worldwide. 1Baylor College of Medicine, Houston, TX, United States, 2Texas Generally associated with tropical and subtropical regions, Leishmania Department of State Health Services, Uvalde, TX, United States, 3University can be found as far north as the United States. Similarly, the vector for of Texas Pan American, Edinburg, TX, United States Leishmania transmission, the sand-fly, has been found within areas as Protozoan pathogen Trypanosoma cruzi is an etiologic agent of Chagas far north as Missouri and Ohio. Leishmania infantum, infects canines as disease, which affects millions of people in Latin America and is an well as humans. In endemic areas dogs serve as the domestic reservoir. emerging public health threat in the United States. Transmission cycle of Within the United States, foxhound hunt populations have developed the parasite involves alternating infection of its vertebrate hosts and insect endemic disease. The first documented foxhound case of Leishmania in vectors. We identified the current vector infection burden and potential the US was reported in 1980. Leishmaniasis is a chronic disease with long range of natural reservoirs of T. cruzi in 11 southwestern counties of Texas latency, diagnosis can be difficult with gold standards limited to invasive by analysis of insects of genus Triatoma collected during the period from techniques including bone marrow aspirate with culture. Serological June 2013 to January 2014. Out of 40 submitted specimens, the vast testing has been used as a diagnostic technique but has cross reactivity majority of the insects were T. gerstaeckeri, with only four samples of T. with Trypanosoma cruzi also found in the southern U.S. To help with sanguisuga, two samples of T. lecticularia, and one sample of T. rubida. diagnosis, a highly sensitive and specific real time quantitative polymerase We found 73% of the insects positive for T. cruzi. Blood meal analysis was chain reaction (RT-qPCR) assay was developed. While this assay is capable performed on the infected triatomines. Blood sources were determined for of identifying parasite DNA within the peripheral blood it lacks the ability all but one of the insects, and included 13 different species of mammals to determine whether the Leishmania is actively proliferating. Over the (mouse, woodrat, squirrel, porcupine, armadillo, cottontail, raccoon, course of the last 8 years, diagnostic testing using PCR increased. Cases fox, coyote, dog, pig, cow, human). Interestingly, 36% of the bugs were of Leishmaniasis can be tracked throughout regions of the US within the identified as having multiple blood sources. Since most of the insects foxhound hunt populations. This study is the first to report changes in were collected in or around residential houses, the most prevalent type of Leishmania prevalence and incidence over a six-year span (2007-2012) blood meal was human (50% of the insects). High infection rate of the within US foxhound hunts. Trends in infection over time and across regions

astmh.org 529 were examined. Leishmania infection has stayed consistent over time with distribution as CL patients in the affected region (CE p = 0.26), but had a point prevalence in 2007 of 3.54 per 1000 foxhounds and 3.23 per a greater proportion of lesions above the waist line (94% in ACL x 33% 1000 foxhounds in 2012. Incidence rates over the 5-year period began at in CL, p = 0.0001) and of failure to antimony treatment (41% in ACL x 4.77 per 1000 foxhounds in 2007 and ended at 3.08 per 1000 foxhounds 0% in CL, p = 0.0006) than CL individuals. Immunologically, ACL showed in 2012. The consistent prevalence and incidence of this infection stresses lower production of TNFα (average 316.5 pg/ml in ACL x 1906.1 pg/ the need for appropriate risk management and disease prevention ml in CL, p=0.0001) and IFNγ (average 747.1 pg/ml in ACL x 4445.9 pg/ techniques in this community. ml in CL, p=0.0002), but higher IL-10 (average 392.8 pg/ml in ACL x 171.9 pg/ml in CL, p=0.0006) and IL-17 (average 218.4 pg/ml in ACL x 69.4 pg/ml in CL, p=0.0008) after in vitro stimulation of peripheral blood 1733 mononuclear cells with leishmania antigen than CL patients. All subjects MATERNAL TRYPANOSOMA CRUZI INFECTION AND INFANT were infected with L. (V.) braziliensis, but parasites from ACL presented GROWTH genotypes that were not found in isolates from CL individuals. Therefore, in the region studied, patients with ACL consist in a more homogeneous Elizabeth J. Howard1, Xu Xiong1, Maria Luisa Cafferata2, group of individuals than originally suspected, and are distinct from Gabriela Cormick3, Alvaro Ciganda2, Pierre Buekens1 classical CL regarding treatment outcome, immune response and causative 1Tulane University School of Public Health, New Orleans, LA, United strain of L. (V.) brazilensis. States, 2La Unidad de Investigación Clínica y Epidemiológica Montevideo, Montevideo, Uruguay, 3Institute for Clinical Effectiveness and Health Policy, 1735 Buenos Aires, Argentina MODELING ECO-BIO-SOCIAL DETERMINANTS FOR Maternal Trypanosoma cruzi infection and subsequent congenital HOUSEHOLD INVASION OF SYLVATIC TRIATOMA DIMIDIATA transmission is a serious, yet neglected, global health issue. Infant growth IN NORTHERN BELIZE assessment can provide an understanding of the health of children and an indirect evaluation of the quality of life for an entire population Angela T. Caranci1, John P. Grieco2, Nicole L. Achee3 afflicted with Chagas disease. The objective of this study was to determine 1Uniformed Services University, Kensington, MD, United States, if maternal T. cruzi infection indirectly affects a newborn’s physical 2Uniformed Services University, Bethesda, MD, United States, 3University of development. Infected mothers and their infants (n=153) were followed Notre Dame, Notre Dame, IN, United States at birth, 4-8 weeks and 10 months post-partum in Tucuman, Argentina from April 2011 until April 2013. Age- and sex-specific estimates of Initial reports have confirmed presence of Triatoma dimidiata, an important infant weight, length, weight-for-length, and head circumference were Chagas disease vector throughout northern Belize. To date, T. dimidiata compared to an international child growth standard. All mean z-scores remains the sole vector species reported from this Central American were between ±1 standard deviations (SD) of the standard. However, the country, yet much of the disease transmission dynamics remain unclear. prevalence of infants falling below -2 SD of the WHO standard peaked at Here, we report updated infection rates of the vector population as well as 16% for weight-for-age for females, 10.9% for length-for-age in males infestation rates for villages in north and central Belize. In order to further and 10.8% for weight-for-length in females, all at visit 1. Infants who characterize the epidemiological risk of human-vector contact, Over 225 experienced growth faltering were more likely to be female and weighed households have been surveyed and characterized with respect to 30 0.8 kg less at birth, 0.9 kg less at 4-8 weeks and 0.5 kg less at 10 months key determinants related to the probability of household infestation by of age. They also were 3 cm shorter and had a 2 cm reduction in head T. dimidiata. These key variables included: presence of domestic animals, circumference. This analysis provides evidence of progressive stunting distance of household to village periphery, and proximity of community over the 10 month period and early failure to thrive with improvement by light sources. The infestation behavior of T. dimidiata in Belize is confirmed 10 months. Nutrition and health interventions, as well as socioeconomic to be distinct from what would classically be designated a domiciliated changes, may be helpful in improving the growth and development of vector population. Risk factors reported here can be used to guide infants from Chagas affected populations. integrated control efforts to reduce infestation and limit human-vector contact. 1734 1736 COMPARISON BETWEEN PATIENTS WITH CLASSICAL AND CHAGAS DISEASE IN MEXICO: SURVEILLANCE AND ATYPICAL PRESENTATIONS OF CUTANEOUS LEISHMANIASIS, PERCEPTIONS OF BURDEN FROM AN AREA OF LEISHMANIA (VIANIA) BRAZILIENSIS TRANSMISSION IN NORTHEAST BRAZIL Ellen Shelly1, Rodolfo Acuna-Soto2, Charles Sterling1, Kacey Ernst1, Heidi E. Brown1 Luiz Henrique Guimarães, Adriano Queiroz, Juliana A. Silva, 1 2 Silvana C. da Silva, Ednaldo L. Lago, Paulo Roberto L. Machado, University of Arizona, Tucson, AZ, United States, National Autonomous Edgar M. Carvalho, Albert Schriefer University of Mexico, Mexico City, Mexico Hospital Universitário Prof. Edgard Santos, Salvador, Brazil Chagas disease, a parasitic disease caused by Trypanosoma cruzi, disproportionately affects the poor throughout Latin America. We describe The purpose of this study was to compare clinical, immunological the spatial and temporal distribution of officially reported Chagas disease and parasitological aspects between patients with atypical cutaneous incidence and mortality in Mexico. The greatest burden appears to occur in leishmaniasis (ACL) and classical cutaneous leishmaniasis (CL) of a region southern states. Incidence rates and deaths were highest in adults (25-44 with high endemicity for American tegumentary leishmaniasis (ATL) years and ≥45 years, respectively). We show increasing temporal trends for in northeast Brazil. Fifty one ACL and an equal number of CL patients incidence (AR(2) p=0.002, 95% CI: 0.040-0.061) and mortality (MA(1) p < were enrolled. ATL was confirmed and parasite species was determined 0.0001, 95% CI: 0.012-0.021). While these results provide insight to the by PCR of lesion biopsies. For each patient clinical data was annotated, changing burden of Chagas in Mexico, under-reporting likely compromises peripheral blood was collected, and parasite isolation was attempted. our capacity to understand the epidemiology of this disease. The reported Cultured parasites were genotyped according to the sequence of a locus 500 new cases and 20 deaths in 2010 are in stark contrast to estimates in chromosome 28, previously shown to be polymorphic in this population of 69,000 new cases and 25,000 deaths per year from seroprevalence of L. (V.) braziliensis. All cases had their living sites geographic coordinates studies. As changes in Chagas surveillance improve our understanding of acquired by GPS then distributions of ACL and CL cases were compared the full burden of this disease, it is likely that the reported and estimated by the Cusick and Edward’s test (CE). Among ACL included there was no pregnant women or HIV positive subjects. ACL presented the same astmh.org 530 incidence will align more closely. This will facilitate understanding the with Giemsa and examined by microscopy. Samples were collected from epidemiology of this disease and result in more focused and successful 318 NHPs on filter paper, FTA cards or EDTA tubes and tested with a control and prevention strategies. nested PCR protocol using primers for the 24S alpha subunit rRNA gene. Primers D75/D76 target the conserved flanking sequences of the D7 alpha 1737 domain in trypanosomatids, while primers D71/D72 target a region in the same domain that is specific to T. cruzi. PCR was used as gold standard TRYPANOSOMA CRUZI AND OTHER TRYPANOSOMATIDS to calculate the sensitivity and specificity of microscopy. Trypanosomatid IN COMMONLY HUNTED WILD MAMMALS FROM REMOTE and T. cruzi prevalences were compared using Chi2 and Fisher’s exact LOCATION OF THE PERUVIAN AMAZON tests. We studied captive NHPs from five families (14 species) and wild NHPs from three families (11 species). Wild NHPs had significantly higher 1 2 3 1 E. Angelo Morales , Pedro Mayor , Mark Bowler , Esar Aysanoa , prevalence of both trypanosomatids (56% vs 27%, p<0.001) and T. cruzi 1 4 1 Julio A. Ventocilla , Jocelyn Perez , G. Christian Baldeviano , (9% vs 3%, p=0.034), compared to captive NHPs. Pitheciidae had the 1 Andres G. Lescano highest trypanosomatid prevalence (18/20, 90%) and Cebidae had the 1U.S. Naval Medical Research Unit - 6, Lima, Peru, 2Autonomous University highest T. cruzi prevalence (14/116, 12%). Captive NHPs from wet markets of Barcelona, Barcelona, Spain, 3San Diego Zoo Global, Iquitos, Peru, (n=38) had very high trypanosomatid (53%) and T. cruzi (13%) prevalence. 4University of Prince Edward Island, Charlottetown, PE, Canada Compared to PCR, microscopy was 83% sensitive and 98% specific. T. Chagas disease is generally transmitted by contact with the feces of cruzi and trypanosomatids are common in Peruvian NHPs and pose a risk triatomine bugs infected with Trypanosoma cruzi, but oral transmission to human and animal health that has not been properly studied. Although has also been documented. We evaluated the prevalence of T. cruzi and microscopy is poorly sensitive compared to PCR, it may still be useful for other trypanosomatids in four orders of mammals hunted for subsistence screening in the field. to better understand the risk of infection associated with human consumption and contact. Blood samples from wild mammal species were 1739 collected on filter papers by subsistence hunters from a remote small community in Loreto, Peru, bordering Brazil. DNA was isolated from filter CHAGAS DISEASE, POVERTY AND BIODIVERSITY IN MEXICO papers and amplified using a nested-PCR targeting the 24S alpha subunit Alejandro Cruz-Reyes, Blanca Camargo-Camargo rRNA gene. Primers D75/D76 and D71/D72 were used to amplify specific regions of trypanosomatids and T. cruzi, respectively. Comparisons of Instituto de Biología, Universidad Nacional Autonoma De Mexico, Mexico prevalence between orders were performed using Chi-Square and Fisher’s City, Mexico exact tests. A total of 142 mammalian blood samples from four orders Since 1928 Chagas disease (ChD) has been studied in Mexico. Official (10 species) were tested: Carnivora (n=34), Edentata (n=24), Artiodactyla data from the National Council for the Evaluation of the Politics for Social (n=28) and Rodentia (n=56). The prevalence of T. cruzi in Carnivora (18%) Development (Consejo Nacional de Evaluación de la Política de Desarrollo was significantly higher (p=0.008) compared to other orders (0% - 4%). Social) [CONEVAL] in 2012 shows that poverty is 50% in Mexican The prevalence of trypanosomatids ranged from 7% in Artiodactyla population out of the 110 M people (2010 national census: National to 27% in Rodentia with no significant differences (p=0.180), possibly Institute of Statistic and Geographical Information) [INEGI], from this, 15% due to the small sample size. Nasua nasua (ring-tailed coati), Dasypus is under extreme poverty. In the Southern region, Mexico has high level novemcinctus (nine-banded armadillo), Agouti paca (spotted paca) and of poverty and the highest biodiversity richness in the country, which is Tayassu tajacu (collared peccary) accounted for 89% of the samples and currently under serious ecological threating. There is evidence that reduced all positive animals. Among these four species the prevalence of T. cruzi biodiversity affects the transmission of infectious diseases in humans and was 19%, 4%, 2% and 0%, respectively (p=0.016); and the prevalence of other animals. This situation matches perfectly in maps with the high trypanosomatids ranged from 9% to 31%. The high prevalence of T. cruzi prevalence of ChD in that region. The importance of this work is related in Nasua nasua, a type of raccoon, suggests the importance of carnivores to the role of multinational control initiatives against ChD. In some way in sylvatic T. cruzi transmission. The multiple hunted species infected with ChD is a neglected disease in Mexico. This situation is also relevant giving trypanosomatids highlights the risk of human infection by consumption of the immigrant phenomenon between Mexico and the US. In this work improperly cooked meat. we present data and geographical evidence that even the great number of Mexican scientists working on ChD, this zoonotic parasitic disease is 1738 still under estimation. Back in 2006 we published information about ChD in Mexico form our data base “CHAGMEX”, now we are working on a PREVALENCE OF TRYPANOSOMATIDS AND TRYPANOSOMA new data base (2004-2014), and so far, the new bibliographic information CRUZI IN WILD AND CAPTIVE NON-HUMAN PRIMATES FROM shows that the number of human cases is increasing considering all clinical PERÚ and epidemiological forms of the disease: vectorial transmission, blood transfusion, congenital transmission. From the 32 species of Triatominae 1 2 3 Esar Aysanoa , Pedro Mayor , A. Patricia Mendoza , E. Angelo identified in Mexico, more than 10 are reported with domestic habits. Also 1 3 4 5 Morales , Carlos M. Zariquiey , Jocelyn G. Perez , Mark Bowler , Trypanosoma cruzi is becoming quite common in domestic dogs, from 1 1 1 Julio A. Ventocilla , G. Christian Baldeviano , Andres G. Lescano urban and rural areas. We think that besides academic research, is urgent 1U.S. Naval Medical Research Unit - 6, Lima, Peru, 2Autonomous University to implement vector control programs by each climatic region of Mexico, of Barcelona, Barcelona, Spain, 3Wildlife Conservation Society, Lima, Peru, along with a wide epidemiological and socio-economical approaches of 4University of Prince Edward Island, Charlottetown, PE, Canada, 5San ChD in Mexico. Diego Global Zoo, Iquitos, Peru Non-human primates (NHPs) can be infected with Trypanosoma cruzi, the etiological agent of Chagas disease, and other trypanosomatids. Primates may act as reservoirs and close contact between humans (traders, owners, hunters, zookeepers, etc.) and NHPs is a potential risk of accidental infection. We compared the prevalence of T. cruzi and other trypanosomatids in wild and captive Peruvian NHPs to assess this risk. Blood samples were obtained from captive NHPs (n=192) at zoos, wildlife rescue centers, wet markets and households in six Peruvian cities, and wild NHP hunted for subsistence (n=126) in two remote communities in the Peruvian Amazon. Blood smears from 88 captive NHPs were stained

astmh.org 531 1740 the need for continuation and even increase of these efforts, especially when recognizing the sizeable health impact that can be gained when MAPPING THE PREVALENCE AND CASE DETECTION RATES achieving the 2020 targets. OF GAMBIENSE HUMAN AFRICAN TRYPANOSOMIASIS 1742 Nick Golding1, Simon I. Hay1, Giuliano Cecchi2, Massimo Paone3, José R. Franco4, Pere P. Simarro4 ANTI-LEISHMANIA ANTIBODIES IN BLOOD DONORS FROM 1University of Oxford, Oxford, United Kingdom, 2Food and Agriculture BRAZIL USING RECOMBINANT L. INFANTUM PROTEIN K39 Organization of the United Nations, Addis Ababa, Ethiopia, 3Food and ELISA Agriculture Organization of the United Nations, Rome, Italy, 4World Health 1 1 Organization, Geneva, Switzerland Maria Carmen A. Sanchez , Marina P. Dornelas , Silvano Wendel2, Silvia M. Di Santi1, Adeiana G. Lima1, Beatriz J. Celeste1, Recorded human cases of gambiense human African Trypanosomiasis Hiro Goto1 (HAT) have undergone a remarkable decline over the past decade, 1 2 prompting plans for elimination of the disease. Effective planning for São Paulo University, São Paulo, Brazil, Sirio Libanês Hospital, São Paulo, this elimination programme will require accurate and reliable spatial Brazil information on the contemporary distribution of the disease. The Atlas In Brazil, visceral leishmaniasis (VL) caused by Leishmania infantum has a of HAT, recently established by WHO and FAO, aims to map the locations wide geographic distribution throughout the country with 3,894 reported of all known HAT cases. These data provide an unparalleled resource for cases in 2011, 47% in the Northeast region. Most of L. infantum-infected spatial risk assessment. However, under-reporting of cases and spatial individuals (around 95%) are asymptomatic and may be undetected and variation in reporting rates complicate their interpretation and reduce accepted as blood donors in endemic areas. Since leishmaniasis can be their utility for continental-scale planning. To overcome these issues, transmitted through blood transfusion, this study aimed to investigate the a collaboration between the Spatial Ecology and Epidemiology Group presence of anti-L. infantum rK39 antibodies in blood samples from Brazil (Oxford), WHO and FAO is developing a spatial modelling framework endemic areas. We used ELISA-rK39 (rK39 antigen kindly provided by to simultaneously map the prevalence of gambiense HAT cases and the Infectious Disease Research Institute, USA) that yielded 95.45% sensitivity, probability of detection of cases through the passive reporting system. In assaying 44 sera from parasitologically confirmed symptomatic VL patients order to construct this framework, a novel Bayesian spatio-temporal joint with positive Direct Agglutination Test (DAT) and 100.0% specificity statistical model has been developed to integrate data from both active assaying 44 healthy endemic DAT negative control samples. The present and passive case detection. The modelling framework will be outlined and study was carried out with 916 blood samples from Brazilian Northeastern preliminary results presented for the first time. states, Bahia (N=604) and Ceará (N=312). Anti-rK39 antibodies were detected in 26 out of 916 samples (2.8%): Bahia (2.8%) and Ceará 1741 (3.0%). The reactivity index (RI = absorbance/cut-off) varied from 1.010 to 6.756. Immunochromatographic rK39 test (ICT) applied to the 26 THE HEALTH IMPACT OF VISCERAL LEISHMANIASIS AND reactive samples showed one positive (from Ceará). Using L. major-like HUMAN AFRICAN TRYPANOSOMIASIS WHEN REACHING THE promastigote antigen (Lm), ELISA-Lm and indirect immunofluorescence 2020 WHO CONTROL AND ELIMINATION TARGETS test (IFT-Lm) detected respectively seven and one out of 26 positive samples. The sample showing RI of 6.756 in ELISA-rK39 was positive also Epke le Rütte, Wilma Stolk, Nico Nagelkerke, Jan H. Richardus, in ELISA-Lm and ICT. Of note, the studied samples had been screened for Sake de Vlas Chagas’ disease using antigen that cross-reacts with Leishmania and has Erasmus MC, Rotterdam, Netherlands been approved for transfusion; however, the present results showed that Visceral leishmaniasis (VL) and human African trypanosomiasis (HAT) this assay using cross-reactive antigen missed those 26 samples with anti- are neglected tropical diseases (NTDs). The London Declaration was Leishmania antibodies that likely result from an asymptomatic L. infantum established in 2012 to support the WHO control and elimination targets infection. As in endemic areas, it is not easy to differentiate transfusion- for ten NTDs by 2020. This initiative intends to globally generate a or vector-mediated transmission; the occurrence of transmission by positive health and socioeconomic impact by decreasing and eliminating transfusion is probably underestimated and raises concerns on blood the disease burden caused by these NTDs. In our study the global health transfusion safety. impact for VL and HAT is calculated for the ideal situation that the 2020 targets are met. The global burden of disease (GBD) study provides 1743 prevalence data for VL and HAT in 1990 and 2010. The 2020 targets, as formulated in the WHO Roadmap, provide the ideal situation: control THE RELATIONSHIP BETWEEN CLIMATIC AND OTHER for VL (100% detection and treatment at global level, and 1/10,000 ENVIRONMENTAL FACTORS AND ANNUAL FLUCTUATIONS new cases at subdistrict level per year on the Indian subcontinent) and IN INCIDENCE OF VISCERAL LEISHMANIASIS IN GEDAREF elimination of HAT by 2030. Linear trends between 1990, 2010, 2020 STATE, EASTERN SUDAN and 2030 provide a simplification of the real situation, representing the 1 2 number of remaining cases with disease, per country, age group and sex. Dia-Eldin A. Elnaiem , Alexandra Sweeney , Andrew Kruczkiewicz2, Caitlin Reid2, François Chappuis33, Koert Ritmeijer4, Continuing the 1990 prevalence until 2030, corrected for demographic 5 2 2 changes based on UNPOP data, serves as baseline situation without Omran Osman , Madeleine Thomson , Ceccato Ceccato interventions. The difference between the baseline and the remaining 1University of Maryland Eastern Shore, Princess Anne, MD, United States, cases results in the number of averted cases. The total number of averted 2International Research Institute for Climate and Society, Columbia years lived with disability (YLD) is calculated by multiplying the number of University, Palisades, NY, United States, 3MSF-Switzerland, Geneva, averted cases with the GBD disability weights. The total number of averted Switzerland, 4MSF-Holland, Amsterdam, Netherlands, 5University of disability adjusted life years (DALYs) between 2010 and 2030 results in Khartoum, Khartoum, Sudan app. 140 and 100 million DALYs for VL and HAT, respectively. The DALYs Visceral leishmaniasis (VL, kala azar), in Gedaref State, eastern Sudan, is are almost completely determined by the number of years of life lost caused by Leishmania donovani, which is transmitted by Phlebotomus (YLLs). The number of averted deaths over these two decades is 2.4 million orientalis sand flies. The endemicity of the disease in this region is and 1.7 million for VL and HAT, respectively. Although there have been characterized by marked annual fluctuations and occasional severe many successful interventions for VL and HAT, it is important to emphasize epidemics that claim the lives of many people. Since no significant interventions are undertaken against the disease in this region, the fall and rise in the incidence of VL in Gedaref state may be due to climate

astmh.org 532 variability, which characterizes the Sahelian region. Previous studies 1745 conducted by our groups and other research teams in Sudan and the Republic of South Sudan (RSS) showed marked variation in spatial DEFICIENCY OF PROLACTIN-INDUCIBLE PROTEIN LEADS TO distribution of kala azar that can be related to a number of environmental IMPAIRED TH1 IMMUNE RESPONSE AND SUSCEPTIBILITY TO and socio-economic factors that may be acting together or independently AN INTRACELLULAR PATHOGEN to increase the vulnerability of specific populations to the disease. Our 1 1 2 1 2 findings supported the previous notion that the vector and the disease Jintao Li , Dong Liu , Anne Blanchard , Ping Jia , Yvonne Myal , 1 are associated with Acacia seyal - Balanites woodland and chromic vertisol Jude Uzonna soils. We used this knowledge to produce a general kala azar risk map 1Institute of Tropical Medicine, Third Military Medical University, based on environmental prediction of the distribution of P. orientalis, the Chongqing, China, 2Department of Pathology and Deprtment of VL vector in Sudan and RSS. However, no attempt has yet been made to Physiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB, correlate annual incidence of kala azar with climatic factors and it is not Canada known whether the flare up in disease incidence is associated with dry The prolactin-inducible protein (PIP) is a secretory protein strategically or wet years. In this study we analyzed VL records of MSF-Switzerland located at several ports of pathogen entry into the body suggesting it and MSF-Holland, form 1996-2004 and 2010-2012, in relation to a might play a role in host defense. To date, no study has addressed the number of climatic and environmental variables, including temperature, contributions of PIP in immunity against infectious agents. Here, we humidity, rainfall and normalized difference vegetation index (NDVI). Our assessed the phenotype and responsiveness of immune cells from PIP results indicated that the incidence of kala azar in this region is related KO mice to polyclonal T cell stimulators and model antigens in vitro and to late onset of the rainy season. Results are discussed in relation to the in vivo. We found comparable numbers of immune cells, (including T, B, epidemiology of the disease. Findings from the study may be used in the natural killer and dendritic cells) in the primary and peripheral lymphoid future to develop an Early Warning System and construct high resolution organs of wild type and PIP KO mice. Further in-depth phenotypic analysis Geographical Information System (GIS) risk-maps for the disease. revealed that PIP KO mice had slightly but significantly lower numbers of CD4+ T cells in their spleens and lymph nodes. CD4+ T cells from PIP KO 1744 mice showed significantly decreased proliferation, IL-2 production and impaired Th1 differentiation in vitro. The impaired in vitro Th1 response CO-STIMULATORY MOLECULES ARE INVOLVED IN ANERGY was confirmed in vivo where CD4+ T cells from OVA-immunized PIP KO IN SYMPTOMATIC VISCERAL LEISHMANIASIS mice showed significantly impaired proliferation and IFN-γ production João Firmino Rodrigues-Neto following in vitro restimulation. Furthermore, PIP KO mice were highly susceptible to Leishmania major infection as evidence by inability to control Federal University of Rio Grande do Norte, Natal, Brazil lesion progression and parasite proliferation. This impaired resistant was The proliferation and differentiation of naive T cells require signals associated with dramatic impairment in IFN-γ and nitric oxide production provided by co-stimulatory molecules on antigen presenting cells (APCs), by splenic and draining lymph node cells from infected mice. Collectively, in addition to antigen-induced signals. In the absence of co-stimulatory our findings implicate PIP as an important regulator of CD4+ Th1 cell signals, T cells become anergic. We hypothesized that co-stimulatory response, and play a critical role in resistance to intracellular pathogens. molecules might be involved in part in the reversible anergy observed during symptomatic visceral leishmaniasis. We analyzed the profile of co- 1746 stimulatory molecules in lymphocytes and in CD14+ monocytes in whole blood collected from subjects with symptomatic visceral leishmaniasis T CELL ACCUMULATION IN THE SPLEEN DURING CHRONIC (sVL) and after their clinical recovery (rVL). An increase in CD8+ T cells PROGRESSIVE VISCERAL LEISHMANIASIS expressing CTLA-4 after stimulation with soluble Leishmania antigens (SLA) was observed (p<0.01) in sVL. An increase in percentage of CTLA-4 Audrie A. Medina, Elvia Y. Osorio, Omar A. Saldarriaga, Peter C. in CD4+ and CD8+ in ex vivo condition was observed, when compared Melby sVL versus rVL (p<0.05). No difference in lymphocytes expressing CD28 University of Texas Medical Branch, Galveston, TX, United States after SLA stimulation was observed in sVL or rVL, but rVL showed a high Visceral leishmaniasis (VL), caused by the protozoan, Leishmania donovani, percentage of CD8+CD28+ in ex vivo condition when compared with sVL is a chronic systemic infection that contributes to half a million new cases (p<0.05). An increase in the percentage of OX-40 in CD4+ T cells after SLA each year. During progressive VL, there is profound expansion of immune stimulation in sVL (p<0.05) was observed, as well as, an increase of ICOS cells in the spleen. Type 1 and type 2 cytokines are increased during VL in CD4+ and CD8+ T cells after SLA stimuli in sVL (p<0.01). Furthermore, in patients, but the T cell population in the spleen during VL has not a high percentage of ICOS in CD4+ in ex vivo condition was observed in been fully characterized. In the Syrian hamster model of progressive VL, sVL (p<0.01). There was no difference in CD40, CD86, CD80, ICOSL and which mimics human VL, we found a significant increase in expression HLA-DR in CD14+ monocytes after SLA stimulation in sVL or rVL. There of the T cell transcription factors Tbet, GATA3 and Foxp3 in spleen cells wasn’t also difference in the median fluorescence intensity (MFI) of CD40, over the course of chronic infection. This suggests the presence of a mix CD80, ICOSL or HLA-DR after SLA stimulation in sVL or rVL observed of Th1, Th2 and Treg cells. In purified splenic CD4 T cells from 28 day in CD14+ monocytes. But, CD86 showed a high expression in rVL after infected hamsters we found an increase in Tbet (p<0.001) and GATA3 SLA stimulation (p<0.05). These data support the role of co-stimulatory (p=0.0087) and Th1-associated chemokine receptors CXCR3 (p<0.0001) molecules in the reversible anergy observed during symptomatic VL and and CCR5 (p<0.001). There was no significant difference in Foxp3 or the might indicate pathways to be explored for immunotherapy against Th2-associated chemokine receptor CCR4 in purified CD4 T cells from leishmaniasis. uninfected and infected animals. These data suggest both Th1 and Th2 cells are present in the spleen during chronic infection, although one would expect Th2 cells to prominently express CCR4. Notably, we also found a significant population of CD4 T cells that expressed both Tbet and GATA3. The increase in Th1 and Th2 cells in the spleen during chronic infection could be due to local proliferation or splenic recruitment by T cell attracting chemokines. We found an array of chemokines (CCL2, CCL4, CCL5, CCL17, CCL22) increased in the spleen of hamsters over a course of disease. This suggests that T cell attracting chemokines may be playing a role in T cell accumulation at the site of infection.

astmh.org 533 1747 expansion in visceral leishmaniasis. L. donovani infection of bone marrow and splenic macrophages resulted in increased cell number, DNA EVALUATION OF THE EFFICACY OF ANTIGEN DELIVERY synthesis (BrDu incorporation) and mitosis (ki-67 antigen expression). BY THE TRANSCUTANEOUS IMMUNIZATION ROUTE IN A The combination of growth factor FGF-2 or IGF-I with IL-4 significantly MURINE MODEL OF CUTANEOUS LEISHMANIASIS increased macrophage proliferation, suggesting that they interact to control the cell cycle. Inhibition of FGFR, IGFR and PI3K significantly 1 1 2 Ines Elakhal Naouar , Nancy Koles , Elaine Morrison , Mangala reduced mitosis indicating that growth factor signaling through PI3K was 2 2 1 Rao , Carl Alving , Naomi Aronson a major contributor to macrophage proliferation. The local amplification 1Uniformed Services University, Bethesda, MD, United States, 2Walter Reed of macrophages in response to chronic infection through the expression of Army Institute of Research, Laboratory of Adjuvant and Antigen Research, type 2 cytokines and growth factors may have broad significance to other Silver Spring, MD, United States chronic infectious diseases. Transcutaneous immunization (TCI) is a novel attractive vaccination method which offers advantages over traditional vaccination routes, 1749 exploiting the abundance of antigen presenting cells in the skin. We provide the first report of Transcutaneous immunization (TCI) is a novel FACTORS CONTRIBUTING TO TISSUE DAMAGE IN HUMAN attractive vaccination method which offers advantages over traditional CUTANEOUS LEISHMANIASIS vaccination routes, exploiting the abundance of antigen presenting cells Sara T. Passos, Taís Menezes, Patrícia Cisneiros, Paulo R. in the skin. We provide the first report of TCI induced immune responses Machado, Juliana Almeida, Viviane Magalhães, Viviane to Leishmania antigens. Leishmania major soluble antigens (SLA) and Magalhães, Lucas P. Carvalho, Edgar M. Carvalho Phlebotomus papatasi salivary gland homogenates (SGH) were delivered Federal University of Bahia, Salvador, Brazil transcutaneously with cholera toxin (CT), a potent adjuvant for developing mucosal immunity. Sixty inbred BALB/c mice were immunized three times Human cutaneous leishmaniasis (CL), due to Leishmania braziliensis at weeks 0, 3 and 6 with the vaccine formulations (different doses of infection, is characterized by intense immune mediated tissue SLA or SGH, SLA+SGH). TCI was well tolerated. Two weeks after the last inflammation and skin ulceration. For many years immunological studies in vaccine boost, we assessed humoral (IgG titer to antigens and CT) and CL patients have focused on the evaluation of host response mainly using cellular immune responses (IFN-γ ELIspot and cytokine levels from splenic peripheral blood cells assays. In this context, high levels of IFN-gamma and cell culture). In contrast to SGH alone, we showed that transcutaneous TNF and low levels of regulatory cytokine, IL-10, is observed in cultures of immunization of mice with SLA resulted in high titers of anti-SLA IgG that peripheral blood mononuclear cells stimulated with soluble Leishmania increased when SLA was combined with SGH antigen. Immunization was antigen. Histopathology studies show mononuclear cells infiltration also associated with high anti-CT IgG titers. A Th1-type immune response and low number of parasites. Our hypothesis is that the inflammatory was demonstrated with high levels of IFN-γ production and lower levels environment helps to control parasitemia but mediates tissue destruction. of IL-10 resulting in a significantly higher IFN-γ/IL-10 ratio compared to In the present study we evaluated cytokine and chemokine profile at the control groups. A high frequency of IFN-γ secreting cells was also lesion site. Cells from CL lesion produced high levels of pro-inflammatory seen in groups of mice immunized with SLA. Altogether, these data are cytokines, TNF, IL-6 and IL-1b in absence of stimuli. To determine the consistent with reported protective immune responses and indicate the contribution of skin epithelial cells to the production of these cytokines strong potential of our TCI strategy to protect against Leishmania major we cultured epidermis and dermis separately. TNF was only produced by infection, with the combined antigen SLA and SGH showing the strongest cells composing the dermis, while IL-6 and IL-1b were produced by dermis responses. Experiments using the same regimen of immunizations and epithelial cells from epidermis. High levels of CCL2, a chemokine that followed by parasite challenge are in progress. Results of lesion evolution recruits mononuclear phagocytes, and CXCL9 and CXCL10, involved in and parasite load along with immune responses pre and post challenge lymphocyte recruitment, were also observed. Metalloproneinase-9 (MMP- will be presented. 9) is a zinc-dependent enzyme that degrades collagen type 4 (present in basal membrane) and has been associated with tissue damage in skin 1748 inflammatory diseases. We found increased production of MMP-9 in CL lesion when compared to healthy skin. TNF is known to induce MMP-9 MECHANISMS OF DISEASE-PROMOTING MACROPHAGE production. To determine the role of TNF in MMP-9 production in CL, we PROLIFERATION IN VISCERAL LEISHMANIASIS cultured CL cells in presence of monoclonal antibodyes anti-TNF. Blockage of TNF decreased MMP-9 production in CL. Ou study contributes to the 1 1 1 E. Yaneth Osorio , Audrie A. Medina , Omar A. Saldarriaga , understanding of immunopathology in CL and revele possible targets for 1 2 1 Bruno L. Travi , Alda M. da-Cruz , Peter C. Melby immunotherapy. 1University of Texas Medical Branch, Galveston, TX, United States, 2FIOCRUZ, Rio de Janeiro, Brazil 1750 Macrophages have classically been viewed as mature non-proliferative cells, which originate from bone marrow monocytes released to peripheral IMMUNOGENICITY AND PROTECTIVE EFFICACY OF blood to infiltrate tissues in response to injury. However, recently it was PVAX-NH36 AS A DNA VACCINE AGAINST CUTANEOUS discovered that macrophages can be locally self-maintained without major LEISHMANIASIS IN A CANINE MODEL contribution of infiltrating monocytes, and that M2 macrophages are Vladimir Cruz-Chan1, Amaru Aguilar-Cetina1, Liliana Villanueva- able to proliferate locally within a Th2 environment. The mechanism(s) Lizama1, Pedro Martinez-Vega1, Maria Jesus Ramirez-Sierra1, that drive this proliferation have not been defined. In a model of chronic Miguel Rosado-Vallado1, Leonardo Guillermo-Cordero1, Jaime progressive visceral leishmaniasis, we found huge expansion of disease Ortega-Lopez2, Eric Dumonteil1 promoting macrophages in the spleen, with signs of Th2-amplified 1 M2 activation, increased STAT-6 activation, and pathological arginase Universidad Autonoma de Yucatan, Merida, Yucatan, Mexico, 2 expression. We discovered that growth factors IGF-I and FGF-2 were CINVESTAV, Mexico, D.F., Mexico key contributors to both STAT-6 activation and arginase expression in L. Leishmaniasis is a major tropical disease affecting 2 million people donovani infected macrophages, and inhibition of growth factor signaling annually, against which there is no effective treatment. Vaccine blocked arginase expression and parasite replication. Since that these development for humans requires preclinical testing in animal models growth factors also drive cellular proliferation and differentiation, and including canines. One of the best vaccine candidates is NH36, a arginase contributes to the cell growth through polyamine production, Leishmania donovani 36 kDa protein. As DNA vaccine NH36 provides we explored the possibility that these factors had a role in macrophage better protection than the recombinant protein or purified fucose-manose

astmh.org 534 ligand. In this study, we evaluated the immunogenicity and protection from a locale. Similar to endemic human infection, infection in these of pVAX-NH36 for the prevention and therapy of L. mexicana infection macaques is thought to have been acquired early in life (age range at in dogs. We first established a model of infection in Beagle dogs. Then, infection 1-15 years) and to persist chronically with an average infection Beagles received 3 doses of 250 microg of pVAX-NH36 as prophylaxis time of 6.4 years. The T. cruzi isolates were obtained from hemocultures of or therapy, while dogs from control group received saline solution. L. 43 out of 64 (67%) macaques. An additional 8 hemocultures were positive mexicana promastigotes were used for infection via intradermal. Immune for T. cruzi DNA by PCR, but failed to yield a culturable line, resulting in response was evaluated measuring antibodies, IFN and IL-10 production, an overall hemoculture detection rate of 80%. This figure is comparable and DTH. Ulcer diameters and parasite burden were evaluated to assess to the frequency of detection of infection by serial PCR (up to 3 samples) protection. Canines receiving pVAX-NH36 showed higher IgG levels of whole blood (81%). Furthermore, parasite isolates were obtained by against NH36 in comparison with the control group. High IFN and low IL- hemoculture from 8 of the 12 PCR-negative animals. The combination 10 levels were produced by PBMC stimulated with NH36 from vaccinated of whole blood PCR and hemoculture +/- PCR confirmed active infection canines. In addition, only vaccinated animals were DTH positive against in 94% of the seropositive animals. Genotyping of hemoculture-isolated recombinant NH36. Finally, some protection was observed based on skin T. cruzi revealed the presence of two lineages: TcI and TcIV, which are parasite burden as two vaccinated animals showed negative results by the most common lineages identified by previous studies in infections qPCR. In conclusion, pVAX-NH36 is safe and immunogenic in dogs, and originating in North America. To date, all tested hemoculture-isolated can confer some protection against L. mexicana. T. cruzi could be converted to metacyclic trypomastigotes and were orally infective in C57Bl/6 and IFN-gamma knockout mice with the 1751 TcIV lineage isolates exhibiting less virulence in both mouse models. All isolates appeared to establish chronic infection in mice and to induce LEISHMANIA SPECIFIC CD4 T CELLS RELEASE IFNΓ THAT immune responses consistent with those observed in infections by long- LIMITS PARASITE REPLICATION IN PATIENTS WITH VISCERAL maintained laboratory-adapted strains of T. cruzi. Future studies will assess LEISHMANIASIS the susceptibility of these fresh isolates to clearance by treatment with benznidazole. Om Prakash Singh1, Rajiv Kumar1, Neetu Singh1, Shalini Gautam1, Kamlesh Gidwani1, Madhukar Rai1, David Sacks2, Shyam Sundar1, Susanne Nylen3 1753 1Institute of Medical Sciences, Banaras Hindu University, Varanasi, India, IMMUNOREGULATORY NETWORKS AND 2National Institute of Allergy and Infectious Diseases, National Institutes of IMMUNOPATHOGENIC PATHWAYS IN VISCERAL Health, Bethesda, MD, United States, 3Karolinska Institutet, Department of LEISHMANIASIS Microbiology Tumor and Cell Biology, Stockholm, Sweden Omar A. Saldarriaga, Fanping Kong, Heidi Spratt, Bruce A. Visceral leishmaniasis (VL) is associated with increased circulating levels Luxon, Elvia Y. Osorio, Bruno L. Travi, Peter C. Melby of multiple pro-inflammatory cytokines and chemokines, including IL-12, IFNγ, and TNF-alpha, and elevated expression of IFNγ mRNA in lesional University of Texas Medical Branch, Galveston, TX, United States tissue such as the spleen and bone marrow. However, an immunological Visceral Leishmaniasis (VL) caused by Leishmania donovani, is an feature of VL patients is that their peripheral blood mononuclear cells important problem in tropical and subtropical areas of the world. VL (PBMCs) typically fail to respond to stimulation with leishmanial antigen. is characterized by a progressive increase in visceral parasite burden, Unexpectedly, it was recently shown that Leishmania specific IFNγ, can cachexia, splenomegaly, pancytopenia and ultimately death. We have readily be detected when a whole blood stimulation assay (WBA) is used. studied the pathogenesis of VL in Syrian hamsters (Mesocricetus auratus) We sought to define the conditions that permit whole blood cells to since it closely mimics active human disease. We demonstrated previously respond to antigen stimulation, and clarify the biological role of the IFNγ that the simultaneous expression of Th1 (IFN-γ) and Th2 (IL-4) cytokines, found to be released by cells from VL patients. CD4+ T cells were found parasite activation of STAT6, and decreased production of nitric oxide to be crucial for and the main source of the IFNγ production in Leishmania are associated with susceptibility to the infection. To fill the gaps in our stimulated whole blood (WB) cultures. Complement, antibodies and red understanding of the pathogenesis of VL we examined global changes blood cells present in whole blood do not play a significant role in the in gene expression in spleen tissue and splenic (adherent) macrophages. IFNγ response. The IFNγ production was reduced by blockade of human We used a novel approach of deep sequencing (RNAseq) coupled with leukocyte antigen (HLA)-DR, indicating that the response to leishmanial de novo assembly of full-length transcripts because the Syrian hamster antigens observed in WB of active VL patients is a classical HLA- T cell does not have an annotated reference genome. Differentially expressed receptor (TCR) driven reaction. Most importantly, blockade of IFNγ in ex- transcripts in L. donovani infected (28 days) vs. uninfected hamsters were vivo splenic aspirate cultures demonstrated that despite the progressive determined by alignment back to the de novo constructed transcriptome nature of their disease, the endogenous IFNγ produced in patients with and cross-species BLAST, after the removal of contaminating parasite active VL serves to limit parasite growth. sequences. Transcriptome analysis confirmed that adherent cells were enriched for expression of macrophage (CD14, CD64 and Mertk) but 1752 not T cells markers (CD4, Gata3 and Tbet). Differentially expressed genes analyzed with IPA software revealed a number of highly enriched ISOLATION AND INITIAL CHARACTERIZATION OF canonical pathways in spleen and splenic macrophages, including hepatic TRYPANOSOMA CRUZI ISOLATES FROM A POPULATION OF fibrosis, pathogenesis of multiple sclerosis, atherosclerosis signaling, CYNOMOLGUS MACAQUES NATURALLY INFECTED IN THE communication between innate and adaptive immune cells, and the UNITED STATES glucocorticoid receptor signaling. Notably, within the differentially expressed transcripts we identified mixed expression of genes associated Phil Yao with classical (M1) and alternative (M2) activation of macrophages University of Georgia, Athens, GA, United States that was confirmed by qPCR. This approach provides a valuable tool to The protozoan parasite Trypanosoma cruzi, causative agent of Chagas overcome the obstacles of working with a non-model organism without disease, is capable of infecting not only humans, but also virtually any a reference genome. With it we can begin to understand the complex mammalian species. Non-human primates housed in outdoor facilities immunopathogenic mechanisms at the site of visceral infection. in the southern United States are known to acquire T. cruzi infection, as is the case for a group of 64 cynomolgus macaques from Texas who are seropositive for T. cruzi infection. This group provided a unique opportunity to investigate natural T. cruzi infection in an entire population

astmh.org 535 1754 contact with backyard birds (OR 2.1, 95% CI 1.4-3.0), and working in agriculture or with livestock (OR 4.3, 95% CI 1.7-10.8). However, exposure ENHANCEMENT OF MURINE VISCERAL LEISHMANIASIS DUE to any kind of animal within the household decreased the odds ratio by TO CROSS REACTIVE LEISHMANIA MAJOR ANTIBODIES half (OR 0.50, 95% CI 0.31-0.81), perhaps indicating that arthropod vectors on birds preferred non-human hosts when they were present, thus Heidi Anderson, Blaise Dondji, Gabrielle A. Stryker diminishing exposure to humans. Age was the only variable associated Central Washington University, Ellensburg, WA, United States with antibody positivity to TFGR (OR 1.03, 95% CI 1.02-1.05). The Leishmaniasis is a global disease found in regions with compatible antibody prevalence to Leptospira was 11.3% in Puerto Maldonado and temperatures for the phlebotomine sandfly vector to survive and lacking 5.8% in Tumbes, with a borderline association with keeping animals in rigorous vector control programs. An estimated 1.3 million new cases and the household (OR 2.5, 95% CI 1.0-6.2). Exposure to Rickettsia, especially 20,000 - 30,000 deaths occur annually due to this parasitic protozoan. SFGR, and Leptospira appears to be frequent in Peru. We plan now to More than twenty different species of Leishmania infect humans with perform testing in domestic animals in some of these sites to determine multiple species occurring in the same geographic areas. Symptoms range the specific reservoirs and vectors for these agents and to obtain pathogen from a minor cutaneous lesion at the bite site due to dermotropic species isolates for identification of the specific species. such as L. major, to life threatening disseminated disease with multiple organ involvement, caused by viscerotropic species such as L. infantum. We have previously shown susceptible BALB/c mice infected with a low/ 1756 self-healing dose of cutaneous L. major and challenged with L. infantum, PROXIMITY TO PIG POPULATIONS AS A KEY RISK FACTOR develop a markedly worsened disease with higher parasite burden, relative FOR JAPANESE ENCEPHALITIS DISEASE; RESULTS OF A to naïve mice. There was little notable difference in the cytokine profiles FIVE-YEAR SURVEILLANCE STUDY FROM NORTHWESTERN between L. major exposed and naïve mice in response to L. infantum. BANGLADESH Cross-reactive antibodies were seen in both groups of L. infantum infected mice regardless of their immune history. Opsonizing antibodies have Henrik Salje1, Salah Uddin Khan2, Abu Hannan3, M. Atiqul been shown to lead to increased disease in visceral leishmaniasis. The Islam4, A. A. Mamun Bhuyan4, M. Ariful Islam4, M. Ziaur Rahman4, present studies focus on exploring the role cross-reactive antibodies may Mahmadur Rahman5, Nazmun Nahar4, M. Jahangir Hossain4, play in exacerbation of visceral disease seen in mice previously exposed Stephen P. Luby6, Emily S. Gurley4 to L. major. Mice receiving passively transferred serum from L. major 1Johns Hopkins School of Public Health, Baltimore, MD, United States, infected mice, 48 hours prior to challenge with L. infantum, developed 2University of Florida, Gainesville, FL, United States, 3Fisheries Ministry, equivalent organ parasitemia to age/L. major-infected matched control Dhaka, Bangladesh, 4International Centre for Diarrhoeal Disease Research, mice. Naïve mice inoculated with control serum did not suffer any Bangladesh, Dhaka, Bangladesh, 5IEDCR, Dhaka, Bangladesh, 6Stanford disease enhancement with L. infantum. We speculate that cross-reactive University, Stanford, CA, United States antibodies are augmenting visceral disease in mice with immunological Japanese encephalitis (JE) is a mosquito-borne virus that causes severe memory to L. major. While L. major is known to produce long lasting disease in humans with 10-20% case fatality. It is the commonest cause of immunological memory and protect against recurrent cutaneous disease, encephalitis in Bangladesh. Humans are dead-end hosts. Pigs, by contrast, antibody enhancement due to inter-Leishmania infection may enhance are viral amplifying hosts, so the distribution of pig populations may be disease in regions with multiple circulating Leishmania-species and particularly important for human disease risk. Previous studies have shown suggests leishmanization might be riskier than previously thought. that the highest incidence of human JE infections occur in northwestern Bangladesh, where JE is also endemic among pigs. The objective of 1755 this study was to explore proximity to pigs as a risk factor for human JE HUMAN SEROPREVALENCE OF LEPTOSPIROSIS AND disease in Bangladesh. We first geocoded the locations of residence for RICKETTSIOSIS IN FOUR REGIONS OF PERU all JE patients identified through hospital-based surveillance in Naogaon, Chapainawabganj, and Rajshahi districts in northwestern Bangladesh Gabriela Salmon-Mulanovich1, Mark Simons1, Maria E. Silva1, between 2007 and 2011. Next, we used data from a 2009 pig census in Bruno M. Ghersi2, Matthew R. Kasper3, Hugo R. Razuri4, Daniel G. these areas to map all pig raising households. To explore the impact of Bausch5, Allen L. Richards6 proximity to pigs as a risk factor for JE disease we compared the odds of 1U.S. Naval Medical Research Unit - 6, Callao, Peru, 2Tulane University, a human JE case living within a set distance of a pig-raising household to New Orleans, LA, United States, 3Armed Forces Health Surveillance that of a randomly selected control population. We identified 81 human Center, Silver Spring, MD, United States, 4McGill University Health Centre, JE cases from throughout the region, with a mean age of 32 years (range: Montreal, QC, Canada, 5Tulane School of Public Health and Tropical 0 - 75 years); 11% died. Disease patterns were highly seasonal with 90% Medicine, New Orleans, LA, United States, 6Naval Medical Research of cases occurring between the months of August and November. Humans Center, Silver Spring, MD, United States infected with JE were 2.7 times more likely to live within 500m of a pig- owning household compared to controls (95% confidence interval [CI] Rickettsia and Leptospira are obligate intracellular bacteria with global 1.3 - 4.6) and 1.7 times more likely to live within 5km (95% CI: 1.0 - 3.7). distributions and a wide variety of animal hosts. Two disease-causing Results from this analysis suggest that proximity to pig populations is an groups of Rickettsia are noted: the spotted fever group Rickettsia (SFGR) important risk factor for human JE disease in northwestern Bangladesh. and the typhus fever group Rickettsia (TFGR). We explored the prevalence JE vaccination is not currently included in the Bangladesh immunization of exposure and risk factors for human Rickettsia infection in four program; therefore, interventions to reduce infections among pigs could ecologically distinct regions of Peru (Lima, Cusco, Puerto Maldonado and be an important strategy for reducing human risk in these areas and Tumbes) and for Leptospira in Puerto Maldonado and Tumbes. In January should be explored. 2012 we randomly collected 2165 serum samples from participants in a surveillance cohort for respiratory disease in the aforementioned sites and tested them for IgG by ELISA (Rickettsia) and microscopic agglutination test (Leptospira). Overall antibody prevalence across the four sites was 10.6% for SFGR (ranging from 6.2-14.0%, with the highest prevalence in Tumbes) and 3.3% for TFGR (ranging from 2.6-6.4%, with highest prevalence in Puerto Maldonado). Factors associated with positive IgG for SFGR on multiple logistic regression analysis were male sex (OR 2.2, 95% CI 1.5-3.3), increasing age (OR 1.02, 95% CI 1.01-1.04 per year),

astmh.org 536 1757 more animal host species in Africa. For several important human-infecting serogroups, multiple animal host species were identified. L. borgpetersenii, ECOLOGICAL NICHE MODELING FOR SYLVATIC RABIES L. interrogans and L. kirschneri were the predominant genetic species TRANSMITTED BY DESMODUS ROTUNDUS IN PERU reported in human and animal populations across Africa, although some local variation was observed. In conclusion, this systematic review 1 2 M. Claudia Guezala Villavicencio , William D. Valerrama , highlights the importance of acute leptospirosis in febrile patients in Africa 1 3 Gabriela Salmon-Mulanovich , Daniel G. Bausch and reveals many areas of uncertainty that remain in our understanding of 1U.S. Naval Medical Research Unit - 6, Callao, Peru, 2Servicio Nacional de this complex, multi-host disease. A ‘One Health’ approach is advocated to Sanidad Agraria, La Molina, Peru, 3Tulane School of Public Health and integrate human and animal studies in future work, and to explore local Tropical Medicine, New Orleans, LA, United States and regional variation in leptospirosis epidemiology in Africa. Rabies is a viral infection endemic in many parts of the world that causes severe and usually fatal encephalitis. In Latin America, a sylvatic cycle 1759 exists in which rabies virus is maintained in several species of wild animals, especially the hematophagous bat Desmodus rotundus. Peru and Brazil CHARACTERIZING EXPOSURE TO BATS AND BAT GUANO report the highest number sylvatic rabies cases, mainly transmitted by AMONG MEN, WOMEN AND CHILDREN IN LAO PDR TO bats. Vaccination of humans and livestock is an effective prevention INFORM INTERVENTIONS FOR REDUCING THE RISK OF method, but vaccination is not usually undertaken until a human exposure ZOONOTIC DISEASE our outbreak in livestock occurs. The ecologic and topographic factors Laura Seckel1, Khounkham Xaymounvong2, Sara Woldehanna1, underlying the distribution of rabies vectors are not well understood. 1 Specifically for sylvatic rabies, there is little data on the factors influencing Susan Zimicki the distribution of hematophagous bats. Such information would help 1FHI 360, Washington, DC, United States, 2FHI 360, Vientiane, Lao People’s define areas and populations at risk for rabies and inform effective Democratic Republic prevention campaigns. We therefore modeled the potential geographic Bats host more viruses per species than other mammals, and are reservoirs distribution of D. rotundus using the ecological niche modeling algorithm of numerous pathogens that pose significant risk to humans. Several MaxEnt. Incorporating climatic, environmental and anthropogenic factors attention-grabbing emerging diseases, such as SARS, Ebola, Nipah and that may relate to the geographic distribution of D. rotundus and rabies- MERS-CoV have been linked to bats. Transmission routes for these, and infected farm animals, we developed a risk map for bat-associated rabies other zoonotic diseases, are extremely varied, and bats are thought to transmission in Peru. D. rotundus occurrence was found to be associated transmit diseases through urine, feces, saliva or via intermediate hosts. with the colder, drier months of the year. In addition, land classification To inform development of interventions to prevent the spread of disease data show that bats prefer firmer, non-flooding low lands. Variables from bats to humans, human exposure to bats and their excrement associated with occurrence of animal rabies included livestock population must be better understood. In Lao PDR, a country of great biodiversity, density, mean diurnal temperature range, and precipitation in the drier people are regularly exposed to bats in a variety of ways, including months of the year. This study offers a first glimpse of the environmental hunting, consumption, and the collection and use of guano. In addition, and bioclimatic factors associated with the distribution of hematophagous both humans and domestic animals are often exposed to bats and their bats and animal rabies using novel techniques that extract the maximum excreta through environmental exposure. We used rapid appraisal and information and offer robust results from the little data available. participatory research methods to characterize bat exposure among men, women and children in four sites in Vientiane and Bolikhamxay provinces, 1758 Lao PDR. We will report findings about human interactions with bats/ excreta, environmental exposure of humans and domestic animals to bats/ THE PROBLEM OF LEPTOSPIROSIS IN AFRICA: REVEALING excreta, and the seasonality of exposure, mediated by gender, location, A NEGLECTED ‘ONE HEALTH’ CHALLENGE THROUGH A ethnicity (Lao-Tai, Hmong, Kammu), age, and occupation and will discuss SYSTEMATIC REVIEW possible preventive interventions. Kathryn J. Allan1, Holly M. Biggs2, Jo E. Halliday1, Rudovick R. Kazwala3, Venance P. Maro4, Sarah Cleaveland1, John A. Crump2 1760 1University of Glasgow, Glasgow, United Kingdom, 2Duke University RISK FACTORS FOR HUMAN EXPOSURE TO WILDLIFE Medical Center, Durham, NC, United States, 3Sokoine University of ZOONOSES IN NIGERIAN HUNTING COMMUNITIES Agriculture, Morogoro, United Republic of Tanzania, 4Kilimanjaro Christian Medical Centre, Moshi, United Republic of Tanzania Sagan Friant, Tony Goldberg, Sarah Paige Recent evidence from Tanzania indicates that leptospirosis is an important University of Wisconsin Madison, Madison, WI, United States cause of non-malarial febrile disease. However, relatively little is known Bushmeat hunting clearly increases the risk of zoonotic pathogen about the incidence and geographic distribution of leptospirosis in Africa transmission. However, limited information exists on the nature and as well as the diversity of infecting Leptospira spp. in human and animal frequency of contact with wildlife in communities that practice bushmeat populations. To examine current knowledge, we performed a systematic hunting, especially with respect to socioeconomic drivers of hunting review of acute human leptospirosis and confirmed Leptospira infection behavior. We surveyed 188 hunters and 139 non-hunters in five rural in animals. We searched eight international and regional scientific communities in Cross River State, Nigeria. Responses were used to: 1) databases using the terms ‘Leptospira’ OR ‘leptospirosis’ AND ‘Africa’ for quantify contact rates with wildlife, 2) identify socioeconomic factors articles published between 1930 and June 2013. Of 630 unique articles predisposing individuals to hunt, and 3) identify specific hunting behaviors identified and reviewed against predetermined inclusion and exclusion that increase frequency of contact with wildlife. Among hunters, 95% criteria, 89 (14.1%) were considered eligible. Eligible articles described hunted rodents, 91% ungulates, 90% carnivores, 78% primates and 35% human and animal Leptospira spp. infection in 26 (44.8%) of 58 countries bats. Hunters used traps (75%), guns (71%), machetes (71%), and dogs included in the UN continental definition of Africa. Prevalence of acute (18%) to hunt animals both day (78%) and night (69%). We constructed leptospirosis in hospital-based cohort studies of patients with non-malarial generalized linear mixed models to examine socioeconomic predictors febrile illness ranged from 2.3% (n=43) to 47.5% (n=59). Estimates of of individual hunting behavior and frequency of contact with wildlife, annual human leptospirosis incidence ranged from 4.1 to 101 cases per especially primates. We found that lower education level (<.01), having a 100,000 based on surveillance studies of island populations. Leptospira father who hunts (p<.0001), and larger household sizes (p<.05) were all spp. infection was also reported in a wide range of animal hosts. 11 out associated with becoming a hunter. Among hunters, high rates of wildlife of 15 human-infecting Leptospira serogroups were isolated from one or contact were associated with high hunting frequency (p<.05), hunting

astmh.org 537 both night and day (p<.05) and with a gun (p<.05); while sleeping in the where inadequate sanitary conditions, high population densities, and forest (p<.0001), hunting night and day (p<.0001) and with a dog (p<.05) frequent contact with animals capable of carrying zoonotic pathogens were associated specifically with high rates of primate contact. Results can exist in both urban slums and rural areas. We present a study to demonstrate that hunters have risky contact with a diversity of wildlife, estimate and compare the prevalence of Giardia infection in rural and and that the decision to become a hunter is deeply rooted in family history urban settings for both human and animal populations residing in coastal and modified by economic necessity. Improved education, reduced family Odisha, India, evaluate if gender or age is associated with Giardia infection sizes, and alternative livelihoods may reduce the risk of zoonotic disease in humans, and estimate fecal loading rates of Giardia cysts from animal exposure in rural hunting communities in Nigeria. Public health programs host populations residing in the study area. From April to May of 2012, aimed at reducing zoonotic transmission of wildlife pathogens in such human fecal samples from 85 diarrhea patients presenting at three settings will be most efficient when they target root socioeconomic drivers diarrhea wards and 111 pooled animal fecal samples across seven host that lead to hunting behavior and risky wildlife contact. species (cattle, buffalo, goat, sheep, chicken, cat, and dog) were collected across urban and rural residential areas served by the wards. Samples were 1761 screened and fluorescent microscopy used to enumerate Giardia cysts and a subset of dog and human samples analyzed by molecular methods to ENVIRONMENTAL RESERVOIRS OF ANTIBIOTIC RESISTANCE identify isolate genotypes. Giardia cysts were detected in 12% of tested ASSOCIATED WITH SMALL SCALE POULTRY FARMING IN diarrhea patients, while 32% of pooled animal samples were positive. No NORTHWESTERN ECUADOR evidence for difference in the presence of Giardia cysts among humans was observed between urban and rural settings, gender, or age groups (<5 1 2 1 Nikolay P. Braykov , Joseph Eisenberg , Marissa Grossman , Lixin years, 5-59 years, >59 years). There was substantial support for a location 2 3 4 4 Zhang , William Cevallos , Karla Vasquez , Diana Muñoz , Carl F. effect on Giardia shedding among animals, with rural animals shedding 2 2 4 1 Marrs , Betsy Foxman , Gabriel Trueba , Karen Levy higher numbers of parasites. Of the seven animal host groups screened, 1Emory University, Atlanta, GA, United States, 2University of Michigan dogs and cattle, both reported to shed zoonotic genotypes of Giardia in School of Public Health, Ann Arbor, MI, United States, 3Universidad Central India, shed decisively more Giardia cysts per gram of feces, as much as del Ecuador, Quito, Ecuador, 4Universidad San Francisco de Quito, Quito, 2-3 orders of magnitude greater than other animal types (adjusting for Ecuador location). Molecular characterization of isolates identified host specific Non-therapeutic use of antibiotics in agriculture poses a threat to human Assemblages in dog samples and a possible zoonotic Assemblage in a health by contributing to an environmental reservoir of antibiotic-resistant human sample. Using current animal populations and observed Giardia (AR) bacteria. Small-scale “backyard” broiler chicken production involving shedding rates, cattle were estimated to contribute >99% of Giardia use of antibiotics for feed promotion is becoming increasingly common animal cysts into the study area environment, followed by dogs as the next in developing countries. We use E. coli antibiotic susceptibility profiles to largest source. This study shows Giardia prevalence is similar for humans assess the potential for transmission of AR from broiler chickens to the living in rural and urban settings, but different for animals and that surrounding environment (water, soil, surface) in the context of small- exposure from infected cattle and dogs may be an important public health scale poultry farming in Northwestern Ecuador. Field work spanned 190 concern in Coastal Odisha. households in 17 villages visited between 08/2010-08/2012. We collected a total of 529 samples from drinking water, soil and food preparation 1763 surfaces, and surveyed water, sanitation and antibiotic use practices. From a subset of 91 households involved in broiler production, we collected 131 COMPARATIVE GENOMIC ANALYSIS OF COCCIDIOIDES AND cloacal samples from production chickens and 66 soil and surface samples RELATED SPECIES from chicken coops. In addition, 153 non-production (“free-range”) birds Christian Olsen, Kashef Qaadri were sampled across all villages, and 54 water samples from local rivers were collected. Up to five E. coli isolates from each sample were tested Biomatters, Newark, NJ, United States against 12 antibiotics using disc diffusion. Zones of inhibition and their Dimorphic fungi Coccidioides immitis and C. posadasii are primary categorical interpretations were compared using mixed-effect models. AR pathogens of immunocompetent mammals, including humans. was more common in broilers than free-range chickens for every antibiotic Coccidioides infection results from environmental exposure, which is tested (p<0.01), particularly tetracycline (76.8% vs 33.1%), sulfisoxazole believed to grow as a soil saprophyte in arid desert environs. To investigate (66.9% vs 19.2%) and streptomycin (61.1% vs 26.7%). A pattern of AR hypotheses about the evolution of Coccidioides, the genomes of several to gentamicin, fluoroquinolones and beta-lactams that was unique to Onygenales, a close, nonpathogenic relative, and a more diverged broilers was also found in coop surfaces and soils, but not in household pathogenic fungus, were compared with those of 13 more distantly samples. The prevalence of this phenotype declined with bird age, related Ascomycetes. This poster aims to identify shifts in gene family implying importation form hatcheries outside the study system. AR was size associated with a host/substrate shift from plants to animals in the more common in coop than household samples. Farming and non-farming Onygenales. Comparison among Onygenales revealed distinct evolutionary households showed no difference in AR profiles. These results suggest changes in Coccidioides that may underlie its infectious phenotype, broiler chickens carry AR and pass it to their immediate environment. coccidioidomycosis. Phylogenetic analysis suggest that Coccidioides species However, transmission to households may operate at different scales. are not soil saprophytes as previously hypothesized. Data indicate that they have evolved to remain associated with their dead animal hosts in soil. 1762 Using a bioinformatics workflow, we show that metabolic pathway genes, membrane-related proteins, and putatively antigenic compounds have FACTORS ASSOCIATED WITH GIARDIA IN HUMANS AND evolved in response to interaction with an animal host. ANIMALS RESIDING IN RURAL AND URBAN AREAS OF COASTAL ODISHA, INDIA AND ENVIRONMENTAL LOADING ESTIMATES FROM ANIMALS Miles Daniels1, Woutrina Miller1, Arpit Shivastava2, Priyadarshi Sahu2, Mitsunori Odagiri1, Marion W. Jenkins1 1University of California Davis, Davis, CA, United States, 2Kalinga Institute of Industrial Technology, Bhubaneswar, India Globally, Giardia lamblia, is one of the most commonly detected intestinal parasites and is particularly relevant in developing countries such as India,

astmh.org 538 1764 and seasonal variation of the bush meat trade as well as how the forms of animals sold (live, freshly dead, large pieces, small pieces, smoked, raw) MODELING BRUCELLA INFECTION DYNAMICS IN and change with the length of time they are in the market. We will discuss PASTORALIST COMMUNITIES: THE ROLE OF HERD the implications of all of these factors for human exposure and risk MANAGEMENT AND SPECIES COMPOSITION IN SUSTAINING BRUCELLA TRANSMISSION 1766 Peter S. Larson, Johannes Foufopoulos, Rebecca Hardin, Annie MASSTAG PCR DETECTION OF EV-D68, RSV-A AND B, AND Wang, Joseph N. Eisenberg MORE, IN CLUSTERS OF UNEXPLAINED ACUTE FEBRILE University of Michigan, Ann Arbor, MI, United States ILLNESS IN CAMBODIA Zoonotic bacterial pathogens present significant threats to human Nary Ly1, Rafal Tokarz2, Nischay Mishra2, Stephen Sameroff2, health and compromise the economic well-being of pastoralist herding Komal Jain2, Agus Rachmat1, Sam An Ung3, Touch Sok3, Steven communities. The dynamics of transmission of pathogens such as Newell1, Ian Lipkin2, Dustin J. Harrison1 Brucella sp., the causative agent of Brucellosis, however, are poorly 1U.S. Naval Medical Research Unit – 2, Phnom Penh, Cambodia, 2Center understood. Prior studies have suggested that transmission of Brucellosis for Infection and Immunity, Mailman School of Public Health, Columbia cannot be sustained in the small to medium size herds characteristic of University, New York, NY, United States, 3National Institute of Public most pastoralist systems. These analyses, however, did not consider the Health, Cambodian Ministry of Health, Phnom Penh, Cambodia joint effects of animal trading between herds and inter-species mixing within herds. Using quantitative and ethnographic data collected in a Fevers of unknown origin constitute a substantial disease burden on pastoralist community in Laikipia, Kenya, we create a deterministic SIR patients in Southeast Asia with the majority of the cases remaining model of Brucella sp. transmission to examine how these transmission undiagnosed. To expand the breadth of possible infectious pathogens, heterogeneities may facilitate disease persistence. We explore dynamics we used MassTag PCR to test for the presence of 20 bacterial and viral of infection given various herd sizes and management strategies observed respiratory agents from 85 patients with unexplained respiratory illness in Laikipia Kenya. Specifically, we model a community that engages in representing six disease clusters that occurred in Cambodia between 2006 multispecies livestock raising, assuming different types and levels of and 2012. We detected potential pathogens in 62 (73%) of 85 total cases, interaction between herds. We find that transmission is unsustainable identifying a virus in 37 patients (44%) and a bacterium in 53 (62%) cases. in small herds of all species when in isolation, confirming prior research. In a cluster from Kandal province from August 2009, we detected a high Multi-species herds which include a high proportion of goats compared frequency of enterovirus 68 and human rhinoviruses. Among 22 cases that with large stock such as cattle can sustain itself for a longer time, but occurred during October 2009 in Kampong Speu province, we detected not indefinitely. Links through sales and purchases or other between- human respiratory syncytial virus B. Finally, a cluster of children < five years herd contact, however, create conditions where Brucella can transmit of age from the Ratanakiri province previously diagnosed with pneumonia, indefinitely. Though it is likely that herding strategies historically revealed infection from human respiratory syncytial virus A. These findings accommodate potentially devastating risks to herd health, it is possible provide insight into the etiologies of previously undiagnosed acute febrile that herders are unaware of threats, both to health and economic illness in Cambodia and point to the utility of multiplexed diagnostics wellbeing, presented by bacterial infections such as Brucellosis. In addition during disease outbreaks. to education programs on specific diseases, herders should be encouraged to cull animals showing obvious signs of illness as soon as possible and 1767 should be discouraged from selling such animals to minimize risks to human health. DIAGNOSTIC VALUE OF CLINICAL FEATURES FOR DIAGNOSING PNEUMONIA IN CHILDREN UNDER FIVE YEARS 1765 OF AGE: A SYSTEMATIC REVIEW AND META-ANALYSIS Clotilde Rambaud Althaus1, Fabrice Althaus2, Blaise Genton1, MARKETS AS HUBS OF RISKY CONTACT BETWEEN HUMANS Valérie D’Acremont1 AND WILD/DOMESTIC ANIMALS: CASE STUDIES FROM 1 2 REPUBLIC OF CONGO (ROC) AND DEMOCRATIC REPUBLIC OF Swiss Tropical and Public Health Institute, Basel, Switzerland, Department of Ambulatory Care and Community Medicine, Lausanne, Switzerland CONGO (DRC) The aim of this review is to assess the diagnostic value of clinical signs and Jariseta Rambeloson Zo1, Henri Banga-Mboko2, Roger Albert symptoms in identifying radiological pneumonia in children under the age Mbété2, Lonna Shafritz1, Tula Michaelides1, Jean-Luc Hornick3, of five and to review the accuracy of WHO criteria for diagnosing clinical Susan Zimicki1 pneumonia in developing countries. Electronic databases (Medline and 1FHI 360, Washington, DC, United States, 2Veterinarian and Zoonotic Embase) and reference lists of relevant studies were searched to identify Research Center, Brazzaville, Republic of the Congo, 3University of Liege, articles assessing clinical predictors of radiological pneumonia in children. Liege, Belgium 1697 potentially relevant studies were identified. Selection was based Traditionally, market studies have focused on elements of market dynamics on: design (diagnostic accuracy studies), target disease (pneumonia), such as the volume of wildlife and domestic animals being sold. We are participants (children below 5 years), setting (ambulatory or hospital care), examining markets through a different lens: to assess them as locations index test (clinical features), reference standard (chest radiography). Quality where humans - consumers, vendors, managers - are at increased risk of assessment was based on the 2011 Quality Assessment of Diagnostic transmission of zoonotic infections via exposure to live animals, animal Accuracy Studies (QUADAS-2) criteria. For each index test, sensitivity and products, and poor biosecurity practices. USAID’s PREVENT Project is specificity were calculated. Meta-analyses with the Bivariate model and studying eight urban markets in Brazzaville and two in Dolisie, ROC, as hierarchical SROC plots were done for index tests assessed in four or well as eight markets in Kinshasa, DRC. We conducted key informant more studies. Eighteen articles were included in the analysis. Age-related interviews, consumer exit interviews and then a household survey in the fast breathing (six studies, pooled sensitivity: 0.62 [95%CI 0.26-0.89]; market catchment areas (as determined by exit interviews) to learn where specificity: 0.59 [0.29-0.84]) and lower chest wall indrawing (four studies, people shop, what they buy and why. In addition, since February 2014 0.48 [0.16-0.82]; 0.72 [0.47-0.89]) showed poor diagnostic performance we have carried out monthly one-week full-day observations of vendor in the meta-analysis. Features with the highest pooled, positive likelihood stalls selling bush meat and/or poultry. This presentation will describe what ratios were: respiratory rate above 50/min (1.90 [1.45-2.48]), grunting we have learned to date about hygiene conditions, infrastructure, and (1.78 [1.10-2.88]), chest indrawing (1.76 [0.86-3.58]), and nasal flaring biosecurity practices in these markets and about the importance, diversity (1.75 [1.20-2.56]). Features with the lowest pooled negative likelihood

astmh.org 539 ratio were: cough (0.30 [0.09-0.96]), history of fever (0.53 [0.41-0.69]), the region of Madre de Dios . Determine the profile of Mycobacterium and respiratory rate above 40/min (0.43 [0.23-0.83]). No single clinical tuberculosis drug resistance in patients admitted to retreat in Madre de feature was sufficient for definitively diagnosing pneumonia. Combining Dios , Peru , 2007-2013 Observational, cross sectional study . Logbook clinical features in a decision tree may improve diagnostic performance, and monitoring of TB patients in retreatment Regional Health Direction but the addition of new point-of-care tests for diagnosing bacterial - Madre de Dios of 2007-2013 , included 111 patients diagnosed with pneumonia would help to reach an acceptable level of accuracy. tuberculosis smear ( +) , they were tested for sensitivity at the National Institute of Health (NIH ) . Frequencies established variables, measures of 1768 central tendency and dispersion in qualitative variables were used. The frequency of monoresistance 33.63 % (n = 37 ) , poliresistencia 9.09 % COMMUNITY ACQUIRED PNEUMONIA IN ADULT HOSPITAL (n = 10 ) , 45.45 % MDR - TB (n = 50) and 12.72% sensitive cases (n = ADMISSIONS NORTHERN VIETNAM; CLINICAL FEATURES 14 ) . The overall frequency of isoniazid resistance was 90.18 % (n = 92) AND ETIOLOGY , rifampicin 79.09 % (n = 87) , Streptomycin 18.18 % (n = 20) Etambutol 11.81 % (n = 13) and pyrazinamide 7.27% ( n = 8).The initial cultures 1 2 2 Behzad Nadjm , Ta Thi Dieu Ngan , Vu Quoc Dat , Rogier van were basciloscopias negative and 44.6% (n = 50) and 32.1% (n = 36) 1 1 2 1 Doorn , Peter Horby , Nguyen Van Kinh , Heiman Wertheim respectively. Only 1.8% (n = 2) showed HIV positive reaction. . In the 1Oxford University Clinical Research Unit, Hanoi, Vietnam, 2National present study we found that there is a high prevalence of resistant TB Hospital for Tropical Diseases, Hanoi, Vietnam in patients admitted to retreat in the Madre de Dios Region where HIV Community acquired pneumonia (CAP) causes an estimated 1 million positive reaction is not related to the presentation. MDR-TB were the most deaths in Asia each year including 160 000 amongst those aged 15 - 60 frequent type of resistance and isoniazid is the drug resistance most often years. Knowledge of local etiology is critical to formulating treatment generated in monoresistencia patients . guidelines and vaccination priorities. Despite this studies of the etiology of CAP in South East Asia have been limited. We conducted a study of 1770 the causes of CAP in adult patients admitted to 3 hospitals in Ha Noi, Vietnam. Adults with an infiltrate on chest radiography and one of: cough, INFLUENZA VACCINE EFFECTIVENESS IN THE TROPICS: dypnoea, fever (≥38.3C) or hypothermia (<36.0C), purulent respiratory MODERATE PROTECTION IN A SURVEILLANCE POPULATION secretions, bronchial breathing or rales on auscultation, leucocytosis IN BANGKOK BETWEEN AUGUST 2009 AND JANUARY 2013 or leucopenia that had not been residing in hospital or a long term Jens W. Levy1, Sriluck Simasathien2, Veerachai Watanaveeradej2, care facility in the 14 days prior to onset were eligible for admission. Piraya Bhoomiboonchoo1, Stefan Fernandez1, Richard G. Consenting patients were enrolled and recieved a standardized clinical Jarman3, Chonticha Klungthong1, Robert V. Gibbons1, Piyarat evaluation. All patients had routine haematology and biochemistry test Suntarattiwong4, In-Kyu Yoon1 results recorded and received blood culture, sputum culture and PCR for the following bacterial pathogens: Streptococcus pneumoniae, 1Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, Mycoplasma pneumoniae, M. amphoriforme, Chlamydophila pneumoniae, 2Phramongkutklao Hospital, Bangkok, Thailand, 3Walter Reed Army C. psittaci, Legionella pneumophila and L longbeachae. PCR was also Institute of Research, Silver Spring, MD, United States, 4Queen Sirikit performed for 14 respiratory viruses on nasal/throat swabs and/or Institute for Child Health, Bangkok, Thailand sputum. Urine was tested for pneumococcal antigen and L pneumophila Influenza in the tropics occurs year round with peaks that correspond serogroup 1. In a selection of cases where acute and convalescent serum variably to temperate regions. However, data on influenza vaccine was available serology for C. pneumoniae, M. pneumoniae, Orientia effectiveness (VE) in the tropics is sparse. We report on the effectiveness tsutsugamushi, Rickettsia typhi and R. prowazekii was also performed. of influenza vaccine to prevent medically attended laboratory confirmed Preliminary results only are available at this time, full results should be influenza from a sentinel surveillance study conducted at a Thai military available in time for presentation. Preliminary results show a case fatality medical facility in Bangkok, Thailand from August 2009 to January 2013. rate (died in hospital or palliative discharge) of 12/116 (10.3%). The Patients ≥6 months old presenting with influenza-like illness underwent rate of positivity for blood culture was low (5/112, 4.5%). Sputum PCR nasal/throat swabs which were tested by influenza RT-PCR. A case test- was positive for S. pneumoniae in 80/125 cases (64%), M. pneumoniae negative study design was used to evaluate VE. Of 2992 samples available in17/125 (13.6%), C. psittaci in 10/125 (8%), M. amphoriforme in 5/125 for analysis, 1058 (35.4%) were PCR-positive (cases) and 1934 (64.6%) (4%), C. pneumoniae and L. pneumophila in 1/125 (0.8%) each and there were PCR-negative (test-negative controls). Five hundred and eight were no cases of L. longbeachea. Clinical findings and outcomes will also (16.9%) of these patients reported being vaccinated within the previous be explored. 12 months. Periods of high and low influenza activity were defined based on publicly available Thai Ministry of Public Health data. Overall adjusted 1769 VE was found to be 51.6% (95%CI: 36.8, 63.1%). Adjusted point estimate for VE was highest in the 18-49 year age group (77.0%) followed PROFILE OF MYCOBACTERIUM TUBERCULOSIS DRUG by 6-23 months (55.1%) and 2-17 years (44.6%). Adjusted estimates were RESISTANCE IN A TROPICAL REGION OF PERU 2007-2013 not done for those ≥50 years of age due to small numbers. VE in patients with underlying disease was 75.5% compared to 48.6% in those without. J. Antonio Grandez-Urbina, J. Gonzalo-Rodriguez, S. Bedoya- VE appeared to be much higher during high versus low influenza activity Vidal, G. Grandez-Castillo, M. Sotelo, A. Santillan-Romero, R. periods. Among those who reported receiving vaccine 14 days-3 months Pichardo-Rodriguez, J. Oscar Tarazona-La Torre, Enver Terrazas prior to illness, VE was 55.8% (95% CI 26.6 to 74.1%), and tended Nuñez to decrease as the interval between vaccination and illness increased Universidad Ricardo Palma, Lima, Peru (46.8% at >3 to 6 months; 48.9% at >6 to 9 months; 31.5% at >9 to Tuberculosis in Peru is a public health problem . In terms of South 12 months). Our findings demonstrate moderate protection by influenza America is the second country in prevalence after Bolivia, and in America vaccination and support the utility of influenza vaccination in the tropics is the third after Haiti and Bolivia . In 2013 , WHO reports that in Peru including in very young children and those with underlying disease. Our Multidrug-resistance Tuberculosis ( MDR - TB ) exceeds the reported study also suggests that booster vaccination may be useful within several cases that year by Colombia , Ecuador , Argentina , Chile and the United months even when the formulation does not change. States throughout its territory , and ranks first in report for MDR - TB and Extreme drug resistance tuberculosis( XDR - TB ) . In Peru the regions with high prevalence of TB are Lima, Callao , Madre de Dios , but the profile of Mycobacterium tuberculosis drug resistance is not known in

astmh.org 540 1771 VF infections, which we followed for lifetime costs. Unreported cases were assumed to be asymptomatic infections with no costs. We included early SEASONAL INFLUENZA’S ASSOCIATION WITH SPECIFIC treatment costs of VF misdiagnosed as community acquired pneumonia. HUMIDITY IN THREE TROPICAL CENTRAL AMERICAN We costed VF by disease categories; (uncomplicated pneumonia [UP], COUNTRIES: HONDURAS, NICARAGUA AND COSTA RICA chronic/diffuse pneumonia with [CD] and without dissemination [CN], chronic pulmonary nodule [PN], and chronic pulmonary cavity [PC]). 1 2 3 Radina P. Soebiyanto , Wilfrido A. Clara , Jorge Jara , Maria L. Direct costs were VF diagnosis, treatment, and follow-up including 4 5 6 6 Matute , Angel Balmaseda , Jenny Lara , Mariel Lopez Moya , physician visits, ER, hospitalization, tests, procedures, and medications. VF 7 7 Marc-Alain Widdowson , Eduardo Azziz-Baumgartner , Richard K. epidemiology data were from literature and expert interviews. Treatment 1 Kiang and utilization were from published guidelines and 2 hour prepared 1NASA Goddard Space Flight Center, Greenbelt, MD, United States, interviews with 5 expert VF physicians. Hospitalizations were from the 2Centers for Disease Control and Prevention, Regional Office for Central 2012 California Patient Discharge Dataset and HCUP prices, medication America Region, Guatemala City, Guatemala, 3Universidad Del Valle de costs from average wholesale price minus 17% for contract pricing, Guatemala, Guatemala City, Guatemala, 4Ministry of Health of Honduras, physician visit were costed using CPT based Medicare estimates. Total Tegucigalpa, Honduras, 5Ministry of Health of Nicaragua, Managua, lifetime costs of 2012 VF incident cases in CA was $212 million (M), Nicaragua, 6Costa Rican Institute for Research and Education on Nutrition $51,743/person. UP accounted for 85% of our population and 31% of and Health, San Jose, Costa Rica, 7Centers for Disease Control and direct lifetime costs ($65.4M), CD 2.5% and 42% costs ($89.2M), CN Prevention, Atlanta, GA, United States 2.5% and only 11% of costs, pulmonary nodules and cavities 10% and Studies have demonstrated the association between seasonal influenza & 16% of costs, respectively, primarily due to cost of differential diagnosis meteorological factors. We previously showed that seasonal influenza in of cancer. Short term work loss costs were $6.4 M, and mortality another Guatemala, El Salvador & Panama was associated with specific humidity. $126 M. VF causes a large cost burden, especially for disseminated cases. In this work, we investigated the association in 6 departments from three Variation exists by geographic regions, insurance status and practice other Central America countries: Cortes & Francisco Morazan in Honduras; patterns. Areas for cost control exist. Managua in Nicaragua; Alajuela, Cartago & San Jose in Costa Rica. As an indicator for influenza activity, we used the weekly proportion of samples 1773 which tested positive for influenza in the period 2008-2013 from each country’s National Influenza Centers. Respiratory samples were collected THE NEED FOR ENHANCED LABORATORY BASED DIAGNOSIS from case-patients presenting with influenza-like illness or severe acute CAPACITY IN THE TESTING AND SURVEILLANCE FOR OTHER respiratory infection. Meteorological factors - rainfall, temperature & RESPIRATORY VIRUSES FROM PATIENTS REPORTING WITH specific humidity (SH) - were obtained from NASA’s satellites & models. We INFLUENZA LIKE ILLNESSES used logistic regression and adjusted for previous influenza activity & co- Nicholas Owor, Barbara Namagambo, Timothy Byaruhanga, circulating viruses (respiratory syncytial virus, adenovirus and parainfluenza John T. Kayiwa, Richard Chiza, Irene Nabukenya, Barnabus virus). We found that SH was proportionally associated (p<.05) Bakamutumaho, Julius Lutwama with influenza activity in all departments (Odds Ratio (OR)=1.2-1.6). Temperature was inversely associated with influenza activity in Alajuela of Uganda Virus Research Institute, Entebbe, Uganda Costa Rica (OR and 95% Confidence Interval=0.7(0.6-0.8)) & Cortes of The capacity of Laboratories to promptly identify particular strains Honduras (OR=0.8(0.7-0.9)). There was no statistical association (p<.05) or subtypes of organisms using modern diagnostic techniques has with rainfall in any locations. Among the meteorological factors, SH had become essential for rapid and efficient response to disease outbreaks the highest contribution (2-15%) to the model in all locations except in and preventing potential epidemic or pandemic spread. In Influenza Cortes. The model estimated influenza activity accurately (R=0.6-0.9) surveillance, patients present with symptoms that match the case for the final 6 months in all countries except Honduras. Time-frequency definition of Influenza Like Illnesses (ILI) and Severe Acute Respiratory analysis using Hilbert-Huang Transform showed that seasonal components Infection (SARI). Up to 24% of the samples collected and tested for of influenza activity was positively correlated with SH (R=0.2-0.6, p<.05), influenza at the National Influenza Centre are positive for influenza further corroborated the SH findings from logistic regression. Our results while the biggest percentage are negative. There are various respiratory highlighted influenza’s proportional association with SH in these countries, viruses and bacteria that affect humans which include adenovirus, human which was consistent with other studies in the tropics. Understanding rhinovirus A, coronavirus OC43, parainfluenza virus 1, parainfluenza of climate role in influenza may help in estimating epidemic timing and virus 3, respiratory syncytial virus B, human metapneumovirus, intensity. respiratory syncytial virus A, parainfluenza virus 2 and coronavirus 229E. Nasopharyngeal and oralpharyngeal swab specimens collected from 1772 patients presenting with ILI and SARI in 8 sentinel sites between December 2011- April 2014 in Uganda were tested for Influenza by RT- PCR, THE ECONOMIC BURDEN OF VALLEY FEVER IN CALIFORNIA subtyping and isolation. Of a total of 5931 samples tested; 645(10.9%) were positive for Influenza with 2(0.3%) co-infections, 405(62.8%) Leslie Wilson1, Jie Ting1, Michael MacLean2, Michael Peterson3, Influenza A and 238(36.9%) Influenza B; though, 5286(89.1%) were Nathan Stockamp3, Robert Libke3, Paul Brown4, Herbert Boro5, negative yet the patients presented with symptoms that match the Harold Lin5 case definition for ILI and SARI. This raises a need to establish baseline 1 University of California San Francisco, San Francisco, CA, United States, information on the prevalence of other respiratory pathogens that cause 2 Kings County Health Department, Kings County, CA, United States, upper and lower respiratory disease in populations through strengthening 3 University of California San Francisco Fresno, Fresno, CA, United States, laboratory diagnostic capabilities for the identification and characterization 4 5 University of California Merced, Merced, CA, United States, Kaiser of infectious agents likely to cause public health emergencies. There is Permanente-Fresno, Fresno, CA, United States need for more-simplified testing systems that enable researchers and Coccidioidomycosis or Valley Fever (VF) is a mycotic disease endemic to clinicians to perform multiplexed molecular diagnostics quickly and southwestern US. In California, where 33% of VF cases nationwide occur, easily. The results would be useful to guide future surveillance and case VF incidence increased from 4.2-10.8/100,000 in the period 2002-2012 management strategies involving other respiratory infections in Uganda. (157% increase). There is no complete study of the economic burden of VF to guide clinical care and public health planning. We estimated the total and per person lifetime direct and indirect cost of VF in 2012 US $. The 4,904 incident VF cases reported in 2012 in California were symptomatic

astmh.org 541 1774 between circulating influenza viruses and recommended A(H3N2) vaccine components for over a decade (1999-2012) in three regions of a southern EPIDEMIOLOGY OF RESPIRATORY VIRAL PATHOGENS FROM hemisphere country with continental dimensions (Brazil). A total of 237 SENTINEL SURVEILLANCE IN WESTERN CAMBODIA NEAR hemagglutinin sequences from Northeast (NE), Southeast (SE) and South THE BORDER WITH THAILAND (S) Brazilian regions were compared against the corresponding vaccine strains recommended annually by the WHO. We used MEGAv5.1 to infer 1 1 2 Ilin Chuang , Ans Timmerman , Melanie Melendrez , Youry nucleotide and amino acid distances between sequences and annual 3 3 1 4 Se , Nou Samon , Nichapat Uthaimongkol N , Stuart Tyner , Rith vaccine prototypes. PhyML was used for phylogenetic reconstructions 5 2 6 1 Sareth , Rick Jarman , Delia Bethell , Nitima Chanarat , Julie by Maximum Likelihood (ML) to infer the antigenic relationship between 7 1 1 Pavlin , Tippa Wongstitwilairoong , Piyaporn Saingam , But Sam viral samples and vaccine composition. We next compared the putative 3 8 8 9 1 El , Sok Touch , Ly Sovann , Lon Chanthap , Stefan Fernandez , effectiveness of the influenza vaccination in the three regions using 5 1 Phillipe Buchy , David Saunders hypothetical vaccination scenarios where alternative vaccine delivery 1Armed Forces Research Institute of Medical Sciences, Bangkok, timing and vaccine compositions (either Southern or Northern Hemisphere Thailand, 2Walter Reed Army Institute of Research, Silver Spring, MD, WHO recommendations) were considered (comparison following method United States, 3Armed Forces Research Institute of Medical Sciences, of Mello et al 2009). We found that, although influenza circulates in Battambang, Cambodia, 4U.S. Army Institute of Surgical Research, most (NE) or all (S,SE) months of year, the current Southern Hemisphere Houston, TX, United States, 5Institute of Pasteur, Cambodge, Phnom Penh, recommendation in Brazil is adequate for these regions. This was less Cambodia, 6Department of Paediatrics, Oxford, United Kingdom, 7Armed expected in the NE region, but we attribute it to the fact that most of the Forces Health Surveillance Center, Silver Spring, MD, United States, samples of the NE actually came from the its southernmost part (Bahia, 8Communicable Disease Control Department, Phnom Penh, Cambodia, at approximately 13oS), where influenza seasonality differ from the 9Armed Forces Research Institute of Medical Sciences, Phnom Penh, equatorial pattern of circulation. Our results show that WHO hemisphere Cambodia vaccine recommendation decisions must consider differences in influenza Little is known about influenza and other common respiratory viruses circulation patterns even within regions of a country. in remote populations along the Thai-Cambodia border in Western Cambodia. Real-time PCR for influenza was performed on combined nasal 1776 and throat specimens from outpatients presenting with influenza-like- illness (ILI) at 4 sentinel sites in Western Cambodia between May 2010 and CONCORDANCE BETWEEN SOLID AND LIQUID CULTURE FOR December 2012; a subset was further characterized by antigenic analysis, ANTITUBERCULOSIS DRUG SUSCEPTIBILITY TEST (DST) IN antiviral susceptibility testing and full genome sequencing for phylogenetic PERU analysis. PCR-negative ILI-specimens for influenza were cultured; cultured Giancarlo A. Valle1, Renato Quispe1, Tatiana Cáceres1, Eduardo negative specimens were then tested with RT-PCR for enteroviruses Gotuzzo2, Carlos Zamudio1, Carlos Seas2 and rhinoviruses (EV/RV) and enterovirus EV71. Among 586 ILI-patients 1 (median age 5 year, range 1-77 years), 168 (29%) tested positive for Instituto de Medicina Tropical “Alexander von Humboldt”, Universidad 2 influenza by RT-PCR and at least 1 respiratory virus was detected in Peruana Cayetano Heredia, Lima, Peru, Departamento de Enfermedades 258 (44%) patients. Influenza strains were highly related and matched Infecciosas, Tropicales y Dermatológicas, Hospital Nacional Cayetano circulating strains and although vaccination coverage was low, most strains Heredia, Lima, Peru matched the vaccine strains. No intrasubtype reassortment was detected. Löwenstein Jensen (LJ) has been the most widely technique for MTB Our Western Cambodian H1N1(2009) isolates were more closely related detection and DST. However, its long incubation and time to detection (based on full genome analysis) to 10 earlier isolates from Cambodia has led to develop alternative methods. Automated Mycobacteria Growth (94.4% genome conservation) compared to 13 Thai isolates (75.9% Indicator Tube (MGIT) constitutes a rapid alternative with comparable genome conservation. Aside from adenovirus (5.74%) and parainfluenza results, but manual MGIT (mMGIT) is favorable in low-resources areas virus (3.8%), detection of non-influenza viruses by viral culture was low because its lower cost. This study evaluates the concordance for DST of (<10%), with no detection of coronavirus, human bocavirus, human LJ and manual MGIT in a country with one of the highest prevalence of metapneumovirus and respiratory syncytial virus. We detected 5.9% of tuberculosis in the Americas. Sputum samples collected from respiratory non-polio enteroviruses among our culture-negative specimens: human TB suspects, enrolled in diagnostic trials during 2007-2011 in Lima Coxsackievirus types A4, A6, A8, A9, A12, B3, B4 and human echovirus (Capital of Peru), underwent LJ and MGIT. Only samples with positive types E6 and E9. We conclude that influenza epidemiology in this sample Capilia test for Mycobacterium tuberculosis (MTB) were included in the of isolates is following similar trends as observed elsewhere in Cambodia. DST analysis. Resistance to Isoniazid (INH), Rifampin (RIF), Streptomycin Further research to clarify the burden of adenovirus and non-polio (SM) and Ethambutol (EMB), as well as resistance to INH plus RIF (MDR) enteroviruses as etiologic agents for acute respiratory infections is needed were evaluated by Proportion Method (PM) in LJ medium and SIRE in Cambodia. system in MGIT. Comparison between performance of MTB detection and susceptibility patterns were assessed by Kappa indices. DST in both 1775 solid and liquid mediums was performed in 319 samples. PM-LJ and SIRE- MGIT detected resistance to INH: 21.6% (69/319) and 20.7% (66/319) DIFFERENT FROM EQUATORIAL BRAZIL, SOUTHERN (Kappa=0.92, p<0.001); RIF: 12.0% (38/319) and 11.0% (35/319) (Kappa HEMISPHERE WHO VACCINATION RECOMMENDATIONS ARE = 0.94, p<0.001); SM: 26.3% (84/319) and 25.1 (80/319) (Kappa = 0.82, ADEQUATE FOR MOST OF SOUTHERN PARTS OF BRAZIL p<0.001); and EMB: 11.9% (38/319) and 9.4% (30/319) (Kappa = 0.77, p<0.001). Furthermore, PM-LJ and SIRE-MGIT found 10.0% (32/319) 1 2 2 Wladimir J. Alonso , Priscila S. Born , Paola C. Resende , and 8.8% (28/319) as MDR cases, respectively (Kappa = 0.89, p<0.001). 3 2 2 Gonzalo Bello , Fernando C. Motta , Marilda M. Siqueira Manual MGIT emerges as a faster DST alternative to LJ in low resources 1Division of International Epidemiology and Population Studies, Bethesda, settings like Peru, with optimal concordance between LJ and mMGIT DST. MD, United States, 2Respiratory Viruses and Measles Laboratory, Rio de Janeiro, Brazil, 3AIDS and Molecular Immunology Laboratory, Rio de Janeiro, Brazil Influenza vaccination is the most important public health measure to prevent severe cases and deaths due to influenza infection. However, influenza viruses are constantly evolving, forcing continuous vaccine reformulation. In this study we investigated the annual genetic matching

astmh.org 542 1777 in an attempt to overcome the challenge of sample collection for the diagnosis of PCP in resource-limited settings. Fifty-one HIV patients COMPARATIVE PERFORMANCE OF WEEKLY TELEPHONE admitted to the hospital for respiratory symptoms in Cochabamba, Bolivia CALLS VERSUS WEEKLY HOME VISITS TO IDENTIFY CASES in 2010 were enrolled in the study. In addition to induced sputum, an OF INFLUENZA-LIKE ILLNESS AMONG A COHORT OF oral rinse, stool, and gastric secretion sample obtained by the string test PREGNANT WOMEN - GUATEMALA, 2013 were collected from each subject. Presence of PCP was evaluated by real- time quantitative PCR in a laboratory in Lima, Peru after the completion Chris Bernart1, Maria R. Lopez1, Marvin Xuya1, Juan C. Moir2, of sample collection. Of the 51 induced sputum samples collected, seven Saad B. Omer3, Jorge Jara1, Eduardo Azziz-Baumgartner4, Joe P. (13.7%) were positive for PCP. The oral rinse and string test each detected Bryan4, John P. McCracken1 PCP in one of the samples. PCP was not detected in any of the stool 1Universidad del Valle, Guatemala, Guatemala, 2Guatemalan Ministry of samples. The mortality rate of those diagnosed with PCP was 42.9% (3/7). Health, Guatemala, Guatemala, 3Emory University, Schools of Public Health Although other studies have reported the ability to diagnose PCP by the and Medicine and Emory Vaccine Center, Atlanta, GA, United States, use of PCR on oral rinse samples, our study was not able to repeat this. 4Global Disease Detection Branch, Division of Global Health Protection, Additionally, we were not able to detect PCP in either stool or gastric Atlanta, GA, United States secretion samples obtained by the string test (as has been used for the Influenza-like illness (ILI) is a leading cause of illness globally. A substantial detection of Mycobacterium tuberculosis in other studies). Our study does proportion of ILI is not detected by health services-based surveillance show that PCP is a cause of substantial morbidity in the HIV population in because not all persons with ILI seek healthcare; this may be exacerbated Bolivia, and it is possible that there were additional cases of PCP that went in settings where healthcare access or utilization is limited. Active undiagnosed in our study because of we were limited to induced sputum. surveillance through household visits can detect most ILI cases, but is costly Thus, improved diagnostic modalities are needed for the detection of PCP and labor-intensive. We followed a cohort of pregnant women in rural in resource-limited settings (delete all in italics). Although the rates of PCP Quetzaltenango, Guatemala for ILI (fever and cough or sore throat) by in our study, and in most resource-scarce settings, are significantly less using either weekly phone calls or home visits. Women with < 20 weeks than tuberculosis, our study shows that PCP is still a cause of substantial gestation were randomized 1:1 to either a weekly phone call or home morbidity and mortality in the HIV population in Bolivia. We were not able visit. Staff attempted up to 3 contacts per week to administer a study to detect PCP in stool samples, and our yield was very low in oral rinse questionnaire for ILI symptoms. Probable ILI cases identified by the call and gastric secretion samples obtained by the string test. It is possible that or visit were then evaluated within 24 hours by a nurse during a home there were additional cases of PCP undiagnosed in our study because we visit. Participants with ILI had a nasopharyngeal swab collected which was were limited to samples of induced sputum. Our study highlights the fact tested by polymerase chain reaction for respiratory syncytial virus (RSV), that improved diagnostic modalities are needed for the detection of PCP in human metapneumovirus, influenza A/B, parainfluenza virus 1/2/3, and resource-limited settings. adenovirus. During May-November 2013, 167 women were enrolled, of whom 85 (51%) were randomized to weekly phone calls and 82 (49%) 1779 to home visits. Surveillance was completed for 864 (63%) of the 1,364 expected person-weeks of follow-up by phone calls versus 1,010 (73%) of HAMSTER WEIGHT PATTERNS PREDICT THE INTENSITY AND the 1,381 expected person-weeks by home visits (p=0.01). Weekly follow- COURSE OF SCHISTOSOMA HAEMATOBIUM INFECTION up identified 9 ILI cases in the phone call group versus 13 ILI cases in the Thien-Linh Le home visit group (p=0.5 for ILI). We detected 3 infections in the phone call Stanford School of Medicine, Stanford, CA, United States group (adenovirus, parainfluenza-2, RSV) versus 6 infections in the home visit group (RSV, flu-B, parainfluenza types 2 and 3) (p=0.8). Although Although Syrian golden hamsters are a widely used host for experimental more costly and time consuming, home visits were more likely to have infection by Schistosoma haematobium, surprisingly little is known about successfully completed questionnaires than phone calls. The sample size the associated intensity and course of infection, making the use of these was inadequate to determine the difference in detection of lab-confirmed animals potentially unreliable. As such, we sought to define inexpensive, ILI cases between visits and calls. The choice between phone calls and simple, noninvasive, and accurate methods for assessing and predicting home visits to identify case-patients will depend on the surveillance the severity of disease in S. haematobium infected hamsters in order objectives and available resources. to prevent premature hamster sacrifice and unexpected morbidity and mortality. Through monitoring the weight and behavior of infected 1778 hamsters, we determined that the weight loss patterns of infected hamsters are highly correlated with commonly used measures of the ATTEMPTED ALTERNATIVE METHODS FOR THE DIAGNOSIS severity of infection (i.e. numbers of eggs passed in the stool and worm OF PNEUMOCYSTIS JIROVECI PNEUMONIA (PCP) IN HIV/AIDS burdens). In contrast, we found no significant correlation between hamster PATIENTS IN RESOURCE-LIMITED SETTINGS weight loss patterns and egg yields from liver and intestinal tissues. Our findings suggest that a more complex relationship exists among Melissa Reimer-McAtee1, Meredith Lora2, Carolina Mejia3, worm burden, fecundity, and egg passage in the feces than previously Monica Pajuelo4, Faustino Torrico3, Daniel Lozano Beltran3, Robert appreciated. Regardless, our data may be useful for workers seeking to Gilman5 optimize harvests of S. haematobium eggs and worm pairs from infected 1Tulane University, New Orleans, LA, United States, 2University of San hamsters for downstream applications. Francisco California, San Francisco, CA, United States, 3Universidad San Simon, Cochabamba, Plurinational State of Bolivia, 4Universidad Peruana Cayetano Heredia, Lima, Peru, 5Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States Despite the increased use of prophylactic therapy and improved access to antiretroviral therapy, Pneumocystis jiroveci pneumonia (PCP) remains one of the most common life-threatening opportunistic infections in HIV- infected patients worldwide. Difficulty obtaining an adequate sputum sample is often a barrier to the diagnosis of PCP in sputum-scarce patients where broncheoalveolar lavage is not available. This study examined the rates of PCP in HIV patients with respiratory symptoms in Cochabamba, Bolivia, and evaluated detection of PCP in samples other than sputum

astmh.org 543 1780 encoding leucine aminopeptidase (LAP) as the target, we refined delivery conditions, identifying electro-soaking (electroporation and subsequent LONG DELAY IN DIAGNOSIS AND HIGH LITHIASIS incubation) as the most efficient method to introduce small RNAs into PROPORTION IN SURGERY SUGGEST UNDERESTIMATION OF the fluke. We observed consistent knock down of LAP with low (2 μg/ HUMAN FASCIOLIASIS IN ARGENTINA ml) dsRNA concentrations. While this low concentration may reduce or obviate off-target effects, it also compromise the tracking of the RNAi 1 2 2 Veronica H. Agramunt , Roberto l. Mera Y. Sierra , Laura Sidoti , incorporation by fluorescent labeling. We also tested the effects of long 1 1 Maria Dolores Bargues , Santiago Mas-Coma and short interfering RNAs. While both long dsRNA and short interfering 1Departamento de Parasitologia, Facultad de Farmacia, Universitat RNA (siRNA) are equally effective at inducing a short-term knock down, de Valencia, Burjassot-Valencia, Spain, 2Catedra de Parasitologia y dsRNA induced more persistent silencing up to 21 days after treatment, Enfermedades Parasitarias, Facultad de Ciencias Veteriarias y Ambientales, suggesting that mechanisms of amplification of the interfering signal can Universidad J.A. Maza, S. Jose, Guaymallen, Mendoza, Argentina be present in this parasite. Persistent silencing from invasive stage for up A retrospective overview of human fascioliasis in Argentina highlighted the to 3 weeks (close to what it takes for the parasite to reach the liver) opens long delay with which many patients were diagnosed. Calculated delay the possibility of using RNAi for the validation of new putative therapeutic average of the time elapsed between the appearance of symptoms and targets confirmation of infection by appropriate diagnosis is very high, of 1262 days, nearly 3.5 years, and there are references about patients having 1782 suffered from symptoms for ten or more years without diagnosis. This suggests either infected subjects not looking for professional diagnosis PREVALENCE OF HAPLORCHIS TAICHUI AMONG HUMANS due to mild symptoms of low fluke burdens and/or misdiagnosis of AND FISH IN LUANG PRABANG PROVINCE, LAO PDR patients due to the non-pathognomonic clinical picture, easily confused Jong-Yil Chai1, Woon-Mok Sohn2, Tai-Soon Yong3, Keeseon with other diseases when the patient attends a health centre not used S. Eom4, Duk-Young Min5, Dongmin Lee4, Bong-Kwang Jung1, to dealing with fascioliasis. Moreover, the number of cases in which a Virasack Banouvong6, Bounnaloth Insisiengmay7, Bounlay surgical procedure contributed to the diagnosis when Fasciola hepatica Phommasack7, Han-Jong Rim8 specimens were unexpectedly found upon liver exploration appeared to be 1Seoul National University College of Medicine, Seoul, Republic of Korea, surprisingly high. In the majority, surgery was indicated due to abdominal 2Gyeongsang National University School of Medicine, Jinju, Republic of pain and biliary obstruction suggestive of lithiasis. The importance of Korea, 3Yonsei University College of Medicine, Seoul, Republic of Korea, intraoperative cholangiography was highlighted in cases in which, even 4Chungbuk National University School of Medicine, Cheongju, Republic of though gallstones were removed, evidence of obstruction observed during Korea, 5Eulji University School of Medicine, Daejeon, Republic of Korea, the cholangiography led to the finding of flukes. In most of these surgical 6Malaria Center, Luang Prabang, Lao People’s Democratic Republic, cases with lithiasis suspicion, the patient inhabited a large city (Buenos 7Ministry of Public Health, Vientiane, Lao People’s Democratic Republic, Aires, Córdoba, Mendoza, Tucuman) as opposed to a rural area where 8Korea University College of Medicine, Seoul, Republic of Korea attending a health care centre is less usual due to economic reasons or at least complicated due to the long journey thas has to be made. This This study confirmed the prevalence of the intestinal fluke Haplorchis additionally suggests a far greater underestimation of the problem in rural taichui (Trematoda: Heterophyidae) among people and fish in Luang areas. There were patients in whom fluke infection was detected only Prabang Province, Lao PDR. Fecal specimens were collected from 559 after a second surgical intervention. Both long delay in diagnosis and high riparian people (229 males and 330 females), residing in 4 Districts (Luang lithiasis proportion suggest that many patients are frequently overlooked Prabang, Xieng Ngeun, Pak Ou, and Nam Bak) and were examined by and pose a question mark about fascioliasis detection in the country. the Kato-Katz fecal smear technique. The overall helminth egg positive rate was 64.9%. The positive rate for small trematode eggs (STE), which 1781 may include H. taichui and other heterophyids, Opisthorchis viverrini, and lecithodendriids, was 15.2%. For recovery of adult helmniths, 10 STE- RNAI OPTIMIZATION IN FASCIOLA HEPATICA NEWLY positive people were treated with 40 mg/kg praziquantel and 15 mg/kg EXCYSTED JUVENILES: LONG DSRNA INDUCE MORE pyrantel pamoate, and then purged. Mixed infections with 3 Haplorchis PERSISTENT SILENCING THAN SIRNA species (H. taichui, H. pumilio, and H. yokogawai), a species of cestode (Taenia saginata), and several species of nematodes including Enterobius Nicolas DellOca1, Tatiana Basika1, Gabriel Rinaldi2, Ileana Corvo1, vermicularis and hookworms were found. The worm load for trematodes Estela Castillo3, Paul J. Brindley2, Jose F. Tort1 was exclusively high for H. taichui with an average of 7691 specimens 1Departamento de Genética, Facultad de Medina, Universidad de per infected person, followed by H. yokogawai (8.3 specimens) and H. la Republica, Montevideo, Uruguay, 2Department of Microbiology, pumilio (4.1 specimens). Out of 207 freshwater fish (17 species) purchased Immunology and Tropical Medicine, and Research Center for Neglected in a market in Luang Prabang District, 138 (67%) harboured H. taichui Diseases of Poverty, School of Medicine and Health Sciences, The metacercariae (metacercarial burden per fish; 520). Lower prevalence George Washington University, Washington, DC, United States, of fish and lower metacercarial density were observed for H. yokogawai 3Sección Bioquímica, Facultad de Ciencias, Universidad de la Republica, (52% and 50 per fish, respectively) and H. pumilio (18% and 3 per fish, Montevideo, Uruguay respectively). STE found in the surveyed population of Luang Prabang The exponential growth of the genomic and transcriptomics knowledge Province were verified to be those of intestinal fukes, particularly H. of parasitic flatworms allows the identification of several novel putative taichui. genes of unknown function. In trematodes RNA interference emerges as almost the only available tool to analyze gene function since classical 1783 genetics or other reverse genetics approaches still remain unavailable. Whereas this approach has been tested in several parasites of this group MOLECULAR MODELING OF ADENYLATE KINASE 1 AND 8 it has been optimized only in schistosomes likely reflecting the difficulties KDA CALCIUM-BINDING PROTEIN OF CLONORCHIS SINENSIS in the establishment of the technology as a routine tool. In this report Tai-Soon Yong1, Tae Yun Kim1, Eun Joo Chung2 we present progress in the optimization of this technology in the liver 1Yonsei University College of Medicine, Seoul, Republic of Korea, 2Korea flukeFasciola hepatica, causative agent of fasciolosis. This disease is National Institute of Health, Seoul, Republic of Korea one of the most problematic infections affecting livestock worldwide, and the increasing appearance of human cases had lead the WHO to Allosteric proteins involved in signal transduction transforms their recognized this disease as a reemeging zoonosis. Using a single copy gene molecular structure by binding co-factors, and thus they have a potential

astmh.org 544 for new drug development. In this study, two allosteric proteins, adenylate 1785 kinase 1 (ADK1) and 8 kDa calcium-binding protein (CaBP) of the Chinese liver fluke Clonorchis sinensis were cloned and modeled. C. sinensis EST IDENTIFICATION, CHARACTERIZATION AND EVALUATION clone Cs63 and Cs296 were cloned and sequenced. To compare them with OF RECOMBINANT ANTIGENS FOR THE DIAGNOSIS HUMAN other proteins of parasites, multiple sequence alignment and phylogenetic PARAGONIMIASIS analysis were performed. For molecular modeling, both sequences were subjected to SWISS-MODEL. Recombinant proteins generated bacterially Peter U. Fischer, Kurt C. Curtis, Kerstin Fischer, Samantha N. were used for their functional analysis. By BLAST search, Cs63 and Cs296 McNulty, Makedonka Mitreva, R. Reid Townsend, Gary J. Weil were confirmed as ADK1 and 8kDa calmodulin-like CaBP, respectively, Washington University School of Medicine, St. Louis, MO, United States and thus they were named as CsADK1 and CsCa8, respectively. Sequence Paragonimiasis is a foodborne trematode infection that affects 23 million and hydrophobicity of them were similar to those identified from parasitic people mainly in Asia. The parasite causes chronic cough with fever and helminthes. Molecular model of CsADK1 contained CORE, LID and NMP hemoptysis, and lung fluke infection is often confused with tuberculosis. domains and expected to transform its structure by binding co-factor-like We used a systems biology approach to identify antigens that might lead AP5. CsCa8 was predicted to have two distinctive EF-handed calcium- to improved diagnostic tests for this infection. Antibodies from patients binding sites by molecular modeling. Calcium ion could bind to each of with Paragonimus kellicotti were used to isolate antigens for proteomic EF-hands of CsCa8 model. Both recombinant proteins were functionally analysis, and RNAseq data from adult worms were used for protein active in biochemical assay. Results obtained from the study provide identification. Among the 22 most abundant identified proteins were structural basis of C. sinensis ADKs and CaBPs for the development of new a number of orthologues to known diagnostic antigen as well as novel anthelminthic drugs. candidates. Sequences for these proteins have 80-90% identity with amino acid sequences for orthologues in P. westermani. We expressed five 1784 P. kellicotti proteins as his-fusion proteins in E. coli, and these were used to raise antibodies in mice. Immunohistology performed with sections of AN OPEN-LABEL, RANDOMIZED, MULTICENTRIC STUDY OF adult worms showed that four of them were localized to the tegument, at TRICLABENDAZOLE FOR FASCIOLIASIS IN CHILDREN FROM the parasite host interface. In contrast, a known egg antigen was absent PERU from the tegument but present in developing and mature eggs. We Vicente Maco1, Luis Marcos1, Jaime Delgado2, Julio Herrera1, evaluated the diagnostic potential of these antigens by Western blot with José Nestares1, Angelica Terashima1, Frine Samalvides1, Eduardo sera from patients with paragonimiasis (from Missouri and the Philippines), Gotuzzo1 fascioliasis, and schisosomiasis and with sera from healthy North American controls. Two recombinant proteins showed high sensitivity and specificity 1 Institute of Tropical Medicine Alexander von Humboldt, Lima, Peru, as diagnostic antigens. Antibodies to the egg protein seemed to be specific 2 Health Care Center of Progreso, Asillo, Azangaro, Puno, Peru marker for patients with mature adult worm infections (with Paragonimus Human fascioliasis is an important public health problem in Latin America ova in stool or sputum). In conclusion, this study has identified and mainly affecting school-aged children in poor areas of the Andean Region characterized promising antigens for diagnosis of human paragonimiasis. (Peru and Bolivia). The optimal therapeutic scheme of triclabendazole (TCBZ), the recommended anthelmintic against the trematode Fasciola 1786 hepatica, has not been well defined (with a 10 mg/kg single dose being the most common regimen recommended), and clinical trials that assess IMPACT OF SCHISTOSOMA MANSONI IN SCHOOL-AGED effectiveness and tolerability in children are scarce. We aimed to evaluate CHILDREN LIVING IN KASANSA, DEMOCRATIC REPUBLIC OF the efficacy and tolerability of 2 therapeutic schemes of TCBZ in different THE CONGO areas of Peru. A total of 84 individuals (mean age ± SD: 9.27± 2.48 years) 1 2 3 with F. hepatica eggs in their stools (chronic infection) were enrolled in Mbuyi M. Kabongo , Sylvie Linsuke , Sylvain Baloji , Wilfried 4 3 1 an open-label, phase II clinical trial from areas located along the Peruvian Mukunda , Raquel Inocêncio da Luz , Christine Stauber , Richard 1 3 5 Andes. Individuals were randomly allocated into 2 groups: 44 received 2 Rothenberg , Jean Pierre Van geertruyden , Katja Polman , 3 2 dosages of TCBZ at 7.5 mg/kg each, with a 12 h interval post-prandially Marleen Boelart , Pascal Lutumba (group I-tested group), and 40 received a single dose of 10 mg/kg, post- 1Georgia State University, Atlanta, GA, United States, 2University of prandially (group II-standard group). The efficacy (parasitological cure) Kinshasa, Kinshasa, Democratic Republic of the Congo, 3University of was evaluated by the presence of eggs in stools at regular intervals and Antwerp, Antwerp, Belgium, 4Programme National de Lutte contre la up to 60 days post-treatment. Tolerability was evaluated by the presence Bilharziose et Parasitoses Intestinales, Kinshasa, Democratic Republic of the of clinical symptoms during the first week after TCBZ administration.A Congo, 5Institute of Tropical Medicine, Antwerp, Belgium parasitological cure was obtained in 100% of individuals from the tested- Schistosomiasis (SCH) is an important public health problem in developing group, and 95.0% in the standard-group (p>0.05). The most common countries and school aged children are the most affected. The aim of adverse event was biliary colic, documented in 25.0% in group II (95% CI= this study was to evaluate the impact of SCH on the population of [Ed.1] 11.9-38.9) on day 2, and in 20.5% in group I (95% CI= 7.8-33.7) school-aged children living in the high endemic area of Kasansa HZ in on day 4, possibly related to the expelling of the adult worms through the terms of malnutrition, anemia and low school performance. The overall biliary tract.In conclusion, the tested scheme was highly efficacious (100% health status of the children was poor with very high prevalence of S. cure rate) and tolerable, and it may be an optimal therapeutic scheme for mansoni infection (89.3%), malaria infection (65.1%), anemia (61.4%) the treatment of fascioliasis in children in Peru. This represents the largest and stunting (61.0%). School performance was also negatively affected series of children treated with TCBZ in a non-hospital setting and the with 54.6% of the children having failed at least one class. Regular largest in Peru. contact with river water was the most significant risk factor related to SCH infection. Anemia was influenced by SCH infection (p=0.003) and weak egg load was associated with stunting (p=0.04). However, due to poverty the causality between chronic malnutrition and anemia can be in either direction, potentially aggravated by SCH. Low school performance was mainly influenced by low income (<1 USD). Poverty exacerbated both health and school performance. Control measures are urgently needed to improve the health status of these children with in depth studies to demonstrate the causalities for each of the diseases in this population.

astmh.org 545 1787 each bladder: 1) the “proximal” urothelium (urothelium overlying the egg granuloma site); 2) the “distal” urothelium (urothelium from the opposite FIELD EVALUATION OF MINI-FLOTAC®, SPONTANEOUS side of the bladder relative to the granuloma site); and 3) granuloma tissue SEDIMENTATION TECHNIQUE IN TUBE OF TELLO, RAPID (subepithelial). Equal areas were harvested for each of the three tissue SEDIMENTATION TECHNIQUE BY LUMBRERAS AND KATO- sites (478,000-1,000,000 µm2/site). RNA was isolated, reverse transcribed KATZ FOR THE DIAGNOSIS OF ENTERIC PARASITES IN THE to cDNA, pre-amplified using the NuGen PicoSL WTA System, and then HIGHLANDS OF PUNO, PERU subjected to qPCR for uroplakin and housekeeping genes. Control vehicle- injected bladders exhibited subepithelial edema, normal urothelium, and Vicente Maco1, Catherine de Beaumont2, Davide Ianniello3, some inflammation. Egg-injected bladders, in contradistinction, featured Giuseppe Cringoli3, Quentin Bickle4, Angelica Terashima1, Eduardo markedly thickened, hyperplastic urothelium overlying areas of significant Gotuzzo1, Luis Marcos1 egg-associated inflammation. The proximal urothelium in egg-injected 1Institute of Tropical Medicine Alexander von Humboldt, Lima, Peru, bladders featured lower expression levels of uroplakin genes relative to 2Division of Infectious Diseases, Albert Einstein College of Medicine, New the distal urothelium. Bladder granulomas, such as those induced by York, NY, United States, 3Department of Pathology and Animal Health, urogenital schistosomiasis, may locally suppress the overlying urothelial University of Naples “Federico II”, Naples, Italy, 4London School of Hygiene barrier. This suppression may be a mechanism by which chronic bladder & Tropical Medicine, London, United Kingdom inflammation results in urothelial-related bladder dysfunction. Future work Enteric parasites show a peculiar distribution along the geography of will further define these mechanisms and their physiologic significance. Peru. Although a low-cost, reliable, local coproparasitological technique such as the Spontaneous Sedimentation Technique in Tube of Tello (SSTT) 1789 has been used over decades this has not been compared to standardized INFLUENCE OF HISTONE MODIFYING ENZYMES AND methods. Here, we compare the performance of SSTT with the mini- FLOTAC®, a coproparasitological technique under validation, in an area MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING where intestinal multiparasitism and fascioliasis are common. Rapid PATHWAY ON SCHISTOSOMA MANSONI SURVIVAL AND Sedimentation Technique (RST) by Lumbreras, and Kato-Katz (KK) were REPRODUCTIVE DEVELOPMENT included. We conducted a stool survey among school-aged children in Marina M. Mourão1, Luiza F. Andrade1, Livia G. Avelar1, Juliana Naupa Pampa, Calapampa, and Progreso, located in Azángaro (3859 m) A. Geraldo1, Fernanda S. Coelho1, Laila A. Nahum2, Raymond J. ® (Puno, Peru). Four techniques, mini-FLOTAC , SSTT, RST, and KK were Pierce3, Guilherme Oliveira1 performed. The sensitivity and negative predictive value (NPV) of each 1Genomics and Computational Biology Group, Instituto Nacional de technique were compared using the combined results of all positive Ciência e Tecnologia em Doenças Tropicais, Centro de Pesquisas René techniques as the “gold standard”. The inter-technique agreement (κ) Rachou, Fundação Oswaldo Cruz – FIOCRUZ, Brazil, Center for Excellence was also evaluated. A p value 10%): Entamoeba coli (77.2%), Blastocystis in Bioinformatics, Belo Horizonte, Brazil, 2Fundação Oswaldo Cruz, hominis (66.5%), Endolimax nana (50.3%), Iodamoeba buetschlii (15.0%), Faculdade Infórium de Tecnologia, Belo Horizonte, Brazil, 3CIIL, Inserm Entamoeba histolytica/dispar (12.0%), Giardia lamblia (11.4%), Fasciola U1019, CNRS UMR8204, University Lille Nord de France, Institut Pasteur hepatica (10.8%), and Chilomastix mesnili (10.2%). The area of study is de Lille, Lille, France hyperendemic for human fascioliasis with high prevalence in Ñaupa Pampa (25%). Mini-FLOTAC® showed a higher performance than SSTT and RST Praziquantel is an efficacious drug against schistosomiasis, however, there for H. nana (13 vs. 11 vs. 8) and F. hepatica (15 vs. 10 vs. 8), and higher is the risk of drug resistance development and therefore new drugs are sensitivities than SSTT, RST and KK for both helminths (p<0.05). The use of necessary. Although, the roles of Histone Modifying Enzymes (HMEs) and multiple techniques is an appropriate approach for highly endemic areas Mitogen-activated protein kinases (MAPKs) are unclear in schistosomes, where intestinal multiparasitism is common. SSTT is highly sensitive for they are increasingly approved as targets for drug development with a enteric protozoa, including when compared to standardized techniques rising number of inhibitors under development. In other organisms, HMEs such as the mini-FLOTAC®. Mini-FLOTAC® holds promise for the and MAPKs influence a number of tissue-specific biological activities such diagnoses of H. nana and F. hepatica in endemic areas. Notably, F. hepatica as cell survival, differentiation and proliferation. Here, we employed RNA infection continues to be highly prevalent among children in Azangaro, interference (RNAi) to elucidate the functional roles of 16 HMEs and 6 thus prompt and realistic field interventions are needed. genes involved in MAPK signaling pathway in S. mansoni. First, the HMEs and ePKs were identified in the predicted proteomes of Schistosoma 1788 mansoni, S. japonicum, and S. haematobium by HMM searches. Genes were annotated and selected regarding their putative function in the GRANULOMATOUS INFLAMMATION OF THE BLADDER parasite. One histone deacetylases (HDAC8), 10 methyltranferases COMPROMISES THE OVERLYING UROTHELIAL BARRIER (HMTs), 5 demethylases (HDM), SmRas, SmERK1, SmERK2, SmJNK, SmCaMK2, and Smp38 were chosen for experimental validation. RNAi Deborah Boyett and pharmacological inhibition were used to elucidate the functional role Stanford University, Stanford, CA, United States of HMEs and MAPK signaling pathway proteins in S. mansoni. Mice were Bladder granulomas can form as a result of retained suture material injected with schistosomula subsequent to RNAi and the development of from surgery, BCG treatment, urinary tuberculosis, and urogenital adult worms observed. The data demonstrate that SmHDAC8 and SmJNK schistosomiasis. We previously demonstrated in a mouse model of contributes to the parasite transformation and survival, whereas HDAC8, urogenital schistosomiasis that multiple urothelial barrier function genes PRMT3, KDM1/KDM2, SmERK, and Smp38 seems to be involved in egg (i.e., uroplakins) were downregulated on a whole bladder level after production as infected mice had significantly lower egg burdens and bladder wall injection with Schistosoma haematobium eggs. Given that female worms presented underdeveloped ovaries. Additionally, SmJNK egg-injected bladders exhibit hyperplasia of the urothelium overlying and Smp38 dsRNA treated worms exhibited tegumental damage. We resulting egg granulomata, we hypothesized that this hyperplastic also observed that Smp38 is involved in the activation of detoxification response was a response to egg inflammation-induced, regional enzymes. Our results help characterize the importance of HMEs and MAPK compromise of the molecular urothelial barrier. Anesthetized mice pathway in the normal development and survival of the schistosome underwent laparotomies and bladder exposure. Mice then underwent parasite and suggest some of these enzymes as useful drug targets to either bladder wall injection with S. haematobium eggs or a vehicle prevent schistosomiasis progression. control. Five days later, mice were sacrificed and their bladders harvested and fixed. Frozen sections of each bladder were stained with Cresyl violet. Laser microdissection was used to harvest RNA from three regions of

astmh.org 546 1790 results that showed that this transcript was induced differentially between juvenile resistant and susceptible parasite-exposed snails. Differential PROTEOMIC ANALYSIS OF BIOMPHALARIA GLABRATA expression of other stress genes, Hsp 90 (12-fold induction), Hsp 83 (40- HEMOCYTES DURING ENCAPSULATION OF SCHISTOSOMA fold induction) and Hsp 68 (5-fold induction) was also detected in these MANSONI SPOROCYSTS IN VITRO BS-90 snails responding to S. mansoni at the permissive temperature. These data, taken together with previous results provides further evidence 1 2 Marília Gabriela dos Santos G. Cavalcanti , Grzegorz Sabat , for the role of stress in the snail- host and schistosome interaction. Xiao-Jun Wu2, Utibe Bickham2, Timothy P. Yoshino2 1Federal University of Paraíba, João Pessoa, Brazil, 2University of Wisconsin, 1792 Madison, WI, United States In order to better understand the effector mechanisms associated IMMUNOMODULATORY PROTEINS OF SCHISTOSOMA with parasitic encapsulation reactions, whole hemolymph containing HAEMATOBIUM hemocytes was extracted from susceptible (NMRI) and resistant (BS-90) Luke F. Pennington1, Debalina Ray2, Chi-Ling Fu1, Yi-Ju Hsieh1, Biomphalaria glabrata strains and incubated in the presence or absence Michael H. Hsieh1 of newly-transformed Schistosoma mansoni sporocysts (Sp). After 18 h of 1Stanford University, Palo Alto, CA, United States, 2University of California incubation at 26 C, hemocyte capsules were isolated, frozen en bloc, cryo- San Francisco, San Francisco, CA, United States sectioned and subjected to laser capture microdissection to yield samples enriched for cells involved in hemocyte/Sp reactions for comparison to Approximately 120 million individuals are infected with Schistosoma hemocyte capsules without Sp. Isolated cryosections were analyzed by haematobium in sub-Saharan African alone. S. haematobium adult nanoLC-ESI-MS/MS for peptide isolation and sequencing. Putative protein worms lay eggs throughout the urogenital tract that induce a pronounced identifications were made by BLAST analyses vs. the non-redundant NCBI inflammatory response. To date very little is known about how S. protein database (db) and a 6-frame translated B. glabrata protein db haematobium is able to produce such a robust immune response while in VectorBase. Preliminary analyses revealed at total of 358 putatively evading immune clearance and immunity in many individuals. One identified proteins of which 71 were from Schistosoma spp. (mainly S. plausible explanation lies in parasite secreted immunomodulatory proteins, mansoni) and 287 were non-Schistosoma sequences. Significantly more however, due to the historical lack of a robust S. haematobium animal larval sequences were identified in NMRI/Sp capsules compared to BS-90/ model, very little is known about the role of these proteins in urogenital Sp samples, consistent with greater killing and larval rejection typically pathology. We have recently cloned a homolog of the IL-4-inducing seen in R hemocyte reactions in vitro. After normalizing the dataset across principle of S. mansoni eggs (IPSE), a protein originally identified in S. samples using total unique peptide counts for actin and tubulin, other mansoni, the sister species of S. haematobium responsible for hepatic notable immune-related observations were made: (a) A greater reduction and enteric schistosomiasis. S. mansoni IPSE has been shown to bind IgE of HSP70 peptides in BS-90/Sp capsules compared to NMRI/Sp (67% BS-90 on the surface of basophils in an antigen independent manner, and drive vs. 37% NMRI); (b) Frep2 was the only Frep identified in all samples, and basophil degranulation. To date it is not clear why it is advantageous only in the BS-90/Sp sample, implying an upregulation of Frep2 during for the parasite to secrete a protein capable of activating effector cells parasite encounters; (c) An upregulation of extracellular matrix/adhesion associated with anti-parasite responses. Intriguingly the S. haematobium proteins (dermatopontin2, HMG1, matrilin2 and α-integrin) only in BS-90/ IPSE homolog shares only 63% identity with its S. mansoni counterpart, Sp or BS-90 control samples suggesting possible roles in immune-related suggesting that the protein may have evolved to suit each species’ cell-parasite or cell-cell adhesion reactions; and (d) Enrichment of MnSOD, infectious niche. Furthermore studies with recombinant S. haematobium a potential effector molecule, in the BS-90 hemocyte capsules compared IPSE and S. mansoni IPSE suggest that the immunoglobulin isotype binding to those of NMRI snails. It is anticipated that continued mining of this rich profiles of IPSE differ across species. Despite IPSE’s sequence divergence dataset will yield valuable insights into hemocyte immune function. and differences in immunoglobulin binding, several important protein features appear to be conserved across species, including a nuclear 1791 localization sequence. Using a novel model of S. haematobium egg- induced pathology we have shown that immunization with IPSE prior to COMPARATIVE ANALYSIS OF GENE EXPRESSION IN S. haematobium egg injection alters urothelial inflammation. Together, SCHISTOSOMA MANSONI-EXPOSED BIOMPHALARIA these results suggest that S. haematobium IPSE also functions as an GLABRATA (BS90 STOCK) SNAILS MAINTAINED AT immunomodulatory protein, and plays an important and distinct role in PERMISSIVE AND NON-PERMISSIVE TEMPERATURES regulating tissue pathology in urogenital schistosomiasis. Matty Knight1, Ramzi Temanni2, Wannaporn Ittiprasert3, Andre 1793 Miller3, Yuan Li2, Najib M. El-Sayed2 1George Washington University, Washington, DC, United States, GENDER DEPENDENCE OF P53-RELATED ABNORMALITIES 2University of Maryland, College Park, MD, United States, 3Biomedical IN MOUSE BLADDER UROTHELIUM DUE TO SCHISTOSOMA Research Institute, Rockville, MD, United States HAEMATOBIUM EGG-INDUCED INFLAMMATION We have shown previously that the refractory phenotype in the Jared Honeycutt1, Olfat Hammam2, Michael H. Hsieh1 Biomphalaria glabrata BS-90 stock is a temperature-dependent trait. Thus, 1Stanford University School of Medicine, Stanford, CA, United States, using mild non-lethal heat (32°C) to induce stress genes such as Hsp70 2Theodor Bilharz Research Institute, Cairo, Egypt and Hsp90 prior to S. mansoni infection rendered these normally resistant snails susceptible. In order to determine differences between transcription The bladder urothelium is dramatically altered during Schistosoma profiles of these snails responding to early parasite infection when they haematobium infection (urogenital schistosomiasis). These alterations are ether resistant or susceptible, RNA samples from Schistosoma mansoni include hyperplasia, ulceration, dysplasia, squamous metaplasia and frank exposed juvenile BS-90 snails, maintained through several generations carcinogenesis. Defining the pathways that drive these urothelial changes either at the permissive (32°C), or non -permissive temperature (25°C) will contribute to a deeper understanding of how S. haematobium egg- were sequenced. Bioinformatic analyses of RNAseq datasets revealed a induced expulsion, hematuria, and bladder cancer develop in humans. preponderance of stress related transcripts in parasite-exposed BS-90 snails Defects in the function of the tumor suppressor gene p53 are evident maintained at the permissive temperature compared to similarly exposed in many cancers, including bladder cancer generally and schistosomal reference stock maintained at room temperature. For example, at 2 hours bladder cancer specifically. To identify any role p53 might play in urothelial post - exposure, a 77- fold induction of Hsp70 transcript was observed alterations due to urogenital schistosomiasis, we employed transgenic in susceptible BS-90 snails maintained at 32°C, corroborating earlier mice with tamoxifen-inducible cre recombinase activity in cells expressing

astmh.org 547 uroplakin-3a, a urothelial-specific gene (Upk3a-GCE mice). We confirmed 1795 specificity of cre expression in Upk3a-GCE mice by crossing them with TdTomato-floxed-EGFP reporter mice and administering tamoxifen to their PATTERNS OF REACTIVITY TO SCHISTOSOMA MANSONI EGG progeny. As expected, these progeny switched from TdTomato to EGFP GLYCAN ANTIGENS IN A POPULATION OF TREATMENT-NAÏVE expression in their bladder urothelium. We then crossed Upk3a-GCE mice KENYAN SCHOOL CHILDREN to p53-floxed mice. The resulting progeny (Upk3a-GCE+/wt;p53fl/wt) 1 2 3 were given tamoxifen or vehicle control to render them urothelial p53- Nina S. Prasanphanich , Jasmine A. Landry , Ryan E. Wiegand , 4 4 5 haploinsufficient or -intact, respectively. We then injected S. haematobium Eric M. Ndombi , Bernard Abudho , Diana K. Riner , Megan 1 4 5 eggs or control vehicle into the bladder walls of these mice. Three months L. Mickum , Diana M. Karanja , Daniel G. Colley , Richard D. 1 3 later, mice were sacrificed and their bladders subjected to histological Cummings , W. Evan Secor analysis (H&E staining). Male p53-intact, egg-injected mice exhibited 1Emory University, Atlanta, GA, United States, 2Macalester College, St. similar histological changes as their p53-haploinsufficient counterparts, Paul, MN, United States, 3Centers for Disease Control and Prevention, including urothelial hyperplasia and ulceration. In contrast, female p53- Atlanta, GA, United States, 4Kenya Medical Research Institute, Kisumu, intact, egg-injected mice featured no urothelial ulceration, whereas Kenya, 5University of Georgia, Athens, GA, United States their p53-haploinsufficient counterparts often had significant ulceration. Children in schistosomiasis-endemic areas develop partial resistance to Additionally, some egg-injected p53-haploinsufficient females exhibited infection as they age. This resistance is associated with immune responses, regions of squamous metaplasia. Thus, intact p53 activity seems to be including IgE and IgG, to parasite antigens. Anti-glycan antibodies can required, in a gender-specific manner, for urothelial homeostasis during kill parasite larvae in vitro and mediate resistance in some animal models S. haematobium infection in this model. Ongoing work includes (1) of helminth infection, but their significance in human Schistosoma examining histological changes in Upk3a-GCE+/wt;p53fl/wt mice beyond mansoni infection is still unclear. Plasma from S. mansoni-infected children 3 months after egg-injection, and (2) measuring alterations in the cell cycle demonstrate specific reactivity with several epitopes on schistosome status of the urothelium as a consequence of schistosomiasis. glycan microarrays. To explore the relationship of such antibodies with naturally-acquired partial immunity, we measured IgG and IgM to 1794 mock- and periodate-treated S. mansoni soluble egg antigen (SEA), and two parasite cross-reactive glycoproteins, keyhole limpet hemocyanin CD4+ T-CELL COUNTS IN WOMEN WITH UROGENITAL (KLH) and horseradish peroxidase (HRP), in plasma from a population of SCHISTOSOMIASIS treatment-naïve Kenyan school children. The ratio of antibody reactivity Elisabeth Kleppa1, Kari F. Klinge1, Kristine Lillebø1, Hashini N. with periodate-resistant (primarily non-glycan) versus total epitopes in Galaphaththi-Arachchige1, Sigve D. Holmen1, Siphosenkosi G. SEA increased, and antibodies to KLH and HRP glycans decreased slightly Zulu2, Birgitte J. Vennervald3, Pavitra Pillay2, Mathias Onsrud1, as children aged. These trends were especially pronounced throughout Svein Gunnar Gundersen4, Myra Taylor2, Eyrun F. Kjetland1 adolescence. The anti-glycan antibodies detected included a variety of IgG subtypes. Our results suggest that immune recognition of the glycan 1Oslo University Hospital, Oslo, Norway, 2University of KwaZulu-Natal, epitopes examined in this study are negatively associated with age, but Durban, South Africa, 3University of Copenhagen, Copenhagen, Denmark, some may warrant further investigation as diagnostics or indicators of 4Sorlandet Hospital, Kristiansand, Norway the length of exposure to schistosomes. Future studies on anti-glycan Schistosoma haematobium causes urogenital schistosomiasis and has been antibodies to other epitopes or of other isotypes/subtypes in naturally- shown to be associated with HIV in cross-sectional studies. Schistosomiasis acquired immunity, and on whether anti-glycan antibodies may be infection has been hypothesized to affect the HIV viral load and HIV involved in resistance to reinfection after praziquantel treatment could be disease progression. Schistosomiasis treatment has been shown to increase informative for vaccine development. CD4 counts in HIV negative individuals. Between May and October 2013, a CD4 count was done in 797 young women who were invited for a 1796 gynecological examination. One urine sample was collected from all, and microscopy for schistosome ova was done. HIV testing was done in 769 CHILDREN WITH CEREBRAL MALARIA LACK women, of which 123 were HIV positive (16.0%). The mean CD4 count SERORECOGNITION OF A DISTINCT PFEMP1 SUBSET was 864 x106 cells / L. It was lower in the HIV negative group than in 1 2 1 the HIV positive group (931 vs. 511 p < 0.001). Urinary schistosomiasis Mark A. Travassos , Amadou Niangaly , Jason A. Bailey , Amed 1 2 3 1 was not associated with a lower CD4 count (862 in urine negative vs. Ouattara , Drissa Coulibaly , Kirsten Lyke , Matthew B. Laurens , 4 4 4 871 in urine positive women, p=0.75). Likewise, there was no significant Jozelyn Pablo , Algis Jasinskas , Rie Nakajima-Sasaki , Andrea A. 1 1 3 association between CD4 counts between women with moderate or Berry , Matthew Adams , Shannon Takala-Harrison , Bourema 2 2 5 severe degree genital schistosomiasis and women without genital lesions Kouriba , Abdoulaye K. Kone , J. Alexandra Rowe , JoAnn 6 2 2 (846 vs. 829, respectively, p= 0.70). Further there was no significant Moulds , Ogobara K. Doumbo , Mahamadou A. Thera , Philip L. 4 1 difference between schistosomiasis positive and negative women were Felgner , Christopher V. Plowe found after stratifying for HIV (data not shown). This cross-sectional 1Howard Hughes Medical Institute/Center for Vaccine Development, study did not show any significant difference in CD4 T-cell counts when Baltimore, MD, United States, 2Malaria Research and Training Center, comparing women with and without urogenital schistosomiasis. Bamako, Mali, 3Center for Vaccine Development, Baltimore, MD, United States, 4University of California Irvine, Irvine, CA, United States, 5University of Edinburgh, Edinburgh, United Kingdom, 6LifeShare Blood Centers, Shreveport, LA, United States Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP1) antigens play an important role in parasite sequestration and host immune system evasion. Acquired antimalarial immunity is at least partially due to antibodies directed against highly variable antigens like PfEMP1 that are present on the red blood cell surface. However, the PfEMP1 antigenic domains that drive this immune-mediated protection have not been identified. We have previously shown that infected erythrocytes from persons with cerebral malaria express a distinct “stealth” PfEMP1 group that do not bind the endothelial receptor CD36. We hypothesized that children with cerebral malaria lack serorecognition to a subset of PfEMP1s

astmh.org 548 that is subsequently recognized in convalescence. A protein microarray 1798 was printed with 171 fragments of PfEMP1s based on the 3D7 reference genome. For comparison, 268 diverse apical membrane 1 (AMA1) RNA-SEQ ANALYSIS OF WHOLE BLOOD FROM MALARIA- fragments, 20 merozoite surface protein 1 (MSP1) fragments, and 30 Rh5 SUSCEPTIBLE AND IMMUNE CHILDREN REVEALS AN fragments were also included on the array, based on sequences derived EARLY PRO-INFLAMMATORY RESPONSE TO PLASMODIUM from field samples. Reactivity was measured in 195 serum samples from FALCIPARUM INFECTION THAT CORRELATES WITH CONTROL Malian children, including 43 cases of cerebral malaria and age-matched OF PARASITE GROWTH: A PROSPECTIVE STUDY IN MALI controls who were healthy or had uncomplicated malaria. Children with cerebral malaria had lower seroreactivity to stealth and non-stealth Tuan M. Tran1, Aissata Ongoiba2, Marcus Jones3, Jeff Skinner1, PfEMP1 antigen variants than both healthy controls and uncomplicated Shanping Li1, Safiatou Doumbo2, Didier Doumtabe2, Younoussou malaria controls. Seroreactivity to AMA1, Rh5, and MSP1 variants tested Kone2, Aboudramane Bathily2, Seydou Dia2, Moussa Niangaly2, did not increase from acute cerebral malaria illness to convalescence, but Abdrahamane Traore2, Charles Dara2, Jules Sangala2, Ogobara seroreactivity to four stealth PfEMP1 fragments increased, suggesting K. Doumbo2, Pratap Venepally3, Kassoum Kayentao2, Boubacar that a lack of immunity to a subset of PfEMP1s may be associated with Traore2, Ewen F. Kirkness3, Peter D. Crompton1 vulnerability to cerebral malaria. 1Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, United States, 2Mali 1797 International Center of Excellence in Research, University of Sciences, Technique and Technology of Bamako, Bamako, Mali, 3Genomic Medicine CYTOKINE RESPONSES TO THE VAR2CSA VACCINE Group, J. Craig Venter Institute, Rockville, MD, United States CANDIDATE IN CORD BLOOD FROM BENINESE NEWBORNS Non-sterile, antibody-mediated immunity that reliably protects from febrile Nadine Fievet1, Komi Gbedandé2, Gilles Cottrell1, Bertin Vianou2, malaria is acquired gradually through repeated Plasmodium falciparum Carine Agbowaï2, Samad Ibitokou2, Marita Troye-Blomberg3, Thor infections; however, the nature of cellular immune responses at the Theander4, Ali Salanti4, Achille Massougbodji2, Nicaise Tuikue onset of clinically apparent versus clinically silent blood-stage infections Ndam1, Philippe Deloron1, Adrian Luty1 in children is unclear. In a prospective study in Mali, we collected whole blood RNA, PBMCs and plasma from healthy, uninfected children aged 1IRD UMR216 “Mère et enfant face aux infections tropicales”, Paris, 6-11 years (n=79) before the 6-month malaria season and from the same France, 2Centre d’Etude et de Recherche sur le Paludisme Associé à la children during their first P. falciparum infection of the ensuing season— Grossesse et à l’Enfance, Faculté des Sciences de la Santé, Cotonou, detected retrospectively through bi-weekly active surveillance by PCR. Benin, 3Department of Immunology, Wenner-Gren Institute, Stockholm We used RNA-seq to compare whole-blood transcriptomes of children University, Stockholm, Sweden, 4Department of International Health, whose clinically silent infections never progressed to fever (immune, Institute of International Health, Immunology and Microbiology, University n=21), children whose infections progressed to fever within 2-14 days of Copenhagen, Copenhagen, Denmark (late fever, n=32) and children who were febrile at the time of infection The STOPPAM consortium conducted 2 longitudinal cohort studies (early fever, n=26). We found that baseline transcription profiles before in pregnant women in Benin and Tanzania in order to evaluate the the malaria season distinguished children whose future P. falciparum immunopathological consequences of infections with Plasmodium infections either progressed to fever or not, including upregulation of falciparum during pregnancy for the newborns. In order to study the B-cell-receptor signaling pathways in immune children. Transcription exposure of the foetal immune system to parasite-derived antigens in profiles induced by the first detected P. falciparum infection of the season utero, we evaluated both, cytokine (IL10, IL12, IL13, IL17, IFN-γ, TNF-α revealed upregulation of pro-inflammatory genes in immune versus late responses and T cell IFN-γ specific responses to the vaccine candidate fever children, despite both groups having similar levels of parasitemia and antigen DBL5 domain of VAR2CSA as a function of placental infection the clinical absence of fever initially. In addition, this early upregulation with P. falciparum. In Come, southwestern Benin, we conducted a of pro-inflammatory genes was associated with slower subsequent longitudinal prospective study of ~1000 pregnant women. Women at parasite growth rates in vivo. In ongoing work, we are testing hypotheses ≤24 weeks of pregnancy were enrolled and followed at each antenatal generated by this study at the protein level and in functional assays visit until delivery. For the immunological sub-study of the cord blood using contemporaneous PBMCs and plasma samples from the same mononuclear cellular (CBMC) responses to VAR2CSA-DBL5 in vitro, a children. Molecular and cellular signatures that correlate with protection group of 200 pregnant women was selected at delivery on the basis of from malaria are yielding novel insights into the mechanisms underlying their history of infection with P. falciparum (uninfected during pregnancy/ naturally acquired immunity to malaria. The resulting datasets may inform infected during pregnancy but uninfected at delivery/infected at delivery). the development of interventions that prevent or mitigate malaria disease. Those harbouring P. falciparum infections at delivery were matched by gravidity and gestational age with mothers with no infection and those with no history of infection earlier in the pregnancy. The amounts of IL10, IL12, IL13, IL17, IFN-γ and TNF-α produced in response to mitogen (PHA) and to VAR2CSA-DBL5 were quantified in supernatants of stimulated CBMC. The ex vivo frequencies of IFN-γ secreting CD4 and CD8 T cells in response to PHA and VAR2CSA domains were also evaluated. At the time of writing, all data have been collected, cytokine concentrations have been evaluated and multivariate analyses are under way. Results will be discussed in the context of cytokine profiles that reflect the in utero acquisition of a specific cellular memory response to the vaccine candidate.

astmh.org 549 1799 1800 IMMUNE CHARACTERIZATION OF PLASMODIUM SINGLE-CELL TRANSCRIPTIONAL ANALYSIS OF MALARIA- FALCIPARUM PARASITES WITH A SHARED GENETIC SPECIFIC CD4+ T LYMPHOCYTES FOLLOWING PFSPZ SIGNATURE: VARIANT SURFACE ANTIGENS AND VAR VACCINATION AND PROTECTION IN HUMANS REPERTOIRES Kailan Sierra-Davidson1, Adam Wheatley1, Patricia Darrah1, Amy K. Bei1, Ababacar Diouf2, Kazutoyo Miura2, Daniel B. Greg Finak2, Madhu Prabhakaran1, Pratip Chattopadhyay1, Mario Larremore3, Ulf Ribacke1, Gregory Tullo2, Eli L. Moss4, Daniel Roederer1, Helen McShane3, Kim Lee Sim4, Stephen Hoffman4, E. Neafsey4, Rachel F. Daniels1, Amir E. Zeituni2, Iguosadolo Raphael Gottardo2, Adrian McDermott1, Robert Seder1 Nosamiefan1, Sarah K. Volkman1, Ambroise D. Ahouidi5, Daouda 1National Institute of Allergy and Infectious Diseases, Bethesda, MD, Ndiaye6, Tandakha Dieye5, Souleymane Mboup5, Caroline O. United States, 2Vaccine and Infectious Disease Division, Seattle, WA, Buckee3, Carole A. Long2, Dyann F. Wirth1 United States, 3Jenner Institute, Oxford, United Kingdom, 4Sanaria, 1Harvard School of Public Health, Boston, MA, United States, 2Laboratory Rockville, MD, United States of Malaria and Vector Research, National Institute of Allergy and Infectious Intravenous immunization with highly purified, radiation-attenuated Diseases, Bethesda, MD, United States, 3Harvard School of Public Health, parasites (PfSPZ Vaccine) is safe, immunogenic and confers high-level Center for Communicable Disease Dynamics, Boston, MA, United States, protection against controlled malaria infection in humans. Protection was 4Broad Institute of Massachusetts Institute of Technology and Harvard, associated with a dose-dependent increase in PfSPZ-specific antibodies, Boston, MA, United States, 5Laboratory of Bacteriology and Virology, CD4+ and CD8+ T cell responses. Heretofore, multi-parameter flow Hopital Aristide Le Dantec, Dakar, Senegal, 6Laboratory of Parasitology and cytometry has been used to characterize the magnitude and quality Mycology, Dakar, Senegal of PfSPZ-specific T cell responses following vaccination or infection. To As malaria transmission intensity has declined in some regions, substantially expand the analysis of such responses, we performed high- Plasmodium falciparum parasite populations are displaying decreased resolution, quantitative transcriptome analysis of PfSPZ-specific CD4+ T clonal diversity resulting from the emergence of many parasites with cells. Accordingly, PfSPZ-specific CD4+ T cells expressing the costimulatory identical genetic signatures. We have monitored genetically identical marker CD154 (CD40L) were sorted following in vitro activation with parasite clusters from 2006-2013 in Thiès, Senegal, and we have sporozoites. We first demonstrate that ~30 percent of PfSPZ-specific characterized the immune response against these parasites. We focus CD154+ CD4+ T cells do not produce IFN-γ, IL-2 or TNFα, the most common on one cluster of identical parasites that was present in 24% of clinical cytokines used to assess T cell responses. This finding highlights the isolates in 2008 and declined to 3.4% of clinical isolates in 2009. We increased sensitivity of the CD154 capture assay for broader assessment of studied the susceptibility of 2 representative common genetic signature antigen-specific responses. Furthermore, isolation of live malaria-specific (CGS) parasites and 1 representative non-CGS parasite and measured the CD4+ T cells permits downstream mRNA analysis using valved microfluidic infected RBC IgG reactivity for 109 individual plasmas distributed between chips from Fluidigm. Quantitative expression of ~100 genes can be rapidly both years by variant surface antigen (VSA) flow cytometry. By VSA flow, analyzed from isolated samples of single antigen-specific T cells. Initial the non-CGS parasites are similary recognized by plasma IgG from 2008 transcriptome analysis of protected subjects revealed that malaria-specific and 2009, but reactivity is increased in 2009 compared to 2008 for the CD4+ T cells express a unique gene expression signature that is distinct CGS parasites. We characterized the var genes expressed by CGS parasites from influenza-specific CD4+ T cells in the same individual. These data by var Ups qRT-PCR and by sequencing using degenerate DBL1alpha will serve as an internal control to compare virus- and parasite-specific domain primers. We observed that the CGS parasites expressed the same responses. We are currently analyzing the gene signature of PfSPZ-specific var Ups classes, and the same dominant var repertoires as identified by CD4+ T cells from vaccinated and protected subjects prior to challenge both DBL1alpha sequence analysis as well as RNAseq. Additionally, we vs. nonvaccinated controls during infection. Overall, this analysis should used network analysis to compare the diversity of the var repertoires advance our understanding of the heterogeneity of parasite-specific CD4+ with that of globally diverse parasites. We generated a var sequence T cell responses at the single-cell level and provide insights into how CD4+ network that shows that the var repertoires of CGS-1 and CGS-2 overlap T cells may influence protection against human malaria infection. substantially, while the repertoire of the non-CGS parasite is unique at the level of globally diverse parasites. Taken together, our work indicates 1801 that these CGS parasites express similar var genes, more than would be expected by chance in the population, and there is year-to-year variation IMMUNOLOGICAL PROFILING AFTER SPOROZOITE in immune recognition of these CGS parasites at the level of surface IMMUNIZATION UNDER CHEMOPROPHYLAXIS IN THE expression of VSAs. We are currently expanding these findings to other CONTROLED HUMAN MALARIA INFECTION MODEL large clusters such as one that emerged in 2009 and persisted at a high Robert W. Sauerwein frequency (26-30%) for multiple years before disappearing from the population in 2013. Radboud University Medical Center, Nijmegen, Netherlands A unique tool to study malaria immunology and efficacy of immunisation strategies form Controlled Human Malaria Infections (CHMI) and has proved to be a reproducible, predictable and safe method of inducing Plasmodium falciparum (Pf) malaria. An efficient method for induction of complete protection in humans was achieved by exposing human subjects to Pf-infected mosquitoes while taking blood-stage suppressive chloroquine prophylaxis. When tested in clinical trials, this protocol induced > 95% clinically and parasitologically sterile protection against a standard challenge infection. Longlasting CPS-induced protection was primarily mediated by immunity to sporozoite and liver stages rather than to asexual blood-stages. This opens opportunities to explore mechanisms of protective immunity, allowing the search for immune correlates/ signatures of protection and clinical development of a whole sporozoite based vaccine. Humoral and cellular immune responses associated with protection to Plasmodium falciparum parasites will be presented.

astmh.org 550 1802 time in the AN group. However AN treatment provided longer protection from gametocytaemic relapse and/or reinfection. P. vivax gametocytes GENE PROFILING IN NAÏVE AND SEMI-IMMUNE COLOMBIAN were found to be cleared very rapidly and along with the asexual blood INDIVIDUALS SUBJECTED TO EXPERIMENTAL CHALLENGE stages upon treatment with AL or AN, highlighting the fundamental WITH PLASMODIUM VIVAX SPOROZOITES differences between the P. falciparum and the P. vivax parasite species. This study represents the first direct comparison of LM, MF and RTPCR on 1 2 Myriam Arévalo-Herrera , Monica Rojas-Peña , Andres F. a large number of field isolates. It provides clear evidence that magnetic 1 1 2 1 Vallejo , Nora Martinez , Greg Gibson , Sócrates Herrera fractionation is superior to light microscopy and can be used to detect 1Caucaseco Scientific Research Center, Cali, Colombia,2 School of Biology, gametocytaemic patients under field conditions with similar sensitivity Georgia Institute of Technology, Atlanta, GA, United States and specificity as RTPCR. Furthermore this study illustrates fundamental In the context of a Plasmodium vivax malaria vaccine program, a total differences between ACT mediated clearing of P. falciparum and P.vivax of 16 Colombian malaria naïve (n=7 from Cali) and semi-immune (n=9 gametocytes and describes differences in the effect of AL and AN on P. from Buenaventura) volunteers were subjected to an experimental falciparum gametocytes. P.vivax sporozoite infectious challenge using direct infected Anopheles mosquito bites (2-4) and followed up to determine the prepatent period, 1804 immune response and clinical outcome of the infection. Volunteers were closely monitored and treated as soon as malaria infection was detected DEVELOPMENT AND EVALUATION OF A SIMPLIFIED by microscopy. The study offered a unique opportunity to assess the MOLECULAR DIAGNOSTIC PLATFORM FOR MALARIA: THE gene expression profile induced by P. vivax malaria infection in naïve DIRECT ON BLOOD PCR-NALFIA SYSTEM and previously infected human volunteers Blood samples were used for Petra Mens, Johanna Roth, Laura de Bes, Henk Schallig immunological analyses and RNA preserved in Tempus tubes was isolated for transcriptomic analyses. We used a Fluidigm nanofluidic qRT-PCR array Royal Tropical Institute, Amsterdam, Netherlands to profile the expression of 92 genes in the 16 individuals across 6 time- Molecular tools allow for specific and sensitive malaria diagnosis, points following infection. The genes were chosen to represent 10 axes but current formats, like PCR with gel-electrophoresis, are difficult to of variation that describe major components of transcriptional variation in implement in resource poor settings. Therefore, a simple, fast, sensitive peripheral blood. Strong covariance of transcript abundance was observed and specific molecular diagnostic platform, direct on blood (db)PCR for 8 of these axes, 2 of which correspond to the first two overall principal combined with nucleic acid lateral flow immunoassay (NALFIA) to detect components of variation. The results show that there is strong up- amplified PCR products of Plasmodium, including species differentiation, regulation of an interferon-response axis at the peak of parasitemia, but a and human GAPDH (internal amplification control) was developed. This down-regulation of the inflammatory response at the same time. Another platform does not require DNA extraction and circumvents complex read- set of transcripts was observed to be significantly up-regulated both in out system. The platforms was evaluated under laboratory conditions, a the naïve samples and at the peak of parasitemia, but across the entire multi country ring trial and in two malaria endemic countries (Burkina Faso experiment the difference between the naïve and pre-immune samples and Thailand). Analytical sensitivity and specificity of the dbPCR-NALFIA in was minor relative to among individual variation. Nevertheless, these a single laboratory evaluation was >95% and the test was able to detect results strongly suggest that whole transcriptome profiling will uncover less than 1 parasite/µl blood. All four laboratories in the ring trial reported a set of genes that respond differently to infection as a function of the ease of use of the system and could successfully perform the protocol. degree of prior exposure to malaria.. Overall laboratory inter-variability was low and the agreement of reported results was high. Overall k-value was 0.89 (95% CI: 0.83 - 0.94; p<0.001). 1803 Overall test sensitivity and specificity was >95% with very small confidence intervals. Field evaluations by local staff without prior training in GAMETOCYTE CLEARANCE IN MELANESIAN CHILDREN performing the dbPCR-NALFIA in malaria endemic countries, Thailand and TREATED FOR PLASMODIUM FALCIPARUM AND P. VIVAX Burkina Faso, were performed. In Burkina Faso (P. falciparum environment) MALARIA WITH ARTEMISININ COMBINATION THERAPY the relative sensitivity was 94,8% and relative specificity 82,4% compared to microscopy and 93,3% and 91.4% compared to RDT. In Thailand 1 2 3 Stephan Karl , Moses Laman , Tamarah Koleala , Anna (P. vivax environment) the relative sensitivity and relative specificity was 4 2 3 2 Rosannas-Urgell , Brioni Moore , Peter Siba , Timothy G. St.Pierre , 93,4% and 90,9 respectively compared to microscopy and 95,6% and 1 2 Ivo Mueller , Timothy Davis 87.1 % compared to RDT. These numbers are an underestimation of test 1Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia, performance as the results are not PCR corrected. The prototype dbPCR- 2The University of Western Australia, Perth, Australia, 3Papua New Guinea NALFIA test will now be moved forward in diagnostic test development Institute of Medical Research, Madang, Papua New Guinea, 4Antwerp (supported by EU funding: www.diagmal.eu) to provide a molecular Institute of Medical Research, Antwerp, Belgium diagnostic test to detect malaria in for example near elimination settings. We present a detailed analysis of gametocyte clearance in Melanesian The final format will include a closed transfer unit to reduce possible children infected with Plasmodium falciparum or P. vivax and treated with workspace contamination with amplicons. Funding: EU FP7 grant 601714: either Artemether-Lumefantrine (AL) or Artemisinin-Naphthoquine (AN). In Translation of the direct-on-blood PCR-NALFIA system into an innovative addition, a detailed comparison of three methods, namely standard light near point-of-care diagnostic for malaria microscopy (LM), magnetic fractionation (MF) and reverse transcriptase polymerase chain reaction (RTPCR) for detection of Plasmodium falciparum and Plasmodium vivax gametocytes is presented. Children (0.5-5 years) from the north coast of Papua New Guinea were randomly assigned to either AL or AN treatment upon presentation at the health centre with either P. falciparum or P. vivax malaria. LM was conducted by 2 trained microscopists with discordant reads judged by an expert microscopist. MF was conducted as previously described on the same day using 200 μL of blood. Samples for RTPCR were placed directly into RNA-later and stored at -80°C until analysis. MF and RTPCR were similarly sensitive and specific, and clearly superior to LM detection of gametocytes. P. falciparum gametocyte clearance characteristics were found to be different between AL and AN, mostly due to a longer predicted gametocyte sequestration

astmh.org 551 1805 millions of people with suspected malaria still do not receive diagnostic tests. As these methods do not rapidly and accurately detect sub- DEVELOPMENT OF A SINGLE NUCLEOTIDE POLYMORPHISM microscopic infections which can also contribute to transmission, high BASED BARCODE FOR THE IDENTIFICATION AND TRACKING throughput molecular assays such as polymerase chain reaction (PCR) OF PLASMODIUM VIVAX are used to detect asymptomatic and/or low-grade infections. Isothermal amplification methods were recently developed to address some of the 1 2 3 Mary Lynn Baniecki , Aubrey Faust , Rachel Daniels , Kevin major shortcomings of PCR. The most common target amplified in these 4 5 6 Galinsky , Marcelo U. Ferreira , Nadira Karunaweera , Elizabeth molecular assays is the conserved small subunit ribosomal RNA 18S 7 8 9 10 Winzeler , David Serre , Peter Zimmerman , Tom Wellems , locus, which in the Plasmodium falciparum chromosomal genome exists 11 11 1 Lise Musset , Stéphane Pelleau , Alexandre Melnikov , Daniel in five to eight copies, depending on the strain. In this study, we report 1 4 2 4 Neafsey , Sarah Volkman , Daniel L. Hartl , Dyann Wirth , Pardis the development and optimization of the apicoplast of P. falciparum 2 Sabeti as a target for molecular diagnosis of malaria using a single step PCR 1The Broad Institute, Cambridge, MA, United States, 2Harvard University, (ssPCR), nested PCR (nPCR) and loop-mediated isothermal amplification Cambridge, MA, United States, 3Harvard School of Public Health, (LAMP) assay. P. falciparum sequences from 15 Gambian isolates and 8 Cambridge, MA, United States, 4Harvard School of Public Health, Boston, laboratory clones were aligned against the PlasmoDB reference sequence MA, United States, 5University of São Paulo, São Paulo, Brazil, 6University (ID: emb|X95275.2|) and primers were designed from a highly conserved of Colombo, Colombo, Sri Lanka, 7The University of California San Diego, region of the consensus sequence, approx. 1.5kb segment of the gene San Diego, CA, United States, 8Cleveland Clinic, Cleveland, OH, United coding for a ribosomal RNA protein (AP|0010:rRNA). The primers were States, 9Case Western Reserve University, Cleveland, OH, United States, validated in silico and mapped unto the consensus sequence with a web 10National Institute of Allergy and Infectious Diseases, Bethesda, MD, based tool. The assays were optimized for temperature and concentration United States, 11Institut Pasteur de la Guyane, Cayenne, French Guiana of primers, deoxyribonucleotides (dNTPs) and magnesium chloride For global eradication of malaria to occur an enhanced understanding (MgCl2). 272 archived DNA samples from across West Africa and S.E Asia of the population structure of Plasmodium vivax is needed. An inability were analyzed against a reference PCR method targeting the 18SrRNA to maintain P. vivax continuously in culture has caused research on this gene. Preliminary results show perfect agreement for ssPCR and nPCR parasite to lag behind that of the other Plasmodium species. Recent compared with the reference PCR method, while Sensitivity of 100 % advances in molecular methods have enabled the P. vivax genome to (95% CI: 94 % to 100 %), Specificity of 84 % (95% CI: 68 % to 94 %), be assayed directly from infected humans, providing the tools needed Positive Predictive Value (PPV) of 91 % (95% CI: 81 % to 97 %), Negative to develop genotyping methods. Single nucleotide polymorphism (SNP) Predictive Value (NPV) of 100 % (95% CI: 89 % to 100 %) and Kappa genotyping provides a robust, inexpensive, field-deployable technology index of 0.86 (95% CI: 0.76 to 0.97) were obtained for LAMP. Based on that allows malaria parasites to be tracked and identified by creating the results, the apicoplast genome appears to be a suitable target for a unique genetic signature or barcode for each parasite. Using our sensitive detection of P.falciparum. experience in developing a SNP genotyping tool with quantitative PCR and High Resolution Melting (qPCR-HRM) for P. falciparum, we have 1807 developed a SNP barcode for P. vivax. The candidate SNPs were selected with a high minor allele frequency (MAF) from available P. vivax genome GEOGRAPHICAL DISPERSION AND GENETIC sequence data and were located at sites including intergenic, intragenic, CHARACTERIZATION OF PFHRP2 NEGATIVE PLASMODIUM or were 4-fold degenerate coding sites. Here, we report a pilot screen of FALCIPARUM PARASITES IN THE PERUVIAN AMAZON: a 95 SNPs by genotyping a set of 89 P. vivax containing clinical samples IMPLICATIONS FOR RAPID DIAGNOSTIC TESTS (RDTS) BASED from geographically distinct parasite populations from the Americas ON DETECTION OF HRP2 (Brazil, French Guiana), Africa (Ethiopia) and Asia (Sri Lanka). Candidate Dionicia Gamboa1, Jorge Bendezu1, Katherine Torres1, Sheila SNPs were winnowed to a 41-SNP barcode based on robustness and 2 2 1 3 reproducibility of the genotyping calls, and the ability to accurately detect Akinyi , Curtis S. Huber , Elizabeth Villasis , Sandra Incardona , David Bell3, Joseph Vinetz4, John W. Barnwell2, Venkatachalam polygenomic infections. The assays are robust with a detection range from 2 10 ng to 0.001 ng with an average assay efficiency of 90% among clinical Udhayakumar samples tested. All 41 assays had an average minor allele frequency (MAF) 1Universidad Peruana Cayetano Heredia, Lima, Peru, 2Malaria Branch, > 0.1. Based on principle component analysis the clinical samples form Division of Parasitic Diseases and Malaria, Center for Global Health, distinct clusters that correspond to their geographic origin. Interesting, Centers for Disease Control and Prevention, Atlanta, GA, United States, these analysis revealed a high level of polygenomic samples among all 3Foundation for Innovative New Diagnostics, Geneva, Switzerland, populations with Brazil (84%), Sri Lanka (95%), Ethiopia (78%), and 4University of California San Diego, San Diego, CA, United States French Guiana (62%). There is prior evidence of Plasmodium falciparum parasites from clinical cases lacking pfhrp2 and pfhrp3 genes in the Peruvian Amazon Region. 1806 As various countries in South America move to introduce malaria rapid diagnostic tests (RDTs) as an alternative for diagnosis, the geographical THE APICOPLAST OF PLASMODIUM FALCIPARUM PROVIDES distribution and genotypic characterization of parasites lacking pfhrp2 A NOVEL TARGET FOR MOLECULAR DIAGNOSIS OF and pfhrp3 genes will have major implications for procurement choices MALARIA USING POLYMERASE CHAIN REACTION AND LOOP for RDTs in the region. Ninety-three P. falciparum samples, collected MEDIATED ISOTHERMAL AMPLIFICATION ASSAYS in different communities from the Peruvian Amazon Region between 2009 and 2010, were used in this study. Genomic DNA was used to 1 2 3 Eniyou C. Oriero , Jan Jacobs , Jean-Pierre Vangeertruyden , amplify 18SrRNA and pfmsp2 to confirm the diagnosis and DNA quality, 1 1 Umberto D’Alessandro , Davis Nwakanma respectively; pfhrp2, pfhrp3, and their flanking genes in order to assess 1Medical Research Council Unit, The Gambia, Fajara, Gambia, 2Institute the frequency of their deletions. Microsatellite analysis was performed of Tropical Medicine, Antwerpen, Belgium, 3University of Antwerp, using seven neutral microsatellites (MS) and seven novel MS loci flanking Antwerpen, Belgium the pfhrp2 gene on chromosome 8 (-41kb, -10kb, -4kb, 1.4kb, 2.5kb, Despite a recent increase in procured rapid diagnostic tests (RDTs), and 5.2kb and 15kb). The data showed deletion of the pfhrp3 gene in 53.76% rate of diagnostic testing in the public sector of the African region, (50/93) and pfhrp2 gene deletion in 33.33% (31/93) of the samples. malaria still remains a global health burden with an estimated 627,000 The proportion of the parasite populations that lacked these genes was deaths worldwide in 2012. WHO recommends all suspected malaria quite variable from community to community. Among the flanking genes, cases to be confirmed by microscopy or RDT prior to treatment however, PF3D7_0831900 (Mal7P1.230) showed the highest deletion frequency,

astmh.org 552 78.49% (73/93). Neutral MS marker analysis revealed the widespread 1809 distribution of P. falciparum hybrid lineages with a hybrid of the A clonal lineage (named AV1) being the most prevalent among parasites lacking IMPACT OF MALARIA RAPID DIAGNOSTIC TESTS ON CARE pfhrp2 and pfhrp3 genes. MS data from loci flanking the pfhrp2 gene OF FEBRILE PATIENTS: CROSS-PROJECT RESULTS FROM THE showed that the haplotypes α and Δ were the most abundant among ACT CONSORTIUM the isolates analyzed. This study confirms that field isolates lacking either pfhrp2, pfhrp3 or their respective flanking genes were still present in Baptiste Leurent, Katia Bruxvoort, Eleanor Grieve, Eleanor the area in 2010. In addition, we identified five P. falciparum hybrid Hutchinson, Joanna Reynolds, Toby Leslie, ACT Consortium lineages circulating in this region. It is possible that certain parasite London School of Hygiene & Tropical Medicine, London, United Kingdom genetic backgrounds (haplotypes) could favor the maintenance and The introduction of malaria rapid diagnostic tests (RDTs) is aimed to expansion of pfhrp2 and pfhrp3 gene deletions in the Peruvian Amazon, increase the proportion of febrile patients tested for malaria, but there however further studies will be required to prove this possibility and also are gaps in knowledge of what happens to patients who receive an to elucidate the genetic basis for the pfhrp2 gene deletion in wild P. RDT compared to those who do not. Multi-project data from the ACT falciparum parasites. Consortium provide an opportunity to examine the impact of RDTs on patient care outcomes under different contexts and settings. Twelve 1808 projects from seven countries contributed data, including projects in public health facilities, private retailers, and community health worker AN EXTERNAL QUALITY ASSURANCE PROTOCOL FOR settings. Each compared different sets of diagnostic methods, resulting in PLACENTAL MALARIA HISTOPATHOLOGY STUDIES 30 arms for analysis, which were grouped into four diagnostic categories: Atis Muehlenbachs1, Miriam K. Laufer2, Tamiwe Tamoka3, presumptive, microscopy, RDT, or enhanced RDT (in which RDTs were Veronica Ades4, Grant Dorsey5, Julie Gutman6 provided alongside supportive interventions). Patient care outcomes such as RDT use, antimalarial prescription, antibiotic prescription, 1Infectious Diseases Pathology Branch, Division of High Consequence referrals, patient satisfaction, and consultation out-of pocket costs were Pathogens and Pathology, Centers for Disease Control and Prevention, summarised for each arm and compared between categories. Preliminary Atlanta, GA, United States, 2Department of , University of results indicate a lower prescription of ACTs and a generally higher Maryland, Baltimore, MD, United States, 3University of Malawi College of prescription of antibiotics where RDTs were used, compared to arms Medicine, Blantyre, Malawi, 4Department of and Gynecology, where only presumptive diagnosis was available. This difference was New York University Langone Medical Center, New York, NY, United more marked in enhanced RDT arms. Referral to higher level care was States, 5Department of Medicine, University of California San Francisco, also more frequent among RDT arms. However, the impact of RDTs was San Francisco, CA, United States, 6Malaria Branch, Division of Parasitic not always consistent and may depend on study setting and design, type Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, of sector, characteristics of patients and providers, and measures of RDT GA, United States implementation and support. Exploration of these contextual factors is Placental histology can contribute to studies on the diagnosis, prevention underway, with complete results expected to be available in August 2014. and treatment of malaria in pregnancy (MIP). Infections prior to delivery While many studies have reported a positive impact of RDTs on patient can be identified by detecting hemozoin deposition, and parasitized outcomes, the effect is complex and likely to vary by setting and context. erythrocytes can be seen in cases where peripheral blood is negative. Further scrutiny is needed to better understand the impact of RDTs on However histology is prone to artifacts, and requires considerable training patient care beyond their role as an important diagnostic tool. in slide reading. There is a general need for external quality control and quality assurance for MIP histopathology studies, where poor preparation 1810 can lead to the appearance of a hemozoin-like material which may be mistaken for evidence of prior malaria infection and debris can be SCHISTOSOME POPULATION GENOMICS USING SINGLE mistaken for parasites, leading to false positive reads. In addition, very low ARCHIVED MIRACIDA level infections may be present in the placentas of semi-immune women, 1 1 2 leading to false negatives if insufficient fields are examined or if the slide Frédéric Chevalier , Winka Le Clec’h , Philip LoVerde , Rafael 3 3 4 reader is inexperienced. We developed a protocol to formally provide Ramiro de Assis , Guillerme Corrêa-Oliveira , Safari Kinunghi , 5 5 6 confidential, blinded, retrospective review of slides with feedback in order Anouk Gouvras , Bonnie Webster , Joanne P. Webster , Aidan 5 5 1 to ensure high quality study data and to work towards building local Emery , David Rollinson , Tim Anderson expertise. A random subset of 10% of negative and 25% of positive slides 1Texas Biomedical Research Institute, San Antonio, TX, United States, are recommended for review by a single, blinded expert reader. Slides are 2Departments of Biochemistry and Pathology, University of Texas Health formally scored on both the presence of parasites and hemozoin in fibrin Science Center, San Antonio, TX, United States, 3Centro de Pesquisas in addition to general quality. Following unblinding, discrepant slides are René Rachou-Fiocruz, Belo Horizonte, Brazil, 4National Institute for re-examined to determine source of discrepancy, and consensus opinion Medical Research, Mwanza Centre, Mwanza, United Republic of Tanzania, with the submitter is attempted by using high quality photomicrographs. 5Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, All slides are returned to the submitter so that they can be used for further Natural History Museum, London, United Kingdom, 6Department of education. Error rates less than 20% are considered standard for passing Infectious Disease Epidemiology, School of Public Health, Imperial College a laboratory proficiency test, and further rounds of QC can be performed Faculty of Medicine, London, United Kingdom as needed. For histology studies of malaria in pregnancy, discordance over Population genomic analyses of schistosomes have not previously been 10% should trigger consideration of additional training and targeted re- possible due to the difficulty of sampling adult worms and the high repeat review of study slides. In pilot studies with experienced readers, there was content and the large size (360-400Mb) of their genomes. We have concordance in 85/94 (90.5%) of submitted cases; discrepancies primarily developed a robust, inexpensive approach for capture and sequencing of included false positives for hemozoin due to artifact, and false negatives the ~15Mb Schistosoma mansoni exome that can be used for single larval due to low levels of hemozoin. We anticipate that a standardized miracidia. The approach uses whole genome amplification of miracidia histopathology QC/QA protocol will be of value to pregnancy malaria preserved on FTA cards, solution-based capture of exome sequences research community, and have potential to strengthen local histopathology using 120bp RNA baits, and Illumina sequencing, and can be extensively expertise. multiplexed to reduce costs and simplify sequence library preparation. To demonstrate the utility of these methods we sequenced exomes from 45 single miracidia collected from a Brazilian location in three lanes of an Illumina HiSeq. We captured >99% of the exome sequences targeted,

astmh.org 553 obtained between 30-80x read depth per miracidia, and robustly called such as microsatellite loci, and analysis of population genetics can be used >70,000 SNPs. The method also efficiently captures exomes from the to monitor changes in parasite populations and the effect of PZQ-reliant related parasite, S. rodhaini, providing outgroup sequences and opening schistosomiasis control on the parasite’s adaptation and evolution. In the up the possibility of detailed dissection of interspecific hybridization Lake Victoria region of Tanzania (Mwanza), as part of a larger treatment within schistosome populations. We are currently using these methods study on intestinal schistosomiasis caused by Schistosoma mansoni, we are to characterize African S. mansoni populations from the SCAN collection collecting parasite genetic material from school children from 16 villages at the British Natural History Museum. The exome data will be used with contrasting PZQ treatment pressures: annual Community-Wide to characterize SNP variation at candidate vaccine and drug resistance Treatment (CWT-highest treatment pressure) and biennial School-Based loci, to examine geographic differentiation in allele frequencies, and to Treatment (SBT-lowest treatment pressure). The infection prevalence at the identify genome regions under strong directional and balancing selection. start of the study was over 25%. In each village larval miracidia samples We believe that this approach will have multiple uses for schistosome were collected prior to treatment from infected school children (n=30). epidemiology, population biology and evolutionary genomics. For baseline (2012) 18,649 S. mansoni miracidia were collected from 263 children in 16 villages. In Year 2 (2013), 4,724 S. mansoni miracidia were 1811 collected from 95 children in the 8 annual CWT villages. Future follow-up collections are planned for May 2014 (16 villages). Samples were collected POPULATION AND COMPARATIVE GENOMICS OF AFRICAN on to FTA cards and stored in SCAN (http://scan.myspecies.info/). Samples SCHISTOSOMA MANSONI are being analysed using a new set of multiplex panels developed from 20 previously published microsatellites for S. mansoni. Comparison of Thomas Crellen1, James Cotton2, Sophia David2, Caroline genetic diversity indices such as number of alleles per locus, allelic richness, Durrant2, Thomas Huckvale2, Nancy Holroyd2, Fiona Allen3, David observed and expected heterozygosity will be made between baseline Rollinson3, Joanne Webster1, Matthew Berriman2 and post treatment samples and between annual CWT and biennial 1Imperial College, London, United Kingdom, 2Wellcome Trust Sanger SBT control intervention strategies. Initial analysis of baseline-collected Institute, Cambridge, United Kingdom, 3Natural History Museum, London, material shows little difference in the genetic diversity indices between United Kingdom the two different treatment arms. Year 2 miracidia samples are currently Schistosomiasis is among the most important parasitic diseases, with over being analysed to determine if there has been a change in genetic 200 million people infected and 300,000 deaths annually across Africa, diversity compared to baseline in the 8 CWT villages. This study utilizes Asia, South America and the Caribbean caused mainly by three closely field-collected material and microsatellite multiplex panels to determine related species. Around 90% of cases are in sub-Saharan Africa, where the differential impact of annual CWT vs. biennial SBT chemotherapy Schistosoma mansoni is one of the two most clinically important species, strategies on parasite clearance and population genetic outcomes. and the principal cause of intestinal schistosomiasis. A draft reference genome is available for S. mansoni and is being actively curated and 1813 improved, based on an isolate from Puerto Rico that has been maintained in research labs for many years. Here, we present genome sequence data REPEATED TREATMENTS ARE REQUIRED TO AFFECT and assemblies from seven adult male S. mansoni that were recently SCHISTOSOME POPULATION STRUCTURE collected from the field with minimal lab passage, including six diverse Lucio M. Barbosa1, Luciano K. Silva1, Eliana A. Reis1, Theomira African isolates - the first genomic data from the region of greatest public M. Carmo1, Claudio R. dos Santos1, Jackson M. Costa1, Peace T. health interest. We confirm that the S. mansoni reference sequence is a Aminu2, Thassila N. Pitanga1, Walter A. Blank2, Mitermayer G. suitable substrate for genomic analysis of African populations. We use Reis1, Ronald E. Blanton2 this genomic diversity data to investigate signatures of natural selection on the S. mansoni genome, and apply two coalescent-based models to 1Oswaldo Cruz Foundation, Salvador, Brazil, 2Case Western Reserve infer the population history of S. mansoni on two continents. Our results University, Cleveland, OH, United States show that the New World strains have smaller past effective population Repeated rounds of praziquantel treatment are able to reduce prevalence sizes (Ne) than African strains, suggesting the possible occurrence of a and morbidity of schistosomiasis, but parasite populations recover within a past population bottleneck. We estimate the divergence time between the few years. To understand the dynamics of this recovery, two rural Brazilian African and New World populations, finding support for the hypothesis communities were surveyed and treated in 3 different years: 2009, 2012 that S. mansoni colonised the New World via the 16-19th century West and 2013. On average, 96% of the residents participated in each year, African slave trade. In the light of this potential population bottleneck, we and those who were positive on at least one of 3 stools were treated. investigate systematic differences between South American populations Schistosoma mansoni eggs were collected from stool and genotyped and African populations in both genome structure (copy number variants) using 11 microsatellite markers. Parasite differentiation was evaluated at and single nucleotide polymorphisms (SNPs). the level of infrapopulations and component populations. Component populations were defined by host characteristics, village of residence 1812 or year of study. During this time, human population demographics changed little. New arrivals were 16% and 5% of the populations in USING MICROSATELLITE MARKERS TO DETERMINE 2012 and 2013, respectively. In both years ~12% moved elsewhere. After SCHISTOSOMA MANSONI GENETIC DIVERSITY UNDER 2 rounds of treatment, prevalence decreased by 64% and intensity by CONTRASTING CHEMOTHERAPY CONTROL STRATEGIES IN 57%. Children 15-20 years old showed the greatest decline, while adults LAKE VICTORIA, TANZANIA between 51-60 showed the least. Reinfection was 34% in 2012 and 18% in 2013, while incidence was 22% and 15%, respectively. The decreasing 1 2 1 Anouk N. Gouvras , Bonnie L. Webster , Fiona Allan , Muriel rates suggest that these treatments have an effect on transmission. 1 3 1 2 Rabone , Safari Kinunghi , Aidan Emery , Joanne P. Webster , Individual infrapopulations were moderately differentiated (D=0.055- 1 David Rollinson 0.077) on reinfection, indicating the pre-treatment multilocus genotypes 1Natural History Museum, London, United Kingdom, 2Imperial College, were not fully reacquired. Differentiation between the 2 villages decreased School of Public Health, London, United Kingdom, 3National Institute for from 0.046 to 0.031, consistent with an increase in gene flow between Medical Research, Mwanza, United Republic of Tanzania them. Parasites from new immigrants were little differentiated from natives National schistosomiasis control programs rely on mass drug administration (D = 0.012). Between consecutive years, there was little differentiation (D using the drug praziquantel (PZQ). The widespread use of a single drug = 0.008), but comparing 2009 to 2013, differentiation increased notably raises concerns of drug tolerance spreading in the parasite population. In (D = 0.014). Population structure began to change only after 2 rounds of the absence of specific markers for PZQ tolerance, neutral genetic markers treatment when total parasite burden decreased by >10 fold. This seems

astmh.org 554 to be the tipping point for producing a genetic bottleneck or reducing other studies we tested the role of several BAs on schistosomes motility. effective population size. Intensive therapy is required to significantly Treatment with both OA and DA caused marked changes in worm impact the parasite’s genetic potential. motility as compared to the control. Next, we performed RNAi targeting proteins predicted to be involved in DA and OA signaling in larvae and 1814 adult schistosomes, and effects of downregulation were assessed. Several of the RNAi-targeted animals showed strong changes in frequency of EPIGENETIC CONTROL OF ENDOGENOUS AND EXOGENOUS body movements and in worm morphology as compared to the control. (RETRO)TRANSPOSABLE ELEMENTS IN SCHISTOSOMA Together these studies highlight the importance of tyrosine derived BAs in MANSONI the control of motor activity in schistosomes. Hong-Bin Yan, Paul J. Brindley, Gabriel Rinaldi 1816 George Washington University, Washington, DC, United States The epigenetic landscape includes modifications on the chromatin MICRORNAS IN THE EXCYSTATION OF FASCIOLA HEPATICA template that establish and propagate patterns of gene expression and METACERCARIAE silencing, not based on differences in the DNA sequence. Four major Jose F. Tort1, Santiago Fontenla1, Nicolas Delloca1, Pablo epigenetic phenomena are involved: 1) DNA methylation; 2) histone Smircich1, Maria del Mar Siles-Lucas2 modifications; 3) nuclear gene repositioning; and 4) regulatory non-coding 1 2 RNAs. Whereas early reports suggested absence of DNA methylation Universidad de la Republica, Montevideo, Uruguay, Instituto de Recursos of the schistosome genome, it has now been shown that cytosine Naturales y Agrobiología, Salamanca, Spain methylation regulates schistosome oviposition and embryo development. Host invasion by the trematode parasite Fasciola hepatica is initiated by the In addition, post-translational core histone modifications in epigenetic activation of the metacercariae usually in the host stomach. The released control of transcription of schistosome genes have been described, as juvenile forms actively transverse the gut wall towards the abdominal have a set of non-coding RNAs -- integral components of the epigenetic cavity and follow their journey to the biliary ducts of the liver. The machinery, including epi-miRNAs, i.e. miRNAs that regulate expression activation is a rapid switch finely tuned by signals from the environment, of enzymes involved in chromatin remodeling and DNA methylation. To that can be easily reproduced in vitro. miRNAs have emerged as relevant evaluate the effect of the DNA methyltransferase inhibitor 5’-azacytidine modulators of gene expression at the post-transcriptional level (either (5’-AzaC) on the expression of both long terminal repeat (LTR) and non-LTR by tranlation blocking or mRNA degradation), playing essential roles in retrotransposons, schistosomules of Schistosoma mansoni were cultured development. We aimed to study the miRNA expressed in the liver fluke in 100 µM and 500 µM of 5’-AzaC. The parasites were harvested 2 or 7 metacercarial activation, and for that purpose we purified and sequenced days after treatment, RNA was isolated and the expression of the Boudicca the small RNA populations expressed at this developmental transition. (LTR-retrotransposon) and SR2 (non-LTR-retrotransposon) multi-copy After filtering the reads with homology to mRNA, repetitive sequences and endogenous mobile genetic elements of schistosomes analyzed by qRT- other non coding RNAs, we ended up with several thousand reads that PCR. The expression level of these retrotransposons was upregulated 10 were compared to miRBase, Rfam and all the miRNA previoulsy identified to 20 times in the presence of 5’-AzaC. In addition, expression of reporter in other flatworms. Within the known miRNAs found, those common to transgenes increased in schistosomules that had been transformed all metazoans and protostomes were the most abundant. Some miRNA with virions of pseudotyped murine leukemia virus following culture in so far only detected in other flatworms were also found, highlighting media supplemented with 5’-AzaC. These findings demonstrated that the existance of flatworm specific miRNA families. Furthermore within endogenous mobile elements, which comprise ~45% of the genome sequences with no homology, novel F. hepatica-specific miRNAs were of this schistosome are controlled by epigenetic marks. In addition, they predicted. We observed subtle differences between dormant and activated indicated a central influence of methylation status on transgene activity, metacercariae and further differences to newly excysted juveniles. While suggesting one avenue forward for enhancing transgenesis of this tropical sequence conservation in mature miRNA is high across the metazoan tree, neglected tropical disease pathogen. we observed that in general flatworm miRNA are more divergent than in other lineages, with strict conservation restricted to seed region. Whether this variability leads to changes in the regulated target genes associated to 1815 the parasitic way of life deserves further investigation. SCHISTOSOMA MANSONI: ROLE OF BIOGENIC AMINES IN NEURONAL CONTROL OF MOTOR FUNCTION 1817 Nelly El-Sakkary, Paula Ribeiro IDENTIFICATION OF SYMBIONTS IN THE REPRODUCTIVE McGill University, Ste. Anne-de-Bellevue, QC, Canada TRACT MICROBIOME OF NATURAL ANOPHELES GAMBIAE POPULATIONS AND IMPLICATIONS FOR NOVEL VECTOR Schistosoma mansoni is the main causative agent of schistosomiasis, a CONTROL METHODS disease which infects over 200 million people worldwide. Treatment of the disease is primarily with praziquantel. With the lack of an available Francesco Baldini1, Nicola Segata2, Julien Pompon3, Perrine alternative, and the widespread use of the drug, there is a fear of the Marcenac4, W. Robert Shaw5, Roch K. Dabiré6, Abdoulaye development of resistance. Biogenic amines (BAs) are the largest family Diabaté6, Elena A. Levashina7, Flaminia Catteruccia4 of classical neurotransmitters in the schistosome nervous system. They 1University of Glasgow, Glasgow, United Kingdom, 2University of Trento, are typically involved in motor control and are important to host infection Trento, Italy, 3University of Strasbourg, Strasbourg, France, 4Harvard School and worm survival. The goal of this study is to determine the role of of Public Health, Boston, MA, United States, 5Imperial College London, BA neurotransmitters in schistosomes, focusing on tyrosine derivatives, London, United Kingdom, 6Institut de Recherche en Sciences de la Santé/ which include phenolamines and catecholamines. Using confocal Centre Muraz, Bobo-Dioulasso, Burkina Faso, 7Max-Planck Institute for immunolocalization, receptors responsive to the catecholamine, dopamine Infection Biology, Berlin, Germany (DA), SmGPR3 and SmD2, were shown to localize to the main nerve cords of the central nervous system (CNS), and the peripheral nervous Identification of symbionts in the reproductive tract microbiome of natural system (PNS), respectively. Both receptors localized to neurons innervating Anopheles gambiae populations and implications for novel vector control worm musculature, indicating a possible role in motility for the receptors. methods Recent findings show that mosquito-microbiota interactions are Octopamine (OA), a phenolamine, was also immunolabeled in the adult crucial determinants of mosquito fitness and vectorial capacity opening parasite, and showed widespread labeling in the main neurons of the novel and promising areas of research in the utilization of symbiotic worm CNS, the first indication that OA is present in schistosomes. In bacteria for the control of vector-borne diseases. Furthermore, recent astmh.org 555 works propose the use of paratransgenesis as a novel vector control tool, 1819 which exploits genetically engineered symbionts to deliver anti-parasite molecules in the vector. In order to identify potential determinants of BREAKING THE LAW OF EFFECTIVE TEMPERATURE: mosquito biology as well as putative candidates for paratransgenesis, we ECOLOGICAL CONTEXT MATTERS FOR DEVELOPMENT RATE have characterized the reproductive microbiome of two major African VARIATION MOSQUITO VECTORS malaria vectors Anopheles gambiae and An. coluzzii from field population 1 2 3 in Burkina Faso. Specifically, we performed high throughput sequencing Jannelle Couret , Ellen Dotson , Mark Q. Benedict of the bacterial 16S gene in the male and female reproductive tracts of 1Emory University, Atlanta, GA, United States, 2Centers for Disease Control mosquitoes. We identified two bacteria genera that are present in all the and Prevention, Atlanta, GA, United States, 3University of Perugia, Perugia, analysed specimens, representing the core taxa of mosquito reproductive Italy organs with possible symbiotic interactions with the vector. Nevertheless, The resurgence and spread of arboviruses in recent years underscores although a general core microbiome was identified, we observed a general the continued need to understand the biology of its mosquito vectors high diversity among different specimens that might indicate that the including, Aedes aegypti and Culex pipiens complex. Despite advances in reproductive microbiome is highly dynamic and might be influenced by the development of vaccines, control of the mosquito vector populations external factors. Indeed, we identified some taxa whose abundance was remains the most effective control measure. In order to understand significantly associated with the environment where the mosquitoes lived. how the environment impacts disease transmission, a thorough Finally, we identified intracellular bacteria that were previously believed not understanding of the impact of environmental variables on mosquito to colonize natural populations of Anopheles. Remarkably, these bacteria biology is needed. Historically, the emphasis has been on temperature are capable of spreading into insect populations and negatively impact and the linear association with insect development rate. This association mosquito vectorial capacity by reducing their lifespan and boosting the is widely observed across ectotherms, and is often referred to as the immune response against parasites. To our knowledge this was the first law of effective temperature. We tested the hypothesis that the law identification of these intracellular bacteria in malaria mosquitoes, which of effective temperature is contingent on the ecological context of the opens new promising opportunities to exploit these organisms as vector larval environment in mosquito development. Through a combination control agents against malaria vectors. These results started to elucidate of statistical modeling of published rearing experimental research and a the composition of malaria mosquito reproductive tract microbiome larval rearing experiment under gradients of conditions in environmental offering novel opportunities to exploit symbiotic bacteria in the fight chambers, we find that intraspecific density and dietary resources mediate against malaria. the importance of temperature in explaining variation of mosquito development rate. Our results support the hypothesis of environmentally 1818 contingent impacts of temperature on mosquito development. These findings have broad implications for the modeling of mosquito population PERIOSTIAL HEMOCYTE AGGREGATION IN ANOPHELES dynamics, climate change and vectorborne disease transmission, and our GAMBIAE OCCURS FOLLOWING DIVERSE IMMUNE STIMULI understanding of nature of mosquito life history evolution. AND IS ACCOMPANIED BY CHANGES IN MOSQUITO HEART PHYSIOLOGY 1820 Julian F. Hillyer, Leah T. Sigle, Tania Y. Estevez-Lao PARASITE CO-INFECTION AND STRAIN DIFFERENCES AS Vanderbilt University, Nashville, TN, United States DRIVERS OF PATHOGENIC VARIATION IN THE CHAGAS Malaria parasites and other mosquito-borne pathogens must traverse DISEASE PARASITE TRYPANOSOMA CRUZI WHEN INFECTING the insect hemocoel prior to being transmitted. This process is affected ITS INSECT VECTOR, RHODNIUS PROLIXUS by hemolymph circulation, as flow influences both pathogen movement 1 1 1 and the movement of mosquito-produced immune factors. Using the Jennifer K. Peterson , Andrea L. Graham , Andrew P. Dobson , 2 African malaria mosquito, Anopheles gambiae, as our study system, we Omar Triana recently identified a novel immune tissue, called periostial hemocytes, 1Princeton University, Princeton, NJ, United States, 2University of Antioquia, that exemplifies the co-adaptation of the insect immune and circulatory Medellín, Colombia systems. Specifically, in response to infection, circulating hemocytes Avoiding the over-exploitation of resources in a food patch while still (immune cells) migrate to the valves of the mosquito heart, where they obtaining all the resources needed to live and reproduce is a common sequester and kill pathogens. We have previously reported the aggregation challenge among all creatures. For many parasites that live inside another of hemocytes on the surface of the heart following Plasmodium and organism this is especially tricky because they do not have the option of Escherichia coli infection, however, little is known about the breadth of moving to a new patch should they be too pathogenic and overexploit this immune response or about how heart physiology changes following their current one. Classical theory suggests that the avoidance of host infection. In the present study we tested whether periostial hemocyte death should favor intermediate levels of parasite pathogenicity (i.e., aggregation occurs following diverse immune stimuli, whether this negative effects) to the host, however that this is not the case: parasites response is uniform across the length of the heart, and whether infection actually exhibit a wide range of pathogenic effects on their hosts. We affects mosquito heart physiology. We found that periostial hemocyte investigated parasite co-infection and strain differences as drivers of aggregation occurs following all types of infections tested, confirming the pathogenic variation in the Chagas disease parasite Trypanosoma cruzi and fundamental role of this immune response. Moreover, periostial hemocyte its sister species, Trypanosoma rangeli when infecting their insect vector, aggregation is not uniform along the length of the heart, as larger Rhodnius prolixus. Using insect survival, reproduction, and parasite load hemocyte aggregates consistently form in abdominal segments 4, 5, and (qPCR amplification of parasite DNA extracted from each insect) as proxies 6. Finally, periostial hemocyte aggregation is accompanied by a decrease in for parasite pathogenicity, we found that T. cruzi-T. rangeli co-infection heart contraction rates. In summary, these data further describe a recently significantly reduces the survival of R. prolixus up to 30 days post-infection, discovered immune tissue in mosquitoes, and demonstrate how the but increases reproduction. We also found that T. cruzi pathogenicity in R. immune and circulatory systems have co-adapted to fight infection. prolixus is highly variable, with R. prolixus death at 90 days ranged from 5-80% depending on T. cruzi strain, and at times was far more pathogenic than T. rangeli, a parasite believed to be highly pathogenic to triatomines. Furthermore, insects with higher parasite loads tended to have higher fecundity, presenting evidence for terminal investment in infected bugs.

astmh.org 556 Our results suggest that the pathogenic variation often found in T. cruzi funestus has been found positive to CSP. These disturbing observations infection of vertebrate hosts extends to its invertebrate hosts as well, with show the capacity of Anopheles to adapt and circumvent strategies aimed strain variability and co-infection with T. rangeli as two of the main drivers. at reducing malaria transmission. In an arms race between malaria control programs and the vector populations, the behaviour change in Anopheles 1821 threatens to thwart the goal of decreasing malaria transmission. THE EFFECTS OF FORCED-EGG RETENTION ON THE BLOOD- 1823 FEEDING BEHAVIOR AND REPRODUCTIVE POTENTIAL OF CULEX PIPIENS (DIPTERA: CULICIDAE) INFECTION OF LABORATORY-COLONIZED ANOPHELES DARLINGI MOSQUITOES BY PLASMODIUM VIVAX AND 1 2 Brian J. Johnson , Dina M. Fonseca TEMPORAL CHANGE IN GENETIC VARIATION IN AN. 1Graduate Program in Ecology and Evolution, Rutgers University, New DARLINGI Brunswick, NJ, United States, 2Center for Vector Biology, Department of Entomology, Rutgers University, New Brunswick, NJ, United States Marta Moreno Leirana1, Carlos Tong2, William Lainhart3, Mitchel 2 2 4 High rates of West Nile virus (WNV) transmission to humans are associated Guzman , Raul Chuquiyauri , Alejandro Llanos-Cuentas , Hugo 5 4 6 1 with exceptionally hot and dry summers. This is paradoxical since the Rodriguez , Dionicia Gamboa , Jan E. Conn , Joseph M. Vinetz , 2 4 eggs of Culex vectors of WNV depend on the persistence of containers Joseph M. Vinetz , Joseph M. Vinetz with water, which decline during droughts. We examined the effects of 1University of California San Diego, La Jolla, CA, United States, forced-egg retention on the reproductive success of female Culex pipiens 2Laboratorio ICEMR-Amazonia, Laboratorios de Investigacion y Desarrollo, as well as behavioral responses, such as likelihood of secondary blood Facultad de Ciencias y Filosofia, Universidad Peruana Cayetano Heredia, meals. As controls we examined the effects of female age and delayed Lima, Peru, 3Department of Biomedical Sciences, School of Public Health, mating. We found that early mating is essential to achieve reproductive State University of New York-Albany, Albany, NY, United States, 4Instituto success and, consistent with an “all-or-none” ovipositing strategy, Cx. de Medicina Tropical “Alexander von Humboldt”, Universidad Peruana pipiens females are able to retain considerable reproductive potential while Cayetano Heredia, Lima, Peru, 5Loreto Regional Directorate of Health, searching for oviposition sites. Specifically, although forced-egg retention Iquitos, Peru, 6Wadsworth Center, New York State Department of Health, resulted in significant decreases in fitness, the decline was moderate Albany, NY, United States for 5 weeks and most can be accounted for by increases in female age. Anopheles darlingi Root is the most important malaria vector in the Consequently, no females took blood more than once per gonotrophic Amazonia region of South America. However, continuous propagation cycle, which eliminates the possibility that heightened vectorial capacity of An. darlingi in the laboratory has been elusive, limiting entomological, due to multiple blood-feedings increases WNV transmission during periods genetic/genomic, and vector–pathogen interaction studies of this mosquito of drought. Instead, our findings suggest that during droughts populations species. We report the establishment and maintenance of an An. darlingi of Cx. pipiens have time to locate the remaining water holes, which are colony (since July 2013) derived from wild-caught mosquitoes obtained in associated with human populations and WNV-competent bird species. the northeastern Peruvian Amazon region of Iquitos in Loreto Department. We demonstrate that the numbers of eggs, larvae, pupae, and adults 1822 continue to rise at least to the F9 generation. In addition, comparison of feeding Plasmodium vivax by artificial membrane feeding of F4-F9 to BEHAVIORAL CHANGE IN ANOPHELES FUNESTUS: AN F1 generation mosquitoes showed the comparable presence of oocysts OBSTACLE TO MALARIA ELIMINATION and sporozoites, with numbers that corresponded to blood-stage asexual parasitemia and gametocytemia, confirming P. vivax susceptibility in the Sougoufara Seynabou1, Diedhiou Mocote Seynabou1, Doucoure colonized mosquitoes. Additionally, analyses of fourteen microsatellites Souleymane1, Trape Jean François1, Sokhna Cheikh1, Sembene markers were performed on a subsample of An. darlingi offspring to Mbacke2, Harry Myriam3, Ndiath Mamadou Ousmane4 detect genetic variation and expected reduction in heterogeneity through 1 2 3 IRD, Dakar, Senegal, University, Dakar, Senegal, LEGS-CNRS, Paris, generations. These results provide new avenues for research on An. 4 France, Institute Pasteur, Bangui, Central African Republic darlingi biology and study of mosquito–Plasmodium interactions and In the long road that leads towards malaria elimination, vector control malaria transmission in the Neotropics, including new genomic analysis was undoubtedly an essential component of success. Two major strategies and assessment of transmission biology of malaria parasites. have marked the vector control: the use of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS). However, the effectiveness of 1824 these tools is being challenged by the emergence of insecticide resistance and behavioural resistance which thwarting the goal of decreasing malaria RATIONAL DESIGN OF OXAMNIQUINE DERIVATIVES THAT transmission. In this study, we focus on behavioural changes of malaria KILL SCHISTOSOMES vectors that can hamper the efficacy of vector control interventions in 1 1 1 Dielmo, a Senegalese rural village where a longitudinal study of malaria Stacey Stahl , Alexander B. Taylor , Xiaohang Cao , Stanton 2 1 3 1 has been conducted. In this village, universal coverage with LLINs was McHardy , P. John Hart , Timothy J. Anderson , Philip T. LoVerde done in July 2008, and in July 2011 all these LLINs were renewed. 1University of Texas Health Science Center, San Antonio, TX, United States, Adult mosquitoes were collected by human landing catches (HLC) from 2University of Texas at San Antonio, San Antonio, TX, United States, 3Texas July 2011 to April 2013 and hourly from 19:00 and 07:00. Collecting Biomedical Research Institute, San Antonio, TX, United States mosquitoes was also done by pyrethrum spray catch (PSC) during this Schistosomiasis, a major cause of morbidity, infects >200 million people period. From January to April 2013, mosquito catches were continued worldwide. Schistosomiasis control is based on a monotherapy consisting until 11:00 and the entomological different parameters were investigated. of repeated doses of praziquantel (PZQ). Drug resistance is a concern, This study shows that Anopheles funestus which have disappeared after especially as it is expected to increase in treatment coverage in sub-Sahara first introduction of LLINs (July 2008) comes back in malaria transmission Africa (250 million doses per year for each of the next 5 years). New in Dielmo. An. funestus remains anthropophilic and endophilic but adopt anti-schistosomal drugs are needed to reduce reliance on a single drug. a behavioural change in biting activity after introduction of LLINs. The A new drug could be used in combination with PZQ to minimize the human biting rate of mosquitoes collected from 07:00 to 11:00 was probability of resistance arising to either drug. The goal of this research is eight times higher than the one from 19:00 to 07:00. So the alarming to modify an existing anti-schistosomal drug oxamniquine (OXA) to make phenomenon is the positive mosquitoes found in the day capture (mean it more efficacious. We recently identified the gene encoding the Sm- CSP rate of 1.28%) while since distribution of LLINs in this village, no An. sulfotransferase (SmSULT) responsible for drug activation and determined

astmh.org 557 the structure of the SmSULT•cofactor•OXA ternary complex at 1.75 Å 1826 resolution. These analyses provide detailed information of the mechanism of action of OXA against Sm, while structural analyses of drug•protein SHORT-TERM ANTIBIOTIC TREATMENT INTERRUPTS interactions direct redesign of OXA. We designed and synthesized 12 THE EXCHANGE OF POLYMORPHIC VESICLES BETWEEN OXA derivatives based on four key design aspects; 1) the structural WOLBACHIA AND THEIR FILARIAL HOST requirements of OXA and its derivatives based on available space in the substrate binding cavity in SmSULT and the key residue interactions from Kerstin Fischer, Peter U. Fischer crystallographic studies, 2) the required ortho-electron withdrawing moiety Washington University School of Medicine, St. Louis, MO, United States necessary for the sulfonation process, 3) the design of analogs that fall Wolbachia endosymbionts are crucial for growth, reproduction, and within favorable “drug-like” physical chemical property ranges and 4) survival of many medically important filarial parasites. Wolbachia have the development of efficient and convergent syntheses, allowing for the a tightly regulated lifecycle within filarial nematodes. While Wolbachia greatest amount of structural diversity and chemical space to establish density is low in microfilariae and vector stage larvae, much higher structure-activity relationships (SAR). In vitro worm killing assays indicated numbers of the endobacteria are observed in developing larvae and young that three of these analogs were as good as or better than OXA itself. adult worms recovered from mammalian hosts. We have recently reported These new compounds with antischistosomal activity have been soaked that Wolbachia release polymorphic outer membrane vesicles (OMV) that into the SmSULT•PAP crystals and their mode of binding elucidated. may be essential for their mutualistic relationship with filarial worms. OMV This information will be used to synthesize the next generation of OXA may transport bacterial products that are required for parasite growth derivatives. and development. Tetracyclines (TET, antibiotics that inhibit bacterial protein synthesis) clear Wolbachia from filarial worms over a period of 1825 weeks. This treatment first sterilizes the parasites and eventually kills them. The present study was performed to elucidate the early effects of TET ROLES OF ATP BINDING CASSETTE (ABC) MULTIDRUG treatment on the morphology of Wolbachia and filarial worms. Gerbils TRANSPORTERS IN SCHISTOSOME PHYSIOLOGY, DRUG with i.p. Brugia malayi infections were treated with TET on days 19 and 20 SUSCEPTIBILITY AND PARASITE-HOST INTERACTIONS post-infection (i.p. injection, 5 mg/kg). Immature female worms recovered Ravi S. Kasinathan, Yue Ning, Robert M. Greenberg on day 21 were studied by transmission electron microscopy using high pressure freezing/freeze substitution fixation. OMV were largely absent University of Pennsylvania, Philadelphia, PA, United States near Wolbachia in TET-treated worms, while about half of the Wolbachia Although its value in the treatment and control of schistosomiasis is well in untreated control worms were associated with OMV. Wolbachia in established, praziquantel (PZQ) has significant limitations. Most notably, treated worms were often surrounded by membranes that appeared to it is largely ineffective against immature schistosomes. It is also essentially come from the endoplasmic reticulum. Lateral chords in treated worms the only drug available for a disease afflicting hundreds of millions. New were heavily vacuolated with increased glycogen granules compared to therapeutics or adjuncts to enhance PZQ activity and overcome possible untreated worms. Thus TET treatment appears to block OMV production drug resistance are urgently needed. We hypothesize that ATP binding by Wolbachia and promote encapsulation of the bacteria by internal host cassette (ABC) multidrug transporters offer attractive candidate targets cell membranes. for new or repurposed drugs that either act as anthelmintics on their own, or that enhance parasite susceptibility to existing anthelmintics. 1827 ABC transporters such as P-glycoprotein (Pgp) mediate efflux of metabolic toxins, xenobiotics, and signaling molecules, and are associated with drug POPULATION GENOMICS OF WUCHERERIA BANCROFTI resistance in many organisms, including parasitic helminths. They exhibit ELIMINATION FROM PAPUA NEW GUINEA broad substrate specificity and are inhibited by several drugs currently in 1 2 2 3 clinical use. ABC transporters are also implicated in a variety of normal Scott T. Small , David Serre , James F. Hester , Lisa J. Reimer , 1 1 physiological activities such as excretion, maintenance of permeability Daniel J. Tisch , Peter A. Zimmerman barrier function, and modulation of immune responses. They transport 1Case Western Reserve University, Cleveland, OH, United States, 2Cleveland many potent signaling molecules with high affinity, including several Clinic, Cleveland, OH, United States, 3Liverpool School of Tropical with immunomodulatory activity. Schistosomes exposed to PZQ increase Medicine, Liverpool, United Kingdom expression of ABC transporters such as Pgp (SMDR2) and multidrug Lymphatic filariasis (LF) is a major threat to human health in the tropics, resistance associated protein (SmMRP1), and worms with reduced PZQ leading to the disfiguring and debilitating conditions hydroceles and sensitivity show higher basal expression of these transporters. PZQ is also elephantiasis. There are approximately 1.2 billion people living in LF both an inhibitor and likely substrate of schistosome Pgp. Disruption of endemic countries where 120 million people currently suffer from the transporter expression (by RNAi) or function (by inhibition) enhances the disease. The parasites that cause LF are separated into two genera, activity of PZQ against adult parasites, and renders PZQ-refractory juvenile Brugia and Wuchereria, with W. bancrofti (Wb) responsible for ~90% worms susceptible to the drug. Schistosome ABC transporters also appear of LF cases. Until recently, Wb has been neglected in genomic studies to be important for normal schistosome egg production. We are currently due to complications in sample collection and preparation, such as lack exploiting molecular and pharmacological tools to understand the of adult stage samples. Here, we report 13 new Wb genomes from mechanism by which schistosome ABC transporters alter PZQ susceptibility the Dreikikir region of Papua New Guinea (PNG). We utilized multiple and to assess the role of these transporters in the parasite’s modulation displacement amplification to amplify and sequence 13 juvenile stage of host immune responses. These experiments could lend important (L3) Wb worms from four patient infections. We report the discovery of insights into schistosome physiology and possibly provide targets for novel 60,000 novel single nucleotide polymorphism (SNPs) and 200 polymorphic antischistosomals. microsatellite loci from the genomes. Within patient infections we find that genetic diversity is high, yet concentrated in specific regions of the genome, with large tracts of intervening homozygous sequence. We also identify candidate regions that harbor genes with extended haplotypes and shifted frequency spectrums, signals of either ongoing or recent positive selection. We discuss our results in the context of the recent mass drug administration, and identify SNP loci ideal for future monitoring of elimination success.

astmh.org 558 1828 liver stage maturation showed that persistence and activation of P. vivax hypnozoites can be further investigated in the model to determine the NOVEL TEGUMENT EXPRESSED KUNITZ TYPE PROTEASE biological basis for liver stage dormancy. Successful evaluation of the INHIBITOR FROM SCHISTOSOMA MANSONI antimalarial drugs with known activities on P. vivax liver stage infection (Primaquine and Atovaquone) confirmed that the FRG huHep/P. vivax Shiwanthi L. Ranasinghe, Katja Fischer, Geoffrey Gobert, infection model could be used as an efficient platform for testing new Donald P. McManus antimalarial drugs in vivo in quest to accelerate the development of QIMR Berghofer Medical Research Institute, Herston, Australia interventions for the radical cure of P. vivax infection. Schistosomiasis is one of the most prevalent and serious parasitic diseases of tropical and subtropical regions with 779 million people being at risk 1830 and 207 million infected. Kunitz type proteins belong to the I2 family of protease inhibitors and are involved in diverse biological processes in CHARACTERIZATION OF COENZYME A BIOSYNTHESIS invertebrates. One secretory type sequence (SmKI) having similarity to PATHWAY REVEAL ESSENTIAL DISTINCTIVE FUNCTIONS Kunitz type protease inhibitors was identified from recent mining of the DURING PLASMODIUM DEVELOPMENT IN BLOOD AND genome of Schistosoma mansoni. Recombinant SmKI was expressed in E. MOSQUITO coli, purified and an antiserum against rSmKI was produced in mice and subsequent immunolocalization and western blotting carried out. Gene Robert J. Hart, Catherine Nation, Panpim Thongsripong, Ahmed expression levels were determined within key lifecycle stages of S. mansoni S. I. Aly using real-time PCR. Serine protease inhibitory assays were also used to Tulane University, New Orleans, LA, United States determine the inhibitory effect of the rSmKI on trypsin, chymotrypsin, Coenzyme A (CoA) is an essential universal cofactor and carrier of neutrophil elastase (NE), pancreatic elastase and Cathepsin G. Real time acyl groups for all prokaryotic and eukaryotic cells. In nearly all non- PCR indicated SmKI is highly expressed in adult worms which reside in the photosynthetic cells, CoA biosynthesis depends on the uptake and mesenteric venules of the definitive host. Immunolocalization showed the phosphorylation of vitamin B5 (pantothenic acid or pantothenate). Kunitz protein is present in the tubercles of the male tegument and along Earlier Studies showed the importance of pantothenate acquisition and the tegument of the female worm. Notably, western blots showed the phosphorylation for Plasmodium survival within erythrocytes. Recently, level of SmKI was higher in the excretory secretory products of adult worm pantothenate plasma membrane transporter (PAT) was functionally pairs than in soluble worm antigens. Further, rSmKI inhibited trypsin, characterized in Plasmodium falciparum. PAT was shown to be refractory chymotrypsin and NE, with the highest inhibitory activity recorded against to deletion and was localized to the parasite plasma membrane. However, trypsin. Initial screens indicated that rSmKI interfere with both intrinsic very little is known about the in vivo cellular functions of CoA biosynthesis and extrinsic blood coagulation pathways as well, indicating another pathway in malaria parasite life cycle stages. We have targeted all enzymes important function. Thus, the SmKI protein may play an important role in of this pathway for deletion in the mouse malaria model P. yoelii. We show schistosome survival in blood by inhibiting NE as well as playing a key role that first enzymes of this pathway are dispensable for asexual and sexual in evading host immune responses. As SmKI is secreted and exposed to the blood stage (BS) development but they are essential for mosquito stages host immune system, we consider that rSmKI may be a useful candidate development and sporozoite production. However, the last enzymes of as novel vaccine target to control schistosomiasis. Assays are underway this pathway are essential for both BS and mosquito stages development. to further understand the function of SmKI and vaccine/challenge These results indicate that the first substrates and intermediate products experiments will be undertaken to evaluate its protective efficacy. of this pathway can be supplemented by alternative novel pathways inside the blood but not inside the mosquito midgut. Collectively, our 1829 data show that CoA de novo biosynthesis is essential for both BS and mosquito stages. This is the first in vivo functional characterization of CoA PLASMODIUM VIVAX LIVER STAGE DEVELOPMENT AND biosynthesis pathway in any protozoan parasite. HYPNOZOITE FORMATION IN THE FRG HUHEP MOUSE MODEL 1831 1 2 Sebastian A. Mikolajczak , Niwat Kangwanrangsan , Ashley A NOVEL RNA APTAMER SYSTEM FOR FUNCTIONAL 1 2 2 M. Vaughan , Wanlapa Roobsoong , Rachaneeporn Jenwithisuk , GENETICS IN PLASMODIUM FALCIPARUM Narathatai Yimamnuaychok2, Matt Fishbauger1, Viswanathan Lakshmanan1, Alexis Kaushansky1, Scott E. Lindner3, John H. Suresh Maddur Ganesan, Jacquin C. Niles Adams4, Jetsumon Sattabongkot2, Stefan H. Kappe1 Massachusetts Institute of Technology, Cambridge, MA, United States 1Seattle BioMed, Seattle, WA, United States, 2Mahidol University, Bangkok, Malaria is a parasitic disease that is widespread in tropical and subtropical Thailand, 3Pennsylvania State University, University Park, PA, United States, regions, and a major cause of human morbidity and mortality. The most 4University of South Florida, Tampa, FL, United States severe form of malaria is caused by the parasite, Plasmodium falciparum. The ability of Plasmodium vivax to form dormant liver stages (hypnozoites) A limited set of antimalarial drugs is used to treat the disease, but drug that can be activated weeks or months after initial infection to cause resistance is an increasing problem. Hence, identification of novel anti- relapse of malaria is of crucial importance for the unprecedented malarial drugs is a high priority. While our understanding of Plasmodium epidemiological success of the parasite. Yet, little progress has been made biology has increased in the post-genomic era, tools for doing functional to understand the unique biology of hypnozoite formation and activation. genetics remain quite limited. This impedes progress in identifying key Due to the parasite’s strong preference for nonhuman primate and human parasite genes and processes that can be prioritized for drug development tissue, the availability of models to study P. vivax liver stages is extremely efforts. To address this need, our laboratory previously developed a limited. Here we report that the FRG KO mouse model transplanted novel small molecule-regulated protein-RNA interaction (TetR-aptamer with human primary hepatocytes (huHep) efficiently supports the system) that facilitates robust and inducible regulation of target gene development of P. vivax liver stages as well as the formation of hypnozoites translation in eukaryotic organisms including Plasmodium. Here, we for Thai isolates of P. vivax. Using a series of P. vivax specific polyclonal present the application of protein engineering approaches to integrate our and monoclonal antibodies, we were able to evaluate the liver stage synthetic control system with native Plasmodium translational regulatory progression and maturation in the infected liver. The ability of the exo- mechanisms. In so doing, we achieve substantially increased regulatory erythrocytic merozoites to establish blood stage infection upon transfusion dynamic ranges (up to 200-fold) compared to a 5-10 fold range of the with human reticulocytes is being currently evaluated. Furthermore, P. original system. With a view to identifying new potential drug targets, vivax infections in the FRG huHep mice carried beyond the time of the we are using this system to study several parasite genes. We envision that

astmh.org 559 this enhancement in regulatory dynamic range will facilitate functional a reduced effective population size. We developed tools to visualize interrogation of larger numbers of parasite genes with greater confidence parasite inter-relatedness by molecular barcode and sequencing and that associated biological outcomes can be readily identified. identified increasing levels of identity by descent in both. SNP genotyping of monogenomic samples showed clusters with identical molecular 1832 barcodes, including several collections where 25-30% of samples shared the same barcode. Whole genome sequence analysis revealed that EXPLORING COMPLEX MALARIA INFECTIONS WITH SINGLE approximately half of the independent isolates shared between 10 and GENOME SEQUENCING 98% of their genomes with other sequenced samples, including one obviously hybrid parasite. To our knowledge, this is the first observation of Shalini Nair1, Standwell Nkhoma2, Karla Gorena3, Benjamin J. increasing identity by descent in an African population. We show evidence Daniel3, Francois Nosten4, Tim J. Anderson1, Ian H. Cheeseman1 of significant parasite population changes undetectable by standard 1Texas Biomedical Research Institute, San Antonio, TX, United States, epidemiological methods. Early identification of population genomic 2Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, changes associated with changes in transmission offers refined criteria for Malawi, 3University of Texas Health Science Center San Antonio, San milestones tracking progress towards elimination. The decreasing costs of Antonio, TX, United States, 4Shoklo Malaria Research Unit, Mae Sot, genomic analysis make this a feasible option for surveillance of malaria Thailand control efforts. We have recently developed a single cell genomics platform capable of generating whole genome sequence from a individual parasitized red 1834 blood cells. This has been extensively validated and can generate SNP calls genome-wide with high (>99%) accuracy, even in species such as P. DENGUE VIRUS NONSTRUCTURAL PROTEIN 1 CONTRIBUTES vivax for which long term culture is not possible. This method provides TO VASCULAR LEAK IN VITRO AND IN VIVO, WHICH CAN BE a powerful approach for empirically determining the composition of BLOCKED BY ANTI-NS1 ANTIBODY multiple genotype parasite infections, and provides information that is not accessible using standard illumina sequencing of parasite infections. We P. Robert Beatty, Sarah Killingbeck, Henry Puerta Guardo, Eva have applied this approach to complex malaria infections by sequencing 17 Harris single genomes from multiple-clone infections of Plasmodium falciparum Division of Infectious Diseases and Vaccinology, School of Public Health, (n=1) or P. vivax (n=2). After stringent quality control we scored an average University of California Berkeley, Berkeley, CA, United States of 62,720 P. vivax SNPs and 61,080 P. falciparum SNPs from each single Dengue virus (DENV) is a mosquito-borne flavivirus consisting of 4 cell sequence allowing us to map within host divergence between single serotypes that causes ~100 million cases of dengue annually. DENV parasite genomes at exceptional resolution. We use this data to highlight nonstructural protein 1 (NS1) is secreted by infected cells and is found how single cell sequencing can be used to reconstruct genome-wide drug at high levels in patient serum during acute infection. We examined the resistance haplotypes from individual infections. Such “phasing” data is protective efficacy of NS1 immunization against lethal DENV2 infection in expected to be of critical importance for determining the outcome of drug a mouse model of vascular leak. Interferon α/β receptor-deficient C57BL/6 treatment, but cannot currently be determined from bulk sequencing of (Ifnar-/-) mice were injected intraperitoneally 3X over 6 weeks with 20 µg infections. Second, we have examined the size of blocks of haplotype of DENV2 recombinant NS1 (rNS1) combined with different adjuvants, sharing between genomes within infections and compared these with including alum, Sigma adjuvant system (SAS), CpG DNA, Addavax and/or population data from single clone infections. We observed that parasite monophosphoryl lipid A (MPLA). Two weeks after the third immunization, genotypes within infections tend to be closely related. Application of mice vaccinated with DENV2 rNS1 with either SAS + CpG or Addavax single cell genomics and can reveal patterns of relatedness at a fine scale, + MPLA were fully protected against lethal peripheral challenge with both within and between malaria infections. DENV2, whereas mice vaccinated with DENV2 rNS1 with alum or CpG DNA alone were not protected. In addition, heterologous cross-protection 1833 was observed, as 75% of mice vaccinated with DENV1 NS1 survived lethal DENV-2 challenge. Because NS1 vaccination blocked DENV pathogenesis, USING GENOMICS TO TRACK PROGRESS TOWARDS we hypothesized that NS1 itself may have direct pathogenic effects. MALARIA ELIMINATION We found that Ifnar-/- mice inoculated intravenously with 10 mg/kg of DENV2 NS1 combined with a sublethal dose of DENV2 succumbed 3-4 Rachel Daniels1, Stephen F. Schaffner2, Noah M. Daniels3, Wesley days post-infection equivalently to mice receiving a lethal dose of DENV2. Wong4, Nicholas Baro4, Hsiao-Han Chang4, Daouda Ndiaye5, Mice inoculated with 10 mg/kg DENV2 NS1 alone exhibited morbidity Daniel E. Neafsey2, Dyann F. Wirth4, Sarah K. Volkman6, Daniel L. but 100% survived, as did control mice receiving a sublethal dose of Hartl1 DENV2. We then tested the direct toxicity of NS1 on endothelial integrity 1 2 Harvard University, Cambridge, MA, United States, Broad Institute, in a trans-endothelial electrical resistance (TEER) in vitro assay. When 3 Cambridge, MA, United States, Massachusetts Institute of Technology, rNS1 was added to cultured human pulmonary microvascular endothelial 4 Cambridge, MA, United States, Harvard School of Public Health, Boston, cells (HPMEC) in a transwell system, the relative TEER value decreased 5 MA, United States, Cheikh Anta Diop University, Dakar, Senegal, compared to untreated or OVA-treated HPMEC cells. We next investigated 6 Simmons College, Boston, MA, United States if the in vivo lethality and in vitro disruption of endothelial integrity caused WHO recommendations for regions of low and moderate endemicity call by rNS1 could be inhibited by NS1-immune serum. Ifnar-/- mice passively for program re-orientation at milestones marked by changes in disease administered anti-NS1 serum after receiving sublethal DENV2 + NS1 prevalence. However, genetic changes in parasite populations may protein were completely protected against death while those receiving occur before changes in prevalence are measurable. To identify these control serum were not. In the HPMEC assay, the disruptive effects of rNS1 genomic changes, we examined Plasmodium falciparum samples collected on TEER were prevented by NS1-immune serum but not serum from OVA- from patients in Thiès, Senegal from 2006 to 2013. We genotyped 24 immunized or control mice. Thus, DENV NS1 appears to directly contribute independent SNPs from across the parasite genome - the molecular to increased vascular permeability, which can be blocked by anti-NS1 barcode - to separate annual collections into monogenomic (single antibody. parasite genome) and polygenomic (multiple parasite genome) infections. We also performed whole genome sequencing on 190 parasites from monogenomic infections. Following increased control efforts beginning in 2008, we observed large changes in population allele frequencies each season, suggesting enhanced random genetic drift expected from

astmh.org 560 1835 pathogenesis remains uncertain. Here we evaluated the role of soluble NS1 (sNS1) in inducing endothelial dysfunction. Cultures of human pulmonary DIFFERENCES IN TYPE I INTERFERON SIGNALING microvascular endothelial cells grown on a transwell permeable membrane ANTAGONISM BY DENGUE VIRUSES IN HUMAN AND NON- system as a model of barrier function in vitro were exposed to sNS1 (0.2- HUMAN PRIMATE CELL LINES 20 µg/mL), and endothelial permeability was examined by continuously measuring the trans-endothelial electrical resistance (TEER). sNS1 induced 1 2 3 Freddy A. Medina , Giselle Torres , Amanda J. Chase , Gilberto a significant dose-dependent increase in endothelial permeability starting 1 1 1 A. Santiago , Juan F. Medina , Luis M. Santiago , Jorge L. Muñoz- 2 hours post-treatment (hpt), at 5 and 20 µg/mL (20 and 50% decrease in 1 Jordán TEER, respectively). This effect persisted for more than 24 h as compared 1Centers for Disease Control and Prevention, San Juan, PR, United States, to the TEER baseline values exhibited by untreated controls and treatment 2University of Puerto Rico Medical Science Campus, San Juan, PR, United with unrelated protein (20 µg/mL OVA). Lower concentrations (0.2 and States, 3Mercer University School of Medicine, Macon, GA, United States 1 µg/mL) showed less dramatic but still significant decreases in TEER that The type I interferon (IFN-α/β) response has been shown to be one of returned to baseline 6-12 hpt. Confocal microscopy revealed concomitant the most regulated systems during dengue virus infections. Moreover, alterations in intercellular junctional proteins. Our findings suggest a new attenuation of IFN stimulated genes has been associated with severe mechanism of sNS1 directly triggering endothelial vascular dysfunction disease. In this study we wanted to address conflicting reports of inhibition that occurs in severe dengue disease. of IFN signaling by dengue viruses and if there were variations in viruses that differed in their pathogenicity. Using a method that combines flow 1837 cytometry and a four-parameter logistic regression model we compared the relative inhibition of IFN-α/β signaling between viruses. Our results MICROVASCULAR AND ENDOTHELIAL FUNCTION IN showed that all dengue virus serotypes were capable of inhibiting IFN PREDICTING CLINICAL OUTCOME OF DENGUE signaling in human cells. A more refined analysis of well-characterized Sophie Yacoub1, Cameron Simmons2, Heiman Wertheim1, DENV-2 clinical isolates from the five DENV-2 genotypes demonstrated Nguyen Van Kinh3, Gavin Screaton4, Bridget Wills2 that all viruses inhibited IFN signaling in human cells, but sylvatic viruses 1Oxford University Clinical Research Unit, Hanoi, Vietnam, 2Oxford displayed a superior ability to inhibit STAT1 phosphorylation. We analyzed University Clinical Research Unit, Ho Chi Minh City, Vietnam, 3National inhibition of STAT1 phosphorylation by sylvatic strains in non-human Hospital for Tropical Diseases, Hanoi, Vietnam, 4Imperial College London, primate cell lines and to our surprise there was no blockage. To determine London, United Kingdom if these observations were specific to sylvatic strains we performed our IFN inhibition assay with a prototypical DENV2 Asian strain and confirmed Dengue can present with a broad spectrum of clinical phenotypes. The that inhibition of STAT1 phosphorylation by dengue viruses does not occur hallmark of severe disease is increased vascular permeability, sometimes in non-human primate cell lines. However, dengue virus was capable leading to hypovolaemic shock. However microvascular/endothelial of inhibiting IFN signaling in both human and Rhesus macaque primary dysfunction are difficult to assess clinically. A prospective observational dendritic cells. IFN-α production was detected in supernatants of dengue study recruiting a) patients presenting to the OPD with fever for <72 hours virus infected Rhesus macaque dendritic cells and contrast to published and a clinical diagnosis of possible dengue, and b) patients hospitalized studies that have suggested that dengue virus can inhibit IFN-α production with warning signs or established severe disease, is ongoing in two in human cells. The observed differences in inhibition of the IFN-α/β hospitals in Hanoi and Ho Chi Minh City, Vietnam. Clinical, laboratory, pathway in human and non-human primate cells may be cell type specific and haemodynamic assessments are performed daily for a maximum or could result from the transformation process. Nevertheless, these of 6 days, and again at follow-up 2 weeks later. Microvascular imaging studies provide awareness of differences in the manipulation of the IFN using Sidestream Darkfield Imaging (SDF) and endothelial function testing system by dengue virus in human and non-human primate cells. using peripheral artery tonometry (EndoPAT) are performed at enrollment, defervescence/hospital discharge and follow-up. To date, 167 patients have been recruited, 92 in the outpatient arm and 75 in the inpatient arm. 1836 The median age is 27 years (range 5-65 years) and 47% are male. In the outpatient arm 29/67 (43%) of the confirmed dengue cases developed DENGUE VIRUS NON-STRUCTURAL PROTEIN-1 (NS1) warning signs and 3/67 (4%) developed shock, while 25/92 (27%) were INCREASES HUMAN PULMONARY ENDOTHELIAL CELL diagnosed as having other febrile illnesses (OFI). At enrolment, the reactive PERMEABILITY IN VITRO hyperaemic index (RHI), a marker of endothelial function, was lowest in Henry Puerta Guardo, Dustin Glasner, Eva Harris the patients who went on to develop severe dengue (median [range] 1. 54 [1.36-1.96]) followed by those who developed warning signs (1.78 Division of Infectious Diseases and Vaccinology, School of Public Health, [1.17-3.5]) and then uncomplicated dengue (2.18 [1.16-2.29]). In the University of California Berkeley, Berkeley, CA, United States OFI category the RHI was 1.63 [1.22-3.38]. Results for the inpatient arm Dengue is the most prevalent arboviral disease in humans and a major showed a similar trend with the lowest RHI seen in severe dengue patients. public health problem worldwide. Systemic plasma leakage leading to The SDF images are being analysed; initial results show microvascular flow profound shock and potentially fatal complications is a critical determinant is impaired in early dengue with a lower proportion of perfused vessels, of dengue severity. Increased vascular permeability without morphological mean flow index and vessel density compared with follow-up. These damage to the capillary endothelium seen in severe dengue suggests preliminary results suggest microvascular and endothelial dysfunction the shock syndrome may be due to endothelial dysfunction. In the are associated with dengue disease severity, and can be detected prior endothelium, dynamic structures including intercellular junctional proteins to severe clinical manifestations. These techniques may prove useful as and the endothelial glycocalyx control the barrier function critical for outcome predictors and/or to monitor endothelium-directed therapies. vascular homeostasis. In certain diseases, e.g., dengue, functional and structural alterations modify the normal architecture of the vessel wall, increasing plasma extravasation. Dengue pathogenesis involves a complex interaction of the virus and host immune response, including cross-reactive antibodies and T cells, complement activation, and elevated levels of cytokines and other soluble mediators that correlate with severe disease. However, the mechanism of vascular dysfunction in dengue disease is still unclear. Secreted and cell-surface-associated dengue virus nonstructural protein 1 (NS1) and anti-NS1 antibodies are implicated in contradictory roles of protection and pathogenesis, and how NS1 contributes to dengue astmh.org 561 1838 of cGAS during DENV infection we evaluated the ability of this protein to be activated and trigger type I IFN during DENV infection. We also MONOCYTE RECRUITMENT TO THE DERMIS AND investigated the role of the NS2B3 protease complex in the inhibition of DIFFERENTIATION TO DENDRITIC CELLS INCREASES type I IFN production induced by cGAS. We have found a novel mechanism THE NUMBERS OF TARGETS FOR EARLY DENGUE VIRUS of type I IFN inhibition by the DENV protease through the interference of REPLICATION the cGAS/STING/IRF3 pathway. Also, over expression of cGAS impaired DENV replication. Alternatively, silencing of cGAS in human dendritic cells Michael A. Schmid, Eva Harris (DCs) resulted in a higher accumulation of DENV RNA after infection. Division of Infectious Diseases and Vaccinology, School of Public Health, These results suggest an active role of cGAS as a sensor during DENV University of California Berkeley, Berkeley, CA, United States infection and confirm the role of DENV protease as a master regulator of The four serotypes of dengue virus (DENV1-4) cause the most prevalent the type I IFN response in DENV infected cells arthropod-borne viral disease in humans. Although Aedes mosquitoes transmit DENV via the skin and studies have identified Langerhans cells as 1840 targets of DENV replication in the epidermis, no information exists about the immune response and DENV infection in the dermis. DENV suppresses DEVELOPMENT OF A NON-HUMAN PRIMATE MODEL FOR the interferon response, replicates, and causes disease in humans but SECONDARY DENGUE VIRUS INFECTION USING MARMOSETS not in wild-type mice. Here, C57BL/6 mice lacking the interferon-α/β (CALLITHRIX JACCHUS): DETECTION OF VIRUS IMMUNE- receptor (Ifnar-/-) had normal frequencies of hematopoietic cells in the COMPLEX USING FCΓ RECEPTOR EXPRESSING CELLS skin, were susceptible to intradermal DENV2 infection, and developed Meng Ling Moi1, Kenji Shirai2, Yasushi Ami3, Chang-kweng Lim1, disease that displayed key features of severe dengue in humans. For the Yuriko Suzaki3, Kazutaka Kitaura2, Masayuki Saijo1, Ryuji Suzuki2, first time, we identified dermal dendritic cells (DCs), macrophages, and Ichiro Kurane4, Tomohiko Takasaki1 monocytes as targets for DENV replication in the dermis of Ifnar-/- mice. 1 We made the following observations. (1) CD103+ DCs and macrophages Department of Virology 1, National Institute of Infectious Diseases, Tokyo, 2 were present in the steady-state dermis and were the first DENV-infected Japan, Department of Rheumatology and Clinical Immunology, Clinical cells in the skin 12 hours post-inoculation (hpi); they then decreased in Research Center for Allergy and Rheumatology, Sagamihara National 3 frequency over time and no longer contributed to DENV replication after Hospital, National Hospital Organization, Kanagawa, Japan, Division of 48 h. (2) Substantial numbers of CD11b+ Ly6C- DCs were present and Experimental Animal Research, National Institute of Infectious Diseases, 4 were continuously DENV-infected between 12 and 72 hpi. (3) Ly6Chigh Tokyo, Japan, National Institute of Infectious Diseases, Tokyo, Japan monocytes were actively recruited to the DENV-infected dermis as early Infection with one dengue virus (DENV) serotype does not offer protection as 12 hpi and, by 48 h, differentiated to Ly6C+ monocyte-derived DCs against secondary infection against a heterologous serotype. Antibodies to (moDCs). Ly6Chigh monocytes and Ly6C+ moDCs became DENV-infected dengue virus (DENV) possess two competing activities: antibody-mediated 48-72 hpi and were then the major subsets for DENV replication in the virus neutralization that leads to protection and infection-enhancement skin. Finally, adoptive transfer of Ly6Chigh monocytes from Ifnar-/- and WT that may cause severe complications. In this study, marmosets (Callithrix mice confirmed recruitment of circulating monocytes to the DENV-infected jacchus) were infected with DENV-2 and subsequently inoculated with dermis, differentiation to Ly6C+ moDCs, and DENV infection of de novo DENV-1, DENV-2 or DENV-3 to evaluate the model utility as an infection recruited cells. Our study identifies dermal DCs and macrophages as the model for DENV infection. Viremia levels were determined by RT-PCR, initial targets for DENV replication in the skin. Further, we establish a novel BHK and FcγR-expressing cell lines. Antibody response was determined by mechanism of how DENV exploits the immune response in the dermis by IgM and IgG ELISA, and neutralizing antibody titers were determined by recruiting monocytes and moDCs, which then become the major targets using BHK cells and FcγR-expressing BHK cells. All marmosets consistently for virus replication in the skin. developed viremia after secondary heterologous infection and primary infection. Viremia was absent during secondary homologous challenge. 1839 As compared to primary infection, viremia during secondary heterologous challenge persisted longer. Higher levels of viremia were detected using DENGUE VIRUS INFECTION INHIBITS THE CGAS/STING/IRF3 FcγR-expressing cells as compared to FcγR-negative cells during secondary PATHWAY IN INFECTED HUMAN CELLS heterologous challenge in marmosets, suggesting presence of infectious virus-immune complex during secondary infection. However, levels of 1 1 2 Sebastian Aguirre , Francise Lamote , Jenish Patel , Ana viremia were similar after primary challenge using FcγR-expressing cells 1 1 2 Maestre , Dabeiba Bernal-Rubio , Adolfo Garcia-Sastre , Ana and FcγR-negative cells. IgM and IgG antibody response in primary and 2 Fernandez-Sesma secondary inoculation were consistent to those of human DENV infection. 1Department of Microbiology and the Global Health and Emerging Marmosets also exhibited thrombocytopenia, leucopenia and increase Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York in AST, ALT and LDH levels during DENV infection. The animal model City, NY, United States, 2Department of Microbiology and the Global also demonstrated enlarged liver and kidney during secondary DENV Health and Emerging Pathogens Institute, Icahn School of Medicine at infection. Neutralizing antibodies were serotype cross-reactive in FcγR- Mount Sinai and Icahn Graduate School of Biomedical Sciences at Mount negative cells but were specific to primary serotype in FcγR-expressing Sinai, New York City, NY, United States cells. During secondary infection, marmosets demonstrate viremia and Dengue virus (DENV) has become the most relevant arthropod-borne virus antibody responses consistent with those of human DENV infection. that affects humans. To productively infect the host, DENV needs to inhibit Strong antibody responses induced after secondary heterologous infection the host innate immune system, particularly the type I interferon (IFN) possess high neutralizing antibody titers against all four DENV serotypes. system. Our group and others have demonstrated that DENV interferes The results suggest the potential of marmosets as a useful animal model with both, the production and signaling of type I IFN pathways through for DENV infection. the expression of viral proteins that specifically target host proteins involved in these essential responses to pathogens. Our group showed that the DENV protease complex NS2B3 is able to interact and cleave the adaptor STING to inhibit the activation of IRF3 in infected cells. A recent report showed an anti flavivirus activity of the newly described pattern recognition receptor cGAS, which after activation generate a second messenger (cGAMP) that in turn activates the adaptor STING and the subsequent induction of type I IFN. In order to investigate the role

astmh.org 562 1841 1960 and March 2014, in which patients were enrolled and treated with antimalarials and where gametocyte data were recorded. Individual MOSQUITO INFECTIVITY AND GAMETOCYTE CARRIAGE patients data from over 100 studies (n>40,000 patients) were collated, AMONG PATIENTS PRESENTING WITH UNCOMPLICATED curated and included in analysis. Data from 21 African and 6 Asian FALCIPARUM MALARIA IN NORTHERN CAMBODIA countries was analysed for gametocyte carriage following treatment with artemether-lumefantrine, amodiaquine-artesunate, dihydroartemisinin- 1 2 2 Jessica T. Lin , Ratawan Ubalee , Chanthap Lon , Michele piperaquine, and mefloquine-artesunate. An apriori data analysis plan 2 1 2 Spring , Sujata Balasubramanian , Ponsa Chanudom , Namtip was developed to identify factors associated with gametocyte prevalence 2 2 2 Trongnitpatt , Worachet Kuntawunginn , Anintita Laoboonchai , and density prior to treatment and following treatment with an ACT. 1 2 3 Christian Parobek , Nitima Chanarat , Char Meng Chuor , Jessica Criteria for the quality of gametocyte assessments have been ascribed 2 1 2 Manning , Steve Meshnick , Charlotte A. Lanteri , David L. to the various studies. In conclusion, the effects of asexual parasite 2 Saunders density, age, transmission intensity and haemoglobin concentration on 1University of North Carolina, Chapel Hill, NC, United States, 2Armed enrolment gametocyte prevalence and density will be presented. The Forces Research Institute of Medical Sciences, Bangkok, Thailand, differential effect of ACTs on post-treatment gametocyte carriage, density 3National Center for Parasitology, Entomology and Malaria Control, Phnom and carriage time will be examined in relation to ACT regimen, parasite Penh, Cambodia clearance time, transmission intensity and human host factors. The results Halting the spread of malaria relies on identifying the transmission of this important study and their relevance for malaria elimination will be reservoir and providing effective transmission blocking therapy. highlighted. Membrane feeding studies have shown that a substantial proportion of Plasmodium falciparum infected African children are capable of infecting 1843 mosquitoes in the absence of smear-detectable gametocytes. Less is known about transmission in Southeast Asia. We sought to determine IVERMECTIN FOR MALARIA CONTROL: INSIGHTS FROM the infectivity of adults with P. falciparum infection in northern Cambodia MODELLING relative to gametocytemia. As part of a therapeutic efficacy study of Hannah C. Slater1, Patrick G. Walker1, Teun Bousema2, Azra C. dihydroartemisinin-piperaquine, half of patients with uncomplicated P. Ghani1 falciparum were randomized to receive a 45mg single dose primaquine 1Imperial College, London, United Kingdom, 2London School of Hygiene & on the 3rd day of dosing. Patient blood was membrane-fed to Anopheles Tropical Medicine, London, United Kingdom dirus mosquitoes prior to treatment and on days 4, 7, and 14 following treatment. At Day 9 after membrane feeding, 50 mosquitoes were Ivermectin (IVM), used alongside mass treatment strategies with an dissected for oocyst detection, while another 50 were saved at Days 9 and artemisinin combination therapy (ACT), has been suggested as a possible 16 for parasite detection by real-time PCR. Among 108 patients studied, 7 tool for reducing malaria transmission. Mosquitoes feeding on humans (6.5%) patients carried smear-detectable gametocytes at baseline (median that have recently ingested ivermectin have a reduced lifespan, meaning 66 gametocytes/μL, range 5-728), and only 2 of 7 successfully infected they have a lower probability of completing sporogony and they complete mosquitoes. Both transmitters had high levels of gametocytemia (705 fewer gonotrophic cycles. We use human pharmacokinetic data and and 728 gametocytes/μL) resulting in high oocyst prevalence (26% and mortality data for mosquitoes taking bloodmeals containing IVM to 70% of mosquitoes with average 2 and 56 oocysts/midgut, respectively). quantify the mosquitocidal effect of IVM. This is incorporated into a Of the remaining patients without smear-detectable gametocytes, only transmission model to estimate the impact of IVM in combination with 1 was infectious to mosquitoes, resulting in 8% oocyst prevalence mass treatment strategies with an ACT on transmission metrics. Adding with an average of 1 oocyst/midgut. These results show a 30-fold IVM increases the reductions in parasite prevalence achieved and delays greater transmission potential in patients with microscopic P. falciparum the re-emergence of parasites compared to mass treatment alone. This gametocytemia. PCR analysis for submicroscopic gametocytemia and transmission effect is obtained through its effect on vector mortality. IVM oocyst positivity is currently in progress. However, our findings suggest that effectiveness depends on coverage with the highest impact achieved in an area with low P. falciparum endemicity where gametocyte carriage is if given to the whole population rather than only those with existing relatively rare, only a small minority of patients with symptomatic malaria detectable parasites. Our results suggest that including IVM in a mass contribute to the bulk of human-to-mosquito transmission. treatment strategy can reduce the time taken to interrupt transmission as well as help to achieve transmission interruption in transmission settings 1842 in which mass treatment strategies alone would be insufficient. . We also investigate the optimal implementation of ivermectin administration THE EFFECT OF ARTEMISININ-COMBINATION THERAPY in a range of intervention scenarios, for example whether it best used TREATMENT OPTIONS ON PLASMODIUM FALCIPARUM alongside dihydroartemsinin-piperaquine or artemether lumefantrine GAMETOCYTE CARRIAGE: A POOLED ANALYSIS OF in a mass treatment intervention, whether there is any benefit of using INDIVIDUAL PATIENT DATA primaquine alongside ivermectin, whether ivermectin could be beneficial if used as a stand-alone drug prior to the peak transmission season, and Teun Bousema1, Chris Drakeley2, Philippe Guerin3, Georgina how the vector ecology and existing vector control interventions in a Humphreys3, Kasia Stepniewska3, Carol Sibley3, Ric Price3 specific region impact the efficacy of ivermectin. Overall, we find that 1Radboud University Medical Center, Nijmegen, Netherlands, 2London including IVM in mass treatment strategies could be a useful adjunct to School of Hygiene & Tropical Medicine, London, United Kingdom, reduce and interrupt malaria transmission. 3WorldWide Antimalarial Resistance Network (WWARN) Gametocyte Study Group, Oxford, United Kingdom The production of gametocytes during malaria is determined by a variety of parasite, human and environmental factors. ACTs rapidly clear the asexual parasite biomass in infected individuals, with potent gametocytocidal activity against early sexual stages of the parasites; they play a critical role in reducing the transmission of malaria and decreasing the spread of drug resistant parasites. We conducted a large pooled analysis of clinical data to examine the differential effect of ACTs on the transmission potential of Plasmodium falciparum. A systematic search of the literature was conducted to identify all studies published between

astmh.org 563 1844 schools and routine health services did not quite maintain the household coverage after one-year of implementation due to its late start almost two MIND THE GAP: ASSOCIATION BETWEEN HOUSE STRUCTURE years after the campaign. Results show, however, that a CD approach is AND MALARIA IN UGANDAN CHILDREN feasible. Lucy S. Tusting1, Humphrey Wanzirah2, Emmanuel Arinaitwe2, 1846 Agaba Katureebe2, Kilama Maxwell2, John Rek2, Christian Bottomley1, Sarah Staedke1, Moses Kamya3, Grant Dorsey4, Steve INCREASE IN INTRA-HOUSEHOLD ACCESS TO AND USE OF 5 W. Lindsay INSECTICIDE TREATED NETS (ITNS) IN SENEGAL 1London School of Hygiene & Tropical Medicine, London, United Kingdom, 1 2 2Infectious Disease Research Collaboration, Kampala, Uganda, 3Makerere Julie I. Thwing , Adama Faye University College of Health Science, Kampala, Uganda, 4University of 1Centers for Disease Control and Prevention, Atlanta, GA, United States, California San Francisco, CA, United States, 5Durham University, Durham, 2Université Cheikh Anta Diop, Dakar, Senegal United Kingdom With the increase in funding to support malaria control, malaria endemic Good house construction can lower malaria risk by reducing house entry countries have increased the availability of insecticide treated nets (ITNs) by vectors. We assessed how house design may affect mosquito house to their populations. Senegal used primarily social marketing until 2008, entry and malaria risk in Uganda. 100 households were enrolled in each when a mass distribution campaign targeting children under 5 was of three sub-counties: Walukuba, Jinja district; Kihihi, Kanungu district; conducted, covering 5 of 11 regions. A nationwide campaign targeting and Nagongera, Tororo district. Light trap collections were made monthly children under 5 was conducted in 2009. A rolling universal coverage in all homes. All children aged six months to ten years were followed campaign started in 2010, in which every sleeping space was counted prospectively to measure parasite prevalence routinely every three months and households received sufficient ITNs to cover every sleeping space, and malaria incidence by passive case detection. Homes were classified as accounting for existing nets in good condition. Between 2010 and early modern (cement, wood or metal walls; and tiled or metal roof; and closed 2013, the universal coverage campaign was implemented in all regions eaves) or traditional (all other homes). We will present the association of Senegal. Over this time, 690,000 ITNs were distributed in 2006; between house design and human biting rate, malaria infection and 1,065,141 in 2007; 1,586,522 in 2008; 2,532,018 in 2009; 1,258,663 clinical malaria and discuss the potential of housing as an intervention in 2010; 2,465,770 in 2011; and 983,725 in 2012. We used nationally against malaria, from low to very high transmission areas. representative survey data to track the evolution of intra-household access to ITNs (a newly recommended indicator), calculated as twice the number 1845 of ITNs divided by the number of persons in the household (not to exceed 100%). Based on this indicator, only 11% and 19% of the population MAINTAINING UNIVERSAL COVERAGE OF LONG had access to an ITN in 2005 and 2006, respectively. In 2008, after the LASTING INSECTICIDAL NETS: IMPACT OF CONTINUOUS subnational distribution to children under 5 years, access was 36%. The DISTRIBUTION ON HOUSEHOLD OWNERSHIP IN EASTERN post-campaign survey in 2009 indicated an increase in ITN access to REGION, GHANA 57%. Universal coverage was completed in four of 14 regions in 2010, resulting in access of 41% nationwide, and 70% in the campaign-covered Celine S. Zegers de Beyl1, Richmond Ato Selby2, Obed E. regions. Access was 63% in 2012, with all but two regions covered. Asamoah3, Joshua K. Ofori4, Constance Bart-Plange5, Hannah National-level household ownership of at least one ITN from these surveys Koenker6, Matthew C. Lynch6, Kojo Lokko6, Albert Kilian7 was 20% (2005), 36% (2006), 60% (2008), 82% (2009), 63% (2010), 1Malaria Consortium, London, United Kingdom, 2Malaria Consortium, and 72% (2012), while use by the general population was 6% (2005), Malaria Consortium Kampala, Uganda, 3Malaria Consortium/NetWorks, 12% (2006), 23% (2008), 34% (2009), 29% (2010), and 41% (2012). Accra, Ghana, 4NetWorks, Accra, Ghana, 5National Malaria Control Two-thirds of those with access to an ITN reported using it the previous Programme, Accra, Ghana, 6Center for Communication Programs, Johns night. Household ITN ownership is an inflated measure of ITN access. Hopkins University, Baltimore, MD, United States, 7Tropical Health LLP and Intra-household access is a more appropriate indicator for assessing Malaria Consortium, Montagut, Spain the gap between ownership and use. Access to nets closely reflects the Distribution of long lasting insecticidal nets (LLIN) is considered a key number of nets distributed annually, and examination of access over time intervention for the prevention of malaria. Mass distribution is required demonstrates the challenge of increasing and maintaining access to ITNs. to rapidly scale up LLIN coverage while continuous distribution systems Additional resources and robust routine distribution strategies are needed are essential to sustain the results achieved. In the Eastern Region (ER), to maintain high access to ITNs during the interim periods between larger the National Malaria Control Programme and implementing partners mass distribution campaigns. supported mass LLIN distributions between December 2010 and April 2011. Continuous distribution (CD) activities were started in October 1847 2012 and included antenatal care services), the expanded program on immunization and primary schools. The outcome was evaluated through BED NET DURABILITY ASSESSMENTS: EXPLORING A cross sectional surveys, conducted at baseline in April 2012, 12-16 months COMPOSITE MEASURE OF NET DAMAGE after the campaign and at endline in December 2013, after one year of Jodi Vanden Eng1, Ana Paula Abílio2, Maria do Rosario Pondja3, CD implementation. For each survey round, a representative sample of 900 Dulcisária Marrenjo3, Jacinta Luciano2, Guilhermina Fernandes3, households in ER was selected using a two-stage cluster sampling design. Samira Sabindy2, Adam Wolkon1, Gabriel Ponce de Leon1, Adeline Household heads were interviewed using a structured questionnaire. Chan1, Juliette Morgan1 Household ownership of at least one LLIN was 91.3% (95%CI 88.4 to 1Centers for Disease Control and Prevention, Atlanta, GA, United States, 93.6) at baseline and fell to 88.4% (85.2 to 91.3) at endline 18 months 2Instituto Nacional da Saúde, Maputo, Mozambique, 3Programa Nacional later but would have been only 81.0% (76.3 to 84.9) without the LLIN de Controlo da Malária, Maputo, Mozambique from CD. Population access to an LLIN within the household decreased from 74.5% (71.1 to 77.6) at baseline to 66.5% (62.9 to 69.9) but would The durability of Long Lasting Insecticidal Nets (LLINs) in field conditions have been 57.4% (53.0 to 61.8) without the CD contribution. Households is of great importance for malaria control programs. Although LLIN bio- reached by any of the CD channels were primarily those who had not efficacy has been investigated, the physical integrity of the net fabric been reached by the campaign with any or sufficient ITN. In addition, the is less well understood making it challenging to determine overall net different CD channels largely complemented each other with little overlap protectiveness. The 2011 World Health Organization Pesticide Evaluation in the first year. The continuous distribution of LLIN through primary Scheme (WHOPES) guidelines provide a simple, standardized method using

astmh.org 564 a proportional hole index (PHI) for assessing net damage. We evaluated 1849 the accuracy and utility of this measure using LLINs collected over three years in Nampula Province, Mozambique following a mass distribution SALMONELLA TYPHI-SPECIFIC EFFECTOR/MEMORY CD8+ campaign in 2008. For each LLIN the type of damage, diameter, and T CELL RESPONSES ELICITED IN A WILD-TYPE S. TYPHI distance from the bottom of the net were recorded for every hole. Holes CONTROLLED HUMAN INFECTION MODEL were classified into four size categories and a PHI was calculated based on 1 1 2 the WHOPES guidelines. The areas of WHOPES defined hole size categories Stephanie Fresnay , Monica A. McArthur , Thomas C. Darton , 2 2 2 were compared to circular and elliptical areas based on actual diameters Claire Jones , Claire S. Waddington , Christoph J. Blohmke , 3 2 1 of each hole; and the PHI was compared to cumulative damaged surface Gordon Dougan , Brian Angus , Myron M. Levine , Andrew J. 2 1 area of the LLIN. The damaged area of small, medium, and large holes was Pollard , Marcelo B. Sztein overestimated and the area of extra-large holes was underestimated using 1University of Maryland, Baltimore, MD, United States, 2University of the WHOPES categories compared to actual measured areas (Wilcoxon Oxford, Oxford, United Kingdom, 3Wellcome Trust Sanger Institute, signed rank test of differences p< 0.0001 for all sizes). Approximating Hinxton, United Kingdom holes as circular overestimated hole surface area by roughly 1.5 to 2 times Salmonella enterica serovar Typhi (S. Typhi) is a human restricted pathogen or more. For a range of hypothetical PHI thresholds associated with a which causes significant morbidity and mortality, particularly in developing “failed LLIN” found in current literature, roughly 75 to 80% of failed LLINs countries. A better understanding of the immune responses which result can be detected by only considering large and extra-large holes (which in protection from S. Typhi infection is imperative for the development are easier to identify and count). Future research studies may refine the of improved attenuated vaccines. Current knowledge is limited due PHI to better approximate overall surface area interrupted. Furthermore, to the lack of appropriate clinical and preclinical models. Recently, a research is needed to identify appropriate PHI thresholds to deem a net no controlled human infection model was re-established in which volunteers longer protective. Once a cutoff is selected, logistically simpler methods received 104 colony forming units of wild-type S. Typhi (Quailes strain) of determining the effective lifespan of LLINs can help guide replacement orally. Twelve volunteers were evaluated for their cell-mediated immune strategies for malaria control programs. (CMI) responses. ex vivo PBMC isolated before and up to 1 year after challenge were exposed to 3 S. Typhi-infected targets, i.e., autologous 1848 B lymphoblastoid cell-lines (B-LCL), autologous blasts and HLA-E restricted AEH B-LCL cells. CMI responses were evaluated using 14-color COMMUNITY ACQUIRED BACTEREMIA AMONG CHILDREN multiparametric flow cytometry to detect simultaneously 5 intracellular IN AREAS OF LOW AND HIGH MALARIA TRANSMISSION IN cytokines/chemokines (i.e., IL-17A, IL-2, IFN-γ, TNF-a and MIP-1b) and RURAL WESTERN KENYA a marker of degranulation (CD107a). Pre-challenge CD107a expression and cytokine production by S. Typhi-specific CD8+ T effector memory Frankline M. Onchiri1, Patricia B. Pavlinac2, Benson O. Singa1, (TEM) following exposure to S. Typhi-infected targets were higher in most Elizabeth A. Odundo3, Grace C. John-Stewart4, Judd L. Walson4 volunteers diagnosed with typhoid (TD) compared to those who were 1Kenya Medical Research Institute and University of Washington-Seattle, not. Direct correlations were observed between the levels of responses Nairobi, Kenya, 2Department of Epidemiology, University of Washington, before challenge and time to disease onset for CD107a, IFN-g and MIP- Seattle, WA, United States, 3Walter Reed Project, United States Army 1b following stimulation with S. Typhi-infected targets. After challenge, Medical Research Unit, Kericho, Kenya, 4Departments of Epidemiology, decreases in immune responses were observed prior to the time of disease Global Health and Pediatrics, University of Washington, Seattle, WA, onset, followed by a sharp increase in most TD volunteers. Multifunctional United States cells (i.e., concomitantly producing 3-5 cytokines/chemokine and/ In many African settings, malaria continues to decline as a cause of febrile or expressing CD107a) were dominant at all time-points. These data illness in children, highlighting the need for improved understanding of suggest that S. Typhi-specific responses prior to challenge, as well as the alternative causes of fever associated with high mortality. We evaluated magnitude, kinetics and quality (“multifunctionality”) of these responses the prevalence, etiologies, and correlates of bacteremia in outpatient might play a critical role in the development of typhoid fever. children at two rural hospitals in Western Kenya. Children aged 6 months-15 years presenting consecutively with fever to Kisii hospital; an 1850 area of low malaria endemicity (entomological infection rates [EIR]<1.5) ACTIVATION OF SALMONELLA TYPHI-SPECIFIC REGULATORY and Homa Bay hospital; a malaria endemic area (EIR≥300), were enrolled between 2012 and 2013. Detailed socio-demographic and clinical data T CELLS IS ASSOCIATED WITH TYPHOID DISEASE IN A WILD- were collected and all children tested for malaria using smear microscopy TYPE S. TYPHI CONTROLLED HUMAN INFECTION MODEL and Paracheck Pf® rapid diagnostic tests, HIV using antibody or PCR Monica A. McArthur1, Stephanie Fresnay1, Thomas C. Darton2, testing, and bacteremia using BACTEC™ 9250 blood culture system. Claire Jones2, Claire S. Waddington2, Cristoph J. Blohmke2, Isolates were identified and tested for antibiotic resistance using MicroScan Gordon Dougan3, Brian Angus2, Myron M. Levine1, Andrew J. Walkaway40®. Correlates of bacteremia were evaluated using multivariate Pollard2, Marcelo B. Sztein1 logistic regression. Overall, 1476 children were enrolled, 742 from Homa 1University of Maryland, Baltimore, MD, United States, 2University of Bay and 732 from Kisii. Children from Homa Bay were younger (mean Oxford, Oxford, United Kingdom, 3Wellcome Trust Sanger Institute, age±SD: 33.9±18.6 vs. 36.8±24.6 months) and more likely to be malaria- Hinxton, United Kingdom infected (49.2% vs. 8.6%), HIV-infected (4.2% vs. 1.2%) or HIV-exposed (19.3% vs. 3.4%) and more severely ill based on presence of ≥1 IMCI Salmonella Typhi (S. Typhi), the causative agent of typhoid fever, causes danger signs (51.5% vs. 16.9%). Only 48 children (3.3%) had bacteremia significant morbidity and mortality throughout the world. Currently (3.1% in Kisii and 3.4% in Homa Bay). Salmonella spp. (19 NTS and 19 available vaccines are only moderately immunogenic. To develop improved typhi) were the predominant cause of bacteremia, accounting for 79.2% vaccines, identification of immunological responses associated with (38/48) of all isolates, and the distribution of pathogens did not differ protection or disease is necessary. This has been hindered, in part, by between sites. Bacteremia was associated with HIV infection (aOR=4.5; the lack of an animal model that faithfully recapitulates human disease. 95% CI: 1.1-19.3) and lower education of caregiver (aOR=2.6; 95% CI: The re-establishment of a controlled human infection model with wild- 1.2-5.7); and inversely associated with malaria infection (aOR=0.4; 95%CI: type S. Typhi has made these critical studies possible. Peripheral blood 0.1-0.9). Bacteremia appears to be a relatively uncommon cause of fever in mononuclear cells were obtained from volunteers (n=10) prior to and 4 outpatient children in Western Kenya. Given the infrequent availability of at multiple time-points after challenge with 10 colony forming units of blood culture, targeted testing of high-risk children, including those with wild-type S. Typhi (Quailes strain). Regulatory T cell (Treg) responses were HIV, may be a useful strategy to reduce mortality among febrile children. measured by flow cytometry and activation status and homing potential of astmh.org 565 S. Typhi-specific T were determined. We identified significantly higher gut reg 1852 homing (integrin-a4b7 expressing) S. Typhi-specific regT prior to challenge in volunteers diagnosed with typhoid (TD) than in those who were not (No INVESTIGATION OF GENOTYPE VARIATIONS IN ORIENTIA TD). At early time-points following challenge, circulating integrin-a4b7 TSUTSUGAMUSHI OBTAINED FROM PATIENTS WITH expressing S. Typhi-specific regT decreased in TD volunteers, indicating MODERATE AND SEVERE SCRUB TYPHUS likely homing and a resulting accumulation in the gut. Additionally, S. 1 2 3 Typhi-specific T from TD volunteers demonstrated up-regulation of Kristin E. Mullins , Daniel Paris , Sabine Dittrich , Stuart reg 2 3 4 activation molecules following challenge, including expression of Human Blacksell , Paul Newton , Allen L. Richards Leukocyte Antigen (HLA)-DR and Lymphocyte function-associated antigen 1Uniformed Services University, Bethesda, MD, United States, 2Mahidol- (LFA)-1/CD11a as early as 1-4 days post-challenge compared to No TD Oxford Tropical Medicine Research Unit, Bangkok, Thailand, 3Wellcome volunteers. Furthermore, significantly higher expression of the chemokine Trust–Mahosot Hospital–Oxford Tropical Medicine Research Collaboration, receptor CXCR3, a molecule associated with homing to sites of active Vientiane, Lao People’s Democratic Republic, 4Naval Medical Research inflammation, was observed on the surface of S. Typhi-specific regT in TD Center, Silver Spring, MD, United States volunteers 1-4 days post-challenge compared to No TD volunteers. Taken Orientia tsutsugamushi is a Gram negative, obligate intracellular bacterium together these results suggest that activation of Treg that home to the that is transmitted by the bite of infected chiggers (Leptotrombidium spp) site of S. Typhi infection may play a role in disease pathogenesis, possibly and is the causative agent of scrub typhus. Scrub typhus presentation through suppression of S. Typhi-specific effector T cell responses. varies from a mild illness to severe disease, including pneumonitis, meningitis, encephalitis, disseminated intravascular coagulation, and in 1851 some cases death. Additionally, O. tsutsugamushi is found in Southeast Asia, the southwestern Pacific Islands, Korea, and parts of Russia, China, CHARACTERIZATION OF CANDIDATUS BARTONELLA Japan, Australia, New Zealand, Pakistan, India, and Afghanistan, where ANCASHI: A NEW AGENT ASSOCIATED WITH CARRION’S over 1 billion persons are at risk for infection and approximately one DISEASE million are infected annually. While the genus Orientia contains only two Kristin E. Mullins1, Jun Hang2, Robert Clifford2, Fatma Onmus- known species O. tsutsugamushi and O. chuto, the former is extremely Leone2, Ju Jiang3, Marina Leguia4, Ciro Maguina5, Richard G. genetically diverse, with >100 genotypes currently recognized. A study Jarman2, David Blazes1, Allen L. Richards3 is currently underway to examine the relationship between disease severity and genotype. 322 clinical isolates were collected from 6,740 1 2 Uniformed Services University, Bethesda, MD, United States, Walter adult patients who presented with suspected scrub typhus in Vientiane 3 Reed Army Institute of Research, Silver Spring, MD, United States, Naval (n=4875), Luang Namtha (n=1335), and Salavan (n=530) provinces of 4 Medical Research Center, Silver Spring, MD, United States, U.S. Naval the Lao People’s Democratic Republic (Lao PDR) from 2004 until 2012. 5 Medical Research Unit - 6, Lima, Peru, Universidad Peruana Cayetano The patients (n=322 ) were divided by disease severity, with 69 patients Heredia, Lima, Peru exhibiting severe disease and 253 patients exhibiting moderate disease. “The genus” Bartonella consists of Gram negative, facultative intracellular, Severe disease included patients with reduced consciousness (Glasgow vector-borne bacteria, which infect a wide range of mammalian Coma Score < 15), shock (systolic pressure < 80 mmHg), jaundice hosts. B. bacilliformis, B. henselae, and B. quintana have long been (clinical observations), meningitis/encephalitis (clinical observations), recognized as pathogens of human importance, while other species, and/or difficulty breathing (respiration rate > 30 breaths/minute). While such as B. clarridgeiae and B. rochalimae, are newly recognized human moderate disease included patients with malaise, fever, headache, and/ pathogens. During a 2003 clinical treatment trial a novel Bartonella or rash, who were sick enough to seek medical attention. Additionally, species, Candidatus B. ancashi, was uncovered. This treatment trial was 30 clinical isolates from various locations, within the endemic area for conducted, in Ancash, Peru (where B. bacilliformis is endemic), to test O. tsutsugamushi, will be used to improve O. tsutsugmaushi genotyping the efficacy of azithromycin as a treatment for B. bacilliformis. During methods. Single Nucleotide Polymorphism (SNPs) analyses will be this trial, two patients were found to be infected with a Bartonella employed to look for differences between the isolates from various species disparate from other Bartonella species based on gltA sequence locations in the endemic region and between the isolates that cause typing. Subsequently, this new agent, Candidatus B. ancashi, was more severe disease and those isolates that cause a moderate illness in Lao completely characterized by 1) observations of in vitro microscopic, PDR. Through this study, we hope to, identify predictors for severe phenotypic, colonial morphology, and growth characteristics, 2) multilocus disease as well as create a more accurate evolutionary phylogeny for O. sequence typing (MLST), multispacer typing (MST), and whole genome tsutsugamushi. analyses, and by 3) the development of species-specific qPCR assays to identify Candidatus B. ancashi’s presence in possible vectors (Lutzoma 1853 spp). Gram-staining and transmission electron microscopy showed the isolates to be small, Gram-negative bacilli with variable expression of SPEEDIER LEPTOSPIRA DIAGNOSIS USING HEMOCULTURE unipolar flagella. Biochemical testing provided a single phenotype for all FLUIDS the isolates, which is consistent with other Bartonella spp. Fully genome 1 2 sequencing and subsequent genome analyses confirmed these isolates to Sabine Dittrich , Phouthasen Hyongvongsithy , Anisone 2 2 2 be genetically identical to one another, yet distinct from other Bartonella Chanthongthip , Michael Knappik , Weerawat Phuklia , David A. 1 1 species. Genome analyses revealed B. bacilliformis to be the closest relative Dance , Paul N. Newton to Candidatus B. ancashi, although unlike B. bacilliformis, the genome 1University of Oxford, Vientiane, Lao People’s Democratic Republic, 2Lao- of Candidatus B. ancashi encodes virulence determinates not seen in Oxford-Mahosot Hospital Wellcome Trust Research Unit, Vientiane, Lao B. bacilliformis. Surprisingly, whole genome mapping showed major People’s Democratic Republic gene rearrangements between the isolates. Additional genome analyses Leptospirosis is a common bacterial zoonosis worldwide, with infections uncovered a possible link between the rearrangements and flagella occurring after exposure to contaminated water. Despite this global expression. Based on the results from these studies, we believe Candidatus problem, diagnosis is difficult with culture results taking up to three B. ancashi is a novel pathogen of human importance. months and Microscopic Agglutination Test (MAT) serology being retrospective by nature. Molecular assays are ~55% sensitive and 90% specific to detect infection on admission blood samples, with low bacterial density complicating detection. Leptospira were shown to survive and multiply in blood culture media and we hypothesised that extracting DNA from incubated hemoculture fluid (HCF) from blood

astmh.org 566 culture bottles, followed by quantitative real-time PCR (qPCR) could 1855 improve the sensitivity and speed of leptospira diagnosis. We assessed this retrospectively, using pre-incubated HCF of leptospira positive MYCOBACTERIUM ULCERANS DISEASE: PERFORMANCE OF (n=109) and negative (n=63) (as determined by culture, PCR directly DIAGNOSTIC TESTS AND CLINICAL OPINION COMPARED on clinical samples and MAT on convalescent serum) febrile patients in TO DIFFERENT REFERENCE STANDARDS IN AKONOLINGA, Vientiane, Lao PDR. After optimization, receiver-operator-characteristics CAMEROON analysis was employed to identify the most suitable qPCR-threshold and corresponding diagnostic values. The finalized method showed promising Yolanda K. Mueller1, Patrick Nkemenang2, Bitoungui Mboua3, sensitivities of 82% (95%CI: 71-90), 66% (95%CI: 55-76) and 59% Sara Eyangoh4, Genevieve Ehounou2, Clotilde Rambaud-Althaus1, (95%CI: 49-68) compared to culture, culture+PCR or culture+PCR+MAT, Eric Comte2, Earnest Njih Tabah5, Jean-Francois Etard1, Laurence as the respective reference standards. The specificities were >95%, for all Toutous Trellu6 three comparisons. This approach may enable the diagnosis of leptospiral 1Epicentre, Paris, France, 2Doctors Without Borders, Geneva, Switzerland, infection without the submission of additional samples and the incubation 3Central Hospital, Yaounde, Cameroon, 4Centre Pasteur of Cameroon, step may further increase the sensitivity without compromising the Yaounde, Cameroon, 5National Program against Buruli Ulcer, Yaounde, specificity. The optimized protocol and its usefulness in a routine laboratory Cameroon, 6Geneva University Hospitals, Geneva, Switzerland setting will be further evaluated prospectively during May-October 2014 In the absence of gold standard, the performance of laboratory test to and these data will be presented. diagnose Mycobacterium ulcerans disease (MU) is often overestimated. We compared the diagnostic accuracy of clinical judgment and laboratory 1854 tests to diagnose MU. Between 2011 and 2013, all individuals presenting at Akonolinga District Hospital, Cameroon, with a skin lesion suspect INNOVATING DIAGNOSIS OF BACTERIAL BLOODSTREAM of new MU were enrolled after consent. Clinical data and clinicians’ INFECTIONS IN MALARIA-ENDEMIC SETTINGS: FROM judgment on probability of MU (four grades) were prospectively collected, DISEASE METABOLOMICS TO RAPID DIAGNOSTIC TESTS before the results of laboratory examination (ZN, PCR, culture and Saskia Decuypere1, Jessica Maltha2, Issa Guiraud3, Bérenger skin biopsy). Photographs of lesions were reviewed independently by Kaboré3, Marc Tahita3, Stijn Deborggraeve2, Malcolm McConville4, two dermatologists, and skin biopsies by two histopathologists. We Stephen Graham5, Halidou Tinto3, Jan Jacobs2, Jonathan constructed a first composite reference standard combining results of Carapetis1 laboratory tests, clinical opinion, and final diagnosis reached by expert consensus, and a second based on WHO definition of at least two positive 1Telethon Kids Institute, Perth, Australia, 2Institute of Tropical Medicine, laboratory tests. The 364 included patients had a median age of 34 years Antwerp, Belgium, 3Clinical Research Unit of Nanoro, Nanoro, Burkina (range 0 to 87), 233 (64%) were males and 66 (19%) were HIV-positive. Faso, 4Metabolomics Australia and Bio21 Institute of Molecular Sciences The 364 patients had a total of 422 lesions, of which 381 (90%) were and Biotechnology, University of Melbourne, Melbourne, Australia, 5Centre ulcerative. Lesion severity was of category 1, 2 and 3 in 32%, 41% and for International Child Health, Melbourne, Australia 26%, respectively. According to expert consensus, MU was diagnosed in The increasing use of malaria rapid diagnostic tests has revealed that 113 (27%) lesions. Main differential diagnoses were vascular ulcers (25%), febrile illnesses are often caused by other pathogens than Plasmodium. other bacterial infections (19%), post-traumatic lesions (7%) and non MU Amongst them, bacterial bloodstream infections (bBSI) are increasingly osteomyelitis (6%). Area under ROC curve (AUC) for clinical diagnosis recognized as an important cause associated with a high mortality, compared to consensus reference standard was 0.84 (95CI 0.80 - 0.88), especially in the African setting. Currently, diagnosis of bBSI is clinical as comparable to PCR (0.84, 95CI 0.80 - 0.89, p=0.98), and 0.82 (95CI 0.69 microbiological culture testing is usually not available and if available, - 0.95) and 0.69 (95CI 0.65 - 0.74) for ZN performed in Akonolinga and takes 2 to 3 days for a result. Missed diagnosis can result in preventable Yaounde, respectively. When using a composite standard of two positive deaths, while overdiagnosis results in inappropriate or unnecessary tests (pending final culture results for 58/422 lesions), AUC for PCR (0.94 use of antibiotics. There is an urgent need to develop rapid diagnostic (95CI 0.92 - 0.97) was superior (p<0.001) while clinical judgment and tests targeting bBSI. We hypothesize that the pathophysiological ZN of both sites were comparable (p=0.47). Clinical judgment is at least processes triggered by bBSI induce characteristic changes in the > comparable to ZN to diagnose MU, while PCR is equivalent or superior 4,000 different blood metabolites. Our objective is to harness these depending on reference standard used. characteristic metabolite features for bBSI diagnosis. We are conducting a first metabolomics study to examine whether blood plasma contains metabolites that could be useful to diagnose bBSI in a malaria-endemic setting. We quantified 1600 polar and lipid metabolites in plasma from 83 children with severe febrile illness admitted to a rural district hospital in Burkina Faso using liquid-chromatography mass-spectrometry. The patients included (i) 12 bBSI cases confirmed by blood culture, (ii) 34 severe malaria cases with a positive thick blood film and (iii) 37 cases with negative blood culture and negative blood film. A distinct metabolite profile was identified in children with culture-confirmed bBSI compared to children with severe malaria. A first diagnostic model including 10 polar metabolites has a sensitivity of 80% (95% CI: 44.4-96.9%) and specificity of 76.5% (95% CI: 62.5-87.2%) to identify culture-confirmed bBSI. Mining of the lipid data is ongoing to fine-tune this diagnostic model for differential diagnosis of bBSI and severe malaria. We will present the predicted diagnosis of the 83 patients by the final metabolite diagnostic model(s), and compare to the results obtained with blood culture, PCR-based SepsiTestTM, malaria thick blood smear and HRP2 rapid diagnostic test. This study demonstrates the potential of plasma metabolites to identify causality in children with severe febrile illness. We will discuss the translation to metabolite-based rapid diagnostic tests and their potential impact on clinical management of severe febrile illness in malaria-endemic settings.

astmh.org 567 1856 1857 OVERCOMING THE CHALLENGES OF CLINICAL DATA LACTIC ACIDOSIS AND RESPIRATORY DISTRESS ARE MANAGEMENT IN LOW AND MIDDLE-INCOME COUNTRIES: FREQUENT IN CEREBRAL MALARIA AND SEVERE MALARIAL A CONTEXT-ADAPTED DATA MANAGEMENT PLAN AND ANEMIA, BUT PREDICT MORTALITY ONLY IN CEREBRAL LIFECYCLE MALARIA Yves Claeys1, James Smedley2, Anish Bhattarai3, Hercule Kalonji4, Nathan R. Brand1, Robert O. Opoka2, Karen E. Hamre3, Chandy Paritosh Malaviya5, Robert Meester6, David Mwakazanga7, Martin C. John3 8 9 10 11 Omello , Sayouba Ouedraogo , Sopheak Sok , Sun Sokleng , 1Columbia University, College of Physicians and Surgeons, New York, NY, 12 13 14 Mary Thiongo , Hiwot Tilahun , Arouna Woukeu , Harry van United States, 2Makerere University, Kampala, Uganda, 3University of 1 2 1 Loen , Brian Faragher , Raffaella Ravinetto Minnesota, Minneapolis, MN, United States 1Institute of Tropical Medicine, Antwerp, Belgium, 2Liverpool School of We investigated the pathogenesis of lactic acidosis (LA) and deep Tropical Medicine, Liverpool, United Kingdom, 3Koirala Institute of Health breathing (DB) and their associated mortality in 249 children with cerebral Sciences, Sunsari, Nepal, 4University of Kinshasa, Kinshasa, Democratic malaria (CM) and 216 children with severe malarial anemia (SMA) in Republic of the Congo, 5IMS/BHU, Varanasi, India, 6Formerly at Amsterdam Kampala, Uganda. Platelet count and hemoglobin, lactate and histidine Institute for Global Health and Development, Amsterdam, Netherlands, rich protein-2 (HRP-2) levels at admission were assessed. Children with a 7Tropical Diseases Research Center, Ndola, Zambia, 8Interact Program, hemoglobin of <5 g/dL were transfused (all SMA, 59 CM). LA was more Uganda, Uganda, 9CRUN, Nanoro, Burkina Faso, 10Sihanouk Hospital frequent in children with SMA (43.1%) than CM (32.1%, P=0.02), and Center of Hope, Phnom Penh, Cambodia, 11National Center for HIV/AIDS, DB was similar in the two groups (CM, 8.8%, SMA, 7.4%, P=0.6), but and STD, Phnom Penh, Cambodia, 12International Center mortality was higher in children with CM (12.5%) than SMA (0.5%, for Reproductive Health, Mombasa, Kenya, 13Armauer Hansen Research P<0.001). In children with CM, mortality was increased in children with Institute, Addis Abeba, Ethiopia, 14Formerly at London School of Tropical LA (odds ratio (OR), 2.2, 95% confidence interval (CI), 1.0, 4.7, P=0.04) Medicine, London, United Kingdom or DB (OR 5.0, 95% CI, 1.9, 13, P=0.001), but in children with SMA More and more non-commercial clinical research is carried out on a neither LA nor DB was associated with mortality. Children with CM had collaborative basis in resource-limited contexts to address priority health higher HRP-2 levels and lower platelet counts than children with SMA, problems in low and middle income countries (LMICs). Despite the while children with SMA had lower hemoglobin levels than children with existence of FDA- and ICH-GCP guidelines for clinical research, there is CM (all P<0.001). In children with CM, both LA and DB were associated a need for a practical approach in all processes including clinical Data with decreased platelet counts and increased HRP-2 levels (all P<0.01), Management (DM) to facilitate the collection and analysis of high-quality while in children with SMA, LA and DB were not associated with platelet data, despite the contextual and financial constraints. Since 2010, counts, and only LA was associated with increased HRP2 levels (P=0.02). the members of the Association for Data Management In the Tropics Conversely, hemoglobin levels were inversely associated with lactate levels (ADMIT:https://admit.tghn.org) share knowledge and tackle common in children with SMA (P<0.001) but not CM. In children with CM and DB, issues experienced like standardization, Electronic Data Capture and Data each natural log increase in HRP-2 levels was associated with an 8.6 fold sharing. One of the first initiatives for ADMIT was to tackle the need for increased risk of mortality (95% CI, 1.1, 67.0, P=0.04). DB and LA in CM uniformity in terms of context-adapted standard operating procedures are associated with parasite sequestration with platelet adhesion, while in (SOPs).We defined the essential DM processes within a research project. SMA they are associated with low hemoglobin. These differences may in The members of the network were assigned authorship to prepare two part explain the high mortality associated with DB and LA in CM, and the SOPs which were presented and peer reviewed during a workshop within lack of DB- or LA-associated mortality in children with SMA who receive a the wider group. A harmonization process was undertaken to ensure blood transfusion. uniform structure, terminology and the level of detail across the suite of SOPs. During the harmonization, the alignment of the individual processes 1858 inspired the creation of an overall lifecycle for DM. As a result, a usable Data Management Plan (DMP) is now available incorporating the suite of WHAT IS AN ASYMPTOMATIC CARRIER IN AN ENDEMIC these SOP’s.(In the oral presentation,) We will present the characteristics AREA? of this DMP and describe how it may help with collaborations on non- Alejandro Llanos1, Miguel Campos2, Mitchel Guzman3, Roberson commercial research projects where DM processes are spread across Ramirez3, Raul Chuquiyauri1, Dionicia Gamboa3, Hugo Rodriguez4, different places. We recommend the DMP to be used to ensure a uniform Joseph Vinetz5 approach to DM, strengthening partnerships and knowledge exchange. We hope that this may be a starting point for standardization in DM in 1Instituto de Medicina Tropical Alexander von Humboldt, Universidad LMICs and possibly to formulate practical recommendations for regulatory Peruana Cayetano Heredia, Lima, Peru, 2Facultad de Ciencias y Filosofia, and GCP guidelines. The next challenge is to look at the definition of the Universidad Peruana Cayetano Heredia, Lima, Peru, 3Laboratorio ICEMR- roles and responsibilities needed to resource these DM activities and the Amazonia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias development of training packages. y Filosofía, Universidad Peruana Cayetano Heredia, Lima, Peru, 4Dirección Regional de Salud de Loreto, Ministry of Health, Loreto, Peru, 5University of California San Diego, San Diego, CA, United States Nowadays there is a greater consensus that is crucial to identify asymptomatic carriers in order to eliminate/eradicate malaria. The methods of evaluating clinical malaria in endemic areas, which include both passive (PCD) and active case detection (ACD), are not very effective in detecting asymptomatic infections, limiting treating asymptomatic carriers. Aggressive infection detection (AID) is other strategy where malaria parasites are searched in people of endemic areas, regardless of the presence of clinical symptoms, using PCR and thick smear (TS). However, applied on a large scale AID would be expensive and impractical. In this perspective, we try to understand who is an asymptomatic carrier in an endemic area through an open population cohort study, conducted in the Peruvian Amazon, and involved 2000 people from 8 communities around

astmh.org 568 Iquitos. AID+ACD and PCD were used. All the subjects were visited once 1860 a week in their homes, and they were given a survey where they were asked systematically for 13 malaria-related symptoms, daily recall for the VALIDATION OF A TUBERCULOUS MENINGITIS CASE past week. A blood smear for TS was taken weekly and a sample for PCR DEFINITION IN MBARARA REGIONAL REFERRAL HOSPITAL, monthly. It was found that the prevalence of any symptom in the group UGANDA with PCR(-) and TS(-) was 21.35% (95%CI:20.67-22.43), for PCRPv(+) 1 2 2 and TS(-) group was 29.87% (95%CI:26.17-33.40), PCRPv(+) and TS(+) Susan A. Adakun , Emily Evans , Bradley Weinbaum , Yap Boum 3 3 4 5 group was 70.5% (95%CI:68.1-72.8) and PCRPf (+) mostly TS(-) group II , Patrick Orikiriza , Mark J. Siedner , Tony Wilson , Christopher 2 was 25.49% (95%CI:21.48-29.51). Fever was the symptom that best C. Moore discriminated infection, but it only achieved 60% specificity and 20% 1Makerere University College of Health Sciences, Kampala, Uganda, sensitivity (40% for the PCRPv(+)/TS(-) group). In addition, 41.4% (95%CI: 2University of Virginia, Charlottesville, VA, United States, 3MSF Epicentre, 38.9- 43.9) of the infections (asymptomatic or no) were negatives to TS. Mbarara, Uganda, 4Massachusetts General Hospital Centre for Global In conclusion, in the Peruvian Amazon study area, 30% of the group Health, Harvard Medical School, Boston, MA, United States, 5Mbarara with sub-microscopic malaria for P. vivax (PCRPv+ and TS-) had at least University of Science and Technology, Mbarara, Uganda one symptom. There are statistical differences between the symptom Tuberculous meningitis (TBM) is a leading cause of death in areas with a prevalence of PCRPv (+)/TS(-) and PCRPv (+)/TS(+). These results suggest high prevalence of both tuberculosis and HIV such as sub-Saharan Africa. that it is feasible to develop a clinical marker score to detect potential A major obstacle to successful treatment of TBM is the inability to obtain asymptomatic carriers that should be treated. a rapid and accurate diagnosis. We evaluated the accuracy of a recently proposed TBM case definition (Marais, et al) which is based on clinical, 1859 CSF, and radiological findings among patients admitted with suspected meningitis to the Mbarara Regional Referral Hospital in Uganda. CSF was ANEMIA AND TRANSFUSION REQUIREMENTS AMONG obtained for routine analysis, bacterial and mycobacterial culture, and CHILDREN WITH SEVERE MALARIA TREATED WITH PCR via GeneXpert® MTB/RIF. Blood was obtained for random blood ARTESUNATE AT A RESOURCE-POOR HOSPITAL IN UGANDA sugar, lactate, malaria blood smear, complete blood count, blood culture, Michael Hawkes1, Andrea Conroy2, Robyn Elphinstone2, Sophie HIV serology and CD4+ count. We determined the diagnostic accuracy Namasopo3, Chandy John4, Robert Opoka5, Kevin C. Kain2 for the TBM clinical score by evaluating the sensitivity and specificity, as well as positive and negative predictive value of each score threshold. 1 2 University of Alberta, Edmonton, AB, Canada, University of Toronto, We used a positive mycobacterial culture of cerebrospinal fluid as a 3 Toronto, ON, Canada, Jinja Regional Referral Hospital, Jinja, Uganda, reference standard. We enrolled 141 participants and the prevalence of 4 5 University of Minnesota, Minneapolis, MN, United States, Makerere TBM was 6%. Patients with higher TBM scores were more likely to have a University, Kampala, Uganda diagnosis of TBM, OR 1.44, p=0.04 CI (1.00-2.06). The ROC curve for the Treatment of non-immune travelers with malaria using parenteral prediction of TBM by the TBM score was 0.75. Of the three case definition artesunate is associated with late-onset hemolysis. Most cases of severe criteria (clinical, CSF, evidence of TB), only the CSF criterion was strongly malaria occur in children in sub-Saharan Africa, where the hematologic associated with TBM (OR 7.73 95% CI (1.04-57.0) p=0.04). For a TBM effects of artesunate have not been well documented. We report a score threshold <7, sensitivity was 100%, specificity 38.7%, PPV 7.1%, prospective case series of 92 children with severe malaria, all treated with NPV 100%. The TBM score has good sensitivity but low specificity for the parenteral artesunate, managed at a resource-poor hospital in Africa, diagnosis of TBM. It has an excellent negative predictive value and may be with detailed longitudinal data on hemoglobin (Hb) levels. The median used to rule out TBM in resource limited settings. (range) age was 2 (1-8) years and 43 (47%) were female. Fifteen patients had tea-coloured urine at admission and 14 were visibly jaundiced. The 1861 median (IQR) admission Hb level was 69 (56-80) g/L and 17 patients (19%) had severe anemia (Hb<50 g/L). During hospitalization, 69 patients (76%) MYOCARDIAL AND HAEMODYNAMIC RESPONSES TO FLUID received one or more transfusions of packed red blood cells or whole MANAGEMENT IN SEVERELY MALNOURISHED AND WELL- blood, for a total of 114 transfusions. The median (IQR) total volume of NOURISHED AFRICAN CHILDREN WITH SEVERE SHOCK AND blood administered was 10.4 (5.6-20) mL/kg. Patients with jaundice at GASTROENTERITIS presentation received significantly larger number (p=0.014) and volume (p=0.043) of transfusions. Fatal outcome in 8 patients was associated with Kathryn Maitland1, Nchafatso Obonyo1, Bernadette Brent2, Peter severe anemia in 6/8 cases. Follow-up Hb measurement was performed on Olupot Olupot3, Julius Nteziyaremye3, Irene Kuipers4, Michael 35 patients (38%) at day 14 after initial hospital admission; the remaining Boele van Hensbroek4, Job van Woensel4 patients had no clinical evidence of anemia (no pallor, tachycardia, 1KEMRI Wellcome Trust Programme, Kilifi, Kenya,2 Imperial College, hyperdynamic circulation, parental report of lethargy, or easy fatigability) London, United Kingdom, 3Mbale Regional Referral Hospital, Mbale, at the follow-up visit. The convalescent Hb was median (range) 90 (60- Uganda, 4Emma Children’s Hospital, Amsterdam, Netherlands 138) g/L, which was significantly higher than the paired admission levels The FEAST (Fluid Expansion as Supportive Therapy) trial, the only (median increase +28 g/L, p<0.001). The day 14 Hb level was higher randomised controlled trial of fluid resuscitation, demonstrated that than any level measured during hospitalization in 22 (63%) patients, but fluid boluses lead to excess mortality in children receiving bolus fluids. decreased or remained the same in 13 (33%). Among children with a Further analysis indicated adverse effects of fluid boluses resulted from decrease in Hb level by day 14, the magnitude of the Hb change ranged cardiovascular events rather than fluid overload. Studies of myocardial from -8 to -66 g/L, but none reached the threshold for severe anemia and haemodynamic responses to boluses are thus warranted in groups (lowest day 14 Hb was 60 g/L). None required transfusion after hospital not included in FEAST (gastroenteritis and malnutrition), where fluid discharge. In this representative cohort of young children with severe resuscitation continue to be recommended in international guidelines. malaria in a hyper-endemic setting treated with artesunate, anemia was We describe in Ugandan children myocardial (echocardiographic and common at admission, required one or more transfusions in a majority of ECG) and haemodynamic responses to fluid bolus (i.e. 0, ½, 1, 1½, 2, 3, patients, and on average was improving by day 14. However, a substantial 4, 8, 12, 16, 20, 24, 32, 40 and 48 hours) pre- and post fluid challenges proportion of children had persistent or worsening anemia at follow-up. recommended by WHO guidelines. Blood and urine samples for analysis Further study is needed to determine whether this effect is attributable to of electrolytes and other markers of myocardial dysfunction were artesunate. collected at admission, 8, 24 and 48 hours. A total of 29 children with severe shock and dehydration (due to gastroenteritis) were studied: 19 had severe malnutrition (SM); 10 were well-nourished (controls). For the

astmh.org 569 SM group receiving WHO guideline resuscitation (15mls/kg over 1 hour, periphery is the induction of co-inhibitory receptors such as CTLA4 and repeated twice if indicated) mortality was 73% (8/11 patients). Following PD1 and their ligands but their role in the immune response in Pf malaria a protocol amendment to slower rehydration (10mls/kg over 1 hour up remains poorly understood. To test the hypothesis that co-inhibitory to a maximum of 50mls/kg) 3/8 died (38%) compared to a mortality of receptors modulate the T cell response in acute malaria, blood samples 2/10 in controls. Echocardiographic and haemodynamic data pre-bolus were obtained from patients with acute uncomplicated Pf malaria showed marked evidence of underfilling. Boluses lead to early rapid shock treated in Hamburg, Germany as well as from healthy volunteers. Flow reversal, in those treated with slower rehydration shock reversal was more cytometric analysis showed high expression of CTLA4 and PD1 on CD4+ protracted. In all study participants we found no evidence of that mortality T cells of malaria patients and the ligands for PD1, PDL1 and PDL2, was due to fluid overload. Fluid boluses administered to children with were upregulated on monocytes, B cells and T cells. We then stimulated SM (per WHO guideline)-resulted in early shock reversal, but this was not PBMCs with Pf-infected red blood cells (iRBCs) to detect antigen-specific associated a survival benefit. Slow rehydration strategy in cases (SM) and cytokine production and proliferation. The majority of antigen-specific effT control patients appeared to be well-tolerated. Further research is required cells were CTLA4+PD1+. IFNg was the most frequently detected cytokine to optimize fluid management and other supportive strategies to inform and >50% of IFNg+ CTLA4+PD1+ T cells simultaneously produced IL10. In future guidelines. some donors T cell proliferation was inhibited by PD1 and blockade of PD1-ligation enhanced antigen-specific proliferation. We further isolated 1862 CTLA4+PD1+CD4+ T cells based on surface expression of PD1 and CTLA4 and investigated their inhibitory function in in-vitro proliferation assays SEVERE MALARIA INFECTIONS IMPAIR GERMINAL CENTRE stimulated with aCD3/28 or iRBCs. CTLA4+PD1+CD4+ T cells suppressed REACTIONS AND INHIBIT EFFICIENT ANTIBODY RESPONSES aCD3/28-induced as well as plasmodial-antigen-specific T-cell proliferation TO INFECTION in a cell-extrinsic manner. In summary, acute Pf infection leads to induction + + of malaria-specific CTLA4 PD1 Teff cells which coproduce IFNg and Diana S. Hansen, Victoria Ryg-Cornejo, Ann Ly, Lisa J. Ioannidis, IL10 while inhibiting CD4+ T cell proliferation in a cell extrinsic manner. Axel Kallies Induction of Teff cells with regulatory function might be an important The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia mechanism to control T cell responses and prevent severe inflammation in Naturally acquired immunity to malaria develops only after many years acute malaria and potentially other acute infections. of repeated exposure to Plasmodium parasites. Protective immunity predominantly targets blood-stage parasites and requires antibody 1864 responses. Despite the key role that antibodies play in protection against malaria, the cellular processes leading to the slow acquisition LOSS AND DYSFUNCTION OF VΔ2+ ΓΔ T CELLS IS of immunity remain unknown. Children in high transmission settings ASSOCIATED WITH CLINICAL TOLERANCE TO MALARIA that experience frequent malaria clinical episodes are characterized by a Prasanna Jagannathan1, Charlie C. Kim1, Bryan Greenhouse1, delayed development of parasite-specific memory B cells, suggesting that Felistas Nankya2, Katherine Bowen1, Ijeoma Eccles-James1, Mary the inflammatory factors contributing to disease hinder these responses. K. Muhindo2, Emmanuel Arinaitwe2, Jordan W. Tappero3, Grant To address that hypothesis we used a severe malaria infection model to Dorsey1, Moses R. Kamya4, Margaret E. Feeney1 investigate the development of germinal centres (GC), memory B cells and 1 plasma cells. C57BL/6 mice were infected with P. berghei ANKA, followed University of California San Francisco, San Francisco, CA, United 2 by treatment with anti-malarial drugs or immunized with equivalent States, Infectious Diseases Research Collaboration, Kampala, Uganda, 3 antigenic loads of irradiated parasites. Reduced numbers of GC B cells and Centers for Diseases Control and Prevention, Atlanta, GA, United States, 4 T follicular helper cells (Tfh) were found in mice experiencing an active Makerere University, Kampala, Uganda infection compared to immunized control animals. Despite normal IL-21 The VΔ2+ subset of γΔ T cells possess intrinsic reactivity to malaria secretion, Tfh cells from infected mice displayed an unusual phenotype antigens, but their role in acquired immunity to malaria is unclear. To characterized by low surface expression of PD-1 and CXCR5, required evaluate γΔ T cell responses in children living in a highly malaria-endemic for their successful localization in GCs. Consistently, confocal microscopy area, peripheral blood mononuclear cells (PBMCs) were obtained from experiments revealed that clinical malaria inhibits the establishment of 78 HIV-uninfected 4-year old children enrolled in a longitudinal cohort GC reactions in the spleen. The frequency of memory B cells and relative study in Tororo, Uganda. The incidence of symptomatic malaria in this antibody affinity of long-lived plasma cells emerging from GCs was also cohort was 5.4 episodes ppy (IQR 3.2-7.0) and peaked at 25 months examined. Unlike immunization with irradiated parasites, active infections of age with a subsequent gradual decline in malaria episodes and a appeared to compromise these processes. Pro-inflammatory cytokines corresponding increase in asymptomatic parasitemia. PBMCs were involved in the induction of severe malaria episodes were found be partly stimulated with Plasmodium falciparum-infected red blood cells (iRBC) responsible for the inhibition of B cell responses. Thus these data indicate or controls and assessed by multiparameter flow cytometry and gene that clinical malaria negatively impact the development of long-term expression microarray. We noted a striking inverse association between humoral immunity by disrupting critical early stages in the development of frequencies of VΔ2+ cells and the prior cumulative incidence of malaria B cell responses. (Rho=-0.39, P=0.003). Repeated episodes of malaria were also associated with decreased cytokine production (Rho=-0.41, P=.0002) and decreased 1863 proliferation (Rho=-0.58, p=0.009) of VΔ2+ cells in response to malaria antigen stimulation, suggesting that children who have survived repeated ACUTE MALARIA INDUCES CTLA4+PD1+ EFFECTOR T CELLS clinical malaria episodes exhibit dysfunction as well as loss of VΔ2 + WITH REGULATORY PHENOTYPE cells. Whole transcriptome analysis of sorted, unstimulated V2+ cells revealed increased expression of immunoregulatory genes in children 1 2 2 Maria S. Mackroth , Annemieke Abel , Christiane Steeg , Julian with heavy prior malaria, including genes encoding Tim-3, BATF, and 1 2 Schulze zur Wiesch , Thomas Jacobs CD57, suggesting that repeated infection may lead to the upregulation 1University Hospital Hamburg-Eppendorf, Hamburg, Germany, 2Bernhard of immunoregulatory pathways that dampen the innate V2 inflammatory Nocht Institute for Tropical Medicine, Hamburg, Germany response. Finally, loss and dysfunction of pro-inflammatory ΔV 2+ γΔ T cells Acute Plasmodium falciparum (Pf) malaria induces proinflammatory was associated with a reduced likelihood of symptoms upon subsequent P. T cell responses which have been shown to confer protection against falciparum infection. Together, these results suggest that repeated malaria malaria but also contribute to the development of severe disease. A infection during childhood results in progressive loss and dysfunction of V 2+ T cells that may facilitate immunological tolerance of the parasite. tight regulation of T effector (Teff) responses is therefore crucial to protect Δ γΔ the host. An important mechanism to fine-tune T cell responses in the

astmh.org 570 1865 malaria exposure is associated with an expansion of atypical memory B cells (MBCs) which are distinguished from classical MBCs by the expression LONGITUDINAL ANTIBODY RESPONSES TO ANTIGENS of inhibitory receptors. A similar subset of B cells has been described in ON THE SURFACE OF PLASMODIUM FALCIPARUM individuals infected with HIV and HCV, yet the origin and function of this B GAMETOCYTE-INFECTED ERYTHROCYTES IN GHANAIAN cell subset remains unclear. We performed a comprehensive investigation SCHOOL CHILDREN of atypical B cells collected from individuals exposed to intense malaria in Mali. Sorted naïve B cells (CD19+ CD21+ CD27-), classical MBCs (CD19+ Bismarck Dinko1, Teun Bousema2, Colin J. Sutherland2 CD21+ CD27+), and atypical MBCs (CD19+ CD21- CD27-) were subjected 1University of Health and Allied Sciences, Ho, Ghana, 2London School of to genome wide expression profiling, VDJ sequence analysis (Ab heavy and Hygiene & Tropical Medicine, London, United Kingdom light chain gene usage and somatic hypermutation rate), KREC analysis Little is known about the immune responses directed at circulating (replicative history), as well as proliferative and cytokine production Plasmodium falciparum gametocytes in humans, knowledge of which analysis following in vitro stimulation. We found that classical and atypical would be useful in the development of intervention strategies to reduce MBCs have distinct expression profiles, but are similar in heavy and light and block malaria transmission. Consequently, antibody responses to chain variable gene usage as well as replicative history. Atypical MBCs surface antigens of P. falciparum gametocyte-infected RBCs (GSA) were have, however, lower levels of somatic hypermutation in heavy and light determined in plasma samples from malaria asymptomatic Ghanaian chain sequences, indicating less antigen-dependent selection compared school children between the ages of 5-17 years. These children were to classical MBCs. We further show how these B cell subsets differ in screened for malaria parasites and treated with dihydro-artemisinin proliferative and cytokine production capacity, and how the expression piperaquine and followed up weekly for one month. Gametocytes were of inhibitory receptors on atypical MBCs impairs their proliferation. This produced from a laboratory adapted parasite line, 3D7 and a recent thorough characterization of B cell subsets in malaria-exposed individuals patient isolate from Kenya (HL1204). From a cohort of 113 children, 56% has generated new hypotheses on how chronic Plasmodium exposure of the children exhibited marked antibody responses to GSA (immune leads to B cell dysregulation and the inefficient acquisition of protective response above the median within the cohort per sampling time) that antibodies. recognized GSA on a proportion of mature gametocyte-infected RBCs of 3D7 by flow cytometry. These responsive individuals were identified 1867 by measuring both the proportion of mature gametocytes recognised by antibodies and the intensity of the antibody binding to GSA. Longitudinal CHARACTERIZATION OF MALARIA PARASITE LINES data provided an additional 10% developing GSA responses during the (PLASMODIUM FALCIPARUM) SELECTED BY LONG-TERM 1 month follow-up. Children with GSA antibodies present at enrolment, CULTURE IN THE PRESENCE OF INHIBITORY ANTIBODIES TO were less likely to develop new gametocytaemia at subsequent visits APICAL MEMBRANE ANTIGEN-1 (AMA1) (odds ratio = 0.29, 95% CI 0.06 - 1.05; P = 0.034). 3D7a is a laboratory Sheetij Dutta, Lisa Dlugosz, J. Kathleen Moch, Adrian Batchelor, adapted parasite line so a selection of positive plasma samples was John D. Haynes tested against mature gametocyte preparations from HL1204 and strong plasma antibody binding was again shown. No binding to the surface Walter Reed Army Institute of Research, Silver Spring, MD, United States of RBCs infected with immature gametocytes of HL1204 was detected. Vaccination has been the most effective medical intervention (after In conclusion, a proportion of malaria infected asymptomatic children sanitation, hygiene, and nutrition) for preventing and eliminating harbour plasma antibodies which strongly recognized antigens on the infectious diseases. The development of resistance to anti-malarial drugs surface of mature gametocyte-infected RBCs. Strong plasma antibody by the parasite has spurred the search for effective vaccines. It remains responses were associated with the control of gametocytaemia in vivo. unclear, however, if the antigenically highly polymorphic malaria parasite Ghanaian GSA responses recognized antigens on both 3D7 and a Kenyan will also evolve resistance to vaccines. Antigenic escape, for example, was parasite line, suggesting that conserved antigenic determinants are present observed for Combination B (MSP2) and FMP2.1 (AMA1) malaria vaccines, on the surface of gametocyte-infected erythrocytes. and the parasite could also shift to alternative invasion pathways that circumvent the requirement of the vaccine antigen. Thus, it is important to 1866 study the effects of long term persistence of inhibitory levels of antibodies induced by a Plasmodium falciparum blood stage vaccine - using in A SYSTEMATIC CHARACTERIZATION OF MALARIA- vitro and if possible in vivo animal models of malaria. We have recently ASSOCIATED ATYPICAL MEMORY B CELLS shown that AMA1 strain-specific antigenic escape could be overcome by inducing broadly inhibitory antibodies using a Quad-allelic formulation 1 1 2 Silvia Portugal , Severin Zinoecker , Youssounou Kone , of AMA1 (QuadVax, or QV: 3D7+ FVO+HB3+W2mef allelic forms) which 1 2 2 Christine E. Schindler , Safiatou Doumbo , Didier Doumtabe , elicited high levels of invasion inhibitory antibodies in rabbits against 3 3 3 Kishore Kanakabandi , Daniel E. Sturdevant , Stephen F. Porcella , not only all four vaccine strains but also against 22 antigenically diverse 2 2 2 Ogobara K. Doumbo , Kassoum Kayentao , Aissata Ongoiba , non-vaccine strains (Dutta et al. 2013, PLOS Pathogens). We now used 4 5 5 Susan Moir , Jean-Nicholas Schickel , Eric Meffre , Boubacar anti-QV antibodies to exert immune pressure on two parasite strains (3D7 2 1 1 Traore , Susan K. Pierce , Peter D. Crompton and W2mef) in long-term cultures. Parasites were maintained in culture 1National Institute of Allergy and Infectious Diseases, National Institutes for six months in the presence of ~50% inhibitory concentration of anti- of Health, Rockville, MD, United States, 2Mali International Center of QV rabbit serum while the control parasites were maintained in parallel Excellence in Research, Bamako, Mali, 3RML, National Institutes of Health, in the absence of antibodies. During the cultures, parasite lines were Hamilton, MT, United States, 4National Institute of Allergy and Infectious frozen at various time-points and at the end of the 6 months of culture Diseases, National Institutes of Health, Bethesda, MD, United States, the parasites were cloned by limiting dilution. Selection pressure was 5Department of Immunobiology, Yale University School of Medicine, New finally removed and anti-AMA1 selected clones were compared to the Haven, CT, United States parental or control selected parasites. Comparative data will be presented Antibodies (Ab) play a critical role in malaria immunity, but Ab-mediated regarding (a) growth and invasion rates in the presence or absence of anti- protection is only acquired after years of repeated infections, leaving AMA1 antibodies, (b) parasite DNA sequences, (c) invasion into enzyme children in endemic areas vulnerable to repeated bouts of febrile malaria. treated red cells, and (d) quantity and location of AMA1 and its proteolytic Many Plasmodium falciparum antigens are diverse and clonally variant, processed products. This study informs an important decision point for contributing to the inefficient acquisition of protective Abs. However, future development of an AMA1 vaccine as well as malaria blood stage mounting evidence suggests that Plasmodium-induced dysregulation vaccine development in general. of B cell function may also play a role. Several studies have shown that

astmh.org 571 1868 157 deaths with an overall mortality rate of 5.4 per 1000 person-years of observation. The median age at the time of death was 31.5 years with ENHANCED MULTIFUNCTIONAL CD4+ T CELL MEMORY the highest (40%) proportion of deaths occurring among women 30-39 RESPONSES TO MALARIA ANTIGENS IN MALIAN CHILDREN years of age. Causes of death were identified in 51% of the individuals. CO-INFECTED WITH SCHISTOSOMA HAEMATOBIUM Maternal deaths as defined by WHO were less frequent compared to non –maternal deaths (7% vs. 93%, respectively; χ²=80.0, p<0.001) in this 1 2 2 Kirsten E. Lyke , Bourema Kouriba , Abdoulaye Dabo , Charles population. Among the non-maternal deaths, 62% were due to infectious 2 1 2 2 Arama , Modibo Daou , Modibo Daou , Issa Diarra , Drissa diseases, with HIV/AIDS associated illness being the leading cause (43%). 2 1 2 Coulibaly , Christopher V. Plowe , Ogobara Doumbo , Marcelo B. Non-communicable diseases were associated with 38% of non-maternal 1 Sztein deaths, of which cancers and cardiovascular disease were common (43% 1University of Maryland, Baltimore, MD, United States, 2University of and 32%, respectively). Communicable diseases were found to be a Sciences Techniques and Technologies, Bamako, Mali major cause of death among women of reproductive age; however, non- Durable immunologic memory to malaria is limited in young children, communicable diseases are increasing in frequency among this population. where repetitive and ongoing exposure to malaria is required to These findings highlight the need to address and reduce the risk of deaths achieve and maintain immunity. We have previously demonstrated that resulting from both communicable and non-communicable diseases, along schistosomiasis-positive (SP) Malian children, aged 4-8 years, are protected with efforts to reduce maternal deaths among women of reproductive from malaria compared to matched schistosomiasis-negative (SN) children. age. The effect of concomitant S. haematobium upon acquisition of memory to malaria antigens is unknown. We examined antigen-specific T cell 1870 frequencies in 48 Malian children aged 4-14 to malaria blood-stage antigens, Apical Membrane Antigen 1 (AMA1) and Merozoite Surface REVISITING THE BURDEN OF TYPHOID FEVER IN LOW AND Protein 1 (MSP1) and to schistosoma antigens, Soluble Worm Antigenic MIDDLE-INCOME COUNTRIES TO INFORM POLICY DECISIONS Preparation (SWAP) and Schistosoma Egg Antigen (SEA) during a malaria Vittal Mogasale, Brian Maskery, R. Leon Ochiai, Jung Seok Lee, episode and at convalescence 6 months later. CD4+ T cell memory Vijayalaxmi V. Mogasale, Enusa Ramani, Young Eun Kim, Jin cytokine (IFN-γ, TNFα, IL2 and/or IL17A) responses specific to schistosoma Kyung Park, Thomas F. Wierzba antigens was measured in 18/23 SP children at one or both time points, International Vaccine Institute, Seoul, Republic of Korea compared to 4/23 SN children (p < 0.0001). At the time of malaria infection, CD4+ T cells from 12/24 SN children and 15/23 SP children Typhoid fever still causes significant burden in low and middle income (p=0.29) stimulated with malaria antigens demonstrated significantly countries where access to safe water and sanitation is compromised. There increased levels of cytokine production. In contrast, 7/23 SN children and have been several efforts to quantify the global burden of typhoid fever, 16/23 SP children (p=0.009) had responses in paired convalescent samples. but the estimates have not considered the heterogeneity in risk levels 46.2% of cytokine-secreting CD4+ T cells expressed a single cytokine after within countries. Since the World Health Organization has recommended stimulation with malaria antigens during the malaria episode. This fell to risk-based use of vaccines against typhoid, we attempted to revisit 40.9% at follow-up with a compensatory rise of multifunctional cytokine the burden on typhoid fever in low and middle income countries after secretion over time (double+: 30.7 to 32.5%, triple+: 20.6 to 23.1%, adjusting to the risk levels at population. The typhoid disease burden was and quadruple+: 2.4 to 3.8%) consistent with memory maturation. The estimated based on community representative typhoid incidence studies majority (53.2%-59.5%) of cytokine responses were observed in CD45RA- applied to 2010 population after correcting for the operational issues CD62L- effector memory T cells with little variation depending upon the related surveillance, limitations of diagnostic tests and risk difference due time point or the study cohort. We conclude that detectable CD4+ T cell to exposure to unimproved water. Incidence estimates, correction factors memory response can be measured against both malaria and schistosoma and mortality estimates were derived from systematic literature review. antigens and that the presence of S. haematobium may be associated with Scenario analyses for risk factors, blood culture sensitivity and case fatality enhanced functional T memory cell induction to malaria antigens. rates were conducted accounting for the uncertainty in these estimates and compared to previous disease burden estimates. Findings: The risk- 1869 adjusted estimate of typhoid fever in low and middle income countries was 11.9 million cases (CI: 9.9 - 14.7 million) and 129,000 deaths (CI: 75,000 CAUSES OF MORTALITY IN WOMEN OF REPRODUCTIVE AGE - 208,000). In comparison, without the risk- adjustment, the burden LIVING IN AN URBAN SLUM (KIBERA) NAIROBI estimate would be 20.6 million cases (CI: 17.5 - 24.2 million) with 223,000 deaths (CI: 131,000 - 344,000). Scenario analyses indicated that the risk 1 2 Beatrice Olack , Leonard Otieno Cosmas , Daniel Ondari factor adjustment and updated diagnostic test correction factor derived 1 2 Mogeni , Joel Mark Montgomery from systematic literature review were the drivers of difference between 1Kenya Medical Research Institute-Centre for Global Health Research and current estimate and past estimates. Interpretation: The risk-factor Centers for Disease for Control and Prevention, Nairobi, Kenya, 2Global adjusted typhoid fever burden estimate is inherently more conservative Disease Detection Branch, Division of Global Health Protection, Center for than previous estimates that did not account for study site selection bias Global Health, Centers for Disease Control and Prevention, Atlanta, GA or fractions of the populations residing in urban slums or rural areas USA, Nairobi, Kenya lacking access to improved water supplies. However, by distinguishing Women of reproductive age (15-49 years) are confronted by dual burden and discriminating the risk differences, it allows better estimation of the of health concerns related to maternal conditions: infectious diseases population level impact and evaluation of cost effectiveness of risk-based and emerging challenges associated with non-communicable diseases. vaccination strategies recommended by World Health Organization. More than 30% of all deaths in resource-limited settings occur in these women, as compared to 15% in resource-rich settings. There is a paucity 1871 of mortality data and causes of death among females of reproductive age in low income countries. We present findings from verbal autopsies among DISABILITY- AND QUALITY-ADJUSTED LIFE YEARS: women of reproductive age living in Kibera, an urban slum in Nairobi, MEASURING HEALTH OR ? Kenya. Verbal autopsies were conducted among women of reproductive Thomas Fürst, Maria-Gloria Basáñez, Lesong Conteh age who were participants in a population-based surveillance system and ⋯ Imperial College London, London, United Kingdom who died between January 2009 and December 2013. Details regarding the death were obtained from close relatives and cause was assigned Disability-adjusted life years (DALYs) and quality-adjusted life years using the InterVA-4verbal autopsy model (version 4.02). We identified (QALYs) have risen to prominence over the past years and are frequently

astmh.org 572 used as denominators in cost-effectiveness analyses. They have become 1873 a powerful “currency” in health economics, health policy and public health decision making. By combining life years lost due to premature FALSIFIED MEDICINES IN AFRICA AND PUBLIC HEALTH - ‘NO mortality with disability- or quality-adjusted life years reflecting morbidity ACTION-TALK ONLY’ and attributing these summary measures to specific health conditions 1 2 3 and interventions, DALYs and QALYs aim at quantifying health losses Paul Newton , Patricia Tabernero , Prabha Dwivedi , Maria 3 3 4 5 and health gains respectively. An explicit assumption is that DALYs Culzoni , Maria Monge , Isabel Swamidoss , Dallas Mildenhall , 4 6 7 and QALYs allow for comparison of different causes of health losses Michael Green , Richard Jähnke , Miguel Santos de Oliveira , Julia 7 8 3 and health gains and that they are therefore suitable to guide global, Simao , Nicholas White , Facundo Fernández national and local decision making on where to invest scarce resources. 1Mahosot Hospital, Vientiane, Lao People’s Democratic Republic, However, based on a literature review, we argue that the DALYs and 2University of Oxford, Oxford, United Kingdom, 3Georgia Institute of QALYs lack a clear definition of the concepts “health”, “disability” and Technology, Atlanta, GA, United States, 4Centers for Disease Control and “health-related quality of life” and therefore also of their disability- and Prevention, Atlanta, GA, United States, 5GNS Science, Twin Hut, New quality-adjustments for individuals’ life years spent in less than perfect Zealand, 6Global Pharma Health Fund, Frankfurt, Germany, 7Inspecção health. Mainly based on the highly topical International Classification of Geral de Saúde, Luanda, Angola, 8Mahidol University, Bangkok, Thailand Functioning, Disability and Health of the World Health Organization, we Poor quality medicines are neglected impediments to improving global developed a conceptual framework to delineate what the DALYs and public health. In June 2012 suspected falsified medicines labelled as the QALYs do, and do not measure. Important similarities and differences antimalarial ‘artemether-lumefantrine’ bearing the Affordable Medicines between the two measures are revealed. Critical questions about the Facility-malaria (AMFm) logo and others labelled as the antihelminthic conceptualization of DALYs and QALYs are discussed and we conclude that ‘mebendazole’ were seized in Luanda, Angola. The tablets were if these questions are not addressed there is a continued risk of inefficient analysed by an array of analytical platforms including high performance decision making and ill-informed advocacy. liquid chromatography, ambient ionization mass spectrometry, Raman spectroscopy, X-ray powder diffraction analysis, nuclear magnetic 1872 resonance spectroscopy, isotope-ratio mass spectrometry, botanical assays and packaging analysis, using the portable counterfeit detection device VALIDATION OF CAUSES-OF-DEATH USING VERBAL CD-3. No artemether or lumefantrine or other active pharmaceutical AUTOPSY DATA COLLECTED FROM NAVRONGO HEALTH ingredients were detected in the ‘artemether-lumefantrine’ tablets. AND DEMOGRAPHIC SURVEILLANCE SYSTEM IN GHANA: Brushite and three different yellow dyes and few pollen grains were 2007-2011 found. No mebendazole was detected in the ‘mebendazole’ formulation,

1 1 1 but calcite and levamisole (270mg/tablet) were present. Both ‘products’ Patrick O. Ansah , Cornelius Debpuur , James Akazili , Timothy showed marked differences in packaging characteristics from genuine Awine1, John Williams1, Nana Akosua Ansah1, Yazoume Ye2, Isaac 1 2 3 1 products. The discovery of falsified artemether-lumefantrine, labelled Osei , Leopoldo Villegas , Osman Sankoh , Abraham Oduro as an AMFm product and without any detectable antimalarial, is of 1Navrongo Health Research Centre, Navrongo, Ghana, 2ICFI, Rockville, MD, considerable concern for malaria control. Presence of levamisole in falsified United States, 3INDEPTH Network, Accra, Ghana ‘mebendazole’ is also of great concern as it has been banned for human Determination of cause-specific mortality rate in developing countries is use. This seizure illustrates many of the current problems regarding a major difficulty due to poor vital registration systems, deaths occurring poor reporting and transparency and inaction. Enhanced collaboration outside health facilities, and deaths not being medically certified. Verbal between African MRAs/police and the authorities in China to stop criminal autopsy conducted on these deaths have proven to be one of the reliable transcontinental trade in falsified essential medicines is urgently needed. methods of compiling cause of death data. We sought to validate the Delays in reporting and action must be reduced by mandatory notification causes of death assigned by physician coders and provide a cause of systems and independent public health risk assessments. Despite multiple death structure to improve estimates on cause specific mortality in Ghana. reports, public health research has failed to stimulate actions required to Longitudinal VA data from the Navrongo Health and Demographic improve the quality of global drug supply. Surveillance System (NHDSS) from 2007-2011 was used. Physicians were retrained on death certification and coding using ICD 10 codes and 1874 Sample Vital Registration with Verbal Autopsy (SAVVY) methods. VA forms were recoded using the SAVVY methods. In all, 7086 VA forms ENHANCING PUBLIC HEALTH RESPONSE TO CLIMATE- were retrieved and recoded using SAVVY methods. Males constituted SENSITIVE INFECTIOUS DISEASE OUTBREAKS IN 56% of total deaths and 60% of deaths occurred outside health facilities. FLOOD-PRONE AREAS OF BANGLADESH: ARE PRIMARY The main causes of neonatal deaths were neonatal sepsis (31.5%), birth HEALTHCARE FACILITIES READY TO RESPOND? asphyxia (18.1%) and low birth weight with prematurity (15.4%). Malaria 1 2 2 (37.0%), diarrhea (13.1%) and acute respiratory infection (11.6%) were Farhana Haque , Mahmudur Rahman , Selina Khatun , 2 2 2 the leading causes of death among children aged 1-11 months. The Mujaddeed Ahmed , M. Mushtuq Husain , Nuzhat Nasreen Banu , 3 3 4 main causes of death for 1-4 year olds were malaria (53.2%), diarrheal Md Saiful Islam , Md Kamal Hossain , Stephen P. Luby , Emily S. 3 5 diseases (9.0%), and unspecified infectious diseases (4.7%). Among Gurley , James D. Heffelfinger children 5- 15 years, the main causes of death were malaria (22.5%), 1International Centre for Diarrhoeal Disease Research, Bangladesh accidental drowning, submersions and falls (14.6%) and meningitis and Institute of Epidemiology, Disease Control and Research, Dhaka, (7.7%). For those above 15 years, unspecified non-communicable diseases Bangladesh, 2Institute of Epidemiology, Disease Control and Research, (14.7%), malaria (6.9%) and cerebro-vascular diseases (6.5%) were the Dhaka, Bangladesh, 3International Centre for Diarrhoeal Disease Research, main causes of death in the districts. Variability between two coders Bangladesh, Dhaka, Bangladesh, 4International Centre for Diarrhoeal using SAVVY method was fair (49.7%; P<0.001) with a higher value for Disease Research, Bangladesh and Centre for Infectious Disease and neonatal deaths compared to adults. Despite the limitations of VA data the Geographic Medicine, Stanford University, Stanford, CA, United States, method provides an understanding of the cause of death structure at the Dhaka, Bangladesh, 5International Centre for Diarrhoeal Disease Research, population level in developing countries comparable with global estimates Bangladesh and Global Disease Detection Branch, Division of Global that is not possible with existing sources of data. Health Protection, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, GA, United States, Dhaka, Bangladesh Global climate change is increasing Bangladesh’s vulnerability to natural disasters including floods and raising the potential for infectious

astmh.org 573 outbreaks. Preparing healthcare for outbreak management is complex and supervision, alone or in combination, had lower effect sizes (typically and requires comprehensive understanding of the systems’ capacities ranging from no effect to +16 %-points). Among 15 studies focused only and challenges. We evaluated preparedness of government primary on community HWs, the strategy with the greatest health impact was care facilities in flood-prone sub-districts to respond to outbreaks. consumer supports (e.g., patient education) + HW training + providing We asked 69 primary care managers to complete a self-administered drugs or equipment (MES = 55 %-points, IQR: 15, 62). Contextual and survey to assess 7 core capacities critical for outbreak response. We methodological heterogeneity made comparisons difficult. Results from determined percentages of facilities reporting each capacity; assessed this review, which will be finalized by the end of 2014, should inform managers’ perceived preparedness level; and conducted 2 focus group decision-making on how best to improve HW performance and health discussions (FGDs) with 24 managers to explore contextual factors driving outcomes in LMICs. preparedness. Qualitative data were analyzed using thematic content analysis. All 69 facilities lacked preparedness plans, emergency stockpiles, 1876 funds and authority to buy drugs or supplies locally; all lacked dedicated evaluation, isolation and vehicles for infectious patients; 58% lacked PREVALENCE OF TRACHOMA IN BRAZILIAN sufficient clinicians and 20% lacked adequate field staff. Rapid response SCHOOLCHILDREN teams and digital communication with referral centers existed in all, Expedito J. Luna1, Maria F. Lopes2, Norma H. Medina3, Maria R. limited infection control practice in 75%, and diagnostic laboratory in Cardoso1 80% facilities. Unlike lower stockpiling, infrastructure and human resource capacities, higher surveillance, communication, infection control and 1Universidade de Sao Paulo, Sao Paulo, Brazil, 2Ministerio da Saude, laboratory capacities were observed. Sixty-three (91%) managers felt their Brasilia, Brazil, 3Secretaria Estadual de Saude de Sao Paulo, Sao Paulo, facilities had limited preparedness. There was moderate (71%; Kappa=0.3; Brazil P=0.001) correlation between measured and perceived preparedness. The aim of the study was to estimate the prevalence and describe the FGDs identified insufficient training of providers, low motivation and rapid distribution of trachoma among schoolchildren in Brazil. We conducted turnover of clinical staff, and lack of epidemiologists posted at facilities a cross-sectional study, using cluster sampling of the schoolchildren as impediments to human resource capacity. Using a mixed methods population, living in Brazilian municipalities with Human Development approach, this assessment identified deficiencies in all capacities, indicated Index lower than the national mean. This prevalence survey was conducted major gaps in stockpiles, infrastructure and human resource as priority by the Brazilian Ministry of Health, in the period 2002-2008. 3,144 areas for investing constrained resources and offered baseline data for schools, with 176,224 schoolchildren, from 1st to 4th grades, located monitoring progress in preparedness. This may be a feasible methodology in 1,491 municipalities, were selected. The selected schoolchildren to evaluate health systems in low-income countries. Development of a underwent an external ocular examination, with a magnifying glass contingency plan that secures emergency financing and improves surge (2.5X), to detect clinical signs of trachoma according to the WHO grading capacity by establishing a system to engage existing field workers in criteria. The prevalence of trachoma, by state and national level, and their outbreaks can sustainably enhance primary care preparedness. respective 95% confidence intervals were estimated. Chi-square and chi- square for trends tests were used to compare categorical variables. 8,526 cases of trachoma were detected, resulting in a prevalence of 5.00% 1875 (95%IC 5.05; 4.95). Most cases were mild (TF prevalence = 4.92%). A SYSTEMATIC REVIEW ON THE EFFECTIVENESS OF Prevalence of intense inflammatory trachoma (TI) and trachomatous STRATEGIES TO IMPROVE HEALTH WORKER PERFORMANCE scarring (TS) was low: 0.03% and 0.05% respectively. There was no significant difference between the sexes. The prevalence of trachoma was IN LOW- AND MIDDLE-INCOME COUNTRIES: PRELIMINARY 10.8% among children under 5 years of age, decreasing as age increased RESULTS ON HEALTH OUTCOMES (chi square for trend p < 0.00001). There was a significant difference in Alexander K. Rowe1, Samantha Y. Rowe1, David H. Peters2, prevalence between urban and rural areas, 4.3% versus 6.2% respectively Kathleen A. Holloway3, John Chalker4, Dennis Ross-Degnan5 (p < 0.001). Cases were detected in 1,189 municipalities (80% of the municipalities in the sample), in all 27 states of the country. In 37% of the 1Centers for Disease Control and Prevention, Atlanta, GA, United States, selected municipalities, the prevalence was higher than 5%. The study has 2Johns Hopkins University, Baltimore, MD, United States, 3World Health shown that trachoma is still endemic in a large proportion of the poorer Organization, New Delhi, India, 4Management Sciences for Health, Brazilian municipalities, contradicting the belief that the disease had been Arlington, VA, United States, 5Harvard Medical School, Boston, MA, controlled in the country. The survey provided a baseline for evaluating United States planned interventions aimed at achieving the goal of certification of Health workers (HWs) play essential roles in delivering health care. In elimination of trachoma as a cause of blindness in Brazil by 2020. low- and middle-income countries (LMICs), however, HW performance is often inadequate. To characterize the effectiveness of strategies to improve HW performance in LMICs, we conducted a systematic review 1877 of 15 electronic databases, 30 document inventories of international BACTERIAL LOAD AND PATHOGEN DIVERSITY IN OCULAR organizations, and bibliographies of 510 articles. We included studies INFECTION WITH CHLAMYDIA TRACHOMATIS IN A meeting accepted criteria for methodological adequacy (e.g., trials with TRACHOMA-HYPERENDEMIC ISLAND SETTING comparison groups) of any strategy on any health topic in any language, published or not. After screening, data from relevant reports were double- Anna Last1, Fengchen Liu2, Travis Porco2, Chrissy Roberts1, Eunice abstracted and entered into a database. This analysis focuses on studies Cassama3, Meno Nabicassa3, Sarah Burr4, Taane Clark1, Francesc that measured health outcomes (morbidity and mortality rates). Effect sizes Coll1, Nick Thomson5, Thomas Lietman2, Martin Holland1, Robin were calculated as percent change over time in the intervention group Bailey1 minus percent change over time among controls. We screened >105,000 1London School of Hygiene & Tropical Medicine, London, United Kingdom, citations, 829 reports met inclusion criteria, and 60 studies measured 2University of California San Francisco, San Francisco, CA, United States, health outcomes (28 on morbidity only, 24 on mortality only, and 8 with 3Ministerio de Saude Publica, Bissau, Guinea-Bissau, 4Medical Research both). Many strategies have been tested, usually with multiple intervention Council The Gambia Unit, Fajara, Gambia, 5Wellcome Trust Sanger components. The median effect size (MES) across all studies was an Institute, Cambridge, United Kingdom improvement of 9 percentage-points (%-points) (interquartile range [IQR]: Trachoma is caused by ocular infection with Chlamydia trachomatis 0, 39). Among 45 studies focused on facility-based HWs, the strategy with (Ct). Acute conjunctival infection may recur and progress to a chronic the greatest health impact was HW training + group problem solving (MES inflammatory process causing conjunctival scarring and blindness. The = 49 %-points, IQR: 24, 77). Often used strategies, such as HW training astmh.org 574 reasons why active and scarring trachoma are so prevalent on the Bijagós 1879 Archipelago of Guinea Bissau are unclear. We collected 1507 population- based conjunctival swabs with corresponding detailed clinical phenotype. REPRODUCTIVE TRACT INFECTIONS AMONG PRIMARY We used droplet digital PCR assay to detect and quantitate Ct DNA SCHOOLGIRLS IN RURAL WESTERN KENYA on swabs. Associations between Ct load and clinical phenotype were 1 1 1 examined using regression models. We used agent-based modeling to Newton Otecko , Emily Kerubo , Elizabeth Nyothach , Alex 1 1 1 1 investigate the role of Ct load in trachoma transmission. Whole genome Morwabe , Harrison Fredrick , Kelvin Oruko , Clifford Oduor , 1 1 1 2 sequence analysis was used to identify variants in putative virulence- Preston Owiti , John Vulule , Frank Odhiambo , Kayla Laserson , 2 3 associated genes/loci. The geometric mean of estimated Ct load in Clement Zeh , Penelope Phillips-Howard clinically normal conjunctivae was 294 copies/swab (95% C.I. 165-524). 1Kenya Medical Research Institute-Centers for Disease Control and In clinically active trachoma it was 8562 copies/swab (95% C.I. 5412- Prevention Research and Public Health Collaboration, Kisumu, Kenya, 13546). In active trachoma Ct load increases with disease severity (for both 2Centers for Disease Control and Prevention, Kisumu, Kenya, 3Liverpool follicular and inflammatory scores). The highest Ct loads were associated School of Tropical Medicine, Liverpool, United Kingdom with the most severe clinical disease and the strongest associations were Reproductive tract infections (RTIs) among adolescent girls remain a great with increasing inflammatory grade (at maximal inflammatory score public health concern in developing countries. However, the burden (P3) OR 30.9, 95% CI 9.39-101.5, p<0.0001). Genotypic differences of RTIs in this key population is insufficiently understood. This study in virulence-associated genes within this population of ocular Ct are presents preliminary analysis of RTIs’ symptoms reporting and laboratory suggested. The association between load and disease severity may be detection rates among girls aged 14-16 (median 14) years, in 30 primary related to Ct strain diversity, where multiple strains are co-circulating. schools, enrolled in a feasibility study on acceptance, use and safety of We used a novel mathematical modeling strategy to investigate the role menstrual products in western Kenya. Vaginal self-swabbing samples of Ct load in trachoma transmission. This is the first application of these were prospectively collected during symptom guided RTI testing (SGT) approaches in understanding the pathogenesis and transmission of Ct (March-October 2013) and cross-sectional end-of-study screening (EOSS) infection, which are fundamental to successful trachoma elimination and (November 2013). Samples were analyzed for Bacterial vaginosis (BV), surveillance strategies. Chlamydia trachomatis (CT), Neisseria gonorrhea (NG), Trichomonas vaginalis (TV) and Candidiasis. Infected girls were referred for treatment. 1878 Data were analyzed using SPSS v.21.0. Overall, 532 girls (SGT: 17, 3.2%; EOSS: 453, 85.2%; and overlap in both: 62, 11.7%) were included. Of a ASSESSING THE BURDEN OF PEDIATRIC ACUTE RHEUMATIC total 79 girls in SGT group, BV 13 (16.5%), Candidiasis 11 (13.9%), TV 5 FEVER AND RHEUMATIC HEART DISEASE --- AMERICAN (6.3%) and CT 2 (2.5%) were test confirmed. None tested positive for NG. SAMOA, 2011-2012 BV was the most common in EOSS, 94 (18.3%), followed by Candidiasis 44 (8.5%), TV 13 (2.5%), CT 13 (2.5%) and NG 3 (0.6%). Of 62 girls in Amanda L. Beaudoin1, Laura Edison1, Camille E. Introcaso2, Lucy both STG and EOSS, RTI detection rates varied (SGT-EOSS) for BV (19.4%- M. Goh3, Chris Van Beneden1 14.5%), Candidiasis (14.5%-12.9%), TV (6.5%-0%), CT remained 1 Centers for Disease Control and Prevention, Atlanta, GA, United States, constant. While only 82 (15.9%) girls reported symptoms for RTI at EOSS, 2 Pennsylvania Hospital, Pennsylvania Center for Dermatology, Philadelphia, laboratory testing showed 146 (28.3%) had at least one RTI. In conclusion, 3 PA, United States, Lyndon Baines Johnson Tropical Medical Center, Pago high detection rate of RTIs was observed among the rural adolescent Pago, American Samoa schoolgirls. Symptom-based diagnosis of RTIs poorly predicted RTI in this In August 2013, LBJ Tropical Medical Center and the American Samoa (AS) population. These findings offer important insights for treatment and health department notified CDC of a perceived high burden of pediatric prevention of RTIs among schoolgirls. acute rheumatic fever (ARF) and rheumatic heart disease (RHD). ARF is an immunologically mediated sequela of inadequately treated group 1880 A Streptococcus pharyngitis and, potentially, pyoderma. Recurrent or severe ARF can cause permanent cardiac damage and RHD. Long-term ACCURACY OF THE INTEGRATED MANAGEMENT OF prophylactic penicillin injections post-ARF diagnosis can prevent RHD. We CHILDHOOD ILLNESS (IMCI) ALGORITHM IN IDENTIFYING aimed to describe pediatric ARF and RHD and prophylaxis in AS and the CULTURE-CONFIRMED DIARRHEAL PATHOGENS REQUIRING pyoderma-ARF association. We used ICD-9 codes and hospital prophylaxis ANTIBIOTIC THERAPY registries from AS’s only medical system to identify all patients aged ≤18 1 1 years with a physician-recorded ARF or RHD diagnosis during 2011-2012. Patricia B. Pavlinac , Frankline M. Onchiri , Grace C. John- 1 1 1 We recorded penicillin compliance and pre-ARF pharyngitis and pyoderma Stewart , Jaqueline M. Naulikha , Denno M. Donna , Elizabeth A. 2 3 1 1 diagnoses (<6weeks preceding) for cases. Two age- and sex-matched Odundo , Benson O. Singa , Lisa E. Manhart , Judd L. Walson control subjects per case-patient were selected from non-ARF/RHD patients 1University of Washington, Seattle, WA, United States, 2United States examined during 2011-2012. We calculated ARF 2011-2012 incidence Army Medical Research Unit, Kericho, Kenya, 3Kenya Medical Research and RHD prevalence by using 2010 U.S. Census data. We used univariate Institute, Nairobi, Kenya statistical tests and conditional logistic regression for case-control Diarrhea is the second leading cause of death in children under 5, with comparisons. During 2013, RHD prevalence was 3.2 cases/1,000 children. most deaths occurring in settings where microbiology facilities are ARF incidence was 1.1 (2011) and 1.5 (2012) cases/1,000. Of 65 children unavailable or cost prohibitive. The World Health Organization (WHO) diagnosed with ARF during 2011-2012, a total of 32 (49%) subsequently developed the Integrated Management of Childhood Illness (IMCI) received RHD diagnoses. Median ARF diagnosis age was 11 (range: 2-18) guidelines to manage sick children based on clinical signs and history. For years. Pharyngitis history was more common among case-patients (18%) children with diarrhea, the guidelines recommend empiric antibiotics for than control subjects (0%; P <0.01), but preceding pyoderma was not. children with suspected shigellosis (presence or history of bloody stool) Post-ARF penicillin prophylaxis compliance (65%) was suboptimal. RHD or suspected cholera (age ≥2 years, severe dehydration, and living in a causes considerable childhood morbidity in AS. Although the pyoderma- cholera endemic area). We assessed the diagnostic performance of the ARF association remains unclear, attempts to curb AS’s RHD burden should IMCI guidelines for diarrhea management as compared to stool bacterial address improved pharyngitis diagnosis and treatment and increased ARF culture. Children aged 6 months to 5 years presenting to two Western prophylaxis compliance. Kenya District hospitals between December 2011 and September 2013 with acute diarrhea were enrolled. Stool samples were tested using standard methods for bacterial culture. Multiplex PCR was used to further classify diarrheagenic Escherichia coli. Among 973 enrolled children,

astmh.org 575 median age was 17 months (interquartile range 10-34), 16.5% were the major health impacts of climate change, but there much uncertainty stunted, and 4.4% were HIV-infected. The most predominate bacterial remains in the estimates associated with the relationship between isolate was EAEC (14.1%), followed by Campylobacter (6.6%), EPEC climatic drivers and diarrheal disease. One source of this uncertainty is (6.2%), Shigella (4.6%), and ETEC (4.4%). IMCI correctly classified 3 of 45 the variety of pathogens associated with infectious diarrhea, each of lab-confirmedShigella cases (sensitivity 6.7%), 2 cases of Shigella flexneri which has different life cycle characteristics such as survival outside of and 1 S. dysenteriae. Among 928 children without shigellosis, IMCI the host. Therefore it is important to understand how the seasonality of correctly classified 871 (specificity 93.9%). Of the 57 children incorrectly different etiological agents of diarrheal disease vary. In this systematic diagnosed with Shigella by IMCI, 73.7% had no other bacterial pathogen review and meta-analysis, we examined the impact of climatic variability identified. Cholera was not detected although 11 (1.1%) children were on three representative diarrheal disease pathogens of different taxa: classified as having suspected cholera based on IMCI criteria (specificity pathogenic E. coli, norovirus, and cryptosporidium. Incidence data for each 98.9%); of these 11, 36.4% had no isolated bacteria. The IMCI guidelines pathogen were taken directly from tables or extracted from graphs in the appear reasonably specific but not sensitive in identifying children published papers. Year-specific monthly temperature and precipitation requiring antibiotic therapy. IMCI guidelines should be adapted to enhance from each location at the time of disease data collection were assembled sensitivity and to account for additional enteric pathogens associated with from publicly available datasets. We examined the relationship between increased morbidity and mortality. climatic variables and incidence of each pathogen for each location using generalized log-linear Poisson regression models, and we also pooled all 1881 datasets for each pathogen to calculate an overall association between monthly cases and mean monthly temperature, using a generalized EVALUATION OF INTEGRATED MANAGEMENT OF estimating equation. We then used the model results to examine what ADOLESCENT AND ADULT ILLNESS DISTRICT CLINICIAN proportion of the total of cases attributable to these three pathogens MANUAL EMPIRIC ANTIMICROBIAL THERAPY would be attributable to any one of the pathogens given increases in RECOMMENDATIONS FOR SEVERE INFECTIONS IN temperature of 1-4°C, in increments of one degree. We found that a one- NORTHERN TANZANIA unit increase in temperature was associated with increases in incidence of pathogenic E. coli (IRR = 1.08, 95% CI = 1.04-1.11) and Cryptosporidium 1 2 1 Matthew P. Rubach , Venance P. Maro , John A. Bartlett , John (IRR=1.03, 95% CI = 1.03-1.04) and decreases in Noroviurs (IRR = 0.92, 1 A. Crump 95% CI: 0.90-0.94). These results highlight the importance examining 1Duke University, Durham, NC, United States, 2Kilimanjaro Christian taxa-specific climate-disease relationships for enteric diseases. As Medical Centre, Moshi, United Republic of Tanzania temperatures increase under future warming scenarios, bacterial and We assessed the effectiveness of Integrated Management of Adolescent protozoan pathogens are expected to represent an increasingly large and Adult Illness District Clinician Manual (IMAI) empiric antimicrobial fraction of the burden of diarrheal disease. This has important implications therapy recommendations for septic shock, severe respiratory distress for development of control strategies. without shock, and severe pneumonia in hospital settings in northern Tanzania. IMAI recommended empiric therapies were retrospectively 1883 evaluated against laboratory-confirmed etiology of illness data for participants in a febrile illness cohort study who met IMAI criteria for the FLY ME TO THE PLUME: VIDEO-TRACKING ANALYSIS OF three clinical syndromes. Therapies evaluated included IMAI emergency ANOPHELES GAMBIAE FLIGHT BEHAVIOR AT HUMAN- antibacterials (ceftriaxone or ampicillin plus gentamicin) for septic shock BAITED BEDNETS and severe respiratory distress without shock, and ceftriaxone plus a Josephine Parker1, Natalia Angarita-Jaimes2, Mayumi Abe1, macrolide for severe pneumonia. Among 423 participants hospitalized Fabian Mashauri3, Jackline Martine3, Catherine E. Towers2, David with febrile illness, 171 cases met IMAI criteria for the three syndromes: Towers2, Philip J. McCall1 25 septic shock, 37 severe respiratory distress without shock, and 109 1 severe pneumonia. Forty-four (10%) of 423 participants died in-hospital. Liverpool School of Tropical Medicine, Liverpool, United Kingdom, 2 Ceftriaxone was the single-most effective agent in all three syndromes, Optical Engineering Group, School of Engineering, University of Warwick, 3 being effective for 12 (48%) septic shock, 5 (14%) severe respiratory Coventry, United Kingdom, National Institute for Medical Research, distress without shock, and 18 (17%) severe pneumonia illnesses. For Mwanza, United Republic of Tanzania each syndrome 17-27% of participants had an etiologic diagnosis non- Understanding how mosquitoes interact with insecticide-treated bednets responsive to ceftriaxone, but responsive to other available antimicrobial (LLINs) is fundamental to advancing the design and performance of regimens, namely amphotericin for cryptococcosis and histoplasmosis; LLINs, and to ensuring they continue to be effective and sustainable anti-tuberculosis therapy for bacteremic disseminated tuberculosis; or tools for malaria prevention. We have developed an innovative video tetracycline therapy for rickettsioses and Q fever. IMAI recommendations system that enables nocturnal flight activity of multiple mosquitoes at a for empiric ceftriaxone to treat septic shock, severe respiratory distress human occupied bednet to be captured at high resolution, and individual without shock, and severe pneumonia are warranted. Etiologies not mosquito flight paths and details of movements to be tracked and explicitly addressed in IMAI guidance for these syndromes, such as analysed over periods of 60 minutes or more. Following initial laboratory cryptococcosis, histoplasmosis and tetracycline-responsive bacterial studies, the system has been deployed in an experimental hut at a field infections, were common. Prospective assessments of IMAI are needed to site in Tanzania, where we are investigating behaviour of local Anopheles confirm these results and improve syndromic management algorithms. sp. populations entering the hut in response to human baits in untreated and insecticide-treated bednets. Analysing flight tracks, we have classified 1882 mosquito activity into four broad types, termed ‘swooping’, ‘visiting’, ‘bouncing’ and ‘resting’. Mosquitoes flew more slowly and flight paths IMPACT OF FUTURE CLIMATIC CONDITIONS ON VIRAL, were more tortuous when nets were baited. Responding to human bait, BACTERIAL AND PROTOZOAN ENTERIC PATHOGENS most activity was spent in flight. The majority of contacts made with the net surface were very brief (duration less than 4 seconds) and activity Karen Levy, Sharia M. Ahmed, Rebecca Philipsborn, Lily M. occurred primarily on the top surface of the net over the sleeper’s torso, Tyndall, Berry J. Brosi with less activity seen at the supine human’s feet. This finding is consistent Emory University, Atlanta, GA, United States with previous studies suggesting that hostseeking mosquitoes orient Understanding the seasonality of infectious diseases is important, in order towards a ‘plume’ of host attractants, funnelled upwards by the ‘chimney’ to prepare for case loads, plan vaccination campaigns, and to anticipate effect of the bednet walls. We compared activity on untreated nets with impacts of climate change. Diarrheal diseases are often cited as one of Permanet 2.0 (deltamethrin-treated LLINs) to examine how treatment

astmh.org 576 influenced mosquito behaviour. Results investigating flight patterns and critical effects of the control measure. While it is usually not possible to visiting patterns at the net surfaces, changes in activity patterns over time simulate large populations with laboratory experiments, it useful to test and LLIN repellency will be presented and implications for current and critical predictions experimentally when such methods can be devised. We future LLIN-based approaches will be considered. will describe experiments conducted to determine whether a previously published model of late-acting lethal transgenic mosquitoes adequately 1884 includes critical biological factors of the technology, specifically effects on development rates and survival. We performed laboratory simulations of SWARMING BEHAVIOR OF ANOPHELES GAMBIAE MALES late-acting lethality and conventional sterile insect technique to determine INCREASES FEMALE INSEMINATION RATE IN CAGED the effect on the development rate and survival of two Aedes species POPULATIONS: AN OPPORTUNITY TO STUDY MOSQUITO larvae. We also considered the results and novel experimental variables in MATING SYSTEMS the context of previous models of control of mosquito populations using late-acting lethals in comparison with SIT. Luca Facchinelli1, Laura Valerio1, Clelia Oliva2, Rosemary S. Lee2, Tania Persampieri1, C. Matilda Collins3, Andrea Crisanti1, Roberta 1886 Spaccapelo1, Mark Q. Benedict1 1University of Perugia, Perugia, Italy, 2Polo d’Innovazione Genomica, ANTIMALARIAL AND ANTI-DENGUE PROPERTIES OF A Genetica e Biologia S.C.a.R.L., Perugia, Italy, 3Imperial College London, NATURAL CHROMOBACTERIUM MOSQUITO MIDGUT London, United Kingdom COMMENSAL Long-term control of Anopheles gambiae, the major malaria vector in Raúl G. Saraiva, Yuemei Dong, Abhai Tripathi, José L. Ramirez, Africa, is difficult to achieve and maintain with traditional control methods. George Dimopoulos Genetically Modified (GM) A. gambiae strains that bear sterility genes are potentially powerful new tools to control the disease. Natural male Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United mating behavior is a key component in GM mosquito control strategies States in order to spread transgenes into the wild type population. Mating Plasmodium and dengue virus, the causative agents of the two most behavior involves the ability of males to form swarms, which is crucial to devastating vector-borne diseases, malaria and dengue, are transmitted by inseminate females in the wild. In spite of its biological importance and the Anopheles gambiae and Aedes aegypti mosquito vectors, respectively. relevance for vector control, swarming is a poorly understood process We have identified a novel Chromobacterium species in adult A. aegypti because it is hard to study in the wild and difficult to stimulate under collected in Panama, Csp_P, that can effectively colonize the midgut of A. laboratory conditions. Here we describe features that promote male A. gambiae and A. aegypti mosquitoes when introduced through an artificial gambiae G3 strain swarming in large cages. In 15.6 m3 cages, a dark nectar meal. We have shown that this isolate exerts entomopathogenic foreground and contrasting illuminated background with a contrasting activity against both of the mosquito species along with in vivo and in mark on the ground stimulated swarm formation during artificial twilight. vitro anti-Plasmodium and anti-dengue activities. Interestingly, the well G3 males have not lost their capability to swarm although this strain has characterized Chromobacterium violaceum does not exert such effect. been colonized since 1975. We asked whether swarming behavior would Upon bioassay-guided fractionation of supernatants of a Csp_P culture, affect mating performance of wild-type (WT) G3 and I-PpoI transgenic A. we were able to map the antiparasitic and antiviral properties to a fraction gambiae sexually sterile males competing for G3 females. We performed significantly enriched in a previously characterized cyclic dehydropeptide competitive matings and recorded female insemination rate and lactone. This bacterial secondary metabolite was previously pursued as proportion of matings by WT and GM males. The presence of swarming an antifungal, being part of a complex of closely related molecules. We stimuli was associated with an increase in mating frequency from 77.4 to have produced n-butanol-based extracts of Csp_P cultures that retain 97.4 %. There was no change in competitiveness by transgenic males as in vitro activity against blood-stage Plasmodium and dengue virus, as a function of swarming stimuli. The increase of mating frequency in the well as against the yeast Saccharomyces cerevisiae. We are currently presence of swarming stimuli highlights the importance of swarming in A. pursuing mass spectrometry analysis to characterize the compounds gambiae mating behavior. Reproducing A. gambiae swarms in controlled behind the antipathogenic activity of our extracts, along with efforts to conditions provides the possibility to dissect the mating behavior of this identify the gene cluster responsible for production of such compounds species and explain the mechanisms controlling it, which is innovative in by means of both comparative genomics and a transposon-mediated mosquito research. We will discuss the results and the possible applications random mutagenesis screening. To our knowledge, this is the first of our findings to investigation of A. gambiae biology and to support identified bacterium that exerts broad spectrum entomopathogenic and vector control strategies antipathogenic activities, thereby rendering it an interesting candidate for the development of novel vector-borne disease control strategies. 1885 1887 NOVEL INSIGHTS INTO GENETIC CONTROL USING EXPERIMENTAL AND MATHEMATICAL SIMULATIONS OF THE EFFICACY OF LONG-LASTING NETS WITH DECLINING LATE-ACTING LETHAL EFFECTS ON POPULATIONS OF AEDES PHYSICAL INTEGRITY MAY BE COMPROMISED IN AREAS SPP. MOSQUITOES WITH HIGH LEVELS OF PYRETHROID RESISTANCE Mark Q. Benedict1, Alessandro Bocci1, Michael A. Robert2 Eric O. Ochomo1, Nabie M. Bayoh2, Edward D. Walker3, Bernard 2 2 1 1 2 O. Abong’o , Maurice O. Ombok , Collins Ouma , Andrew K. University Perugia, Perugia, Italy, University New Mexico, Albuquerque, 4 4 5 6 NM, United States Githeko , John M. Vulule , Guiyun Yan , John E. Gimnig 1 2 Mathematical models have been used to predict that late-acting lethal Maseno University, Kisumu, Kenya, KEMRI/Centers for Disease Control 3 transgenic mosquitoes provide enhanced control of target mosquito and Prevention, Kisumu, Kenya, Michigan State University, East Lansing, 4 populations by maintaining competition in the larval stage. The expected MI, United States, Kenya Medical Research Institute, Kisumu, Kenya, 5 6 effect is to diminish larval survival among the wild-type larvae, and in University of California, Irvine, CA, United States, Centers for Disease comparison with situations in which such competition is not present (e.g. Control and Prevention, Atlanta, GA, United States conventional sterile insect technique - SIT), improved control. Mathematical Long-lasting insecticide-treated mosquito nets (LLINs) are a primary simulations often include simplifying assumptions that might not reflect malaria prevention tool in sub-Saharan Africa but emergence of insecticide biological realities and yet provide useful frameworks for predicting resistance threatens their effectiveness. Cross-sectional surveys of LLINs the relative value of various control approaches - that is, if they include were conducted in houses of seven and four villages in Gem and Bungoma

astmh.org 577 Districts in western Kenya, respectively in May 2013. LLIN condition 1889 (number and area of holes), number and species of mosquitoes resting inside, and insecticidal activity of LLINs were quantified. Mosquitoes FACTORS MEDIATING MATING SUCCESS IN MALE collected inside nets were allowed to lay eggs and the progeny were ANOPHELES GAMBIAE MOSQUITOES tested for susceptibility to deltamethrin and permethrin, pyrethoids 1 2 1 commonly deployed in LLINs in western Kenya. In Gem, 83.3% of LLINs Adam South , Paolo Gabrieli , Evdoxia Kakani , Flaminia 1 were less than three years old and 32.4% had at least one hole of any Catteruccia size; while in Bungoma, 92% were less than three years old and 48% had 1Harvard School of Public Health, Boston, MA, United States, 2Università at least one hole. No anopheline and five Culex spp. mosquitoes were degli Studi di Perugia, Terni, Italy found resting inside LLINs in Gem (N=216) regardless of the number and Human malaria, a major public health burden in tropical and subtropical size of holes, while 552 Anopheles gambiae s.l., five An. funestus s.l. countries, is transmitted exclusively by female Anopheles mosquitoes. and 137 Culex spp. were found inside LLINs (N=216) in Bungoma. The Malaria control strategies aimed at inducing sexual sterility in natural number of mosquitoes resting inside LLINs increased with hole areas >50 vector populations are an attractive alternative due to increasing levels cm2 in Bungoma. In WHO resistance assays, f1 offspring of fed or gravid of insecticide resistance. However, the development of these strategies females collected in nets in Bungoma had 6% and 35% mortaltiy to is hampered by a profound lack of knowledge regarding the most basic deltamethrin and permethrin, respectively. LLINs from Bungoma retained elements of Anopheles mating ecology. Females mate only once, and strong activity against a susceptible laboratory strain achieving >90% the suite of mating induced physiological and behavioral changes are mortality in all bioassays (N=99), but mortality of f1 offspring of field- predicated in large part upon the transfer of a mating plug containing collected An. gambiae s.s. in cone tests was <60% in all assays (N=99) All steroid hormones (SH). Here we report mechanisms of pre and peri- An. gambiae s.s. samples collected in LLINs were homozygous for the kdr copulatory sexual selection in An. gambiae mosquitoes and a role for SH genotype L1014S. In conclusion, LLINs develop holes within three years as a possible selective mechanism. High-speed video analysis of mosquito of distribution. In areas with pyrethroid resistance, mosquitoes are able to mating swarms revealed definitive evidence of female choice, as females enter LLINs and survive. LLINs with >50cm2 of damage were more likely employ specific rejection and acceptance behaviors. Furthermore, video to harbour mosquitoes than nets with no holes. The data indicate that a analysis revealed behavioral mechanisms of male competition. We small amount of damage could compromise the protective efficacy of nets show that successfully mating males are not only larger, but through in areas with high levels of pyrethroid resistance. an ELISA assay we demonstrate that they have significantly higher SH titers in their reproductive accessory glands relative to their unsuccessful 1888 counterparts. The mechanisms behind female discrimination is currently being investigated. Additionally, females mated to males with reduced INDOOR USE OF ATTRACTIVE TOXIC SUGAR BAIT (ATSB) SH levels have lower fecundity and fertility compared to females mated FOR CONTROL OF MOSQUITOES AND FOR RESISTANCE with controls. Given that previous work has demonstrated the importance MANAGEMENT of male SH in female reproductive phenotypes, fitness in both sexes of An. gambiae appears at least partially SH dependent. This work provides Mark Rowland1, Zachary Stewart1, Richard Oxborough1, Patrick critical insights into the mating ecology of a major disease vector and Tungu2, Matthew Kirby1, Seth Irish1 implicates SH as a key factor determining fitness across sequential episodes 1 London School of Hygiene & Tropical Medicine, London, United Kingdom, of sexual selection. Moreover, these results extend our understanding of 2 National Institute for Medical Research, Tanga, United Republic of swarming and monogamous insect mating systems. Tanzania Attractive toxic sugar bait (ATSB), a mixture on insecticide and sugar 1890 solution, sprayed onto vegetation has been successful in controlling Anopheles mosquitoes outdoors. Indoor application of ATSB has yet to STAGE-SPECIFIC, STRUCTURAL PROTEOMES AND THE be explored. This study determined whether ATSB stations positioned NODULAR SECRETOME FROM ONCHOCERCA OCHENGI, inside the home would kill host-seeking mosquitoes and constitute a new THE CLOSEST RELATIVE OF THE HUMAN RIVER BLINDNESS approach to control of malaria. Classes of insecticides new to malaria PARASITE vector control were mixed with sugar solution and tested as toxic baits 1 1 2 against Anopheles in feeding bioassay tests. The most promising ATSB Stuart D. Armstrong , Germanus S. Bah , Henrietta F. Ngangyung , 1 3 4 2 candidates were then trialed in experimental huts in Tanzania against Dong Xia , Samuel Wanji , Peter A. Enyong , Vincent N. Tanya , 5 5 1 free flying, host seeking mosquitoes. The ATSB stations were hung from David W. Taylor , Mark L. Blaxter , Jonathan M. Wastling , 1 ceilings of huts next to untreated mosquito nets occupied by human Benjamin L. Makepeace volunteers. In feeding bioassays, chlorfenapyr (a pyrrole), boric acid and 1University of Liverpool, Liverpool, United Kingdom, 2Institut de Recherche tolfenpyrad (a mitochondrial electron transport inhibitor), mixed in a guava Agricole pour le Développement, Ngaoundéré, Cameroon, 3Research juice-based bait, each killed more than 90% of pyrethroid-susceptible An. Foundation for Tropical Diseases and Environment, Buea, Cameroon, gambiae s.s. and pyrethroid-resistant An. arabiensis at less than 1% w/v. In 4Tropical Medicine Research Station, Kumba, Cameroon, 5University of the experimental hut trial, the mortality rates of the three ATSB treatments Edinburgh, Edinburgh, United Kingdom were comparable to long lasting insecticidal nets (LLINs) tested against the The bovine filaria Onchocerca ochengi is the closest extant relative same species in the same area. Indoor ATSB constitute a novel application of the human river blindness parasite, Onchocerca volvulus, and has method for insecticide classes that act as stomach poisons and have been used as a natural model of onchocerciasis for two decades. The not been exploited for mosquito control hitherto. Combined with LLIN, close parallels between these species include the propensity of the indoor use of ATSB has the potential to serve as a strategy for managing adult worms to form collagenous nodules and their obligate symbiosis insecticide resistance. with supergroup C strains of Wolbachia. To date, in-depth proteomic analysis of filarial structural proteomes has been restricted to a single species, Brugia malayi, which has a fundamentally different lifestyle to O. volvulus. Here, we report stage-specific proteomes of O. ochengi from intrauterine microfilariae, vector-derived L3, and adult female and male worms; alongside host and parasite excretory-secretory products identified in nodule fluid ex vivo. We applied a combination of anion exchange fractionation and geLC-MS with interrogation of a draft O. ochengi genome assembly to identify >4,600 filarial proteins and 176

astmh.org 578 proteins from Wolbachia strain wOo (33% and 27% of their theoretical 1892 proteomes, respectively). Of the filarial proteins, 1,038 (22%) were common to all stages, whereas microfilariae exhibited the greatest number VACCINATION WITH BRUGIA MALAYI-103 AND BRUGIA of stage-specific proteins (~920), despite direct harvesting of this material MALAYI-RAL-2 CONFER SIGNIFICANT PROTECTION AGAINST from adult female uteri. Proteins identified by geLC-MS alone accounted SUBCUTANEOUS CHALLENGE OF B. MALAYI INFECTIVE for <20% of the total for any single stage, but showed enrichment for LARVAE IN MONGOLIAN GERBILS membrane transporters, polyubiquitin and respiratory chain components. Preliminary analyses suggested that the relative abundance of galectins, Sridhar Arumugam1, Bin Zhan2, Junfei Wei2, David Abraham3, calponins, myosins and antioxidant proteins varied between lifecycle Danielle Ward1, Sara Lustigman4, Maria Bottazzi2, Peter Hotez2, stages. In nodule fluid, >2,000 proteins were identified (77% bovine, 23% Thomas Klei1 filarial, 0.1% bacterial), with strong representation of bovine antimicrobial 1Louisiana State University, Baton Rouge, LA, United States, 2Baylor College proteins and filarial transthyretin-like proteins. These data provide a rich of Medicine, Houston, TX, United States, 3Thomas Jefferson University, resource for comparative analyses of filarial protein expression throughout Philadelphia, PA, United States, 4New York Blood Center, New York, NY, the lifecycle, as well as supporting research efforts directed at the United States development of a filarial vaccine, new drugs and diagnostic biomarkers. Two Brugia malayi proteins, Bm-103 and Bm-RAL-2, are orthologous proteins of confirmed vaccine candidates of Onchocerca volvulus. The 1891 Ov-103 was identified first as a microfilariae surface associated protein, but later was found to be also expressed in the cuticle, hypodermis and TOWARDS IDENTIFICATION AND VALIDATION OF mutivesicular bodies of infective stage larvae of both filarial parasites. BIOMARKERS FOR THE QUANTIFICATION OF LOA LOA Ov-RAL-2 and Bm-RAL-2 are immunodominant proteins expressed in the MICROFILARIAE (MF) USING PROTEOMIC ANALYSES OF hypodermis of all stages. Bm-103 was cloned and expressed in Pichia BODY FLUIDS FROM MICROFILAREMIC LOA-INFECTED pastoris and Bm-RAL-2 was cloned and expressed in Escherichia coli. INDIVIDUALS These recombinant proteins were tested for their efficacy as a vaccine in the B. malayi - Mongolian gerbil animal model of lymphatic filariasis. Papa M. Drame1, Sasisekhar Bennuru1, Zaojing Meng2, Jesica A. Vaccination was via 3 intraperitoneal injections separated by 2 week Herrick1, Zhou Ming2, Thomas B. Nutman1 intervals. Animals were challenged subcutaneously with 100 infective 1 2 National Institutes of Health, Bethesda, MD, United States, National larvae third stage larvae (L3) and worm recovery was performed 42 or Institutes of Health, Frederick, MD, United States 90 days post infection. Vaccination with Bm-103 administered with Among parasitic helminths, Loa loa (Ll) presents a challenge for the alum showed 40% worm reduction in comparison to alum controls. mass drug administration programs in areas co-endemic for Wuchereria Vaccination with Bm-RAL-2 showed 43% worm reduction in comparison bancrofti and Onchocerca volvulus because of the severe adverse events to controls. A fusion protein of Bm-103 and Bm-RAL-2 was created, (SAE’s) in cases of very high Ll microfilaraemia. To identify microfilarial- cloned and expressed in E. coli. Vaccination of gerbils with the Bm-103- derived Ll-specific biomarker(s) that could provide the basis for a Bm-RAL-2 fusion protein induced a 51% worm reduction in comparison microfilarial quantitative immunoassay, we characterized the excretory/ to controls. Vaccination of gerbils with the two antigens, Bm-103 and secretory (E/S) proteome of Ll mf as well as the Ll-specific proteins found Bm-RAL-2, each injected separately resulted in worm reduction of 69%. in urine and plasma of Ll-infected and -uninfected individuals using LC The development of embryograms to study the fecundity of female worms MS/MS. From 20 x 106 mf purified from the blood of Ll-infected patients harvested from control and vaccinated gerbils are currently underway and and cultured in vitro, 1273 proteins (representing 8.2% of the Ll putative will bring insights on impact of vaccination on fertility of female worms. proteome) were identified. Among the most abundant proteins identified In all vaccination experiments, a strong antigen-specific IgG response was were endochitinase, cyclophilins, and a phosphatidyl ethanolamine detected by ELISA to the recombinant proteins. Moreover, in vitro killing binding protein. In addition several hypothetical proteins unique to Ll assays using peritoneal exudates cells (PEC) in the presence of gerbil anti- were identified. To further identify if any of these ES proteins were present serum against Bm-103 and Bm-RAL-2 showed active killing of L3 larvae in in body fluids, proteomic analyses of urine and plasma of Ll-infected comparison to L3 larvae cultured with appropriate controls sera suggesting individuals (depleted of the top 12 to 20 human abundant proteins in that an antibody dependent cell mediated cytotoxicity (ADCC) maybe a plasma) resulted in the identification of 18 (from urine) and 29 (from potential mechanism of protection. The results suggested that further plasma) Ll proteins found only in Ll-infected individuals that were identified experiments using these proteins alone or in combination are warranted. by having at least 2 unique peptides. 4/18 antigens found in urine and 13/20 found in plasma have been selected for biomarker validation based 1893 on limited homology to other filarial species and, specific reactivity to polyclonal antibody raised to Ll mf ES. In addition, 9 of these tested to A COMPARATIVE STUDY OF POST-DIETHYLCARBAMAZINE date were found to be immunogenic in humans (based on antigen-specific TREATMENT REACTIONS IN ONCHOCERCIASIS AND LOIASIS

IgG4 reactivity by serum from mf+ Ll-infected plasma (n=30) and not by 1 1 those from uninfected plasma (n=20)). Development and testing of rapid Senbagavalli Prakash Babu , Jacqueline Herold , Fanny Legrand1, Jesica Herick1, Eric Ottesen2, Joseph Kamgno3, Raceline antigen capture immunoassays are underway to provide an alternative to 3 1 1 more standard methods of mf quantification. Gounoue , Thomas Nutman , Amy D. Klion 1National Institutes of Health, Bethesda, MD, United States, 2The Taskforce for Global Health, Decatur, GA, United States, 3Center for Filariasis and Other Tropical Disease Research, Yaounde, Cameroon, South Africa Severe post-treatment reactions occur following diethylcarbamazine (DEC) treatment of filarial infections, including onchocerciasis and loiasis. Although release of the intracellular bacteria, Wolbachia, has been implicated in the pathogenesis of these reactions, Loa loa does not contain Wolbachia. The aim of this study was to compare eosinophil, neutrophil and cytokine responses post-DEC treatment in subjects with L. loa infection to those in patients infected with Onchocerca volvulus. The study included three groups: Group I [LOA-NIH], subjects with microfilaremic (MF+) loiasis treated with DEC (8-10 mg/kg/day for 21 days) at NIH; Group II [LOA-CAM], subjects with MF+ loiasis treated with DEC

astmh.org 579 (8 mg/kg in a single dose) in Cameroon; Group III [ONCHO], subjects with 1895 MF+ onchocerciasis treated with DEC (200 mg/day for 7 days) in Ghana. Complete blood counts and previously collected serum were available at THE IMPACT OF MATERNAL HELMINTH INFECTIONS ON TH2 0h, 4h, 8h, 1-7d and 14d post-initiation of treatment for LOA-CAM and RESPONSES AND ATOPIC SENSITIZATION OF INDONESIAN ONCHO and at variable time points for LOA-NIH. The early pattern of YOUNG CHILDREN eosinophilia post-DEC (a decrease from baseline during the first 24 hours 1 1 2 followed by a significant increase over the next 3-5 days) was similar in all Yenny Djuardi , Taniawati Supali , Erliyani Sartono , Maria 2 3 groups and the rise was preceded by a transient increase in serum IL-5 Yazdanbakhsh levels. In contrast, the % baseline ANC increased significantly post-DEC 1Department of Parasitology, Faculty of Medicine University of Indonesia, only in the ONCHO group (P<0.05 at days 1,2 and 3 compared to LOA- Jakarta, Indonesia, 2Department of Parasitology, Leiden University Medical CAM and LOA-NIH. Serum IL-10 levels increased transiently in all 3 groups, Center, Leiden, Netherlands reaching peak values at 1-2 days post-DEC. Although serum TNF-alpha, Immune responses to helminth infection and allergy are both characterized levels increased at 1-2 days post-DEC in all subjects in the ONCHO group, with TH2 responses. Manifestation of in children of low-to- there was no consistent pattern in the subjects with loiasis. To conclude, middle income countries were much lower compared to those of affluent parasite antigen release and the resultant Th2-driven eosinophilia may be countries. We aimed to investigate the impact of maternal helminth a major driver of post-DEC reactions in both onchocerciasis and loiasis. infections and other factors on child’s development of type 2 responses The increased TNF-alpha and neutrophilia seen post-DEC in onchocerciasis and atopic sensitization at 4 years of age in an area endemic for filaria and is likely due to the concomitant release of Wolbachia during microfilarial soil-transmitted helminths. Data were collected from pregnant mothers killing. on helminth infections, total IgE and Ascaris-specific IgE, education and socioeconomic status (SES). Total IgE and IL-5 in response to mitogen, 1894 and helminth antigens were measured in children at 2, 5, 12, 24 and 48 months of age. Ascaris and allergen-specific IgE and skin prick testing (SPT) ENDOTHELIAL CELLS RELEASE SOLUBLE FACTORS THAT were determined at 4 years of age. Strong TH2 responses were seen at 5 PROLONG THE SURVIVAL OF FILARIAL WORMS IN VITRO months of age and increased with time. Child’s helminth-antigen specific Holly Evans, Edward Mitre TH2 responses increased significantly with age and were associated with maternal filarial infection, while the increasing of child’s general TH2 Uniformed Services of the Health Sciences, Bethesda, MD, United States responses with age were more associated with higher maternal total A major barrier for current mass drug administration (MDA) efforts to and Ascaris-specific IgE, as well as with low maternal education or SES. control lymphatic filariasis is the inability of current medications to kill Child’s Ascaris-specific IgE were both associated with child’s general and adult worms when given as a short course. The development of novel helminth-specific TH2 responses. At 4 years of age when allergen reactivity drugs is complicated by the inability to maintain worms for long periods was assessed by SPT, the high general TH2 responses did not translate into of time in vitro, making effective screening of new drugs difficult. In an higher SPT. The risk factor for SPT reactivity was low maternal education attempt to improve in vitro culture methodology for filarial worms, we which decreased the risk of SPT positivity to allergens (adjusted OR, 0.32; have conducted a series of experiments using microfilariae (MF) obtained 95% CI, 0.12 - 0.87) independently of maternal filarial infection which from gerbils infected with Litomosoides sigmodontis, a filarial parasite of tended to reduce the child’s risk for being SPT positive (adjusted OR, 0.35; rodents. While the culture of L. sigmodontis MF in Dulbecco’s Modified 95% CI, 0.07 - 1.70). In conclusion, young children living in areas endemic Eagle Medium supplemented with 10% FBS results in an average survival for helminths developed a strong TH2 responses which was influenced by of only 7 days, co-culturing MF with a mouse endothelial cell line (EOMA) maternal or child’s exposure to helminth infections, but did not translate expanded survival to 40 days. Not all cell lines have this property, as MF to a higher SPT reactivity to allergens. This result might explain why the co-cultured with a rat basophilic cell line (RBL-2H3) survived for only 5 prevalence of allergies in low-to-middle income countries were much days. Culturing EOMA cells in transwell plates extended MF survival to the lower compared to the more affluent countries. same degree as direct co-culture, suggesting that the factors microfilariae require are soluble in nature. Heat inactivation of EOMA conditioned 1896 media at 56°C reduced MF survival by approximately 50%. However, heat inactivation at 100°C reduced survival to 3 days, signifying that MF TRANSCRIPTIONAL PROFILE OF THE DIROFILARIA IMMITIS require both heat labile and heat stable factors. EOMA cells require FBS LIFE CYCLE to produce these factors, as conditioned media collected from EOMA 1 2 2 cells grown in the absence of FBS fail to prolong survival. Importantly, Ashley N. Luck , Christopher C. Evans , Molly D. Riggs , Laurence 1 1 2 these findings also pertain to adult worms. Both rodent L. sigmodontis Ettwiller , Jeremy M. Foster , Andrew R. Moorhead , Barton E. 1 3 and human Brugia malayi adult worms also show significantly extended Slatko , Michelle L. Michalski survival when cultured in EOMA conditioned media. We are poised to 1New England Biolabs, Ipswich, MA, United States, 2University of Georgia, begin biochemical and comparative analyses to elucidate the chemical Athens, GA, United States, 3University of Wisconsin Oshkosh, Oshkosh, nature of these essential factors. Identification of such factors will advance WI, United States our ability to cultivate filarial pathogens in vitro and may provide insights Dirofilaria immitis (Di), or canine heartworm, is a filarial nematode for the development of new anti-filarial compounds. evolutionarily related to those responsible for human parasitic diseases. The D. immitis genome, along with the genome of its obligate endosymbiont, Wolbachia (wDi), was recently completed and published. We initiated a series of transcriptional profiling experiments to better understand the temporal transcriptional activity of Di and wDi throughout the nematode life cycle. Over 215 million single-end 50 bp reads were generated from total RNA from five Di life cycle stages. Based on hierarchical clustering of expression data, nearly 60% of all Di genes display stage-specific transcriptional patterns. Pairwise comparison of adult male (AM) and adult female (AF) samples reveals that over 9,000 genes display sex-biased transcriptional patterns. The L3 to L4 transition, which occurs upon entering the mammalian host, is critical to the Di life cycle and a potential point of intervention. Among all five life cycle stages examined, a significant portion of Di genes are L4-

astmh.org 580 associated (3525 transcripts), whereas only 65 transcripts show L3-biased 1898 expression. Pairwise comparison of the L3 and L4 stages reveals 3157 significantly differentially expressed genes (1170 L3 upregulated, 1987 L4 MOLECULAR BASIS FOR SIALIC-ACID DEPENDENT RECEPTOR upregulated) and provides important information regarding transcriptional RECOGNITION BY THE PLASMODIUM FALCIPARUM changes required for this transition. As anticipated, significantly fewer INVASION PROTEIN ERYTHROCYTE-BINDING ANTIGEN-140 reads mapped to wDi genes than to Di genes in each life cycle stage. (EBA-140/BAEBL) Interestingly, synthesis of the critical metabolite, heme, by wDi appears to be synchronized with the production of heme-binding proteins in Di in a Brian Malpede, Dan Lin, Niraj Tolia stage-specific manner. Comparative analysis to human filarial nematodes Washington University School of Medicine, St. Louis, MO, United States provides further information on the evolutionary biology of these parasites, Erythrocyte-binding antigen 140 (PfEBA-140/BAEBL) is a Plasmodium while also highlighting opportunities for further drug targeting initiatives. falciparum erythrocyte invasion ligand that engages Glycophorin C (GPC) A better understanding of how these genomes function in concert with on host erythrocytes during malaria infection. PfEBA-140 is a member one another is required for unraveling the complex relationship of the of the erythrocyte-binding ligand (EBL) family, which contains the four nematode with its endosymbiont, Wolbachia, as well as with its canine sialic acid dependent invasion proteins utilized by P. falciparum. Each of and mosquito hosts. these ligands recognizes a different erythrocyte receptor despite being composed of a highly conserved domain architecture. To elucidate the 1897 foundations of receptor specificity within the EBL family and define the structural basis of GPC engagement, we determined two crystal EXPERIMENTAL PLASMODIUM FALCIPARUM GENETIC structures of the PfEBA-140 minimal binding domain unbound and in CROSSES IN HUMAN LIVER-CHIMERIC MICE complex with a glycan containing the essential sugar component of GPC Ashley M. Vaughan1, Nelly Camargo1, Matthew Fishbaugher1, that is recognized during erythrocyte engagement. The two domains Lisa A. Checkley2, Richard S. Pinapati2, Sebastian A. Mikolajczak1, composing the minimal binding region contain unique structural elements Michael Ferdig2, Stefan H. Kappe1 that are likely determinants of receptor specificity. Two glycan binding pockets were observed, one per domain, and the bound sialic acid was 1Seattle Biomedical Research Institute, Seattle, WA, United States, modeled into each site. Erythrocyte binding experiments elucidated 2University of Notre Dame, Notre Dame, IN, United States important glycan contact residues and identified distinct functional roles Forward genetic studies using experimental genetic crosses are an for the individual sugar binding sites. Our studies provide a structural incredibly powerful tool to pinpoint the genetic basis of phenotypic framework for GPC recognition, form a foundation for future studies of variation and are typically more powerful than population analysis. the interaction between PfEBA-140 and erythrocytes, and offer insight into For Plasmodium falciparum however, only three experimental crosses deficient receptor binding and putative receptor switching described for have been carried out due to the hurdles associated with the process polymorphisms in PfEBA-140. Preventing erythrocyte engagement is an – for ethical reasons humans can not be used and until recently, only excellent opportunity to inhibit merozoite invasion. Our results will thus aid splenectomized chimpanzees allowed for the transition from the in the design of rational therapeutics and vaccines that target erythrocyte sporozoite stage to the asexual blood stage of the life cycle necessary for invasion ligands. progeny amplification and analysis. Nevertheless, the recombinant progeny from the three crosses led to the discovery of genetic determinants for 1899 drug resistance, erythrocyte invasion and blood stage parasite growth. The recent decision by the NIH to cease chimpanzee biomedical research EVOLUTION BEFORE OUR EYES: GENOME MUTATION IN suggests further forward genetic studies are impossible. However, we PLASMODIUM FALCIPARUM have shown that a mouse harboring human hepatocytes (the FRG KO huHep mouse) infused with human erythrocytes can support P. falciparum William L. Hamilton, Antoine Claessens, Mihir Kekre, Thomas D. sporozoite infection, the completion of liver stage development and Otto, Adnan Faizullabhoy, Julian C. Rayner, Dominic Kwiatkowski the transition to asexual blood stage replication. This thus suggests Wellcome Trust Sanger Institute, Hinxton, United Kingdom that P. falciparum experimental crosses can be achieved in the FRG KO Genetic mutation is a central process in evolution, and is involved in the huHep mouse and here we show here that this is indeed possible and emergence of drug resistance and the generation of antigen sequence thus this mouse model can replace the previously essential chimpanzee. diversity in the malaria parasite Plasmodium falciparum. We studied P. To achieve our goal, we generated P. falciparum gametocytes from the falciparum mutation by regularly sub-cloning parasites cultured in vitro to NF54 chloroquine sensitive and GB4 chloroquine resistant parasite lines isolate single infected red blood cells, so that mutations arising in asexually and used these in mixed feeds to mosquitoes to generate recombinant dividing cells could be detected using whole genome sequencing. We sporozoite progeny – produced after zygote formation and sexual produced the most comprehensive description of P. falciparum mutation recombination. Sporozoites were injected into FRG KO huHep mice to date, studying four lab strains, artemisinin resistant Cambodian field harboring human erythrocytes and after the liver stage-to-blood stage isolates and parasites with experimentally induced mutations in DNA repair transition, blood stage parasites were maintained in vitro. Parasite cloning genes. In total, we analysed >300 genomes from parasites cultured for a and downstream microsatellite analysis revealed the presence of unique total of >1,000 days, capturing hundreds of mutations. We found that: recombinant progeny. Furthermore, drug selection demonstrated the (1) Point mutations are distributed throughout the genome and occur at a creation of recombinant progeny with unique drug resistance patterns similar rate between strains regardless of the drug sensitivity status of the not shared by the parental populations. Thus we provide evidence of line, contrary to previous studies suggesting that drug resistant parasites successful experimental genetic crosses. This methodology should allow are hypermutable; (2) There is a strong mutation bias with G/C to A/T for P. falciparum “systems genetics” – the study of complex genetic traits transition mutations over-represented. We estimate this would equilibrate in which genomic data and clinical phenotypes are obtained using global at a similar AT ratio to that observed in the P. falciparum genome (~80%); “omic” technologies. (3) InDels occur predominantly in AT rich low-complexity regions at a higher rate than point mutations, likely due to DNA polymerase slippage events; (4) Structural variation is focused in and around var genes, which encode highly polymorphic PfEMP1 surface-expressed antigens, and this mitotic recombination generates sequence diversity by producing mosaic var genes. With 10^10 parasites in a single infected individual, our data indicate that every nucleotide in the P. falciparum genome will undergo point mutations and millions of new mosaic var genes will be produced

astmh.org 581 every 48-hour life cycle. In summary, we have produced a comprehensive that result in fully infectious sporozoites with very low levels of vector, catalogue of P. falciparum mitotic genome mutation at all scales from bacterial, and fungal contamination. These approaches have enabled the point mutations to interchromosomal translocations, adding considerably most comprehensive total proteomics of two human-infective malaria to our understanding of parasite genomics and evolution. species (Plasmodium falciparum and P. vivax), as well as the model rodent- infective species P. yoelii. Moreover, these purified sporozoites are also 1900 sufficiently devoid of soluble mosquito material to permit the assessment of the surface-accessible proteome of the salivary gland sporozoite. This IDENTIFYING NOVEL TRAFFICKING COMPONENTS OF THE was done using an amine-reactive crosslinker bearing a cleavable biotin PLASMODIUM FALCIPARUM VIRULENCE FACTOR PFEMP1 group, which enables high affinity purification and yet leaves a covalent THROUGH QTL ANALYSIS modification of accessible lysines for high confidence identification. We have developed an extensive and stringent washing strategy to minimize 1 2 1 Aaron T. Neal , Lisa C. Ranford-Cartwright , Richard T. Eastman , the binding of non-specific proteins, which has yielded a greatly expanded 3 1 Chris I. Newbold , Rick M. Fairhurst surface-accessible proteome above and beyond our previously published 1National Institutes of Health, Rockville, MD, United States, 2University of list. Several of these candidates have been confirmed with transgenic Glasgow, Glasgow, United Kingdom, 3University of Oxford, Oxford, United parasites and the generation of specific antisera. Finally, we have treated Kingdom purified sporozoites with molecular mimics of mammalian body conditions Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is the to observe any differences in protein accessibility or secretion onto the main parasite virulence factor due to its central role in mediating the parasite surface in response to these stimuli. Taken together, these cytoadherence of infected red blood cells (iRBCs) to the host microvascular characterizations provide a sizeable list of surface accessible proteins that endothelium. Directly targeting PfEMP1 as a therapeutic strategy is may be valuable new targets for antibody-based interventions. greatly limited due to the protein’s hypervariable nature, which gives rise to approximately 60 different variants. However, interfering with 1902 the trafficking of PfEMP1 to the iRBC surface is an attractive approach, as hypervariability becomes inconsequential and previous studies have AGE SPECIFIC INCIDENCE RATES OF MALARIA SUGGEST demonstrated that reduced surface PfEMP1 levels significantly weaken DIFFERENT RATES OF NATURALLY ACQUIRED IMMUNITY TO cytoadherence, likely lessening the severity of malaria symptoms and MALARIA ACROSS HUMAN HOST GENOTYPES permitting parasite clearance by the spleen. Interestingly, the in vitro Oscar A. Nyangiri1, Meghna Desai2, Godfrey Bigogo3, Deron culture-adapted parasite line 3D7 is inherently defective in exporting Burton4, Billy G. Ogwel3, Allan Audi3, Joel M. Montgomery5, PfEMP1 to the iRBC surface. Presuming that PfEMP1 export from Simon Kariuki3, Thomas N. Williams6 the parasite to the iRBC surface is controlled at the genetic level, we hypothesized that 3D7 harbors one or more genetic determinants of 1Kenya Medical Research Institute-Wellcome Trust Research Programme, impaired PfEMP1 trafficking. To test this possibility, we examined the Kilifi, Kenya,2 Malaria Branch, Division of Parasitic Diseases and Malaria, surface PfEMP1 levels of 17 progeny clones from the genetic cross Centers for Disease Control and Prevention, Atlanta, GA, United States, between 3D7 and the ‘trafficking-competent’ parasite line HB3. This 3Kenya Medical Research Institute-Centers for Disease Control and was assessed using Western blotting and a two-color, triple-layer flow Prevention, Kisumu, Kenya, 4National Center for HIV/AIDS, Viral Hepatitis, cytometry assay with plasma from malaria-immune Malian adults. STD and TB Prevention, Office of Infectious Diseases, Centers for Disease Normalized to HB3, we found that 3D7 displays 75% less PfEMP1 on Control and Prevention, Atlanta, GA, United States, 5Global Disease the iRBC surface, with progeny phenotypes ranging from 37% more to Detection Branch, Division of Global Health Protection, Center for Global 88% less PfEMP1. QTL analysis using 3,597 genome-wide SNP markers Health, Centers for Disease Control and Prevention, Atlanta, GA, United identified a significant locus with a LOD score of 4.963 on chromosome States, 6Imperial College, London, United Kingdom 12 that explains approximately 50% of the phenotypic variance. This locus In endemic areas, the incidence of clinical malaria declines with age, contains a single gene, Pf3D7_1245600, encoding a putative kinesin. The reflecting the role of naturally acquired immunity (NAI) in modulating role of this gene in the trafficking of PfEMP1 is being confirmed in allele- malaria. Asembo is a malaria endemic area in western Kenya. We exchange experiments, where the defect is rescued in 3D7 and introduced investigated whether NAI develops at different rates among children in HB3. The results of this study may strengthen our understanding carrying a range of malaria-associated genes by comparing age-specific of malaria pathogenesis and provide new targets for much needed malaria incidence in children, stratified by genotype. We recruited a birth therapeutics. cohort in March 2012-December 2013 (n=700) from a population under passive surveillance for clinical malaria since August 2006. Additionaly, 1901 we recruited birth cohort member siblings <12 years of age into a sibling cohort (n=780) for a combined cohort of 1480 children. Clinical PROTEOMIC COMPOSITION AND SURFACE ACCESSIBLE malaria was defined as fever with a malaria-positive blood film in the TARGETS OF PLASMODIUM SPOROZOITES absence of bacterial co-infections causing fever. Participants were typed for polymorphisms in 40 malaria-associated genes. Age-specific malaria Scott E. Lindner1, Kristian E. Swearingen2, Lirong Shi3, Anke incidence was significantly lower for sickle cell trait (AS) compared to sickle Harupa4, Ashley M. Vaughan4, Robert L. Moritz2, Stefan H. normal (AA) individuals (P<0.001), marginally significant for homozygous(- Kappe4, Photini Sinnis3 α/-α) compared to normal (αα/αα) alpha thalassemia individuals 1 2 Pennsylvania State University, University Park, PA, United States, Institute (P=0.04) and not significant forαα /αα compared to heterozygous(-α/αα) for Systems Biology, Seattle, WA, United States, 3Johns Hopkins University, individuals . Incidence rates peaked earlier in AA (4-5 years, 0.77 episodes Baltimore, MD, United States, 4Seattle Biomedical Research Institute, per child per year)compared to AS children (9-10 years, 0.8episodes per Seattle, WA, United States child year). For alpha thalassemia, incidence rates peaked earlier in -α/αα The transmission of salivary gland sporozoites from an Anopheline (2-3 years, 0.73 episodes per child year), compared to -α/-α individuals mosquito initiates the malarial infection of a new mammalian host, which ( 6 years, 1 episode per child per year) and αα/αα individuals (4-5 years, has prompted the development of various therapeutic interventions 0.79 episodes per child year). Analyses for the 40 malaria susceptibility targeted against sporozoites. While several sporozoite proteins have genes are ongoing. Preliminary results show shifts in the peaks of age- been historically well studied, systematic proteomic analyses have been specific incidence rates by genotype. These findings suggest different rates hampered by the presence of overwhelming amounts of proteins, nucleic of NAI among children of different genotypes. Results may be useful in acids, and lipids derived from the dissected mosquito vector. We have understanding genotype-specific effects of interventions such as malaria overcome this obstacle by developing streamlined purification methods vaccines that can modulate NAI.

astmh.org 582 1903 FOI accounting for unobserved heterogeneity using data collected from August 2011 to August 2013 in a cohort of children aged less than 11 QUANTIFYING THE INDEPENDENT EFFECTS OF AGE AND years in three sites in Uganda (Tororo, Kanungu and Jinja) with variable EXPOSURE ON TWO COMPONENTS OF MALARIA IMMUNITY: malaria transmission intensities. We applied the statistical methodology RESTRICTION AND TOLERANCE using linear and nonlinear mixed effects models to estimate both a constant and time-dependent FOI at each site, while accommodating for 1 2 Isabel Rodriguez-Barraquer , Emmanuel Arinaitwe , Prasana individual heterogeneity in the acquisition of malaria, and accounting 3 2 2 Jagannathan , Isaac Ssewanyana , Agaba Katureebe , Moses for re-infections. Differences in the FOI were more pronounced between 4 5 3 1 Kamya , Sarah Staedke , Grant Dorsey , David L. Smith , Bryan households (variance=2.25) than between children (variance=0.67). The 3 Greenhouse FOI did not vary with time, but differed between the three study sites with 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, higher risk in Tororo (FOI=4.0, 95%CI: 3.4 - 4.6), followed by Kanungu United States, 2Infectious Diseases Research Collaboration, Kampala, (FOI=0.6, 95%CI: 0.4 - 0.8), and by Jinja (FOI=0.2, 95%CI: 0.1 - 0.3). The Uganda, 3Department of Medicine, University of California San Francisco, FOI was also higher in children above five years of age (FOI=1.5, 95%CI: San Francisco, CA, United States, 4Department of Medicine, Makerere 1.2 - 1.7), those with symptomatic infection (FOI=0.7, 95%CI: 0.6 - 0.9) University, Kampala, Uganda, 5London School of Hygiene & Tropical and those with anemia (FOI=2.2, 95%CI: 1.7 - 2.7). Therefore, housing Medicine, London, United Kingdom structure, individual differences, location of an area, age, symptomatic While epidemiologic data consistently suggest that clinical immunity to status and anemia are important factors to consider when estimating the Plasmodium falciparum develops over time in exposed children, lack of burden of malaria. accurate measures of exposure and protection have limited our capacity to understand the development of immunity to infection and disease. 1905 The objective of this study was to define the acquisition of immunity against P. falciparum malaria among children living in endemic areas SEVERE MALARIAL THROMBOCYTOPENIA: A RISK FACTOR of varying malaria transmission. In particular, we were interested in FOR MORTALITY measuring the development of clinical tolerance (lack of symptomatic Daniel A. Lampah1, Tsin Yeo2, Michael Malloy3, Enny Kenangalam4, disease given parasitemia, evaluated here by parasite density at which Nicholas M. Douglas2, Paulus Sugiarto5, Julie A. Simpson3, subjects developed objective fever) and parasite restriction (ability to Nicholas M. Anstey2, Nicholas M. Anstey2, Ric Price2 kill or otherwise control the growth of parasites, evaluated here by 1Papuan Health and Community Development Foundation, Timika, parasite density). We used data from representative cohort studies being Indonesia, 2Menzies School of Health Research, Darwin, Australia, conducted in 100 households from each of three sub-counties in Uganda: 3Melbourne School of Population and Global Health, Melbourne, Australia, Walakuba (aEIR= 3.3), Kihihi (aEIR=31.5) and Nagongera (aEIR=315). The 4Mimika District Health Authority, Papua, Indonesia, 5Rumah Sakit Mitra study comprises continuous passive surveillance, active surveillance every Masyarakat, Papua, Indonesia 3 months, and monthly mosquito collections in all households. Thus, the dataset used for this analysis included data on over 3400 episodes of The significance of thrombocytopenia to the morbidity and mortality of clinical malaria and 1400 episodes of asymptomatic parasitemia occurring malaria is poorly defined. We compared the platelet profiles of patients in 739 children aged 6 months to 11 years of age over two years of with and without malaria in southern Papua, Indonesia. Between April follow up. Results from generalized additive models, allowing for flexible 2004 and December 2012 data were available on patient demographics, interactions between variables of interest, are consistent with strong malaria diagnosis, haematological investigations and clinical outcome independent effects of both age and exposure on the development of in a referral hospital. Of 922,120 patient episodes a total of 215,479 both tolerance and restriction. Tolerance develops gradually beginning (23.4%) were associated with a platelet measurement, of whom 66,421 early in life and is not strongly modified by variable exposure. In contrast, (30.8%) had clinical malaria. Patients with Plasmodium falciparum restriction starts to develop later in life (4-6 years of age) and depends monoinfection had the lowest platelet counts with an adjusted odds strongly on cumulative exposure. Further analyses will explore the role of ratio (AOR) for severe thrombocytopenia (platelet count <50,000 µl-1), recent and persistent exposure on both of these components of immunity. compared to those without malaria, of 6.03 [95% Confidence Interval These findings provide unprecedented insight about the roles of age and (CI) 5.77-6.30]. The corresponding risks were 5.4 [95% CI 5.02-5.80] for exposure on the development of immunity to malaria. mixed infections, 3.73 [95% CI 3.51-3.97] for P. vivax and 2.16 [95% CI 1.78-2.63] for P. malariae; p<0.001. In total 1.3% (2,701/215,479) 1904 of patients died. Compared to patients with neither severe anemia nor severe thrombocytopenia, those with severe anemia alone had an AOR for ESTIMATING MALARIA FORCE OF INFECTION ACCOUNTING death of 5.21 [95%CI 4.53-5.98], those with severe thrombocytopenia FOR HETEROGENEITY IN THE RISK OF INFECTION alone had an AOR of 4.65 [95%CI 4.10-5.28] and those with both risk factors an AOR of 16.44 [95%CI 13.70-19.74]; p<0.001. In conclusion, 1 2 3 2 Levi Mugenyi , Steven Abrams , Sarah Staedke , Niel Hens severe thrombocytopenia is associated with malarial related mortality. 1Infectious Diseases Research Collaboration, Kampala, Uganda, 2Centre Prospective studies are warranted to define its utility in defining the clinical for Statistics, Interuniversity Institute for Biostatistics and Statistical management of patients with malaria. Bioinformatics, Hasselt University, Diepenbeek, Belgium, 3London School of Hygiene & Tropical Medicine, London, United Kingdom The burden of malaria and the intensity of its transmission have been estimated using clinical incidence, parasite rate, and entomological inoculation rate. Less emphasis has been placed on the rate at which individuals become infected, or the force of infection (FOI), the number of new infections per person per unit of time. The above parameters are often estimated without accounting for the intrinsic variability among individuals in the risk of infection. However, such heterogeneity exists due to variability in risk factors, including proximity of residence to mosquito breeding sites, housing structure, density of mosquitoes and frequency of biting, use of prevention measures, differences in surface area between adults and children and in human sweat components, and antimalarial immunity. Here we have proposed approaches for estimating malaria

astmh.org 583 1906 and SMC awareness. We estimated the partial SMC coverage (proportion of under-fives receiving at least one of the four monthly doses) and full EXTENDING THE AGE RANGE FOR SEASONAL MALARIA coverage (proportion of under fives receiving all doses), and identified CHEMOPREVENTION (SMC): EFFECTIVENESS OF SMC IN factors associated with coverage using multivariable logistic regression CHILDREN UNDER 10 YEARS OF AGE DELIVERED THROUGH models. 176,281 doses of SP-AQ were distributed over four months. The THE DISTRICT HEALTH SERVICE IN SENEGAL survey collected data from 5,291 children and 3,206 caregivers in 3,079 households. Adjusted partial coverage was significantly higher via door-to- Elhadji K. Ba1, B. Cisse2, C. Pitt3, J. L. Ndiaye2, M. Ndiaye2, J. F. door distribution (86.5%) than via health facility (46.7%) or private sector Gomis2, M. Cairns3, B. M. Greenwood3, C. Sokhna1, O. Gaye2, P. J. (27.9%). Full coverage was also highest in the door-to-door delivery arm Milligan3 (56.3%) compared to health facility (19.4%) and private sector (12.2%). 1Institut de Recherche pour le Developpement, Dakar, Senegal, 2Universite Children 1-4 years old were significantly more likely than those <1 year old Cheikh Anta Diop de Dakar, Dakar, Senegal, 3London School of Hygiene & to receive SMC (p<0.001), and child use of an insecticide-treated bed net Tropical Medicine, London, United Kingdom was significantly associated with partial coverage (OR=1.4). Door-to-door Seasonal Malaria Chemoprevention (SMC) is recommended for malaria delivery achieved the highest coverage although a substantial population control in children under 5yrs of age where transmission is highly seasonal. did not receive SMC. The findings are informing plans for 2014 SMC scale- In some areas the disease burden may justify extending the age range for up across Kano State including community mobilization and sensitization SMC. We investigated the effectiveness of SMC in children under 10 years strategies and other fixed-point distribution opportunities. of age in Senegal. SMC with sulfadoxine-pyrimethamine plus amodiaquine was introduced in three districts in a step-wedge design. Fifty-four health 1908 posts were randomized to implement SMC starting in 2008, 2009, or 2010, or to remain without the intervention. A surveillance system was THE EFFECTIVENESS OF INSECTICIDE TREATED BEDNETS IN established to record all deaths and all malaria cases diagnosed at health HAITI: RESULTS FROM A CASE-CONTROL STUDY facilities, and a pharmacovigilance system was put in place to detect Laura Steinhardt1, Yvan St. Jean2, Kimberly Mace1, Daniel adverse drug reactions. A poisson regression model was used to estimate Impoinvil1, Ryan Wiegand1, Jean Seme Fils Alexandre3, Joseph the effectiveness of SMC in reducing malaria incidence in treated children, Frederick2, Roland Oscar2, Jean Frantz Lemoine2, Emery with a random effect to account for variation in incidence between Nkuruziza4, Samuel Jean4, Brian Wheeler3, John Barnwell1, Ellen health posts. To determine whether SMC was able to reduce malaria Dotson1, Laurence Slutsker1, S. Patrick Kachur1, Michelle Chang1 transmission, incidence of malaria in age groups too old to receive SMC, 1Centers for Disease Control and Prevention, Atlanta, GA, United States, was compared between health posts in which SMC was delivered to 2Programme National de la Control de la Malaria, Port-au-Prince, Haiti, children, and health posts without SMC, using random-effects poisson 3Centers for Disease Control and Prevention, Port-au-Prince, Haiti, 4PSI, regression. SMC was administered to about 14,000 children under 5yrs Port-au-Prince, Haiti in 2008, 90,000 children under 10yrs in 2009, and to 155,000 children under 10 yrs in 2010. No serious adverse events attributed to SMC were Insecticide-treated bednets (ITNs) are a cornerstone of malaria prevention, detected despite a high level of surveillance. Where SMC was delivered, but little evidence exists on their effectiveness in Haiti where the primary the number of malaria cases in children under 10 years was reduced vector, Anopheles albimanus, has been documented as exophilic and with by 69% (95%CI 65%,72%). Malaria incidence in older age groups variable biting times. We conducted a case-control study to assess ITN was reduced in areas where SMC was delivered to children, by 29% effectiveness in Haiti, following a national ITN distribution campaign in (21%,35%). In conclusion, in some regions of the Sahel and sub Sahel, 2012. Patients presenting to outpatient departments were systematically the age distribution of malaria may justify extending the age range for screened for fever or history of fever. Eligible patients were administered a SMC. Including older children in SMC programmes is safe and effective, brief questionnaire and blood was collected for a malaria rapid diagnostic and may contribute to reducing transmission. test (RDT) and dried blood spots. From September 2012-February 2014, 9,318 patients, including 379 (4.1%) RDT-positive patients, were enrolled 1907 across 17 health facilities in five departments in Haiti. Retrospectively matching up to four RDT-negative controls per RDT-positive case by age COMPARISON OF SEASONAL MALARIA CHEMOPREVENTION group, sex, location of residence, and enrollment period yielded 365 COVERAGE IN NORTHERN NIGERIA VIA DOOR-TO-DOOR, cases and 1,204 RDT-negative controls. Slightly more than half (57.1%) of HEALTH FACILITY AND RETAIL SECTOR DELIVERY patients reported owning any bednet, with no difference among matched cases and controls. We found no difference in the proportion of cases and Abigail Ward1, Justin C. Graves2, Omowunmi F. Omoniwa2, Nasir controls who reported using any bednet (34.5% vs. 32.9%, p=0.39) or a Mohammed Bashir2, Kathleen Maloney1, Leslie Emegbuonye2, campaign ITN (21.9% vs. 19.5%, p=0.30) the previous night, or always Arnaud Le Menach1, Caitlin F. Dolkart3, Kabir Masokano4, Owens using a campaign ITN in the two weeks before their illness (18.4% vs. Wiwa2 18.5%, p=0.84). In a multivariabe conditional logistic regression model, 1Clinton Health Access Initiative, Boston, MA, United States, 2Clinton consistent use of a campaign ITN was not related to RDT positivity. Health Access Initiative, Abuja, Nigeria, 3Clinton Health Access Initiative, The only variable related to RDT positivity in the model included body Nairobi, Kenya, 4Kano SMOH, Kano, Nigeria temperature (Odds Ratio = 1.40, 95% Confidence Interval: 1.25, 1.57 In 2012, WHO recommended seasonal malaria chemoprevention (SMC) per one-degree Celsius increase). Additional entomologic investigation with sulfadoxine-pyrimethamine and amodiaquine (SP-AQ) to prevent found that all Anopheles mosquitoes tested from the study areas were malaria in children under five in the Sahel sub-region. As the National susceptible to permethrin, the insecticide used on campaign ITNs. Our Malaria Elimination program in Nigeria plans to apply the recommendation results based on RDT status do not provide evidence to support ITNs as to protect 6 million children in its 9 northern states, identifying the most an effective malaria prevention strategy in Haiti. Additional results using a effective ways to deliver the drug is critical. From August-November PCR-based case definition will be presented. 2013, SP-AQ was delivered monthly to under-fives in 3 local government areas of Kano State using 3 delivery mechanisms: door-to-door using community drug distributors, at public health facilities (fixed points), and for a low cost in the private sector. A household survey was conducted in December 2013, one month after the final round of SMC distribution, to assess the coverage achieved through each distribution method. Data were collected on demographics, malaria knowledge, treatment practices,

astmh.org 584 1909 not contribute to antiviral defense during early arboviral infection in the midgut while it is protective at later stages in the systemic compartment; AEDES AEGYPTI FEMALE MOSQUITOES WITH ALANINE (iii) and implicate the siRNA pathway in evasion of immunity by malaria AMINOTRANSFERASE DEFICIENCY FACE A STRESSFUL parasites in the midgut. These results expand our understanding of the METABOLIC CHALLENGE DURING BLOOD DIGESTION small RNA immunity machinery in an important African vector. Jun Isoe1, Stacy Mazzalupo1, Virginia Belloni2, Patricia Y. 1911 Scaraffia2 1The University of Arizona, Tucson, AZ, United States, 2Tulane University, STRUCTURAL DIVERGENCE OF HETEROCHROMATIN New Orleans, LA, United States BETWEEN INCIPIENT SPECIES OF ANOPHELES GAMBIAE We have recently evaluated the exposure of A. aegypti females to Atashi Sharma, Maria V. Sharakhova, Igor V. Sharakhov L-cycloserine (LCS), a well-known inhibitor of alanine aminotransferase University of Virginia Tech, Blacksburg, VA, United States (ALAT) in animals. Our results indicated that 10 mM LCS interferes with Aedes aegypti blood metabolism causing motor impairment and a 35% The major African malaria vector Anopheles gambiae is known to of mortality with an acute effect during the first 6 hours after treatment. be undergoing incipient speciation into two molecular forms – M Interestingly, only 11% of the total mortality was observed between and S – recently named An. coluzzi and An. gambiae, respectively. 24 and 72 hours after feeding. In order to follow up this finding, the Heterochromatin plays a vital role in several important biological functions, expression pattern of two genes encoding ALAT (1 and 2) was first and is known to be associated with longevity and individual fitness of analyzed in sucrose- and blood-fed A. aegypti tissues by qRT-PCR. ALAT1 organisms as well as with postmating reproductive isolation between and ALAT2 transcript levels exhibited a distinct expression pattern in species. The genome of Anopheles gambiae was first sequenced in 2002 mosquito tissues dissected during a gonotrophic cycle. Next, RNAi- and has been updated since but still contains important information gaps mediated gene silencing was used to knock down endogenous levels with regards to the repetitive DNA content in heterochromatin. In order of each transcript in mosquito tissues. Injection of female mosquitoes to achieve a better understanding of differentiation within An. gambiae, with either dsRNA-ALAT1 or dsRNA-ALAT2 or both (dsRNA-ALAT1/2) it is essential to develop a physical map containing information about significantly decreased the expression of ALAT1 or ALAT2 or ALAT1/2 the repetitive DNA and determine the differences in heterochromatin in fat body (FB) and Malphigian tubules (MT) at 24 hours after blood between the incipient species. Using multiple strains of M and S forms, we feeding, when compared to dsRNA-firefly luciferase-injected control. mapped repetitive DNA sequences including satellite DNA and ribosomal As expected, the expression of ALAT1 was not modified in tissues from DNA (rDNA) with respect to bands of pericentric heterochromatin of dsRNA-ALAT2-injected females and vice versa. Western blot analysis mitotic chromosomes. Satellite DNA probe Ag53A hybridized to the demonstrated that the protein levels of ALAT were also significantly pericentric heterochromatin/rDNA locus junction in both forms. However, reduced in tissues of dsRNA-ALAT-injected females when compared to unexpectedly, satellite DNA AgY53B hybridized at the base of proximal control mosquitoes. Moreover, the knockdown of A. aegypti ALAT1 or band in the M form but at the tip of the band in the S form, indicating a ALAT2 or ALAT1/2 caused unexpected phenotypes such as a delay in blood possible shift or inversion in the satellite DNA position during divergence digestion, a massive accumulation of uric acid in the midgut posterior of these forms. Satellite DNA AgY477 depicted a similar pattern, region, and a significant decrease in nitrogen waste excretion during the hybridizing to different positions between the forms. Idiograms based first 48 hours after blood feeding. Concomitant with these results, the on above information were prepared for the M and S forms as well as expression of genes encoding both the ammonia transporter and xanthine other members of the An. gambiae complex, serving as a tool in better dehydrogenase were significantly increased in FB and MT of dsRNA- understanding of evolution of the repeat rich regions in the An. gambiae ALAT-injected females. These findings highlight the efficient and complex genome. Our results revealed that the rapid evolution of heterochromatin mechanisms that blood-fed mosquitoes use to avoid ammonia and free is not restricted to species with the postmating reproductive isolation. The radical toxicity. structural reorganization of satellite DNA observed between the M and S forms suggests a possible role of heterochromatin in initial diversification 1910 of malarial vectors. ANOPHELES GAMBIAE SMALL-RNA PATHWAYS IMMUNITY 1912 TO DIFFERENT PATHOGENS X-CHROMOSOME LOCALIZED RECOMBINATION HOTSPOTS Guillaume Carissimo1, Emilie Pondeville1, Emmanuel Bischoff1, UNDERMINE EXISTING MOLECULAR DIAGNOSIS OF 1 1 Catherine Bourgouin , Anna-Bella Failloux , Christophe ANOPHELES GAMBIAE AND AN. COLUZZII UNDER HIGH 2 3 1 Antoniewski , Alain Kohl , Kenneth D. Vernick HYBRIDIZATION 1Institut Pasteur, Paris, France, 2Université Pierre et Marie Curie - UPMC, 1 1 1 Paris, France, 3University of Glasgow - MRC Center for Virus Research, Beniamino Caputo , Verena Pichler , Emiliano Mancini , Marco 1 2 3 3 Glasgow, United Kingdom Pombi , Josè Vicente , Amabelia Rodrigues , Joao Dinis , Vincenzo Petrarca4, Joao Pinto5, Alessandra della Torre1, David Weetman6 Many mosquito species are vectors of human pathogens, such as 1 viruses responsible for Dengue or Chikungunya, or malaria parasites. Department of Public Health and Infectious Diseases, Università di Roma 2 Mosquitoes from the Aedes genus mainly transmit viruses, whereas the “La Sapienza”, Italy, Rome, Italy, Centro de Malária e outras Doenças Anopheles genus is exclusively responsible for transmission of human Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de 3 malaria. The basis for these pathogen-vector specificities is currently Lisboa, Lisboa, Portugal, Instituto Nacional de Saúde Pública, Ministério 4 unknown. Mosquito small RNA pathways have many functions such as da Saúde Pública, Bissau, Guinea-Bissau, Department of Biology and 5 regulation of genes in the development or immunity. Previous studies in Biotechnology, Università di Roma Sapienza, Rome, Italy, Centro de An. gambiae have shown that (i) regulation of anti-Plasmodium immunity Malária e outras Doenças Tropicais, Instituto de Higiene e Medicina 6 by microRNA was essential for correct mosquito protection, and that Tropical, Universidade Nova de Lisboa, Lisboa, Portugal, Vector Group, (ii) the siRNA pathway is essential for the control of arboviral infection Liverpool School of Tropical Medicine, Liverpool, United Kingdom after intrathoracic inoculation. Using next generation sequencing and Anopheles gambiae and An. coluzzii are defined based on SNPs in the functional genomics to examine Anopheles gambiae mosquitoes infected multi-copy IGS chromosome-X linked rDNA region. In most of their range with O’Nyong Nyong virus and Plasmodium parasites, we were able to (i) of sympatry, where they rarely hybridize, multiple areas of the genome discover new Anopheles microRNAs and characterize specific microRNAs are differentiated, with highest divergence found toward centromeres. that are regulated upon infection; (ii) show that the siRNA pathway does However, genomic divergence varies markedly with levels of introgression.

astmh.org 585 In a putative secondary contact region in Guinea Bissau, where hybrid 1914 females have been observed at frequencies >20%, differentiation is largely limited to a region proximal to the X-centromere. This region may play a MICROSATELLITE AND DNA SEQUENCE POPULATION critical role in the speciation process, but the evolutionary forces acting GENETICS EVALUATION OF THE SOUTHWEST PACIFIC upon it are poorly understood. The aim of our work was to evaluate inter- MALARIA VECTOR ANOPHELES KOLIENSIS (OWEN) specific genomic differentiation and frequency of recombination along the 1 2 3 X-centromere in the Guinean hybridization region, focusing the analysis Nigel W. Beebe , Robert D. Cooper , Weixin Xu , Sarah L. 4 3 on X-chromosome hemizygous males to permit unambiguous haplotype Fordyce , Luke Ambrose analysis. We genotyped 263 males for the IGS diagnostic locus and for 1School of Biological Sciences, University of Queensland and CSIRO, two additional markers about 1 Mb from it: i) the insertion of a SINE Brisbane, Australia, 2Australian Army Malaria Institute, Brisbane, Australia, retrotransposable element specific for An. coluzzii, which is widely used 3School of Biological Sciences, University of Queensland, Brisbane, as a species diagnostic; and ii) a 57 bp-insertion in intron 4 of cytochrome Australia, 4Department of Forensic Medicine, University of Copenhagen, CYP4G16 (CYP) gene specific for An. gambiae. Moreover, using Illumina Copenhagen, Denmark and Sequenom genotyping we characterised almost 800 SNPs (34 of The southwest Pacific malaria vector Anopheles koliensis Owen is one of which are species-specific and located in the X centromeric region) in 59 13 cryptic members of the Anopheles punctulatus group that can only be males. We observed: i) lack of inter-specific differentiation in the overall identified by molecular tools. Its distribution in Papua New Guinea (PNG) genome, with the exception of chromosome-X centromere; ii) intra- has only recently been described and it is found predominantly throughout individual mixed IGS-arrays in 12% of the whole male sample, suggesting inland lowlands and river valley flood plains below 300m and is common the occurrence of introgression events; iii) unexpected recombination throughout the continual wet Sepik, Ramu, and Markham River valleys on among IGS, SINE and CYP in 24% of the males, and between SINE and the north side of PNG’s central ranges as well as throughout the northern CYP in 13% of them despite the close proximity of these 2 loci (7 Kb). and southern lowland region of PNG’s Papuan Peninsula. An. koliensis Moreover, results from SNP-genotyping showed: i) some, although low, utilizes both natural larval habitat (ground pools and swamps), as well as levels of recombination in the X-centromere; ii) introgression in the IGS- human modified habitat (vehicle wheel tracks and drains). In this study, region in the absence of recombination in the X-centromere; iii) a hot-spot we drill into the population genetic of An. koliensis in PNG - a region of of recombination nearby the SINE-insertion. The results highlight (1) the incredible biogeography - to detail the spatial and genetic connectivity of likely importance of reduced recombination in maintaining integrity of this malaria vector species. We evaluate nuclear and mitochondrial DNA the X-genomic island of divergence with high gene flow, consistent with sequence as well as develop and analyse 12 microsatellites. We find a genetic-hitchhiking-based speciation models and (2) the poor reliability species with overt genetic and geographic population structure that can of existing diagnostics (IGS, SINE) for the two species in the secondary be explained, in most cases, by natural barriers. We do not find evidence contact region. to support the existence of intraspecific rDNA genotypes previously described but we do find An. koliensis to be a single species with a long 1913 history in New Guinea. AN INTEGRATED CHROMOSOME, GENETIC LINKAGE AND 1915 GENOME MAP FOR THE SOUTHERN HOUSE MOSQUITO CULEX QUINQUEFASCIATUS CHARACTERIZATION OF MALE REPRODUCTIVE FACTORS ESSENTIAL FOR MATING SUCCESS IN THE LIFE CYCLE OF Anastasia N. Naumenko1, Vladimir A. Timoshevskiy1, Becky S. deBruyn2, David W. Severson2, Igor V. Sharakhov1, Maria V. ANOPHELES GAMBIAE MOSQUITOES Sharakhova1 Evdoxia Kakani1, Sara Mitchell2, Paolo Gabrieli1, Enzo Mameli1, 1Fralin Biotechnology Center, Virginia Tech, Blacksburg, VA, United States, Ainhoa Mariezcurrena3, Flaminia Catteruccia2 2 Department of Biological Sciences, Eck Institute for Global Health, 1University of Perugia, Perugia, Italy, 2Harvard School of Public Health, University of Notre Dame, Notre Dame, IN, United States Boston, MA, United States, 3Imperial College, London, United Kingdom Culex quinquefasciatus, a southern house mosquito, is a member Mating is a vulnerable step in the lifecycle of Anopheles gambiae of geographically widespread mosquito complex species with high mosquitoes, as females of this species mate only once. Seminal variation in behavioral patterns and ability to transmit diseases, fluids produced in the male accessory glands (MAGs) and transferred including lymphatic filariasis and West Nile fever. Only 10.4 % of the during mating are likely to induce permanent refractoriness to further 579 Mb genome is currently assigned to the chromosomes based on insemination in females. Moreover, these factors are likely to play a key genetic linkage mapping. Although cytogenetic maps for the polytene role in other post-mating processes, including egg laying and fertility. chromosomes for this mosquito were developed, their utilization for However, the identity of these male molecular triggers is largely unknown. the genome mapping remains difficult because of the low number As changes in levels of individual semen components after mating may of high quality spreads in chromosome preparations. We constructed indicate factors crucial for mating success, we performed a time course idiograms for mitotic chromosomes of Cx. quinquefasciatus based on transcriptional analysis of MAG genes at 3 time points (3h, 12h, 24h) after their banding patterns at early metaphase. These idiograms represent mating, representing the period between 2 mating events in the field. the first cytogenetic map developed for mitotic chromosomes of Cx. Mated tissues were compared to those from virgin males in 4 replicates quinquefasciatus. Genetic contigs associated with 14 major genetic using whole-genome microarrays. Surprisingly, a total of 4,319 genes markers, 18S rDNA and 10 largest contigs were anchored to the exact were differentially expressed after mating (p<0.05 FDR). Gene enrichment positions on Cx. quinquefasciatus chromosomes using fluorescent in analysis revealed a number of functional groups significantly enriched situ hybridization. The order of genetic markers was consistent with in the dataset. During early time points after mating, genes associated the previously developed genetic linkage map. Some new insights were with RNA transcription, translation and post-translational modifications provided into chromosome evolution in mosquitoes. For example, FISH were enriched, suggesting an induction of pathways essential for the result of 18S rDNA suggests an inverted position of the ribosomal locus in replenishment of MAG content. At the latest time point enrichment was chromosome 1 of Cx. quinquefasciatus compared with Ae. aegypti. This observed in genes involved in protein export, indicative of males preparing locus was mapped close to the centromere above the heterochromatin for the next mating event. Interestingly, genes involved in hormone band in Cx. quinquefasciatus but in the middle of the 1q arm below biosynthesis were also highly enriched, supporting previous findings that the heterochromatin band in Ae. aegypti. Our study in progress male hormones may play a critical role in An. gambiae reproduction. We linked chromosome and genetic linkage maps with 4.8% of the Cx. then investigated the function of male hormones that are replenished quinquefasciatus genome. after mating. Tampering with the synthesis of these hormones in males

astmh.org 586 had dramatic effects on the reproductive physiology of mated females, high malarial transmission to prevent low birthweight (LBW), neontal which showed strongly reduced fecundity and fertility. These results reveal deaths and maternal anemia. The incidence of malaria below which previously unknown pathways that are key to mosquito reproductive IPTp-SP no longer provides a cost-benefit is unknown, but it is important physiology, and offer novel targets for vector control efforts aimed at to estimate as countries make progress towards malaria elimination. reducing mosquito reproductive success. We conducted a review of IPTp-SP studies and matched the protective efficacy of IPTp-SP against LBW to a proxy measure of malarial incidence 1916 in women of the same studies, the prevalence of malaria in children. The latter was calculated in two ways. We first extracted prevalence estimates AN INPUT ON PREVENTIVE STRATEGIES FROM THE FIELD: in children from the Malaria Atlas Project (MAP) database and, for the IRON LEVELS AND INTERMITTENT PREVENTIVE TREATMENT second measure, we selected and then pooled the point prevalence data (IPTP) CALENDAR ARE ASSOCIATED WITH PLASMODIUM that underly the MAP estimates using random-effects models. We then FALCIPARUM PARASITEMIA DURING THE FIRST YEAR OF LIFE applied meta-regression models of the protective efficacy of IPTp-SP against LBW to the estimates of malarial prevalence in children calculated 1 2 Violeta Moya-Alvarez , Smaila Ouedraogo , Florence Bodeau- in both ways, and stratified results by gravidity. Among multigravidae, the 3 2 4 Livinec , Gilles Cottrell , Michel Cot protective effect of IPTp-SP against LBW was no longer significant in areas 1Institut de Recherche pour le Développement, EHESP, Paris/Benin, France, where the malarial prevalence in children was < to 9% when we applied 2Institut de Recherche pour le Développement, Cotonou, Benin, 3Ecole des MAP estimates, and < to 8% using our pooled estimates. The latter Hautes Etudes en Santé Publique, Rennes, France, 4Institut de Recherche analysis showed a significant linear trend (P=0.043). Malarial transmission pour le Développement, UPMC, Paris, France intensity could not explain variations in the efficacy of IPTp-SP among Malaria is the disease with the highest infant mortality and morbidity paucigravidae. IPTp-SP no longer protects against the incidence of LBW worldwide. In 2012 the WHO reported over 207 million cases and more among multigravidae in geographical areas of 20 sub-Saharan countries than 627,000 deaths. In Benin malaria is the leading cause of mortality where the parasite prevalence among children is < 8%. In contrast, our (23%) among children under 5 years. There are significant differences analysis among paucigravidae suggests that two or more doses of SP is in P. falciparum parasitemia among children during the first year of life. protective against LBW in transmission settings that are below the current We aim at describing the factors contributing to malaria episodes and to recommendation set by the WHO. high parasitemia during the first year of life in Benin. Therefore we have investigated the association of maternal exposure to the vector in utero 1918 (determined by pregnancy associated malaria (PAM) and the intermittent preventive treatment (ITPp)) to infant parasitemia analyzing as well MISSED OPPORTUNITIES FOR DELIVERING PREVENTIVE nutritional, environmental and socio-economic risk factors. 1000 pregnant TREATMENT FOR MALARIA IN PREGNANCY DURING women and 400 of their children were followed during pregnancy and the ANTENATAL CARE AND COMPARISON WITH DELIVERY first year of life in Allada (Benin), between 2010 and 2012. At inclusion OF NEONATAL TETANUS PREVENTION: AN ANALYSIS OF socio-demographic status and gyneco-obstetric history were investigated. HOUSEHOLD SURVEYS IN SUB-SAHARAN AFRICA Extensive medical and biological exams were realized with both doses of 1 2 3 2 IPTp and at delivery for the mothers and at 6, 9, and 12 months for the Kathryn Andrews , Michael Lynch , Erin Eckert , Julie Gutman infants. Further exams were realized at each emergency consultation. 1Harvard School of Public Health, Cambridge, MA, United States, All patients were treated in case of disease. Random coefficient models 2Centers for Disease Control and Prevention, Atlanta, GA, United States, assessed the relationship between the different parasitemia measures 3President’s Malaria Initiative, United States Agency for International in infants and other variables. A novel approach consisting in pathway Development, Washington, DC, United States analysis was used to analyze the evolution pattern of parasitemia. Coverage of intermittent preventive treatment for malaria during Maternal age at both IPTp doses, infant weight, mother parasitemia at pregnancy (IPTp), a potentially lifesaving intervention, remains low in delivery, number of emergency consultations and total body iron were Africa, despite high antenatal care attendance among pregnant women correlated with infant parasitemia. Placental malaria was not correlated and recommendations for IPTp to be given at each antenatal care visit with infant parasitemia when adjusting for mother parasitemia at delivery. after the first trimester. To highlight areas of potential improvement, we We find for the first time that IPTp has not only an effect on LBW but also assessed trends in IPTp coverage over time, and identified the missed on infant parasitemia. Therefore IPTp calendar should cover extensively the opportunities to deliver IPTp at antenatal clinics. Data from 66 national pregnancy and protect both the mother and the infant. Total body iron is household surveys conducted in 31 sub-Saharan African countries also correlated with infant parasitemia. WHO recommends supplements between 2000 and 2012, with relevant questions on antenatal care (ANC) with iron and folic acid when the prevalence of anaemia exceeds 40%. were used to determine the coverage of ANC visits, IPTp and tetanus However the Pemba study and a Cochrane review conclude to an toxoid (TT). Missed opportunities for IPTp were calculated by comparing increased risk for malaria among supplemented children in the context the number of IPTp doses received to the number of ANC visits during of limited malaria coverage. Our results confirm the association between which IPTp could be given. To account for visits occurring in the first iron and malaria and plead for protective measures in the context of iron trimester when IPTp is not given, one visit was subtracted from the total supplementation number of visits for women who reported having their first visit in the first trimester. The median proportion of pregnant women receiving at 1917 least 2 doses of IPTp was 1.0% (IQR 0-10.3%) during 2000-2007 and 27.2% (IQR 13.9-42.1%) during 2008-2012. Missed opportunities for MALARIA-TRANSMISSION INTENSITY AND THE PROTECTIVE IPTp delivery occurred in a median 99.2% (IQR 90.2-100%) of ANC EFFECT OF INTERMITTENT PREVENTIVE TREATMENT (IPTP): visits from 2000-2007 and 76.8% (IQR 65.6-92.9%) from 2008-2012. POLICY IMPLICATIONS FOR THE ANTENATAL CARE OF The median proportion of primigravid women receiving at least 2 doses PREGNANT WOMEN IN SUB-SAHARAN AFRICA of TT is much higher: 50.2% (IQR 34.4-64.0%) during 2000-2007, and 59.2% (IQR 48.6-64.9%) during 2008-2012. With the exception of R. Matthew Chico, Jorge Cano, Cono Ariti, Timothy Collier, two countries, the proportion of primigravid women receiving at least 2 Daniel Chandramohan, Brian Greenwood doses of IPTp is lower than the proportion receiving at least 2 of TT: the London School of Hygiene & Tropical Medicine, London, United Kingdom median absolute difference is 41.7% from 2000-2007 (IQR 29.2-54.1%) The World Health Organization (WHO) recommends the provision of and 36.7% (IQR 23.7-40.6%) from 2008-2012. Although IPTp coverage intermittent preventive treatment with sulphadoxine-pyrimethamine has increased slightly over time, levels remain disappointingly low, and (IPTp-SP) to pregnant women resident in areas of moderate (stable) or missed opportunities for IPTp occur at the majority of ANC visits. Although

astmh.org 587 both are delivered through the ANC, delivery of IPTp occurs much less recurrent malaria (i.e. with a history of P. vivax) and their infants (at least frequently than delivery of TT, suggesting that barriers to IPTp delivery 28 days old) are being recruited for detailed pharmacokinetic study. Prior could be overcome. Further work is required to determine the specific to enrolment, G6PD deficiency is excluded by rapid qualitative fluorescent factors that are driving the surprising discrepancies between IPTp and TT spot test and G6PD genotype from PCR spot. Anemic patients are treated coverage, with an eye toward improving IPTp coverage through potential and enrolment is delayed until normal HCT is established. Hemoglobin linkage with the TT administration infrastructure. typing is analyzed and fetal hemoglobin in infant blood is quantified. Primaquine is administered to eligible mothers at a dose of 0.5 mg/kg/ 1919 day and is directly observed. Primaquine and carboxyprimaquine levels are measured by liquid chromatography coupled with tandem mass PHARMACOKINETICS OF ARTEMETHER-LUMEFANTRINE spectrometry (LC-MS/MS) on venous and capillary plasma, urine, saliva IN PREGNANT AND NON-PREGNANT WOMEN WITH and breast milk samples from the mothers, as well as capillary plasma UNCOMPLICATED PLASMODIUM FALCIPARUM MALARIA IN samples from the infants. There have been no drug-related adverse events WESTERN KENYA to infants, though several women have experienced mild to moderate methemaglobinemia (not requiring treatment). Preliminary data shows low 1 1 2 Elizabeth A. Juma , Bernhards R. Ogutu , Florence Oloo , Tara but measurable levels of primaquine in both breast milk and infant plasma. 2 3 Barwa , Rashid Aman The final results of this study could have profound impacts on malaria 1Kenya Medical Research Institute, Nairobi, Kenya, 2Centre for Research control and women’s health in the tropics. in Therapeutic Sciences, Nairobi, Kenya, 3African Centre for Clinical Trials, Nairobi, Kenya 1921 Pregnancy increases the vulnerability to malaria infection in all women living in areas of malaria risk. The World Health Organization recommends SAFETY OF ARTEMETHER-LUMEFANTRINE EXPOSURE IN the use of artemisinin-based combination therapies (ACTs) for treatment of EARLY PREGNANCY: AN OBSERVATIONAL COHORT acute uncomplicated falciparum malaria in the second and third trimesters Dominic Franklin Mosha of pregnancy. The pharmacokinetic properties of antimalarial drugs are often affected by pregnancy, resulting in lower drug concentrations Ifakara Health Institute, Dar es salaam, United Republic of Tanzania and a consequently higher risk of treatment failure. While artemether- There is limited data available regarding safety profile of artemisinins in lumefantrine is used in Kenya and most Eastern Africa countries as first early pregnancy. They are therefore not recommended by WHO as a first line treatment for malaria in pregnancy in the second and third trimesters, line treatment for malaria in first trimester due to associated embryo-foetal its pharmacokinetics in pregnant African women with malaria is not well toxicity in animal studies. The aim of the study was to assess birth outcome characterized. This study evaluated the population pharmacokinetics of among pregnant women inadvertently exposed to AL during first trimester artemether, dihydroartemisinin, lumefantrine and desbutyl-lumefantrine in comparison to those of women exposed to other antimalarial drugs or in 45 pregnant and 25 non-pregnant women with uncomplicated malaria no drug at all during the same period of pregnancy. Pregnant women with in Western Kenya. All patients were treated with the standard fixed gestational age < 20 weeks were recruited from Reproductive and Child dose artemether-lumefantrine 20/120mg tablets over 3 days. Frequent Health (RCH) clinic or from monthly house visits (demography surveillance), venous blood sampling was obtained over the treatment period for and followed prospectively until delivery. A structured questionnaire was pharmacokinetic evaluation. Estimates for pharmacokinetic and variability used to interview participants. 2167 pregnant women were recruited parameters will be obtained through nonlinear mixed effects modeling. and 1783 (82.3%) completed the study until delivery. 319 (17.9%) used Simultaneous modeling of parent drug and metabolite will be used antimalarials in first trimester, of whom 172 (53.9%) used artemether- for both artemether and lumefantrine. Absorption and clearance of lumefantrine (AL), 78 (24.4%) quinine, 66 (20.7%) sulfadoxine- artemether-lumefantrine in pregnant compared with non-pregnant African pyrimethamine (SP) and 11 (3.4%) amodiaquine. Quinine exposure in first women with uncomplicated malaria and the implications of findings will trimester was associated with an increased risk of miscarriage/stillbirth (OR be presented. 2.5; 1.3 - 5.1) and premature birth (OR 2.6; 1.3 - 5.3) as opposed to AL with (OR 1.4; 0.8 - 2.5) for miscarriage/stillbirth and (OR 0.9; 0.5 - 1.8) for 1920 preterm birth. Congenital anomalies were identified in 4 exposed groups namely AL only (1/164 [0.6%]), quinine only (1/70 [1.4%]), SP (2/66 A STUDY OF THE PHARMACOKINETICS OF PRIMAQUINE IN [3.0%]), and non-antimalarial exposed group (19/1464 [1.3%]). Exposure LACTATING WOMEN AND BREASTFED INFANTS FOR THE to AL in first trimester was more common than to any other antimalarial RADICAL TREATMENT OF UNCOMPLICATED MATERNAL drugs. Quinine exposure was associated with adverse pregnancy outcome, PLASMODIUM VIVAX which was not the case following other antimalarial intake. Since AL and quinine were used according to their availability rather than to disease Mary Ellen T. Gilder1, Joel Tarning2, Niklas Lindegårdh severity, it is likely that the effect observed was related to the drug, and (posthumous)2, Germana Bancone1, Warunee Hanpithakphong2, not to the disease itself. Detailed information on developmental milestone Hilda Moo1, Tun Tun Win1, Nicholas White3, François Nosten3, up to 12 months is ongoing to rule out any adverse effect on infancy as Rose McGready3 a result of AL exposure in first trimester. Even with this caveat, a change 1Shoklo Malaria Research Unit, Mae Sot, Tak, Thailand, 2Mahidol Oxford of policy from quinine to AL for the treatment of uncomplicated malaria Research Unit, Bangkok, Thailand, 3Centre for Tropical Medicine, Nuffield during the whole pregnancy period could be already envisaged. Department of Medicine, University of Oxford, Oxford, United Kingdom Women of reproductive age in malarial areas of the world suffer due to lack of evidence on the safety of effective antimalarials in pregnancy and lactation. Plasmodium vivax recurrences are more common during pregnancy but the only widely available medication for radical treatment of P.vivax, primaquine, is contraindicated in pregnancy. The postpartum period presents a key opportunity for radical treatment of P. vivax, but there are no studies quantifying primaquine excretion in breast milk and the dose that breastfed infants would be exposed to is unknown. We are conducting the first-ever study of the pharmacokinetics of primaquine lactating women and their breastfed infants during a 14-day radical treatment of P.vivax. Twenty-four healthy lactating women at risk for

astmh.org 588 1922 was evolving from bad to worse. We will describe here the early laboratory investigations that contributed to the identification of the aetiological SAFETY AND EFFICACY OF FOUR ARTEMISININ-BASED agents of the outbreaks that recently occurred in the Pacific, notably based COMBINATION TREATMENTS IN AFRICAN PREGNANT on the use of filter paper-spotted serum and saliva collected on cotton WOMEN WITH MALARIA swab as a source of viral RNA. Based on phylogenetic data we will discuss how these viruses were introduced from continental regions into the 1 2 3 Umberto D’Alessandro , Halidou Tinto , Linda Kalilani Phiri , Pacific and how they spread from one Pacific island country to another. 4 5 2 Michael Nambozi , Harry Tagbor , Maminata Traore , Innocent We will also discuss the particular features of these outbreaks, notably in 2 3 2 4 Valea , Victor Mwapasa , Gertrude Kalanda , Modest Mulenga , the occurrence of unusual clinical manifestations, like observed in French 6 6 6 Joris Menten , Raffaella Ravinetto , Yves Claeys , Maaike De Polynesia during the ZIKV outbreak. Crop6, Theonest Mutabingwa7 1Medical Research Council, Fajara, Gambia, 2Clinical Research Unit, 1924 Nanoro, Burkina Faso, 3College of Medicine, University of Malawi, Blantyre, Malawi, 4Tropical Diseases Research Centre, Ndola, Zambia, POTENT ANTI-MERS COV (MIDDLE EAST RESPIRATORY 5Kwame Nkrumah University of Science and Technology, Ku, Ghana, SYNDROME CORONAVIRUS) FULLY HUMAN ANTIBODIES 6London School of Hygiene & Tropical Medicine, London, United Kingdom, FROM TRANSCHROMOSOMIC BOVINES FOR PASSIVE 7Hubert Kairuki Memorial University, Dar es Salaam, United Republic of IMMUNOTHERAPY Tanzania Thomas Luke1, Kanakatte Raviprakash1, Eddie Sullivan2, Stanley Pregnant women are at increased risk of Plasmodium falciparum malaria Perlman3, Jincun Zhao3, Hua Wu2, Jin-An Jiao2, Lisa Hensley4, Reed which is associated with increased maternal, foetal and neonatal morbidity Johnson5, Gabriel Defang1, Peter Jahrling4, Maya Williams1 and mortality. Thus, malaria-infected pregnant women need prompt and 1 2 effective treatment. Artemisinin-based combination treatments (ACT) are Naval Medical Research Center, Silver Spring, MD, United States, SAB 3 recommended for pregnant women in their second and third trimesters Biotherapeutics, Sioux Falls, SD, United States, University of Iowa, Iowa 4 of pregnancy though information on their safety and efficacy in African City, IA, United States, Integrated Research Facility, National Institute of 5 pregnant women is limited. A Phase 3, non-inferiority, multicentre, Allergy and Infectious Diseases, Fort Detrick, MD, United States, Emerging randomized, open-label clinical trial compared the efficacy and safety Viral Pathogens Section, National Institute of Allergy and Infectious of four ACTs, namely amodiaquine-artesunate, dihydroartemisinin- Diseases, Fort Detrick, MD, United States piperaquine, artemether-lumefantrine, and mefloquine-artesunate, in No specific treatments of proven effectiveness for Middle East Respiratory women with malaria and in the second or third trimester of pregnancy. A Syndrome Coronavirus (MERS-CoV) infections are currently available. The total of 3,423 pregnant women were recruited in Burkina Faso, Ghana, International Severe Acute Respiratory & Emerging Infection Consortium Malawi and Zambia. After being treated with one of the 4 ACTs at day (ISARIC) identified passive immunotherapy with neutralizing antibodies as 0, 1 and 2, women were reviewed at days 3, 7, 14, 21, 28, 35, 42, 49, a treatment approach that warrants priority study. A platform technology 56 and 63, and whenever they were sick. There were 3 early treatment using transchromosomic bovines (Tc-bovines) that produce fully human, failures, 2 in Malawi and 1 in Zambia. Eight hundred twenty five women antigen specific, polyclonal IgG antibody (Tc-pAb) of all subclasses (24.1%) had a recurrent infection during the follow up, 81 (2.4%) of them following immunization has been developed. The quantity of Tc-pAb that identified as recrudescences after genotyping. No major safety problems can be derived from each animal after plasmapheresis ranges from 150 to were observed during the follow up. This is the largest trial on ACT use 300 grams per month. Purified Tc-pAb for intravenous or intramuscular during pregnancy ever done in sub-Saharan Africa. Its preliminary results administration has extremely low quantities of bovine proteins and no are reassuring. evidence of adventitious agents. The Tc-bovine platform can also rapidly produce a diverse repertoire of fully human monoclonal antibodies. Two 1923 experimental anti-MERS CoV Tc-pAb immunoglobulins were produced in Tc-bovines hyperimmunized with inactivated whole virion Jordan strain EPIDEMIOLOGICAL AND MOLECULAR FEATURES OF virus (clade A) or a recombinant spike protein derived from an Al-Hasa DENGUE, ZIKA AND CHIKUNGUNYA CONCURRENT strain (clade B). Both Tc-pAb immunoglobulins, termed SAB-300 and SAB- OUTBREAKS IN THE PACIFIC, 2014 301, demonstrated 50% plaque reduction neutralizing antibody titers > 10e4/ml and cross neutralized other MERS-CoV strains. SAB-300/SAB-301 1 1 1 Van-Mai Cao-Lormeau , Claudine Roche , Anita Teissier , Eric J. were evaluated in recombinant mice expressing the DPP-4 receptor (5 mg/ 2 1 Nilles , Didier Musso kg and 25 mg/kg IP as a single dose 12 hours before intranasal challenge) 1Institut Louis Malardé, Papeete - Tahiti, French Polynesia, 2World Health and SAB-300 in marmosets (80 mg/kg in 4 divided doses IV starting 24 Organization, Division of Pacific Technical Support, Suva, Fiji hours after intratracheal challenge). Control infected mice had a lung viral

During more than a century dengue has been the only mosquito-borne titer of ~6.0 log10 PFU/mg through day 5 post inoculation but treated mice virus considered as major public health concern for Pacific nations. approached, or were below, the limit of detection (2.0 log10 PFU/mg) by However, during the past 5 years the epidemiology of arboviruses in 24-72 hours and displayed no toxicity. Treated marmosets displayed no the Pacific region has shown terrific changes. The situation gradually toxicity and the clinical/virologic data will be presented. Because of these switched from the predominant circulation of a single dengue virus encouraging pre-clinical findings, an IND application is in development. (DENV) to active transmission of multiple DENV serotypes and genotypes as observed in French Polynesia from 2013 and in New Caledonia and Fiji since the beginning of 2014. In the mean time, Chikungunya virus (CHIKV) appeared for the first time in New Caledonia with autochtonous cases sporadically reported from 2011 up to 2013, and large outbreaks occurring in Papua New Guinea in 2012, Yap Island in 2013 and in Tonga in 2014. Another unexpected event was the emergence of Zika virus (ZIKV) in French Polynesia at the end of 2013. ZIKV caused in French Polynesia the largest outbreak ever documented, and in a context of active circulation of DENV serotypes 1 and 3. At the beginning of 2014, ZIKV outbreaks also emerged in New Caledonia and Cook Islands. As of April 2014, outbreaks of “dengue-like illnesses” were under investigation in several other Pacific islands suggesting that the situation

astmh.org 589 1925 mosaic virus to express TMV coat protein. The production of in planta nanoparticles was verified by transmission electron microscopy and these CLINICAL STUDIES OF DNA VACCINES FOR HEMORRHAGIC yielded an immune response in vaccinated mice that was superior to that FEVER WITH RENAL SYNDROME of in vitro assembled nanoparticles. In this study, we validated the use of in planta encapsidated RNAs as an immune activator in the absence of 1 1 2 Connie Schmaljohn , Jay W. Hooper , Drew Hannaman , adjuvants. We can now use this system to create sturdy and inexpensive 3 Kristopher M. Paolino vaccines for tropical infectious diseases. 1United States Army Medical Research Institute for Infectious Diseases, Fort Detrick, MD, United States, 2Ichor Medical Systems, San Diego, CA, United 1927 States, 3Walter Reed Army Institute of Research, Silver Spring, MD, United States SEROPREVALENCE OF NGARI AND BUNYAMWERA VIRUSES Hemorrhagic fever with renal syndrome (HFRS) is endemic in Asia, Europe IN SELECT PARTS OF RIFT VALLEY AND NORTHERN KENYA and Scandinavia, and is caused by infection with the hantaviruses Hantaan Collins O. Odhiambo1, Marietjie Venter2, Robert Swanepoel2, (HTNV), Seoul (SEOV), Puumala (PUUV), or Dobrava (DOBV) viruses. We Rosemary Sang1 developed candidate DNA vaccines for HFRS expressing Gn and Gc genes 1 of HTNV or PUUV and evaluated them in an open-label, single-center International Centre for Insect Physiology and Ecology, Nairobi, Kenya, 2 Phase 1 study. Three groups of nine subjects each were vaccinated on days University of Pretoria, Pretoria, South Africa 0, 28 and 56 with the DNA vaccines for HTNV, PUUV, or mixture of both Ngari and Bunyamwera viruses are among the few mosquito-borne human vaccines using the Ichor Medical Systems TriGridTM Intramuscular Delivery pathogens in the Orthobunyavirus genus, family Bunyaviridae, associated System (TDS-IM). All vaccinations consisted of a total dose of 2.0 mg with febrile illness. Ngari virus has been associated with hemorrhagic fever DNA in an injected volume of 1 mL saline. For the combined vaccine, the during Rift Valley fever outbreaks in Africa. Ngari virus is a reassortant mixture contained equal amounts (1 .0 mg) of each DNA vaccine. There virus composed of the S and L segments from Bunyamwera and the M were no study-related serious adverse events (SAEs). Neutralizing antibody segment from Batai virus. While isolations of both viruses have been responses were detected in 5/9 and 7/9 of individuals who completed all made from mosquito and tick vectors in the transmission foci in Kenya, three vaccinations with the HTNV or PUUV DNA vaccines, respectively. no human serosurveys have been conducted. We report findings from In the combined vaccine group, 7/9 of the volunteers receiving all three a retrospective serosurvey of febrile ill patients attending three health vaccinations developed neutralizing antibodies to PUUV. The three facilities located in Sangailu, Kotile (in Garissa) and Naivasha in Kenya. strongest responders to the PUUV vaccine also had strong neutralizing Bunyamwera and Ngari virus specific antibodies were detected by plaque antibody responses to HTNV. These results demonstrate that the HTNV and reduction neutralization tests in 84 (24.3%) of 345 persons tested; PUUV DNA vaccines delivered by TDS-IM separately or as a mixture are Prevalence rates were 11.9% for Bunyamwera virus and 15.9% for Ngari safe. In addition, both vaccines were immunogenic, although when mixed virus. Multivariable analysis revealed age and location as risk factors for together, more subjects responded to the PUUV than to the HTNV DNA Bunyamwera and Ngari virus infections. Patients presenting with febrile vaccine, suggesting immunological interference. Consequently, we have illness in identified endemic regions should be vigorously investigated to developed an optimized HTNV DNA vaccine that shows no interference determine the public health impact of these infections especially during in hamsters when mixed with the PUUV vaccine. A Phase 2a clinical study seasons of high mosquito abundance. will be initiated in 2014 to assess dose and schedule with the combined, optimized HTNV and PUUV DNA vaccines. An additional Phase 1 study is 1928 being planned to compare intradermal and intramuscular delivery of the mixed DNA vaccines. ITAYA VIRUS: A NOVEL ORTHOBUNYAVIRUS ASSOCIATED WITH HUMAN FEBRILE ILLNESS IN PERU 1926 Carolina Guevara1, Jesus Silvas2, Robert Hontz1, Wilma Casanova3, Nikos Vasilakis2, Douglas Watts4, Tadeusz Kochel1, Julia AN INEXPENSIVE SYSTEM FOR PRODUCING STRUCTURALLY S. Ampuero1, Hideki Ebihara5, Patricia V. Aguilar2 STABLE REPLICATIVE RNA VIRUS-BASED NANOPARTICLE 1 2 VACCINES U.S. Naval Medical Research Unit - 6, Lima, Peru, Department of Pathology, Institute for Human Infections and Immunity, University of Yiyang Zhou, Christopher Kearney Texas Medical Branch, Galveston, TX, United States, 3Direccion Regional 4 Baylor University, Waco, TX, United States de Salud Loreto, Iquitos, Peru, University of Texas at El Paso, El Paso, TX, United States, 5Laboratory of Virology, Division of Intramural Research, Tropical infectious disease vaccine development requires particular National Institute of Allergy and Infectious Diseases, National Institutes of attention to cost-efficiency and minimized storage conditions to be Health, Rocky Mountain Laboratories, Hamilton, MT, United States optimally useful in third world countries. In this study, the use of a plant- produced, trans-encapsidated non-pathogenic RNA virus as a vaccine The Orthobunyavirus genus in the family Bunyaviridae comprises more vector addresses these issues. Plant-based manufacturing allows large than 170 viruses, at least 30 of them associated with human disease. scale production with greatly reduced expense. Transencapsidation with Caraparu virus, a member of the group C (Orthobunyavirus genus), was Tobacco mosaic virus (TMV) coat protein provides superior chemical first isolated from a sentinel monkey in Brazil and subsequently isolated and environmental protection to any RNA sequence containing a from febrile patients in Bolivia, Brazil, Peru and Trinidad. Febrile surveillance TMV encapsidation site. A transencapsidated RNA virus that expresses studies conducted by the U.S. Naval Medical Research Center Unit No. vaccine antigens in human cells would be highly attractive for vaccine 6 identified Caraparu virus (and other group C viruses) as an important use due to its ability to induce innate immune activation pathways to cause of febrile illness in the Amazon region of Peru. We conducted aid immunogenicity of its antigen payload. To avoid the potential safety genetic analyses of previously uncharacterized bunyavirus strains isolated concerns using a human virus, we have used the insect RNA virus, Flock from febrile patients in Peru, and identified a novel reassortant virus House virus (FHV), which replicates in both human and plant cells but containing the S and L segment of Caraparu virus and the M segment of is not pathogenic to either. The RNA2 of the bipartite FHV genome an unidentified Group C virus. Neutralization test using mouse antisera codes for FHV coat protein and is not necessary for replication. We prepared against the prototype Caraparu strain BeAn 3994 and the novel created an FHV RNA1/eGFP vector and noted strong expression of reassortant virus showed that there was more than a 4-fold difference eGFP in inoculated Nicotiana benthamiana plants. This was followed by in titer between these viruses, indicating that the new reassortant was inserting the TMV encapsidation site into the FHV RNA1 and inoculating serologically distinct from the prototype Caraparu strain. Serological plants in combination with a high expression vector derived from Foxtail analyses also confirmed that the novel reassortant was antigenically

astmh.org 590 distinct from Peruvian Caraparu strains. This new reassortant virus, which N-isopropylacrylamide nano-particles functionalized with trypan blue were we named Itaya virus, was first isolated during 1999 from a 25-year-old synthesized by precipitation polymerization and characterized with photon male febrile patient in Iquitos, Peru, and subsequently isolated during correlation spectroscopy. We evaluated the ability of the nanoparticles to 2006 from a 59-year-old male febrile patient in Yurimaguas, another city capture, concentrate and preserve T. cruzi antigens. Urine samples from within the Amazon region of Peru. Geographical distance between these congenitally infected and uninfected infants were then concentrated using two cases indicates that Itaya virus may be widely distributed within the these nanoparticles. The antigens were eluted and detected by Western Peruvian Amazon. The recognition of a new Orthobunyavirus human Blot using a monoclonal antibody against T. cruzi lipophosphoglycan. The pathogen in the Amazon region of Peru reinforces the need to continue nanoparticles concentrated T. cruzi antigens by 100 fold (western blot and expand viral disease surveillance activities in tropical regions of South detection limit decreased from 50 ng/ml to 0.5 ng/ml). The sensitivity of America. Chunap in a single specimen at one month of age was 91.3% (21/23, 95% CI: 71.92%-98.68%), comparable to PCR in two specimens at 0 and 1 1929 month (91.3%) and significantly higher than microscopy in two specimens (34.8%, 95% CI: 16.42%-57.26%). Chunap specificity was 96.5% (71/74 MAPPING THE ZOONOTIC NICHE OF EBOLA VIRUS DISEASE IN endemic, 12/12 non-endemic specimens). Particle-sequestered T. cruzi AFRICA antigens were protected from trypsin digestion. Chunap has the potential to be developed into a simple and sensitive test for the early diagnosis of David Pigott1, Nick Golding2, Adrian Mylne2, Zhi Huang2, Andrew congenital Chagas disease. Henry2, Daniel Weiss2, Oliver Brady2, Moritz Kraemer2, David Smith2, Catherine Moyes2, Samir Bhatt2, Peter Gething2, Peter Horby2, Isaac Bogoch3, John Brownstein4, Sumiko Mekaru4, Andrew Tatem5, 1931 3 2 Kamran Khan , Simon Hay A THERAPEUTIC NANOPARTICLE VACCINE AGAINST ADepartment of Zoology, 1University of Oxford, Oxford, United Kingdom, TRYPANOSOMA CRUZI IN A MOUSE MODEL OF CHAGAS 2University of Toronto, Toronto, ON, Canada, 3Boston Children's Hospital, DISEASE Boston, MA, United States, 4University of Southampton, Southampton, United Kingdom Meagan A. Barry, Qian Wang, Coreen M. Beaumier, Kathryn M. Ebola virus disease (EVD) is a complex zoonosis that is highly virulent in Jones, Brian P. Keegen, Maria Elena Bottazzi, Michael J. Heffernan, humans. The largest recorded outbreak of EVD is ongoing in West Africa, Peter J. Hotez outside of its previously reported and predicted niche. We assembled Baylor College of Medicine, Houston, TX, United States location data on all recorded zoonotic transmission to humans and Ebola Chagas disease is a neglected tropical disease of great importance in virus infection in bats and primates (1976-2014). Using species distribution the Americas, with 7-8 million people infected. The causative agent is models these occurrence data were paired with environmental covariates to Trypanosoma cruzi, and results in an acute febrile illness that progresses predict a zoonotic transmission niche covering 22 countries across Central to chronic chagasic cardiomyopathy in 30% of patients. In endemic areas, and West Africa. Vegetation, elevation, temperature, evapotranspiration Chagas disease is the leading cause of cardiovascular death between ages and suspected reservoir bat distributions define this relationship. At-risk 30-50. Current pharmacological treatments are plagued by significant areas are inhabited by 22 million people, however the rarity of human side effects, poor efficacy, and are contraindicated in pregnancy. There outbreaks emphasises the very low probability of transmission to humans. is an urgent need for new treatment modalities. A therapeutic vaccine Increasing population sizes and international connectivity by air since the for Chagas disease has potential advantages that include cost savings, first detection of EVD in 1976 suggest that the dynamics of human-to- reduced adverse effects, and the potential to be used as a replacement for human secondary transmission in contemporary outbreaks will be very current therapies or when paired with chemotherapy. Prior work in mice different to those of the past. has identified an efficacious T. cruzi antigen (Tc24). To elicit a protective cell-mediated immune response to the Tc24 protein, we have utilized a 1930 nanoparticle delivery system in conjunction with CpG motif-containing oligodeoxynucleotides (ODN) as an immunomodulatory adjuvant. When USE OF A NOVEL CHAGAS URINE NANOPARTICLE TEST tested in a BALB/c mouse model, a dose response study demonstrated (CHUNAP) FOR DIAGNOSIS OF CONGENITAL CHAGAS a positive relationship between dose of vaccine and Tc24-specific IFN-γ DISEASE response. Our nanoparticle vaccine, comprised of Tc24 and CpG ODN encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles, produced Yagahira E. Castro1, Robert Gilman2, Gerson Galdos-Cardenas2, the most robust T 1-mediated CD8+ T cell immune response. When tested Lisbeth Ferrufino3, Gerardo Sánchez1, Edward Valencia Ayala1, H for therapeutic efficacy in T. cruzi infected BALB/c mice, improved survival Lance Liotta4, Caryn Bern5 was seen in the vaccine group compared to the control groups. Additionally, 1Universidad Peruana Cayetano Heredia, Lima, Peru, 2Johns Hopkins there was a significant reduction in the number of parasites in the cardiac University, Bloomberg School of Hygiene and Public Health, Baltimore, MD, tissue of the vaccine group compared to the PBS sham vaccine group, United States, 3Hospital Universitario Japonés, Santa Cruz, Plurinational indicating protection from parasite-driven cardiac damage. The mice that State of Bolivia, 4Center for Applied Proteomics and Molecular Medicine, survived to the end of the study had almost undetectable numbers of George Mason University, Manassas, VA, United States, 5Global Health parasites in the cardiac tissue. These data demonstrate the immunogenicity Sciences, Department of Epidemiology and Biostatistics School of Medicine, and efficacy of a Tc24/CpG ODN nanoparticle vaccine and are convincing University of California San Francisco, CA, United States evidence for a potential new therapeutic vaccine against Chagas disease. Detection of congenital Trypanosoma cruzi transmission is considered one of the pillars of control programs of Chagas disease, because 25% of new infections occur by this route with an estimated of 15,000 infected infants per year in Latin America. Current programs to detect congenital Chagas disease in Latin America utilize microscopy early in life and serology after 6 months. These programs suffer from low sensitivity by microscopy and high loss to follow-up later in infancy. We developed a Chagas urine nanoparticle test (Chunap) to concentrate, preserve and detect T. cruzi antigens in urine for early, non-invasive diagnosis of congenital Chagas disease. This is a proof-of-concept study to provide an initial indication that Chunap allows for the early diagnosis of congenital Chagas disease. Poly

astmh.org 591 1932 these issues within a phenotypic screening setting will be discussed. The current set lead compounds within this series are being evaluated for pre- ASSEMBLING NEW CHEMICAL BOXES AS AN OPEN SOURCE candidate selection. OF STARTING POINTS FOR DRUG DISCOVERY AGAINST KINETOPLASTID PARASITES CAUSING NEGLECTED TROPICAL 1934 DISEASES CLINICAL EVALUATION OF CL DETECT™ RAPID TEST Julio J. Martin1, Emilio Alvarez1, Ana I. Bardera1, Juan Cantizani1, FOR CUTANEOUS LEISHMANIASIS: PERFORMANCE Gonzalo Colmenarejo1, Jose M. Fiandor1, Albane Kessler1, Maria P. CHARACTERISTICS WHEN COMPARED TO SMEAR 1 2 1 2 Manzano , Julio Alonso-Padilla , Imanol Peña , Ana Rodriguez MICROSCOPY AT MULTIPLE TEST SITES 1GlaxoSmithKline, Tres Cantos, Spain, 2New York University School of 1 1 1 Medicine, New York, NY, United States Afif Ben Salah , Nabil Bel Haj Hamida , Amor Zaatour , Adel Gharbi1, Jihene Bettaieb1, Wissem Ghawar1, Amina Khedher1, Neglected Tropical Diseases (NTDs) are a group of infectious diseases Aicha Boukthir1, Nathalie Ben Messaoud1, Evelyne Guedri1, Nathalie categorized by the particular neglect they have suffered in terms of Ben Mbarek1, Nasri Mbarek1, Rabiaa Harrabi1, Chadhi Dawahi1, investment in control measures, when compared with malaria and Hongjing Chen2, Stan Morkowski2, James Needham2, Annabel tuberculosis, “the big two” within the diseases. The NTDs encompass a Tsai2, Yvonne Stevens2, Syamal Raychaudhuri2, Stuart Kahn2, Janet broad range of viral, bacterial, and parasitic infections. Kinetoplastids are Ransom3, Charles Scott3, Giselle Singer4, Danielle Baum4, Shelby a group of flagellated protozoans that include the species Leishmania Jim On4, Mark Richardson4, William McCarthy5, Max Grogl5, Mara and Trypanosoma, some of which are human pathogens with devastating Kreishman-Deitrick5, Jeanne Norwood5, Raymond Houghton2, Mark health and economic effect. The most common human diseases caused Lebwohl4 by kinetoplastids are included within the list of 17 core NTDs declared by 1 2 WHO. They are human African trypanosomiasis, caused by subspecies of T. Institut Pasteur de Tunis, Tunis, Tunisia, InBios International Inc., Seattle, 3 brucei; Chagas disease, caused by the infection with T. cruzi; and various WA, United States, Fast-Track Drugs and Biologics, LLC, North Potomac, 4 clinical manifestations of leishmaniasis, caused by more than 20 species MD, United States, Icahn School of Medicine at Mount Sinai, New York, 5 of Leishmania. All NTDs have been categorized as “tool ready,” yet also NY, United States, Office U.S. Army Medical Materiel Development Activity, “tool deficient” because many of these tools (i.e. drugs and diagnostics) Fort Detrick, MD, United States and implementation strategies are inadequate to achieve the desired goals. This study focused on establishing the performance characteristics of the New effective, safe, and affordable drugs, preferably oral, are needed. The CL Detect™ rapid immunoassay when compared to smear microscopy. general neglect that these diseases have encountered by the pharmaceutical The assay is intended for the diagnosis of Cutaneous Leishmaniasis in far- industry has meant that basic research findings have not found their way forward, rugged environments. The test is based on the detection of the into a drug discovery pipeline. In this paper we present an integral approach thiol specific antioxidant protein (TSA, Peroxidoxin) present in amastigotes to the early drug discovery for the three major kinetoplastid NTDs, i.e. and promastigotes of L. major and other Leishmania spp. It uses a capture visceral leishmaniasis, Chagas disease and sleeping sickness. The GSK 1.8 polyclonal antibody to TSA in combination with a gold conjugated million compounds diverse collection has been screened phenotypically monoclonal antibody directed to L. major amastigotes but reactive with against their causative parasites, respectively L. donovani, T. cruzi and TSA. Testing was performed on samples from skin lesions collected with T. brucei, using the state-of-the-art methodologies available in high a dental broach device. In each case dipstick reactivity was compared to throughput screening. As a result of this effort, three anti-kinetoplastidal the lesion parasite load determined by microscopy and quantified using boxes of approximately 200 compounds each have been assembled, the WHO scale. A total of 168 patients ranging in age from 18-79 years which represent all the chemical and biological diversity identified and with suspected CL lesions were enrolled with written informed consent are intended to serve as an open source of starting points for further lead at 2 sites endemic for L. major infections in central Tunisia (Sidi Bouzid, discovery programs. Gafsa). 149 were positive by CL Detect™ and microscopy while 16 were negative by both tests and 3 were positive by dipstick but not microscopy. 1933 Of these three, 1 was positive by culture. Of the 16 negatives by dipstick and microscopy 2 were positive by culture. In the Icahn School of Medicine, A FULLY INTEGRATED PARTNERSHIP PERFORMING DRUG Mount Sinai specificity study, 150 samples were tested by CL Detect™ and DISCOVERY TOWARDS VISCERAL LEISHMANIASIS: PART 1 microscopy. These included patients ranging in age from 18-92 years with other skin lesions and non-CL infections. In the specificity study 144 of 150 Paul G. Wyatt were true negatives for parasites by both CL Detect™ and microscopy for a University of Dundee, Dundee, United Kingdom specificity of 96.0%. Six samples negative by microscopy were low positive GSK Kinetoplastid DPU and the Drug Discovery Unit, University of Dundee by rapid test but negative by microscopy. Cross reactivity studies with other have formed a partnership to conduct drug discovery within kinetoplastid bacteria, parasites, viruses and fungi confirmed specificity for Leishmania diseases (Visceral Leishmaniasis, Chagas disease and human African spp. Interference, stability and reproducibility studies indicate that CL trypanosomiasis). The collaboration, with support from the Wellcome Trust Detect™ is a robust assay. The pairing of this test with a safe and easy to has made significant progress, in building new methods and infrastructure use drug treatment has the potential to greatly enhance the management to carry out drug discovery for these parasites. Our advances has resulted of CL patients in far-forward rugged environments. in the identification of a lead optimisation series for Visceral Leishmaniasis through phenotypic optimisation. Estimates suggest that Visceral Leishmaniasis worldwide causes 51,000 deaths per year. The current drugs are not fit for purpose, suffering from many issues including poor efficacy and unacceptable levels of toxicity. Part 1, by Paul Wyatt from the Drug Discovery Unit, will describe the transition of a T. brucei GSK3 kinase inhibitor series into a series that fulfils lead optimisation criteria for Visceral Leishmaniasis. This novel series is one of the few reported globally to show oral efficacy in an acute in vivo mouse model against Visceral Leishmaniasis. Part 2, by Tim Miles from GSK, will concentrate on the lead optimisation and progression of this series. As a number of issues were highlighted through critical path screening that have been overcome (i.e. solubility and exposure). Hence a discussion of medicinal chemistry strategies to solve

astmh.org 592 1935 until September 2014. These preliminary results suggest that the selected antigens are immunogenic in humans and may thus be good candidates for ACCESS TO DIAGNOSIS AND TREATMENT FOR CHAGAS further development of a Chagas disease vaccine DISEASE IN THE UNITED STATES: A HEALTH SYSTEMS PERSPECTIVE 1937 Jen Manne-Goehler1, Michael R. Reich2, Veronika J. Wirtz3 EARLY IL-10 PRODUCTION BY CD4+ T CELLS IN THE SKIN IS 1Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, FUNCTIONALLY SUPPRESSIVE MA, United States, 2Department of Global Health and Population, Harvard David Sanin, Catriona T. Prendergast, Adrian P. Mountford School of Public Health, Boston, MA, United States, 3Boston University Center for Global Health and Development, Boston, MA, United States Centre for Immunology and Infection, Department of Biology, University of York, York, United Kingdom Chagas disease, caused by infection with Trypanosoma cruzi, is a vector- borne disease with an estimated 300,000 cases in the United States (US). Communities in endemic areas affected by Schistosomiasis, a parasitic Screening of blood donors for infection with T. cruzi was started in 2007 disease caused by Schistosoma mansoni, are repeatedly infected when in the US. Currently, benznidazole and nifurtimox are available to treat they come into contact with contaminated water. S. mansoni cercariae this infection through direct request from the US Centers for Diseases penetrate the skin and elicit a strong inflammatory response, which after Control and Prevention (CDC) under compassionate use protocols. This repeated exposure becomes both exacerbated and skewed towards type study analyzed epidemiological trends in diagnosis and treatment for 2 immunity, with high levels of IL-10, IL-4, eosinophilia and alternative Chagas disease in the US and assessed national and state barriers to activation of macrophages. In compass with this phenomenon at the site of access. Data on the distribution of cases of Chagas disease identified in infection, local skin draining draining lymph node (sdLN) cells progressively blood donors and drug releases were obtained from the AABB and CDC lose their ability to respond to S. mansoni cercarial antigens, becoming respectively. Semi-structured in-depth interviews were conducted with 30 unable to proliferate or produce cytokines. Our evidence shows that the key informants at the national level and in 6 high-burden states (CA, FL, effect multiple infections have on sdLN responsiveness is mediated by IL- VA, NY, MA, and TX) where treatments were provided. Interview responses 10, which is found at significantly higher levels in the skin after repeated were analyzed according to the health system’s dimensions of regulation, exposure to the parasite. sdLN cells from IL-10 deficient mice retained their financing, payment, organization, and persuasion. Data indicate that 1,908 ability to respond to schistosomula antigens, while the immune response in cases were identified in the blood donation system from 2007-2013 and the skin was significantly more pro-inflammatory. After the initial exposure + + high that CDC provided 422 courses of benznidazole or nifurtimox during this to the parasite, CD4 T cells and F4/80 MHC-II monocytes produced + period. Interview data revealed that local ad-hoc procedures were used by most of the IL-10 in the skin. Strikingly, CD4 T cells in the skin made IL-10 individual physicians with an interest in the disease to increase access to as early as day 1 after the initial exposure. This initial response was directed medicines for Chagas disease, especially through cross-financing of patient against commensal antigens that would penetrate the skin during cercariae + care activities using grants and donations. The primary barriers to access at invasion. However, by day 4 after the first exposure, non-regulatory CD4 T the national level include limited diagnostic and institutionalized referral and cells respond to S. mansoni antigens, expanding considerably after multiple care processes (Organization), lack of financing for patient care activities infections and accounting for most of the detected IL-10. Furthermore, + in most states (Financing), and limited awareness and training among IL-10 producing CD4 T cells from the skin have the ability to inhibit the + + physicians and patients (Persuasion). This study demonstrates that access to proliferation of sdLN CD4 cells. In summary, CD4 T cells in the skin treatment for Chagas disease in the US is limited. The lack of licensing for produce IL-10 and prevent cells in the lymph nodes from responding to the two medications used in treatment was only one of several barriers to repeated infections with the parasite, whilst they contribute to a type 2 access, highlighting the need for a health systems perspective when scaling immune response environment in the skin. Skin commensals are partly up access to these essential medicines. responsible for this type of response, as they penetrate the skin when S. mansoni cercarieae invade the tissue. 1936 1938 IMMUNOGENICITY OF TRYPANOSOMA CRUZI VACCINE A CENTRAL ROLE FOR TYPE I IFN IN THE INDUCTION OF TH2 CANDIDATE ANTIGENS TSA-1 AND TC24 IN MEXICAN RESPONSES BY DENDRITIC CELLS CHAGASIC PATIENTS Lauren Webb1, Rachel J. Lundie2, Jessica G. Borger2, Peter C. Liliana Villanueva-Lizama, Amaru Aguilar-Cetina, Vladimir Cruz- Cook1, Sheila L. Brown1, Lucy H. Jones3, Alexander T. Phythian- Chan, Maria Jesus Ramirez-Sierra, Miguel Rosado-Vallado, Eric Adams1, Andrew S. MacDonald1 Dumonteil 1Manchester Collaborative Centre for Inflammation Research, University of Universidad Autonoma de Yucatan, Merida, Yucatan, Mexico Manchester, Manchester, United Kingdom, 2Institute of Inflammation and Chagas disease affects 8-10 million persons worldwide, and at least Infection Research, University of Edinburgh, Edinburgh, United Kingdom, 1-2 millions in Mexico. Current drugs have a limited efficacy and the 3Institute of Inflammation and Infection Research, University of Edinburgh, development of a preventive and/or therapeutic vaccine is an important Edinburgh, United Kingdom goal. Based on previous work in mouse and dog models, trypomastigote Although dendritic cells (DCs) are critical for induction of Th2 immunity surface antigen-1 (TSA-1) and 24-kDa-trypomastigote secretion/excretion against helminths or allergens, relatively little is known about how they antigen (Tc24) are good candidates for a vaccine against Trypanosoma become activated and function in response to Th2-polarizing antigens. cruzi. We evaluated here the recall immune response against these We have discovered a previously unrecognized role for Type I IFN (IFN-I) in vaccine antigens in Chagas disease patients, as a first step to assess their the optimal activation and function of DCs following exposure to strongly immunogenicity in humans. We used peripheral blood mononuclear cells Th2-polarizing antigens from the parasitic helminth Schistosoma mansoni. (PBMC) from chagasic patients (n=8) and healthy controls (n=8) that were To date, IFN-I has primarily been associated with anti-viral immunity, and stimulated in vitro with TSA-1 and Tc24 recombinant antigens. After 120 its role in Th2 settings is currently unclear. DCs lacking the IFN-I receptor hours of stimulation, we evaluated cell proliferation, identified CD4+ and displayed a dramatically impaired ability to induce Th2 cytokines in vivo, CD8+ memory cell subpopulations, and IFN-gamma and IL-10-producing but unimpaired ability to support Th2 polarization in vitro. Further, Th2- cells by flow cytometry. We observed a specific proliferative response promoting DCs depended on IFN-I signaling for efficient migration to to TSA-1 and to a lesser extent to Tc24, with a central memory T cell the draining LN. We are now investigating whether IFN-I is also required phenotype and antigen-specific INF-gamma and IL-10 production in several for effective localization within the draining LN and interaction with LN- Chagas disease patients. Additional patients and controls will be enrolled astmh.org 593 resident T cells. Together, our data suggest a key role for IFN-I to enable Th2 membrane nanotubes are the structures with which trypanosomes produce induction by DCs against helminths in vivo. Future work will address the EVs. This demonstrates that T. brucei is capable of cellular communication wider role of IFN-I in Th2 inflammation, including during helminth infection, and may have significant impact to understanding infection, immune and the activation of allergic responses in the airways. evasion, and differentiation of this parasite. 1939 1941 VSG-SEQ: A QUANTITATIVE METHOD FOR TRACKING MONOCYTE-DERIVED ALTERNATIVELY ACTIVATED THE IN VIVO DYNAMICS OF ANTIGENIC VARIATION IN MACROPHAGES RETAIN PLASTICITY AFTER ACTIVATION AND TRYPANOSOMA BRUCEI DIFFERENTIATION Monica Mugnier, George Cross, Nina Papavasiliou Natasha Girgis, Uma Mahesh Gundra, Michael A. Gonzales, P’ng The Rockefeller University, New York, NY, United States Loke Using a repertoire of over 2000 different variant surface glycoprotein New York University School of Medicine, Department of Microbiology, New (VSG) genes within its genome, Trypanosoma brucei, the causative agent York, NY, United States of African sleeping sickness, changes its dense VSG surface coat to avoid Alternatively activated macrophages (AAM) induced by type 2 responses detection by the immune system of its mammalian host. The dynamics of promote clearance of helminth parasites, wound healing, and tissue antigenic variation in T. brucei during an infection, however, are poorly remodeling. AAM can accumulate through recruitment and differentiation understood. How many variants appear over the course of an infection? of monocytes, or through proliferation of tissue resident macrophages.

Is there a pattern to VSG expression over time? Although some of these We have recently shown that AAM derived from monocytes (monoAAM) are questions have been broached using Sanger sequencing of VSG cDNA, phenotypically and functionally distinct from AAM that arise from tissue high technical limitations have prevented a high-resolution, quantitative study macrophages (tissAAM). tissAAM are F480 and express the mitochondrial of VSG expression during T. brucei infection. Here we present VSG-seq, protein Ucp1, but low levels of MR1 and PDL2, while monoAAM were the first method for quantitatively examining the diversity of expressed F480int and expressed high levels of MR1 and PDL2. Here, we find that

VSGs in a population of trypanosomes, isolated either from culture or from monoAAM recruited to sites of inflammation remain plastic after activation blood. This next-generation sequencing approach requires very little input and differentiation. In short-term transfer experiments, monoAAM donor material and is quite sensitive, detecting VSGs expressed on less than 0.1% cells transferred into naïve recipients retain expression of MR1, but lose of a population of trypanosomes. Using samples isolated from mouse expression of PDL2. In long-term transfer experiments, monoAAM can convert high infections, expressed VSG sequences can be assembled accurately de novo, to F4/80 macrophages with a phenotype similar to tissAAM. Hence, demonstrating that this approach can be used for the high-resolution monoAAM remain plastic after activation, which is dependent on Stat6 study of VSG expression in any strain of T. brucei, whether in the lab or in signaling in donor AAM, as well as accessory cells in the recipient mice. We the field. We have used VSG-seq to study the kinetics of VSG populations have also found that AAM in the hepatic granulomas of mice infected with throughout T. brucei infections. These studies reveal more complex the parasitic helminth Schistosoma mansoni accumulate primarily through switching dynamics than previously expected and hint at the possibility of the recruitment of inflammatory monocytes. Future experiments will new mechanisms for increasing antigenic diversity in vivo. determine if these monoAAM will further adopt the phenotype of tissAAM in the livers of infected mice after long-term residence in the granulomas 1940 1942 BLOODSTREAM FORM TRYPANOSOMA BRUCEI MEMBRANE NANOTUBES AND EXTRACELLULAR VESICLES MEDIATE METABOLIC REGULATION OF TYPE 2 IMMUNITY CONTROLS INTERCELLULAR INTERACTIONS AND HOST ANEMIA TISSUE REPAIR Anthony Szempruch1, S. E. Sykes1, J. M. Harrington1, A. C. Wildaliz Nieves, Taylor Oniskey, Li-Yin Hung, De’Broski R. Herbert Becker1, A. Gartrell1, E. Arigi2, I. C. Almeida2, S. L. Hajduk1 Division of Experimental Medicine, Department of Medicine, University of 1Department of Biochemistry and Molecular Biology, University of Georgia, California at San Francisco, San Francisco, CA, United States Athens, GA, United States, 2Department of Biology, University of Texas El Host metabolism is profoundly affected by gastrointestinal (GI) nematodes. Paso, El Paso, TX, United States Many GI nematodes feed upon the tissues and blood of their hosts, Trypanosoma brucei cycles between an insect vector and a mammalian resulting in anemia, malnutrition, and generalized immunosuppression. host, causing human African sleeping sickness and Nagana in cattle. We It is debatable whether these features of worm infection are due to identified extracellular vesicles (EVs) from wild type bloodstream form parasite and/or host-derived factors. In this study, we investigated whether (BF) cells by electron microscopy. Purified vesicles have been shown to adenosine monophosphate kinase (AMPK) controlled the outcome of GI propagate a distinctive morphological phenotype when added to wild type nematode infection through controlling the inflammatory response. AMPK cells. Fluorescently labeled EVs bind to the trypanosome flagellar pocket is a heterotrimeric enzyme complex of αβγ subunits that restores cellular and are subsequently endocytosed. We have developed an RNAi cell line energy through oxidative phosphorylation. Given that AMPK activity is that can be induced to produce an excess of EVs. When used in transwell regulated by phosphorylation of the catalytic α subunit, we generated separation experiments, these cells showed EVs mediate phenotype transfer. CD11cCre x AMPKα1flox/flox (DC-AMPK-/-) mice to study the importance of Proteomic analysis of EVs from both wild type and RNAi cells revealed ~50 AMPK in alveolar macrophage and dendritic cell function. DC-AMPK-/- mice shared proteins. In addition, we observe that purified EVs are capable of infected with the hookworm Nippostrongylus brasiliensis (N.b.) generated membrane fusion and transferring variant surface glycoprotein to human abnormal Type-2 immune responses and failed to regenerate areas of red blood cells. This fusion and protein transfer alters the physical properties hookworm-damaged tissue 9 days post-primary infection. In comparison of the red blood cell membrane, potentially leading to anemia seen during to littermate controls, DC-AMPK-/- mice were unable to generate intestinal infection. Imaging of T. brucei cells reveal the formation of long membrane goblet cell metaplasia and failed to expel adult worms from the intestine. nanotubes at the posterior end of the cells that are able to bind other Moreover, N.b.-induced lung injury was more severe in DC-AMPK-/- mice trypanosomes. These membrane nanotubes originate from budding of the and the restoration of pulmonary function was significantly delayed flagellar membrane and form a helical wrapping structure that resembles compared to controls. Dysregulated responses generated in DC-AMPK-/- “beads on a string.” These “beads” closely resemble the structure of cell- mice were associated with increased Type-1 responses (IL-12, iNOS), greater associated EVs. In addition, live cell imaging suggests that these nanotubes numbers of TH17 cells, and defects in the generation of alternatively can disassociate into what appear to be free vesicles. We hypothesize that activated macrophages. Taken together, our data are consistent with an

astmh.org 594 important role for host metabolism in shaping inflammatory responses Our studies with blood-stage parasite lines expressing these biosensors during helminth infection. Thus, AMPK activity within myeloid antigen indicate that micromolar concentrations of labile heme are maintained in presenting cells regulates host protection against GI parasites. the parasite cytosol throughout development. Furthermore, exposure to chloroquine, but not pyrimethamine, leads to accumulation of cytosolic 1943 labile heme, thus directly linking heme dysregulation to the in situ effects of aminoquinolines. We believe these studies will advance our understanding THE TOXOPLASMA DENSE GRANULE PROTEINS GRA17 AND of how heme perturbation is linked to antimalarial drug potency, and help GRA23 MEDIATE THE MOVEMENT OF SMALL MOLECULES to mechanistically inform future drug development efforts. BETWEEN THE HOST AND THE PARASITOPHOROUS VACUOLE Dan Gold1, Aaron D. Kaplan2, Agnieszka Lis2, Glenna C. Bett2, 1945 Emily E. Rosowski1, Kimberly M. Cirelli1, Alexandre Bougdour3, CONDITIONAL EXPRESSION OF PFRIPR CONFIRMS ITS 4 5 6 7 Saima M. Sidik , Josh R. Beck , Sebastian Lourido , Pascal F. Egea , ESSENTIALITY FOR PLASMODIUM FALCIPARUM MEROZOITE 5 3 2 Peter J. Bradley , Mohamed-Ali Hakimi , Randall L. Rasmusson , INVASION INTO HUMAN ERYTHROCYTES Jeroen P. Saeij1 1Massachusetts Institute of Technology, Department of Biology, Cambridge, Alan Yap, Alan F. Cowman MA, United States, 2Physiology and Biophysics, The State University of New Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, York, University at Buffalo, Buffalo, NY, United States, 3Centre National de Australia la Recherche Scientifique, UMR5163, Laboratoire Adaptation et Pathogénie Plasmodium falciparum merozoites actively invade human erythrocytes des Microorganismes, Grenoble, France, 4Whitehead Institute for Biomedical during blood stage malaria, the stage that causes clinical symptoms. Research, Cambridge, MA, United States, 5Department of Microbiology, Invasion of erythrocytes is a complex process that involves a cascade of Immunology and Molecular Genetics, University of California Los Angeles, protein-protein interactions between merozoite ligands and erythrocyte Los Angeles, CA, United States, 6Université Joseph Fourier, Grenoble, receptors. To date, many merozoite ligands have been described as essential France, 7Department of Biological Chemistry, University of California Los based on the inability to knockout the proteins in vitro. One of these is P. Angeles David Geffen School of Medicine, Los Angeles, CA, United States falciparum Rh5 interacting protein (PfRipr). Here we present evidence for Toxoplasma gondii is a widespread protozoan pathogen in the phylum the efficient knockdown of PfRipr using a dimerizable Cre recombinase Apicomplexa that resides intracellularly within a parasitophorous vacuole (DiCre) system. PfRipr is an essential invasion protein that forms a complex (PV) that is selectively permeable to small molecules through an unknown with P. falciparum reticulocyte binding-like homologues 5 (PfRh5). Previous mechanism. We have identified GRA17 as a novel Toxoplasma-secreted studies have shown that the PfRipr/PfRh5 complex plays a critical role protein, which localizes to the parasitophorous vacuole membrane (PVM) in merozoite attachment and invasion as anti-PfRipr antibodies block and is conserved across PV-residing apicomplexans. GRA17 mediates the merozoite invasion. We generated parasites expressing DiCre, which is passive transport of small molecules across the PVM. The PVs of GRA17- activated by the addition of rapamycin leading to the deletion of Pfripr. deficient parasites have aberrant morphology, reduced permeability to small Knockdown of gene and protein levels up to 90% within one cycle of the molecules, and structural instability. GRA17-deficient parasites proliferate blood stage (about 48 hours) was achieved. This led to a growth reduction slowly and are avirulent in mice. GRA17 functions synergistically with a in the following cycles, which relates to a reduction in invasion efficiency as related protein, GRA23. Exogenous expression of GRA17 or GRA23 alters determined by flow cytometry, live-imaging and superresolution microscopy. the membrane conductance properties of Xenopus oocytes in a manner In summary, conditional regulation of PfRipr confirms the essential role of consistent with a large non-selective pore. GRA17 and GRA23 provide the this protein and further elucidates its functions. first molecular basis to explain the PVM permeability to small molecules. 1946 1944 HSP101/PTEX MEDIATES EXPORT OF DIVERSE MALARIA QUANTIFYING LABILE HEME IN LIVE MALARIA PARASITES EFFECTOR PROTEINS INTO THE HOST ERYTHROCYTE USING NOVEL FRET BIOSENSORS Josh Beck, Vasant Muralidharan, Anna Oksman, Daniel E. James Abshire, Christopher J. Rowlands, Peter T. So, Jacquin C. Goldberg Niles Washington University School of Medicine, St. Louis, MO, United States Department of Biological Engineering, Massachusetts Institute of To mediate its survival and virulence the malaria parasite Plasmodium Technology, Cambridge, MA, United States falciparum exports hundreds of proteins into the host erythrocyte. To enter The heme cofactor is centrally important in malaria parasite physiology. On the host cell, exported proteins must cross the parasitophorous vacuolar one hand, it is a byproduct generated in large quantities during parasite- membrane (PVM) within which the parasite resides, but the mechanism mediated hemoglobin degradation during the blood stage of malaria remains unclear. A putative Plasmodium translocon of exported proteins infection. Parasites must detoxify excess heme to limit oxidative damage, (PTEX) has been suggested to be involved for at least one class of exported and do so by polymerizing liberated heme into nonreactive hemozoin. proteins; however, direct functional evidence has remained elusive. Several successful antimalarial drugs such as the aminoquinolines inhibit Here we show that export across the PVM requires heat shock protein this process, presumably leading to the toxic accumulation of labile heme, 101 (HSP101), a ClpB-like AAA+ ATPase component of PTEX. Using a although this has not been directly demonstrated or quantified. On the chaperone auto-inhibition strategy, we achieved rapid, reversible ablation other hand, the parasite genome encodes a complete heme biosynthetic of HSP101 function, resulting in a nearly complete block in export with pathway that is essential for other stages of parasite development. This substrates accumulating in the vacuole in both asexual and sexual parasites. suggests heme is a necessary cofactor that is toxic in excess. Despite the Surprisingly, this block extended to all classes of exported proteins, revealing central role of heme to parasite metabolism and its link to antimalarial drug HSP101-dependent translocation across the PVM as a convergent step potency, little is known about heme dynamics in normal parasite physiology in the multi-pathway export process. Under export-blocked conditions, or how these dynamics change under stresses imposed by heme-interacting association between HSP101 and other components of the PTEX complex drugs. To address these questions, we have developed and characterized was lost while association with exported substrates was maintained, a family of novel, genetically-encoded FRET biosensors for quantifying suggesting that HSP101 first recognizes proteins destined for export before labile heme in live parasites. In vitro spectroscopic characterization of feeding them into the translocon. Our results demonstrate an essential and the purified protein sensors demonstrates their ability to reversibly bind universal role for HSP101 in protein export and provide strong evidence for heme, and to exhibit significant heme-dependent changes in FRET. PTEX function in protein translocation into the host cell.

astmh.org Abstract Author Index A-595

The number(s) following author name refers to the abstract number.

Acosta, Luz 71, 1727, 56, 1144 Agramunt, Veronica H. 1780 Aktar, Amena 35, 433, 434 A ACT Consortium 1809 Agudelo Garcia, Olga M. 408 Akter, Aklima 433 Abakar, Fayiz M. 1113 Acuna-Soto, Rodolfo 1736 Agüero, Fernán 1201 Akter, Nasrin 48 Abanda, Prosper 1547 Adade, Kangni 481, 506 Aguero, María J. 202 Akue, Adovi 613 Abange, William B. 524 Adaje, Austin 1475 Aguiar, Joao 608 Akullian, Adam N. 1260 Abbas, Ali K. 927 Adaji, Justice 264 Aguiar, Joao C. 1603 Akuoku, Jonathan 1032 Abbas, Salah F. 1419 Adak, Tridebis 132 Aguiar, Julia 1655 Akweongo, Patricia 296 Abdalal, Shaymaa 979 Adakun, Susan A. 1860 Aguiar-Alves, Fábio 1658 Akyeampong, Andrea 607, 1574 Abdoulaye, Diabate 775 Adamou Bathiri, Salissou 62, 1084 Aguiar Barros, Cláudia 1658 Al-Abbasi, A M. 1688 Abdul, Muhammad K. 1139 Adamowicz, Beth M. 352 Aguilar, David 698 Alaii, Jane 333 Abdulgadir, Mohammed J. 1074 Adams, David 89, 499, 1325, 1468, Aguilar, Patricia V. 1423, 1928 Alam, Ashraful 1110 Abdulla, Salim 374, 743, 1478, 204 Aguilar-Cetina, Amaru 1750, 1936 Alam, Masud 1288 1482, 1602, 670, 876 Adams, John H. 231, 419, 890, Aguirre, Paula 140 Alam, Md. Murshid 588 Ab-dullah, Husam E. 1419 898, 976, 994, 1465, 1585, Aguirre, Sebastian 1839 Alam, Mohammad S. 891 Abe, Eniola M. 1069 1829 Agunyo, Stella 593, 1085 Alam, Mohammed A. 1654 Abe, Mayumi 1883 Adams, Matthew 270, 324, 380, Agyapong, Jeffrey 409 Alam, Shafiul 1359 Abebe, Almaz 87 389, 895, 984, 1235, 1596, Ahmad, Abdullahi 314 Al-Amin, H. M. 1359 Abebe, Dawit 509 1796 Ahmad, Rushdy 50 Alano, Pietro 288 Abebe, Yonas 1556 Adams, Yamila 1416 Ahmad Shanizza, Azzy Izyati 1342 Alava, Freddy 959, 966 Abegaz, Debritu M. 215 Adaui, Vanessa 538, 1095 Ahmed, Ammar A. 114 Alava, Wieslawa 807 Abeku, Tarekegn A. 705, 1499 Addiss, David 496, 596 Ahmed, Arwa M. 475 Alba, Milena 538, 1095 Abel, Annemieke 1863 Addy, Ebenezer 476 Ahmed, B. U. M. Wahid 1101 Al-Badry, Saad H. 1686 Abel, Lucy 370 Ade, Fredrick 1220 Ahmed, Fayaz 954 Albers, Anna 1704 Abenakyo, Vicky 1536 Adebiyi, Akindele 1024 Ahmed, Khalil 54 Albonico, Marco 642 Abernethy, Neil 1293 Adedire, Elizabeth B. 566, 1680 Ahmed, Mohamed E. 437 Albrecht, Letusa 233 Abeynayake, Janaki 1058 Adegbola, Richard A. 1139 Ahmed, Mohammed 446 Albregtsen, Fritz 1155 Abeyratne, Gaveshika 1381 Adegle, Richard 1092 Ahmed, Mujaddeed 1874 Alcoba, Gabriel 1103, 1425 Abílio, Ana Paula 1847 Adegnika, Akim 21 Ahmed, Sharia M. 1882 Alegana, Victor 653 Abiola, Annie 394 Ademowo, Olusegun G. 133 Ahmed, Tahmeed 585, 634, 1654 Alemayehu, Abreham T. Aboagye, Frederick 1093 Adeniji, Adekunle J. 175 Aho, Celestine 53 Alemayehu. 1070 Aboderin, Oladipo A. 424 Adeniji, Johnson A. 172 Ahorlu, Collins 1035, 1548 Alencar, Aline 677 Abolude, Olukemi O. 1175 Ades, Veronica 1808 Ahouidi, Ambroise D. 234, 1450, Alera, Maria T. 8, 1274 Abong’o, Bernard O. 1887 Adesiji, Yemisi O. 1108 1799, 1443 Alexander, Neal 1472 Abot, Esteban 1603 Adesokan, Hezekiah K. 436 Aina, Oluwagbemiga O. 362, 932, Alexander, Tiffany 93 Abot, Steve 1005 Adetifa, Jane 671 932 Alexandre, Catarina 1153 Aboud, Said 722 Adewusi, Funto 1363 Aith, Fernando M. A. 1230 Alexandre, Jean Seme Fils 1908 Abraham, David 488, 1264, 1892 Adeyemi, Mitchell 1139 Aiyenigba, Bolatito 265, 1576, 264 Al-Fartosi, Khalid G. 1686 Abrams, Steven 1904 Adeyemo, Adebowale 1286 Ajayi, Ikeoluwa 365 Alfonso, Catalina 756 Abreu, Cláudia B. 1315 Adjei, George 1067 Ajayi, Ikeoluwapo O. 566, 1680 Alfonso-Parra, Catalina 749 Abshire, James 1944 Adjei, Ohene 1207 Ajayi, Toyin 637 Ali, Abdulla S. 368 Abuaku, Benjamin 1035, 1548 Adjeloh, Poukpessi 1136 Ajibaye, Olusola 932 Ali, Abdullah S. 868, 927, 929, Abu Bakar, Sazaly 1342 Adjobimey, Tomabu A. 30, 1713 Ajiji, Joseph 950 1040 Abubakar, Aisha 853 Adnan, D. 1141 Ajumobi, Olufemi 365, 988, 1680, Ali, Alyaa H. 1061, 1685 Abudho, Bernard 1795 Adoke, Yeka 893 357, 865 Ali, Asad 54, 954 Abuelmaali, Sara 1706 Adolfi, Adriana 1186 Akala, Hosea 306 Ali, Doreen 879, 1253 Abugri, James 1445 Adongo, Emmanuel 468 Akanbi, Ibikunle M. 436 Ali, Mohammad 1433 Abu Sayeed, Abdullah 1101 Adu, Eric 465 Akanbi, IfeOluwapo O. 436 Ali, Mousab S. E. 1229 Acácio, Sozinho 50, 338 Aebig, Joan 668, 1008, 1621 Akano, Kazeem 253, 256 Ali Ber Lucien, Mentor 1300 Accrombessi, Manfred M. K. 635, Affara, Muna 238, 373, 376 Akatu, Johnstone 1014 Alidina, Zainab 1287 1257 Afolabi, Muhammed 671, 720 Akazili, James 1872 Alifrangis, Michael 239, 310, 394, Acebo Falcón, Selene 1657 Afrane, Yaw A. 120 Akhund, Tauseef 954 897, 1227 Achan, Jane 1457, 1458, 1577 Afrin, Sadia 433 Akhwele, Willis 949 Aliota, Matthew 198 Achata, Jorge 1432 Agaba, Bosco B. 866, 1536 Akinjeji, Adewale 81 Aliota, Matthew T. 163 Achee, Nicole 797, 1324, 1191, Agaba, Steven 358 Akinkunmi, Ezekiel O. 428 Alique, Matilde 610 1369, 1735 Agbenyega, Tsiri 21 Akinyede, Akinwumi A. 1475 Alirol, Emilie 692, 1679 Achilla, Rachel 559 Agbo, Eddy C. 882 Akinyemi, Oluwaseun 1024 Aljayoussi, Ghaith 502 Achoki, Tom 1618 Agbo, Ugochi J. 450 Akinyi, Sheila 911, 1807 Aljayyoussi, Ghaith 504 Acholonu, Alex D. W. 534 Agbowaï, Carine 1557, 1797 Akizili, James 1299 Al-Kohlani, Abdulhakim A. 1154 Achonduh, Olivia 884 Agier, Isabelle 1403 Akkoyunlu, Mustafa 230 Allan, Fiona 1812 Ackerman, Hans 1452 Agomo, Chimere O. 308, 362, 256, Akogbeto, Martin 118, 134, 769, Allan, Kathryn J. 1758 Acosta, Angela 1030, 1031 932 770 Allegretti, Gina 493 Acosta, Belsy 857, 1416 Agomo, Philip 256 Akpasa, Aniefiok 264 Allen, Fiona 1811 Agossadou, Didier 1655 Akpatou, Bertin 659 Allen, Henrietta 84, 905 astmh.org A-596 Abstract Author Index The number(s) following author name refers to the abstract number.

Allen, Jeff 1606 Ames, Abbe 91 Andujar-Perez, Doris A. 1402 Araújo Cardoso, Claudete 1658 Allen, Jonathan E. 1397 Ameur, Btissam 1370 Añez, Germán 160, 1383 Arauz, Maria Jose 745 Allen, Koya C. 1398 The AMFm Independent Evaluation Angarita-Jaimes, Natalia 1883 Arboleda, Sair O. 793 Allers, Claudia 1125 Team 1459 Angelo, Michael A. 822, 1275 Arca, Bruno 962, 964 Allman, Windy 230 Ami, Yasushi 1840 Angov, Evelina 1586, 1595 Archer, Julie 1533 Al-mafazy, Abdul-wahid 368 Amiel, Olga 1087 Angrisano, Fiona 623 Archer, W. Roodly 1163 Al-mafazy, Abdul-wahiyd 927 Aminu, Peace T. 1813 Angulo, Cecilia 1095 Aregger, Fabian Cedric 21 Al Majed, Marow 1061 Amlaga, Kwame C. 1136 Angulo, Victor M. 1232 Arencibia, Amely 857, 1416 Almazor, C. Patrick 37 Ammah, Naa O. 1093 Angus, Brian 1849, 1850 Arévalo, Andrea 1268 Almeida, Anne 340 Amman, Brian 861 Anh, Chu X. 249 Arévalo, Jorge 538, 1095 Almeida, Gabriel M. F. 858 Amoa-Bosompem, Michael 1092 Ankarlev, Johan 1441 Arévalo-Herrera, Myriam 225, 334, Almeida, Juliana 1749 Amoako, Douglas 565 Annan, Francis K. 267 379, 930, 1562, 1802 Almeida, I. C. 1940 Amoako, Nicholas 1445, 1637 Annison, Lawrence 1036 Arguello, D. Fermin 1415 Almeida, Sonia R. 1230 Amoo, George 276, 280, 880, 881, Annoura, Takeshi 1594 Arias, Jorge R. 100 Almendinger, Katherine 50 1523, 1549 Annuzaili, Dhekra A. A. 1154 Arias, Luis 643 Al-Nuiemi, Zahraa W. 1068 Amorim, Leila 1265 Anopheles Genomes Cluster Arias, Luis M. 514 Alonso, Pedro 50, 338, 1280, 1602 Amos, Ben 532 Consortium, The 1355 Arichabala, Ana 916 Alonso, Wladimir J. 54, 1775 Amós Macuvule, Sónia 470 Anova, L. 280 Arigi, E. 1940 Alonso-Padilla, Julio 1932 Amour, Caroline 585 Anova, Lalaine 276, 880, 1523, Arik, Anam 750 Alout, Haoues 982, 1635 Amouzou, Agbessi 731 1549 Arinaitwe, Emmanuel 26, 257, 402, Alphey, Luke 2 Amoyaw, Frank 1637 Ansah, Evelyn K. 296, 1471 946, 1569, 1844, 1864, 1903 Alphonse, Traoré 1019 Ampuero, Julia S. 1420, 1928 Ansah, Nana Akosua 1872 Aristide, Hien 775 Alphonsus, Kal 950 Amuasi, John H. 1459 Ansah, Patrick 1484, 1872 Ariti, Cono 1917 Al-Tamimi, Manal B. 1686 Amuge, Pauline 242 Anstey, Nicholas 219, 22, 1905, Arlian, Larry 1331 Altamiranda, Mariano A. 140 Amukoye, Evans I. 528 1905 Armien, Blas 663 Altcheh, Jaime 1201 Amwayi, Samuel 1424 Antani, Sameer 1453 Armoo, Samuel 479 Alter, Galit 630 Amza, Abdou 1048, 1066 Anthony, Andrew 458, 1198 Armstrong, Stuart D. 1890 Althaus, Fabrice 1767 Anagnostou, Nick 671, 986, 1590 Anticona, Cynthia F. 51, 457 Arney, Leslie A. 572, 1630 Alvarado, Luisa 696, 1379, 1672, Anandaraja, Natasha 732 Anticona Huaynate, Cynthia F. 1296 Arnold, Catherine 1046 1405, 1407 Anantapreecha, Surapee 1414 Antierens, Annick 1103, 1425 Arnold, Fred 1579 Alvarado-Domenech, Luisa I. 1402 Anantatat, Tippawan 1214 Antillon, Marina 1649 Arnold, Ludovic 1238 Álvarez, Álvaro 225 Anchang-Kimbi, Judith 918 Anto, Francis 594, 1302 Aroian, Raffi 1724 Alvarez, Carlos 397 Andagalu, Ben 306, 666, 1239 Antoine, Sanon 117 Aroian, Raffi V. 520, 1267 Alvarez, Danilo 1115, 1665 Andemel, Naissem 1539 Antoniewski, Christophe 1910 Aronson, Naomi 1747 Alvarez, Emilio 1932 Anderios, Fread 1442 Antonio, Martin 431, 1139 Arredondo, Jose Luis 577, 578 Alvarez, Maricruz 598 Anders, Katherine 1384 Antoshechkin, Igor 1267 Arriens, Sandra 1704 Álvarez, Natalí 1358 Anderson, Alicia D. 661 Antwi, Gifty D. 1053 Arruda, Sérgio 1118 Alves, Eliana B. 390 Anderson, Benjamin 1566 A-Nuegoonpipat, Atchareeya 1414 Arsanok, Montri 912 Alves, Tânia M. A. 1094 Anderson, Charles 248, 668, 1008 Anupindi, Ravi M. 1630 Artimovich, Elena M. 321, 888 Alving, Carl 1747 Anderson, Heidi 1754 Anwar, Asif 1495 Artino, Anthony R. 1673 Aly, Ahmed S. 979, 1830 Anderson, Jennifer M. 150, 298, Anyan, William K. 1092, 1093 Arumugam, Sridhar 1892 Alyko Chaffa, Evelyne S. A. 1551 1371 Anyanti, Jennifer 934 Arvelo, Wences 562, 598, 1424, Al-Zadawi, K. M. 1141 Anderson, Katy 500 Anyanwu, Maureen O. 365 1643 Alzua, Maria Laura 709 Anderson, Marc 285 Anyona, Samuel B. 967, 968, 1553, Aryeetey, Richmond 1302 Amado, Idali 815 Anderson, Michael P. 1193, 1195 420 Arze, Cesar 1600 Amado, Inês F. 1498 Anderson, Roy 1715, 1722 Anyorigiya, Thomas 883 Asamoah, Obed 1032, 1845 Amambua-Ngwa, Alfred 124, Anderson, Tara 601 Aonuma, Hiroka 126 Asante, Kwaku P. 1067, 1637, 1520 1443, 376, 1239 Anderson, Tavis 842 Aparecido Da Silva, Thiago 549 Aseffa, Abraham 316, 1286, 1690 Aman, Rashid 1919 Anderson, Tim J. C. 906, 983, Apeanyo, Sena 1718, 1720 Asensio, Norberto 1107 Amanfo, Seth A. 381 1832, 1824, 908, 1236, 1810 Apinjoh, Tobias 1239 Ashley, Elizabeth 904, 906, 908, Aman-Oloniyo, Abimbola F. 854 Anderson, Troy D. 1190 Apiwattanakul, Nopporn 183 1236 Amaratunga, Chanaki 298, 904, Andersson, Maria 1040 Aponte, John 338, 1478, 1567 Ashley, Elizabeth A. 268 1237 Andrade, Luiza F. 1789 Appawu, Maxwell 1622 Ashong, Yvonne A. 519, 1717 Amarista, Manuel 820 Andrade Filho, Jose D. 1346 Apperson, Charles S. 109 Asiedu, Kingsley 1655 Amato Neto, Vicente 718 Andrews, Benetta 465 Appiah-Opong, Regina 1093 Asiimwe, Grace K. 1112 Amaya, Moushimi 157 Andrews, Elizabeth S. 154, 789, Arama, Charles 1868 Aslan, Hamide 1202 A. M. Brito, Marcelo 340 1374 Aranda, Maria 1405 Asoala, Victor 883, 1637 Ambrose, Luke 1914 Andrews, Howard 1197 Arang, Nadia 17 Aspeling-Jones, Harvey 1443 Ambuel, Yuping 182 Andrews, Jason R. 66 Arango, Santiago 361 Asque, Elizabeth M. 492 Ambuludi, Mariano 916 Andrews, Kathryn 1918 Arango Florez, Eliana M. 408 A.S. Romero, Gustavo 340 Amenga-Etego, Lucas 1239 Andrews, Ross 597 Araújo, Dorival 1118 Assadou, Mahamadoun H. 668, Amenga-Etego, Seeba 1067 Andrianaivoarimanana, Voahangy Araujo, Emilia 1263 1009, 1598, 796 Amenya, Dolphine 1186 1660 Araújo, Marcelo C. 1666 Assefa, Ashenafi 1690 astmh.org Abstract Author Index A-597

The number(s) following author name refers to the abstract number.

Assefa, Samuel 1442, 1445 Azad, Abdu F. 685 Balabaskaran Nina, Praveen 407, Barker, Christopher M. 141, 839, Assogba, Benoit 776, 1192 Azam, Syed Iqbal 54 618 1376 Assoum, Mohamad 518 Azcarate-Peril, M. Andrea 586 Balassiano, Ilana 1058 Barnes, Karen Astete, Helvio 153, 787 Azevedo, Lucas R. 393 Balasubramanian, Sujata 913, 1841 Barnes, Karen I. 678, 917, 897 Astorga, Nestor G. 540 Azevedo, Luiz S. 718 Balazova, Miriam 1517, 1518 Barnes, Kayla G. 1188 Aswa, Daniel 370 Azimzadeh, Philippe 389 Baldeviano, Geral C. 1699, 1737, Barnes, Samantha J. 994, 1585 Atawurah, Henry 1210 Aziz, Fatima 54 1738, 277, 397, 604, 545 Barnett, Elizabeth D. 1282 Athanase, Badolo 117 Aziz, Nabil 1706 Baldini, Francesco 1625, 1817 Barnett, Eric 913 Athrey, Giridhar 1351, 1355 Azman, Andrew 632, 1650 Baldwin, Susan L. 1595 Barnor, Jacob 848 Atia, Atia Abdallah 1103, 1229 Azman, Andrew S. 38 Ballantyne, Christie 698 Barnwell, John 621, 1908, 911, Atieli, Francis 760 Azziz-Baumgartner, Eduardo 1115, Ballard, Sarah-Blythe 636, 852 1807 Atkinson, Barry 211 1266, 1771, 1777 Ballén, Diana 1150 Baro, Nicholas 320, 1833, 930, Atkinson, Peter 653 Balmaseda, Angel 191, 192, 806, 1628 Atkinson, Peter M. 1290 1273, 1275, 1771 Barogui, Yves T. 1655 Attah, Simon K. 1716 B Balogh, Beatrix 1630 Barral, Aldina 1203 Attaher, Oumar 1539 Baloji, Sylvain 1786 Barral-Netto, Manoel 1555 Ba, El hadji 302, 658, 359, 330, Attiogbe, Gédéon 1136 Balsinger, Jack 880, 280 Barratt, Michael J. 1654 1906 Atuguba, Frank 883 Baltazar, Palmera 56, 71 Barratt-Boyes, Simon M. 818 Ba, Fatou 246 Atwal, Sharan 348 Baltzell, Kimberly 1040 Barrera, Roberto 810 Ba, Mady 246, 278, 1250, 1456 Atwell, Jessica E. 1119 Balu, Bharath 231, 890 Barreto, Mauricio L. 1265 Baas, Lisette Baas 358 Auburn, Sarah 958 Bamadio, Modibo 25 Barrett, Alan 802 Baba, Ebenezer 265 Audi, Allan 824, 1902 Bamrungtrakul, Thavisakdi 1120 Barrett, Leah 1639 Baba, Marycelin M. 859 Audi, Allan O. 1045 Banajee, K. 106 Barrie, Assiatu 1177 Baba-Moussa, Lamine 1192 Audibert, Martine 703 Banajee, Kaikhushroo H. 104, 113, Barrientos, Franklin P. 51 Baber, Ibrahima 796, 1371, 1509 Auma, Ann 402 1217 Barrios, Ana L. 1115 Babineau, Denise 1560 Austin, Amy L. 107 Bancone, Germana 328, 940, 1920 Barron, Alexander M. 1554 Babji, Sudhir 585 Austin, Laura 17 Banda, Tamara 9, 651, 1271 Barron, Ellen J. 1126 Babu, Josephin Justin 1195 Auvergne, Stephane 830 Bando, Hironori 126 Barry, Aïssata 1590 Babu, Narasimhan P. 29 Avelar, Livia G. A. 1789 Banea, J. P. 1256 Barry, Amadou 1539 Bacher, John 613 Avendaño-López, Adrian E. 812 Banek, Kristin 243 Barry, Amanda 1087 Backers, Sharone 1259 Avery, Vicky 1498 Banerjee, Rajdeep 16 Barry, Meagan A. 1931 Badara Ly, Alioune 1694 Avila, Mario 663 Banga-Mboko, Henri 1765 Bartelt, Luther A. 1178 Badgandi, Hemant 750 Avilés, William 725 Bangirana, Paul 24 Bartholomay, Lyric 490, 492 Badolo, Athanas 1019 Avril, Marion 222 Bangs, Derek J. 822, 1275 Bartholomeu, Daniella C. 1263 Badolo, Athanase 126 A Wali, Abdul R. A. 1419 Bangsberg, David 1487 Bartkovjak, Marian 531, 1518 Baea, Manasseh 1212 Awandare, Gordon 1443, 1445, Baniecki, Mary Lynn 1805 Bartlett, John A. 1881 Baer, Alan 166 409, 1404 Banjo, Olajide 256 Bart-Plange, Constance 1845 Baeza Garcia, Alvaro 387 Awando, Janet A. 1045 Banla, Meba 481, 506 Barusya, Chris 946 Bagdasarian, Michael 1368 Aweeka, Francesca 26, 257, 1457, Banouvong, Virasack 1782 Barwa, Tara 1919 Bagiella, Emilia 1197 1458 Bansal, Geetha P. 404, 1000, 1586 Basanez, Maria Gloria 518, 1232, Bah, Germanus S. 1890 Awine, Timothy 883, 1872 Bansil, Pooja 506 1330, 1871 Bahita, Ashenafi Assefa 591 Awino, Norbert 666 Banu, Nuzhat N. 1874 Bashir, Nasir Mohammed 1907 Baiatone Alencar, Ronildo 1327 Awiti, Alphonce 1071 Baquerizo, Valerie 1295 Bashir Aamir, Uzma 54 Baiden, Frank 1299 Awodele, Olufunsho 1475 Baqui, Abdullah H. 1312 Basika, Tatiana 1781 Baidjoe, Amrish Y. 41, 700, 1521 Awolola, Taiwo S. 124 Baragana, Beatriz 287 Basilico, Nicoletta 288, 1461 Baig, Farrukh 1347 Awor, Phyllis 1577 Barahona, Martha 201 Basnyat, Buddha 1641 Bailey, Adam L. 665 Awuondo, Ken 876, 1556 Baraka, Vito 318 Bassat, Quique 50, 340, 470, 1280, Bailey, Charles 157 Axen, Heather 1337 Barban, Veronique 835 1482 Bailey, Jason 389 Ayala, Eduardo R. 453 Barber, Bridget E. 22 Bastos, Maria de Lourdes S. 1118 Bailey, Jason A. 1605, 1796 Ayala, Ramses 1606 Barbosa, Danielle R. Lima. 1563 Bastos, Melissa 1224 Bailey, Jeffrey A. 1454 Ayanniyi, Aderonke 1475 Barbosa, Fernando S. 1263 Basu-Roy, Upal 610 Bailey, Robin L. 1048, 1877 Aydin-Schmidt, Berit 868 Barbosa, Lúcio M. 1179, 1813 Batchelor, Adrian 1867 Bailey, Trevor 730 Aye, Kyin Hla 1235 Barbosa, Susana C. 1224 Batchelor, Joseph D. 995 Baird, J. K. 360 Ayede, Adejumoke I. 253 Barcellos, Christovam 730 Batcho, Wilfrid 1078 Baird, Sarah 64, 467, 473 Ayers, Tracy 582, 601, 1261 Bardají, Azucena 338 Batengana, Bernard 768 Bakajika, Didier 476, 1063, 1716 Ayi, Irene 409, 468, 472, 570, Bardera, Ana I. 1932 Bates, Imelda 1053 Bakalar, Matthew 1208 1092, 1093 Barends, Marion 268 Bates, Paul 1092 Bakamutumaho, Barnabus 1773 Ayisi-Boateng, Nana K. 1210 Barett, Alan D. T. 205 Bathily, Aboudramane 962, 1552, Baker, Bill J. 1695 Ayorigiya, Thomas 1445 Bargues, Maria Dolores 1780 1572, 1798 Baker, Julia M. 54 Aysanoa, Esar 1737, 1738 Baric, Ralph 575, 576, 626, 625, Batista, Camilla L. 1224 Baker, Mark 675, 1490 Ayukenchengamba, Bate 918 628 Batista, Izabella C. A. 858 Baker, Stephen 430, 1641, 1642 Ayyoub, Amal 1486 Barillas-Mury, Carolina 682, 759, Batsa, Linda 1207, 1210, 1713 Balabanidou, Vasilia 1186 Azad, A. 106 1008 Battle, Katherine E. 360 astmh.org A-598 Abstract Author Index The number(s) following author name refers to the abstract number.

Batwala, Vincent 268 Beltrán, Efraín 569, 916, 1411 Bhatt, Samir 360, 943, 944, 945, Blake, Rachel A. 71 Batzloff, Michael 1042 Benavidez, Yoldy 225 1929 Blanc, Patricia R. 830 Baudin, Elisabeth 1487 Benca, George 530, 1517 Bhattacharyya, Arindam 386 Blanchard, Anne 1745 Bauleni, Andy 953, 1522 Benca, Juraj 1518 Bhattarai, Achuyt 927, 929, 1035 Blanco-Tuirán, Pedro 1408 Baum, Danielle 1934 Bendezu, Jorge 1807 Bhattarai, Anish 1856 Blaney, David 477 Baum, Elizabeth 937, 1567, 1223 Benedict, Mark Q. 7, 1819, 1884, Bhongo Mavoungou, Lelia Blank, Walter A. 1813 Baum, Jake 623 1885 Chadlene 977 Blankenship, D’Arbra 604 Bausch, Daniel G. 167, 184, 803, Benezra, Amber 1654 Bhoomiboonchoo, Piraya 1770 Blanton, Ronald E. 1179, 1813 852, 1284, 1295, 1417, 1420, Benitez, David 200 Bhuiyan, Taufiqur R. 35, 434, 583 Blaxter, Mark L. 1890 1755, 1757 Benjamin, Tinkitina 1160 Bhullar, Vinod 1064 Blay, Emmanuel A. 468, 472 Bausell, Loren 1627 Ben Mbarek, Nathalie 1934 Bhutta, Waqaas 421 Blazes, David 1851 Bautista, Christian T. 1304 Ben Messaoud, Nathalie 1934 Bhutta, Zulfiqar A. 421 Blazes, David L. 184, 803 Baxter, Peter 1290 Bennett, Adam 1470, 1507 Bhuyan, A. A. Mamun 1756 Blessborn, Daniel 257, 311 Bayih, Abebe G. 285 Bennett, Andrew J. 1431 Bianco, Simone 1414 Blessbron, Daniel 301 Baynesagn, Mekonen Getahun 87 Bennett, Jason W. 252 Bicalho, Kelly A. 61 Bliss, Carly 671, 1590 Bayoh, Nabie M. 760, 1887 Bennett, Shannon 798, 805 Bich Chau, Tran Nguyen 783 Blitvich, Bradley J. 649 Bayot, Bonny 201 Bennuru, Sasisekhar 29, 488, 1213, Bickham, Utibe 1790 Blizzard, John 1671 Bazeyo, William 599 1891 Bickle, Quentin 1787 Bloch, Kenneth 1487 Bazira, Joel 1283 Benoit-Vical, Francoise 1237 Bidii, Ngala 306 Blohmke, Christoph J. 1849, 1850 Beall, Bernard 1064 Ben Salah, Afif 1934 Bieler, Sylvain 461 Blum, Laura 477 Beaso, Delma 1212 Bentsi-Enchill, Adwoa D. 1306 Biemba, Godfrey 637, 706 Blum, Lauren S. 711, 712 Beatty, P. Robert 1834 Berg, Maya 688 Bierwert, Lou Ann 1725 Boakye, Daniel A. 409, 468, 1092, Beaty, Barry 137 Berger, Jacques 1721 Biggs, Holly M. 661, 1758 1093, 1248, 1330, 594, 1114, Beau De Rochars, Madsen V. E. Bergman, Lawrence W. 980 Bigira, Victor 24, 26, 257, 903, 1207, 1526, 1622 1566 Bergmann-Leitner, Elke 1595 1569 Boamah, Ellen A. 1067 Beaudoin, Amanda 1215, 1878 Bergqvist, Yngve 257 Bigogo, Godfrey 824, 1902, 1045 Boateng, Richard 1302 Beaudoin, Jennifer 286 Berhe, Nega 316 Bijker, Else M. 992, 1602 Boaz, Mark 575, 576, 577, 578, Beaumier, Coreen M. 1931 Berkley, James A. 347 Bilak, Hana 1532 831, 833 Bebrevska, Lidiya 287, 675 Bern, Caryn 548, 564, 716, 1102, Bilgic, Fatma 1236 Bobanga, Thierry L. 1463, 1469, Bechir, Khalid 1527 1930 Bilgo, Etienne 757 1538, 1547, 1633, 1633, 1634 Beck, Hans-Peter 1550 Bernabeu, Maria 223 Biligui, Sylvestre 1498 Bobbili, Naveen 228 Beck, Josh R. 1943, 1946 Bernal Garcia, Sebastian 361 Billeter, Sarah A. 105 Bobogare, Alby 872 Becker, A. C. 1940 Bernal-Rubio, Dabeiba 1839 Billingsley, Peter F. 670, 876, 1556, Bocci, Alessandro 1885 Becker-Dreps, Sylvia 586 Bernard, Samuel 1163 1602, 1608, 1610, 1620, 669, Boccia, Delia 1278 Beckwith, Colin L. 1075 Bernart, Chris 1643, 1777 1592 Bock, Ronnie A. 1502 Bedenbaugh, Rachel 1163 Berrardi, Victor 1216 Billman, Zachary P. 270 Bockarie, Moses 591, 594, 1701, Bedoya-Vidal, S. 1769 Berrie, Eleanor 671, 672 Binh, Nguyen T. 1519 1708, 1690, 1696 Beebe, Nigel W. 872, 1914 Berriman, Matt 547, 488, 1811 Birbeck, Gretchen L. 20 Bodeau-Livinec, Florence 635, Beer, Netta 927 Berry, Andrea A. 270, 389, 1605, Biritwum, Nana-Kwadwo 1689 1257, 1916 Beeson, James G. 606 1796 Birrell, Geoffrey 249 Bodhidatta, Ladaporn 585 Begashaw, Kalkidan Mekete 591 Berry, Neil G. 484 Bischoff, Emmanuel 1910 Bodoor, Khaldon 813 Begum, Sharmin 1426 Bertollo, Caryne M. 1094 Biselli, Joice M. 194 Boehm, Alexandria 1058 Begum, Yasmin 432 Bertuso, Arlene G. 1349 Bishop, Henry S. 1182 Boelaert, Marleen 522, 1157, 1233 Behrman, Jere 600 Besansky, Nora J. 680 Bishop, Richard P. 1175 Boelart, Marleen 1786 Bei, Amy K. 234, 1450, 1799 Beshir, Khalid B. 323, 1493 Biswas, Hope 192 Boele van Hensbroek, Michael 1861 Beido, Nassirou 1066 Bessette, Richard E. 1060 Bita Kwenti, Tayong Dizzle 503 Boer, Kimberly R. 358 Beisel, Uli 1326 Bessong, Pascal 585, 1639 Bitek, Austine O. 660 Boete, Christophe 1326 Bejar, Vilma 1284 Best, Abigail 1689 Bittencourt, Valéria 1118 Boggild, Andrea K. 466, 538, 1095 Bejarano, Eduar E. 110 Best, Shannon E. 1437 Biya, Oladayo 633 Bogoch, Isaac 1929 Bejon, Philip 671, 1556, 1590 Bethell, Delia 299, 1494, 1774 Bizimana, Jean Pierre 358 Bogoch, Isaac I. 66 Bekele, Abyot 215 Betson, Martha 124, 1109 Björkman, Anders 655, 868, 927, Boillat, Noémie 1065 Belay, Kassahun 893 Bett, Andrew 197, 580, 1378 929, 1040, 324 Boisson, Sophie 710 Belemvire, Allison 781, 1551 Bett, Glenna C. 1943 Black, Carolyn 1064 Boivin, Michael J. 24, 635, 1256, Belenchia, Matthew 499 Bettaieb, Jihene 1934 Black, Robert E. 1312 460 Bel Haj Hamida, Nabil 1934 Betuela, Inoni 1531 Black, IV, William C. 137, 779, 837 Bojang, Kalifa 671, 720, 1544 Bell, David 461, 871, 1514, 1807 Betzana, Zambrano 577 Blackburn, Brian G. 66 Bolay, Fatorma K. 982 Bello, Felio J. 1338 Bever, Caitlin A. 667, 1542 Blackburn, Jason 663 Bolick, David T. 1178, 1639 Bello, Gonzalo 1775 Bewa, Christopher 950 Blackmore, Carina 1390 Bolton, Jessica 608, 1603 Belloni, Virginia 1909 Beyene, Berhane 87 Blacksell, Stuart 1852 Bonanno, Daniela 643 Belmonte, Arnel 1005, 1603 Bezabih, Belay 332 Blackwell, Jenefer 691 Boncy, Jacques 34 Belmonte, Maria 1005 Bezerra, Rita 718 Blagborough, Andrew M. 284, Bond, Craig V. 226 Belofsky, Gil 515 Bezerra-Fernandez, Carmen 233 1029 Bonfoh, Bassirou 659, 1114 Beltran, Davis 663, 1423 Bhat, Geeta 276 Blair, Patrick J. 733, 737, 1429 Bonhoeffer, Sebastian 896 astmh.org Abstract Author Index A-599

The number(s) following author name refers to the abstract number.

Bonizzoni, Mariangela 758 Bowman, Natalie M. 548 Brosi, Berry J. 1882 Burkett-Cadena, Nathan D. 1695 Bonnabau, Henri 795 Bowyer, Georgina 671, 1590 Brouwer, Kimberly C. 329 Burkhardt, Martin 1008, 1621 Bonne-Annee, Sandra 1264 Bowyer, Paul 1443 Brown, Arthur 1434 Burkot, Thomas R. 872 Bonnet, Emmanuel 1403 Boyce, Ross 873 Brown, Charles A. 1036 Burnett, Jennifer 1316 Bonnet, Maryline 1283 Boyett, Deborah 1788 Brown, Heidi E. 1736 Burnett, Sarah M. 275 Bonney, J. H. Kofi 1404 Boyko, Ryan H. 519 Brown, Joelle M. 348, 1509 Burns, James M. 228 Boone, Katelyn 1165 Boyle, David 1318 Brown, Kimberly 880, 280 Burns, Matthew 1504 Bopda, Jean 65, 1208 Boyle, Michelle 402, 1569 Brown, Lisa D. 1217 Burns, Stephanie 1669 Bopp, Cheryl A. 582, 584 Bozo, Ricardo 1102 Brown, Paul 1772 Burr, Sarah 1877 Bopp, Selina 18, 617 Bozzi, Adriana 61 Brown, Sarah 888 Burrell-Saward, Hollie 1088 Borbor, Mercy 201, 1411 Bracken, Tara C. 224 Brown, Sheila L. 1938 Burrows, Jeremy N. 673 Borbor-Cordova, Mercy J. 569 Brackney, Doug E. 837, 982 Brown, Tyler S. 1605 Burt, Austin 969 Borchert, Jeff 1415 Bradley, John 774 Brown, William 842 Burton, Deron 1064, 1902, 1045 Borger, Jessica G. 1938 Bradley, Peter J. 1943 Brownstein, John S. 1422, 935, Burton, Robert 339, 1318 Borghini-Fuhrer, Isabelle 1486 Bradshaw, Thomas A. 1462 1929 Buscaglia, Carlos A. 1201 Borin, Khieu 721 Brady, Oliver 1929 Bruce, Nigel 1258 Bustamante, Dulce M. 1337 Borland, Erin 9, 861, 1271 Brady, Oliver J. 832 Bruder, Joseph T. 1604 Bustinduy, Amaya 1183 Borlon, Céline 229 Brady, Rebecca 39 Brugnara, Carlo 415 Bustos Arriaga, Jose 828 Born, Priscila S. 1775 Braga, Cynthia 1399 Bruhse, Laura 692 Butler, Elissa K. 78 Boro, Herbert 1772 Braganza Menezes, Darryl 1101 Brumeanu, Teodor-Doru 607, 1574, Butler-Dawson, Jaime 1164 Borodo, Musa M. 713 Braibant, Bertrand 286 1601 Butraporn, Piyarat 315 Borooah, Shyamanga 1101 Braide, Ekanem I. 1069 Brunetti, Enrico 439, 571, 639, Button-Simons, Katrina A. 890, Borrero, Elizabeth 1676 Brand, Nathan R. 1857 1668 898, 899 Borrmann, Steffen 1492 Brasil, Patricia 892, 1409 Bruniera-Oliveira, Robson 723 Buza, Joram 318 Bosco, John 5 Brault, Aaron C. 645, 646, 837, Brusic, Vladimir 1389 Bwanika, John Baptist 871 Bosco-Lauth, Angela 646, 845, 837 845 Brusich, Macy 1324 Byakika, Pauline 1577 Bose, Anuradha 634 Brawn, Jeffrey 842 Bruxvoort, Katia 28, 255, 1578, Byamukama, Edson 1693, 1693 Bosompem, Kwabena M. 472, Braykov, Nikolay P. 1761 1579, 1809 Byarugaba, Frederick 1283 1147, 1181 Brazier, Andrew Jay 222 Bryan, Brad B. 97 Byaruhanga, Timothy 1773 Bosquet, Nathalie 835 Bredenbeek, Peter J. 649 Bryan, Joe 1643, 1665, 1777 Byers, Anthony 576, 575, 833 Boström, Stephanie 964 Breiman, Robert F. 582, 584 Bryan, Julie 1096 Bygbjerg, Ib 254, 897 Bottai, Matteo 405 Breitling, Frank 806 Bryson, Lindsay 1536 Bottazzi, Maria Elena 517, 1931, Breitling, Rainer 688 Búa, Jacqueline 544 1892, 698 Brent, Bernadette 1861 Buathong, Nillawan 299, 912, 1481 C Bottomley, Christian 1844 Brenyah, Ruth C. 780 Bucala, Rick 387 Cabada, Miguel M. 427, 1173 Boubacar, Kadri 1066 Brewer, Jonathan 1044, 1671 Bucheton, Bruno 1100 Caballero-Garcia, Maria de Lourdes Boudet, Florence 835 Brewer, Timothy 935 Buchy, Phillipe 1774 1682 Boudova, Sarah 1226, 1559 Briand, Valerie 251, 936 Buckee, Caroline 83, 415, 44, 616, Cabarrus, Rita L. Vizcaino. 135 Bougdour Alexandre 1943 Brice, Gary 1429 1799 Cabello de Quintana, Maritza 202 Bougma, Roland 68 Brichard, Julie 9, 177, 651, 1271 Buckeridge, David 935, 1293 Cabezas, Cesar 397, 1284 Bougouma, Edith C. 1590 Bridges, Daniel J. 136, 922, 924, Budachetri, Khemraj 1340 Cabrera, Ana 221 Boukthir, Aicha 1934 925, 1504 Budge, Philip J. 505 Cabrera, Lilia 1279 Boulos, Marcos 901 Brieger, William 882, 1317 Buekens, Pierre 546, 1733 Cabrera-Mora, Monica 218, 603 Boum, II, Yap 1487, 1860, 1283 Brien, James D. 648 Bueno, Lilian 1263 Cabrerra, Lilia 1684 Boumediene, Farid 1196 Briët, Olivier 42 Buettner, Marcelle 1207 Cáceres, Abraham G. 1332 Bounds, Callie E. 13 Briet, Olivier J. T. 914 Bufano, Meagan K. 433, 588 Caceres, Lorezo 206 Bourgouin, Catherine 1910 Brindley, Paul J. 1781, 1814 Buff, Ann M. 237 Cáceres, Tatiana 1129, 1776 Bourguinat, Catherine 511 Brito, Miguel 1153, 1700 Buffet, Pierre A. 1498 Cafferata, Maria Luisa 1733 Bourhy, Pascale 1056 Brito, Patricia Kellen Martins Oliveira Bugoro, Hugo 872 Cairns, M. 1906 Bourne, Nigel 802 551, 552 Bulimo, Wallace 558, 559 Cairns, Mathew 1067, 254, 1520, Bousema, Teun 41, 325, 348, 700, Britton, Collette 486 Bull, Peter C. 1556 28, 1578 1018, 1509, 1521, 1842, 1843, Brnova, Jaroslava 530, 531, 1516, Bulo, Helder 1478 Cairo, Cristiana 1559 1865 1518 Bumoko, B. M. 1256 Cajal, Pamela 514, 643, 695 Boussari, Olayidé 122 Brock, Patrick M. 1029 Bumoko, Guy M. M. 460 Calcina, Juan F. 451 Boussinesq, Michel 65, 592, 1208 Brockley, Sarah 668 Bun, Rathvichet 912 Calderón, Enrique J. 1280 Bouyer, Donald H. 103 Broder, Ryan 833 Bunk, Sebastian 21 Calderon, Jose 746 Bouyou Akotet, Marielle Karine Brogdon, William 131 Bunleng, Sam 1334 Calderón, Maritza 414 977, 1473 Bronowski, Christina 489 Burgerhof, Hans 202 Calderón-Arguedas, Olger 778 Bouyoukou Hounkpatin, Larissa 401 Bronzan, Rachel 1136 Burgess, Timothy 471, 1051 Calderwood, Stephen B. 35, 432, Bowen, Katherine 402, 1569, 1864 Brooker, Simon J. 242, 508, 1690, Burhenne, Jürgen 1492 433, 588, 1651 Bowen, Richard A. 646, 845 591, 879, 1253 Burja, Kurt 1721 Calle, Alvaro 916 Bowler, Mark 1737, 1738 Brooks, Abdullah 801 Burke, Donald 1414 Calvo, Manuela 459 Bowman, Leigh 200, 763 Brooks, Tim 211 Burke, Rachel M. 741, 1645 astmh.org A-600 Abstract Author Index The number(s) following author name refers to the abstract number.

Calzada, Jose E. 930, 1328, 1436 Carbajal, Faustino 789 Castanha, Priscila M. S. 1399 Chan, Jo-Anne 606 Calzavara-Silva, Carlos Eduardo Cárdenas, Danitza 1128 Castanys, Santiago 688 Chan, Kuan Rong 180, 804 61, 858 Cardenas, Jenny C. 790 Castellanos-Gonzalez, Alejandro Chanarat, Nitima 299, 1774, 1841 Cama, Vitaliano 481, 1079, 1711, Cardenas, Lucio D. 790 690, 1173 Chancey, Caren 1383 1712, 1692, 1694 Cárdenas-Jaramillo, Luz María 1133 Castelli, Ilaria 1361 Chanda, Emmanuel 767 Camacho, Daria 202 Cardenas-Vegas, Brianda 515 Casteras, Jessica 1241 Chanda, Javan 767 Camacho, Leslie 696 Cardoso, Maria R. A. 1876 Castillo, Alex 663 Chandler, Clare I. R. 243, 885 Camacho-Burgos, Erwin 1408 Careagabarja, Julio 252 Castillo, Andreina 970 Chandler, James 798 Camara, Mamady 1100 Carey, Cristiam 807, 1025 Castillo, Elizabeth 185 Chandonait, Peter J. 1636 Camarda, Grazia 288 Carias, Lenore 1455 Castillo, Estela 1781 Chandramohan, Daniel 243, 348, Camargo, Nelly 1897 Caridha, Diana 252, 282, 283, 291 Castillo, Giancarlo 1134 720, 1067, 1227, 1466, 1520, Camargo-Camargo, Blanca 1739 Carissimo, Guillaume 1910 Castro, Carlos J. 412 1917 Camargo-Paredes, Carolina 1099 Carlsen, Eric D. 1205 Castro, Fanny 144 Chandrappa, Kavitha 177 Cameron, Ewan 943, 944, 945 Carmargo, Nelly 1018 Castro, Márcia C. 1224 Chandrasekera, Ruvani M. 1647 Camilli, Andrew 34 Carmo, Theomira M. A. 1813 Castro, Yagahira E. 875, 1106, Chandre, Fabrice 118, 122, 1192 Campbell, Carl H. 1530 Carmolli, Marya 181, 579, 1288, 1930 Chandyo, Ram K. 634 Campbell, Karen M. 1412 583 Castro Rodriguez, Raul 1388 Chang, Howard 602, 1169 Campbell, Scott 794 Carmona, Santiago J. 1201 Casulli, Adriano 439 Chang, Hsiao-Han 930, 1833 Campbell, Simon 287 Carmona Fonseca, Jaime 408 Catteruccia, Flaminia 681, 1625, Chang, Michelle 893, 1908 Campbell, Suzy 597 Carn, Gwenaelle 1483 1817, 1889, 1915 Chang, Sandra P. 1447 Campetella, Oscar 1201 Carneiro, Claudia M. 1263 Cauchemez, Simon 45, 195, 196 Changalucha, John 123, 743, 1466 Campino, Susana 1239 Carneiro, Ilona 1227 Caulfield, Laura E. 634 Chann, Soklyda 299, 912, 1481 Campo, Joe 338 Carneiro, Jr., Nivaldo 1230 Cavalcante, Paloma A. 1648 Chan Quang, Luong 200 Campo, Joseph J. 1567, 1604 Caro, Nicolas 643 Cavalcanti, Marília Gabriela dos Chan Sophal, Ngan 1334 Campos, Karen 1699 Caroll, Lauren 1293 Santos G. S.. 1790 Chansue, Nantarika 1137 Campos, Miguel 1858 Carpenter, David 599 Cavanagh, David 381 Chantha, Ngan 1334 Campos, Ronald B. 51 Carpio, Arturo 1197 Cavasini, Carlos E. 1224 Chanthap, Lon 1774 Campos Ponce, Maiza 1721 Carr, Steve 50 Ceccato, Ceccato 1743 Chanthongthip, Anisone 1046, Campos-Rodríguez, Rafael 1133 Carrasquilla, Gabriel 577, 578 Cecchi, Giuliano 1740 1853 The Canadian Coalition of Global Carrera, Claudia 388 Cecilio-Elfa, Dulce 739 Chanudom, Ponsa 1841 Health Research International Carrera, Jean Paul 663, 206, 1423 Ceesay, Serign 373, 314, 1544, Chao, Chien-Chung 1664 Mentorship Working Group Carrillo-Tripp, Jimena 649 1544 Chaorattanakawee, Suwanna 299, 1301 Carrington, Lauren B. 784 Celeste, Beatriz J. 1742 912, 913, 1454 Canavati, Sara 337, 1499 Carrington, Mary 630 Celestino, Daniela 1206 Chapagain, Moti L. 647 Canedy, Jordan C. 676 Carrion, Jessica 1415 Celis, Violeta 1428 Chapleau, Gina 62 Canelo-Aybar, Carlos 883 Carroll, Ryan W. 1487 Cerami, Carla 1444, 1451 Chapman, Colin A. 665 Canepa, Gaspar 682 Carter, Bryan 963 Cerpas, Cristhiam 1275 Chaponda, Michael 956, 957 Cangalaya, Carla M. 455 Carter, Dariyen 613 Cesard, Nicolas 1326 Chapotera-Kalanda, Gertrude 678 Canier, Lydie 657, 926, 1514, 1626 Carter, Darrick 1595 Cevallos, William 1761 Chappuis, François 692, 1679, Cano, Jorge 1917 Carter, Emily 84, 731 Cevenini, Luca 288 1103, 1229, 1743 Cantelena, Louis 1481 Carter, Tamar E. 978 Chacky, Frank 351, 1287 Charabati, Jirias 1337 Cantey, Paul 493 Carvalho, Andrea T. 61 Chagla, Zain 93, 1675 Chard, Anna 602, 1169 Cantey, Paul N. 1711 Carvalho, Augusto M. 1203 Chai, Jong-Yil 1782 Chard, Anna N. 1136 Cantey, Paul T. 1692, 1694, 1712 Carvalho, Edgar M. 1118, 1203, Chaiwan, Jintana 315 Chareonviriyaphap, Theeraphap Cantizani, Juan 1932 1206, 1734, 1749 Chakir, Ismael 454 1324 Cao, Jun 128, 958 Carvalho, Eunice B. 1648 Chakma, Sumit 1359 Charland, Katia 935 Cao, Xiaohang 1824 Carvalho, Lucas P. 1203, 1206, Chakraborty, Poulomy 82 Charles, Richelle C. 34, 35, 36, 432, Cao, Yuanyuan 128 1749 Chakraborty, Sounak 1303 433, 588 Cao-Lormeau, Van-Mai 1923 Carvalho, Maria E. 1540 Chakravarty, Jaya 691 Charman, Nikki 877, 1528 Capobiano, Marcela P. 391, 393, Carvalho, Maria da Gloria 1064 Chakravarty, Sumana 1005, 1594, Charman, Susan 287, 673 396 Carvalho, Noemia B. 718, 1230 1610 Charnay, Christophe 835 Cappello, Michael 519, 1526, Carvalho-Dantas, Ive M. 1200 Chalfein, Ferryanto 219 Charron, Brigitte 1473 1717, 1720 Casanova, Wilma 1928 Chalker, John 1875 Chartrel, Nathalie 1498 Captain-Esoah, Millicent 142 Casapia, Martin 1269 Chambela, Mayara C. 536 Charunwatthana, Prakaykaew 1120 Caputo, Beniamino 1352, 1367, Casares, Sofia 607, 1574, 1601 Chambi, Marcos 1134 Chase, Amanda J. 1835 1912 Casenghi, Martina 1653 Chamnan, Chhoun 1721 Chatterjee, Gangadhar 611 Caputo, Francesco Paolo 1352 Casey, Joanne L. 1585 Chamot-Rooke, Julia 1194 Chatterjee, Soumya 1277 Carabali, Mabel 745, 1403 Casimiro, Danilo R. 1378 Champagne, Deborah B. 236 Chattopadhyay, Pratip 1800 Caraballo, Elba V. 1386 Cassama, Eunice 1877 Chan, Adeline 1847 Chau, Nguyen V. V. 1384 Carabin, Helene 1193, 1195, 1196 Cassiano, Gustavo C. 391, 393, Chan, Chim 655, 949 Chaudhry, Suchita 1559 Caranci, Angela T. 1735 396, 1563 Chan, Ernest R. 621, 622 Chaudhury, Sidhartha 1595, 1597 Carapetis, Jonathan 1854 Cassidy, Andrew 483 Chan, Grace J. 740, 1312 Chauhan, Virander S. 1002 Caravedo, Maria A. 427 Castañeda, Benjamin 51 Chan, Hwang Ching 965 Chaurasiya, Narayan D. 674 astmh.org Abstract Author Index A-601

The number(s) following author name refers to the abstract number.

Chauty, Annick 1655 Chinh, Nguyen T. 249 Cisse, B. 1906 Colgate, Elizabeth R. 583 Chavarría, Johnny 201 Chinnawirotpisan, Piyawan 1391 Cissé, Badara 254, 302, 330, 359, Coll, Francesc 1877 Chavchich, Marina 249 Chintala, Ramesh 1378 394, 658, 986, 1520 Coller, Beth-Ann G. 197, 580, 1378 Chaves, Ana Thereza 61 Chipeta, James 313 Cisse, Kadidia B. 1539 Colley, Daniel G. 57, 1795 Chaves, Luis F. 655, 1328 Chipwaza, Beatrice 169 Cisse, Moustapha 1250, 1456 Collier, Timothy 1917 Chaves Martins, Yuri 612 Chirwa, Brian 136, 767 Cisteró, Pau 338 Collins, C. M. 1884 Chawla, Nitesh 797 Chirwa, Gowokani 731 Claessens, Antoine 1899 Collins, Frank H. 872, 1341 Chawla, Swati 1027 Chisenhall, Daniel M. 790, 792, Claeys, Yves 1856, 1922 Collins, Katharine 671 Chayapum, Kwanta 1137 823, 1393 Clapham, Hannah 1392, 1395, Collins, Katharine A. 1587 Cheah, Phaik Yeong 1308 Chisha, Zunda 922, 924, 925, 1504 816, 1380 Colmenarejo, Gonzalo 1932 Chebon, Lorna J. C. 300 Chitadze, Nazibrola 1304 Clara, Wilfrido A. 1771 Coloma, Josefina 188, 725, 726 Checkley, Lisa 908, 1240, 1897 Chitnis, Chetan E. 672, 338 Clare, Rachel 483, 487, 495 Colombo, Tatiana E. 194 Checkley, William 51 Chitnis, Nakul 41, 1511 Clark, Brent S. 1462 Colón-González, Felipe J. 1497 Cheelo, Sanford 922, 924 Chiza, Richard 1773 Clark, Daniel 548 Colpitts, Tonya 176 Cheeseman, Ian H. 906, 983, 1832, Chizema, Elizabeth 1708 Clark, Erin 1016 Colquechagua Aliaga, Fabiola D. 1236 Cho, Cho 864 Clark, Eva H. 1102 636 Cheick, Sangare P. O. 1527 Choi, Jeong 548 Clark, Martha A. 1444, 1451 Colson, Katherine E. 1618 Cheikh, Sokhna 1822 Chokephaibulkit, Kulkanya 183 Clark, Michael 797 Comach, Guillermo 820 Cheke, Robert A. 1330 Chokheli, Maiko 1304 Clark, Roger 495 Combescure, Christophe 1679 Chellappan, Savitha 407 Chotivanich, Kesinee 268 Clark, Taane G. 958, 1877 Comer, Eamon 286, 292 Chema, Celia 80 Chotiwan, Nunya 1276 Clarke, Sian E. 25, 885 Comte, Eric 1653, 1855 Chen, Edwin 995, 1585 Chou, Pei-Yun 513 Clasen, Thomas 708, 710 Conceição, Luciana M. 391, 393, Chen, Gong 673 Chounna Ndongmo, Winston Clavijo-Ramirez, Carlos 541 396 Chen, Hongjing 1934 Patrick 503 Cleaveland, Sarah 77, 664, 1758 Conn, Jan E. 149, 149, 1823 Chen, Hua-Wei 1664 Chowdhury, Anwarul H. 1426 Clemente, Isabel 1153 Conrad, Melissa D. 903 Chen, I-Tzu 513 Chowdhury, Fahima 35, 433 Clements, Archie 518, 521, 597 Conroy, Andrea L. 722, 49, 1485, Chen, Nanhua 887 Chowwiwat, Nongnud 328, 940 Clerk, Christine 339 1859 Chen, Rubing 1397 Choy, Milly M. 199 Clermont, Miliane 1163 Constantini, Carlo 1373 Chen, Shicheng 1368 Christensen, Bryan 1258 Cliffe, Matthew 1689 Contamin, Benedicte 454 Chen, Tien-Huang 207 Christofferson, Rebecca C. 10, 165, Clifford, Robert 1851 Conteh, Lesong 1232, 1871 Chen, Wei-June 207 843, 1393 Clish, Clary C. 907 Conteh, Solomon 248, 1593 Chen, Wilbur H. 39 Christophides, George K. 1335 Co, Mary 630 Conti, Nico 210 Chen, Xiaoguang 758, 1353 Chu, Brian 1259 Coalson, Jenna E. 953, 1225, 1522 Contreras, Arnol 110 Chen, Yao-Shen 513 Chu, Cindy S. 1670 Coates, Craig J. 1356 Contreras, Eliza 1134 Chen, Yu-Shin 513 Chu, Haiyan 11, 182, 802 Cobos, Daniel 1581 Contreras-Mancilla, Juan Jose 959, Chen, Zhongyuan 156 Chu, Yui Yin 787 Coelho, Fernanda S. 1789 1541, 966 Cheng, Changde 680 Chua, Tock Hing 139 Coelho, Giovanini 200 Conway, David 1352, 1442, 1443, Cheng, Qin 887, 958 Chuang, Ilin 1494, 1774 Coelho-dos-Reis, Jordana 1606 1445 Cheng, Xixi 1458 Chuang, Ting-Wu 809 Coetzee, Maureen 766 Conway, Michael J. 210, 214 Cheng, Yang 864, 993 Chuanshan, Zhang 440, 442 Cohee, Lauren 1226, 1225, 1522 Cook, Darren A. N. 483, 485, 495 Cherif, Mahamoud S. 400, 985 Chuansumrit, Ampaiwan 183 Cohee, Lauren M. S. 953 Cook, Jackie 655, 868, 927, 929 Cherry, Sara 648 Chubinidze, Marina 1304 Cohen, Elizabeth 69 Cook, Peter C. 1938 Cheruiyot, Nancy J. 303 Chukwuocha, Uchechukwu M. Cohen, Justin M. 45, 886, 1536, Cooley, Gretchen 1087 Chery, Laura 910 260, 529 344, 652, 878 Cooper, Michael J. 52 Chesnais, Cédric B. 65, 592, 1208 Chung, Eun Joo 1783 Cohen, Mitchell B. 39 Cooper, Philip J. 1265, 1268 Chevalier, Frédéric 1810 Chung, Ida H. 107 Cohn, Jennifer 1653 Cooper, Robert D. 872, 1914 Chhorvann, Chhea 733 Chuor, Char Meng 912, 913 Cohuet, Anna 284 Cooper, Roland 291, 319 Chi, Benjamin 963 Chuor, Meng C. 298 Coker, Richard 721 Coosemans, Marc 657, 926, 1033, Chiaravalloti Neto, Francisco 827 Chuquiyauri, Raul 144, 227, 329, Colacicco-Mayhugh, Michelle G. 1514, 1624, 1626 Chico, Martha E. 1265, 1268 414, 1025, 1510, 1823, 1858 1369 Coppellotti, Olimpia 119 Chico, R. Matthew 1466, 1917 Churcher, Thomas S. 45, 284, Colaco, Rajeev 699 Coppens, Delphi G. M. 1298 Chiduo, S. 280, 276, 880, 1523, 1018, 1029, 999, 1488 Colantuoni, Elizabeth 1496 Corbel, Vincent 118 1549 Chusri, Sarunyou 1039 Colanzi Zeballos, Ronny P. 548 Corbett, Mark 101 Chies, José A. Bogo. 1563 Cianci, Fiona 1581 Colares, Jeová K. B. 1406 Corbett, Yolanda 288 CHIKV RNA Reference Reagent Cicéron, Liliane 1498 Colborn, James 1535 Cordeiro, Marli T. 1399 Working Group 160 Ciesla, Jamie 1533 Colborn, Kathryn 1535 Cordon-Rosales, Celia 1115, 1665 Chikviladze, Tamar 1304 Ciganda, Alvaro 1733 Coldren, Rodney 559 Cordova Rojas, Marisol 1104 Childs, Lauren M. 44 Cirelli, Kimberly M. 1943 Cole, Andrew O. 1556 Corey, Kristin 370 Chimbiya, Nelson 953, 1225, 1522 Cimino, Ruben 514, 643, 695 Cole, Donald C. 1301 Corey, Victoria C. 294 Chimello Ferreira, Aline 827 Ciraiz, Rafael 1665 Coleman, Jane 1316 Cormick, Gabriela 1733 Chimuna, Tiyese 879, 1253 Cisneiros, Patrícia 1749 Coleman, Michael 767 Corrales, Homer 1408 Chin, Stephanie 1048 Cisneros, Julio 663 Coleman, Sylvester 1622 Correa, Malena 51, 457, 1296 Ching, Wei-Mei 1664 Cissao, Yacouba 1299 Colgate, E R. 1288 Correa, Margarita M. 140, 1358 astmh.org A-602 Abstract Author Index The number(s) following author name refers to the abstract number.

Corrêa-Oliveira, Guillerme 1810 Crowe, James 575, 576 Daly, Thomas M. 980 Davies, D. Huw 1567 Correa-Oliveira, Rodrigo 61, 536 Crowe, James B. 626 Dama, Souleymane 902, 1492, Davies, Douglas R. 1038 Cortes, Margarita 577, 578 Crudder, Chris 339 1527 Davies, Huw 329, 1510, 1552, Cortes-Vecino, Jesús A. 1338 Crump, John 467, 473 Damasceno, Camila 135 1562, 1223 Corujo, Alfredo 426 Crump, John A. 1758, 1881 Damasio, Marcos P. 536 Davies, Jill 502 Corvo, Ileana 1781 Cruz, Diego 514, 643 D’Ambrosio, Michael 1208 Davis, Chris B. 606 Cosgrove, Cormac 630 Cruz, Isabelle F. S. 1315 Damen, James 950 Davis, Emma L. 1723 Cosmas, Leonard 819, 1869, 1260 Cruz, Oswaldo G. 1409 Damiano, Silvia 119 Davis, Justin K. 843, 1372 Cosme, Luciano 1351, 1356 Cruz-Chan, Vladimir 1750, 1936 Damiens, David 920 Davis, Kaitlin A. 841 Costa, Fabio T. M. 233 Cruz-Reyes, Alejandro 1739 Dance, David A. B. 1046, 1853 Davis, Timothy 1803 Costa, Jackson M. 1179, 1813 C. Silva, Joana 1596 Danforth, Mary E. 141 Davy, Taing 912 Costa, Maria Selma Neves 577, 578 Cucunubá, Zulma M. 1232, 1234 Daniel, Benjamin J. 983, 1832 Dawahi, Chadhi 1934 Costa, Monica R. F. 390 Cuéllar, Victoria M. 598 Daniel, Gabriel 1077 Dawit, Asrat 509 Costa-Nascimento, Maria J. 901, Cui, Liwang 371, 382, 937, 941, Daniel, Simone 135 Dawood, Fatimah 1300 1540, 874 993, 1223, 1537 Daniel-Ribeiro, Cláudio T. 677, 892, Dawson, Nikiah 1621 Cot, Michel 251, 635, 936, 1257, Culzoni, Maria 1873 390 Day, Nicholas J. 1107 1916 Cummings, Derek A. T. 816, 829, Daniels, Miles 1762 Dayan, Anthony 675 Cotter, Chris 654, 915, 1513 1272, 1380, 1392, 1395, 1400, Daniels, Noah M. 1833 Dayan, Gustavo H. 577, 578, 831 Cottingham, Matthew 1587 1414 Daniels, Rachel 320, 331, 904, 930, De, Sai Lata 1003 Cotton, James 488, 547, 1811 Cummings, James F. 738, 744, 1628, 1805, 1833, 1799 de Almeida, Mariana 287 Cotton, Rachel N. 29 1494 Dankwa, Selasi 1450 de Alwis, Ruklanthi 625, 1275 Cottrell, Gilles 1557, 1797, 1916 Cummings, Richard D. 1795 Danner, Rebecca 1603 de Araujo, Fernanda F. 536, 1090, Coulibaly, Boubacar 762 Cundill, Bonnie 884, 897, 1472 Danon, Leon 1723 1105 Coulibaly, Drissa 380, 389, 619, Cunha, Rivaldo 577, 578 Dao, Francois 1493 Dearden, Kirk A. 600 984, 1605, 1796, 1868 Cunningham, Jane 871 Dao, Sounkalo 1495 Deason, Nicholas A. 872 Coulibaly, Jean T. 1719 Cupp, Eddie W. 1695 Daou, Modibo 380, 619, 1868, De Beaudrap, Pierre 1487 Coulibaly, Mamadou B. 796, 1371 Curriero, Frank C. 701, 957, 938, 1868 de Beaumont, Catherine 1787 Coulibaly, Michel E. 589, 1697, 1529 Dara, Antoine 323, 380, 619, 1527, Debella, Asfaw 509 1698 Curtis, Kurt C. 1785 1596 de Bes, Laura 1804 Coulibaly, Moctar 1493, 1527 Cusick, Sarah E. 232, 1439 Dara, Charles 962, 1798 DeBess, Emilio E. 97, 1044 Coulibaly, Sam 1590 Cuypers, Bart 688 Dara, Nianwalou 1492, 1493 Deborggraeve, Stijn 1100, 1854 Coulibaly, Siaka Y. 1697, 1698 Czerkinsky, Cecil 423 Dara, Niawalou 1527 Debpuur, Cornelius 1872 Coulibaly, Yaya I. 589, 1696, 1697, Daré, Abdoulaye 1078 Debrabant, Alain 547, 1105 1698, 491 Darling, Anne Marie 722 Debrah, Alexander Y. 1207, 1210, Couret, Jannelle 1819 D Darrah, Patricia 1800 1689, 1713 Coursen, Jill 962, 1571 Darton, Thomas C. 1849, 1850 Debrah, Linda B. 1689 Da, Dari F. 284 Cowden, Jessica 666, 1595 Das, Smita 146 deBruyn, Becky S. 1913 Dabiré, Roch K. 1817, 920, 119, Cowman, Alan F. 385, 1945 Das, Subash 11, 802 de Caravalho, Adelaide 811 982, 1373, 1403 Cox, Jonathan 41, 310, 700, 1109, Dasch, Gregory A. 105 de Cassan, Simone C. 672 Dabo, Abdoulaye 1868 1521 da Silva, Andréa L. Soares. 1563 de Castro Guimaraes, Ana 502 Dacheux, Mélanie 1237 Cox, Momodou 671 da Silva, Silvana C. 1734 Dechering, Koen J. 1018 da Costa, Luis M. 811 Coyle, Christina M. 458, 459, 1198, Dasilva, Alexandre 98 Decosterd, Laurent 1238 da-Costa Vroom, Baaba 1302 1729 da Silva Figueiredo, Adrya 1327 DeCotiis, Mark 1008, 1621 D’Acremont, Valérie 1065, 1767 Coyne, Philip 471 Dassi, Christelle 659, 1114 De Crop, Maaike 1922 da-Cruz, Alda M. 1748 Crabtree, Mary 861 Daszak, Peter 1116 Decuypere, Saskia 1854 Dac Tho, Nguyen 1384 Crabtree-Ide, Christina 48 Dat, Vu Quoc 1768 Deekshit, V K. 1108 Dadzie, Samuel 1248, 1622 Craft, David W. 1170 Datta, Dibyadyuti 404, 1000, 1586 Defang, Gabriel 1924 Dagur, Pradeep K. 1202 Crainey, Lee J. 1330 Datta, Shivani 93 de Gier, Brechje 1721 Dahir, Saidi 651 Crannell, Zachary 1173 Daubenberger, Claudia A. 670, de Gier, Nicole 1316 Daijogo, Sarah 197 Cravo, Pedro 901 1175 Degner, Ethan 756 Daily, Johanna 289, 615, 1441 Creek, Darren 26, 257 Daugschies, Arwid 446 De Grave, Dany 576 Dairo, Magbagbeola David 523 Crellen, Thomas 1184, 1811 Dauner, Allison 1396 De Groot, Anne 1713 Dale, Martin 1528 Cringoli, Giuseppe 1787 Davenport, Gregory 420 Deichsel, Emily L. 427 D’alessandro, Alessandro 571 Crisanti, Andrea 1884 Davenport, Miles P. 1437, 1489 Deik, Amy 907 D’Alessandro, Umberto 229, 261, Crispin, Luis 1134 Daver, Roshni G. 431 Deitz, Kevin C. 1355 274, 314, 373, 720, 894, 900, Cristal, Juqueline 1203 Davey, Dylan 1526 Dejene, Seyoum 310 959, 966, 1806, 1922, 376, Cromer, Deborah 1437, 1489 Davey, Gail 591, 1286, 1290, 1690 Dek, Dalin 298 1222, 1299, 1519 Crompton, Peter D. 405, 1552, David, Lawrence 432 DeKoster, Gregory T. 995 D’Alessandro, Sarah 288 1555, 1567, 1571, 1572, 1798, David, Sophia 1811 de la Fuente, Laura 338 Dalhat, Mahmood M. 713 1866, 962, 398 David, Weetman 129 de la Rua, Nicholas 1337 Dalipanda, Tenneth 724 Crookston, Benjamin 600 Davidson, Kailan-Sierra 1609 de la Vega, Patricia 1594 Dalrymple, Ursula 943, 944, 945 Cross, George 1939 Davidson, Leslie L. 635 Del Bello, David P. 493 Dalsgaard, Anders 570 Crowe, Andrew 240 Davidson, Silas A. 1446, 1610 Delcher, Arthur L. 619 astmh.org Abstract Author Index A-603

The number(s) following author name refers to the abstract number.

Delgado, Jaime 1784 de Vasconcelos, Janaina M. 1563 Dietsch, Kirk 1441 Dolo, Amagana 668, 669, 1009, Delgado, Yosnaida 202 Devine, Gregor 1242, 1619, 121 Dietze, Reynaldo 577, 578, 1124 1592 Delgado-Ratto, Christopher 959, de Vlas, Sake J. 63, 67, 1741 Dieudonne, Soma D. 775 Dolo, Housseini 1697, 1698 1519 de Vries, Henry 1097 Dieye, Baba 1477 Domingo, Gonzalo J. 481, 506 Delimini, Rupert K. 1404 De Vries, Peter J. 358 Dieye, Tandakha 986, 1799, 1450 Dominguez, Claudia 663 della Torre, Alessandra 1352, 1367, Dey, Ayan 423, 1433 Díez, Luisa 361 Domínguez Trejo, Pablo 1657 1912 Dey, Ranadhir 1202 Diggle, Peter J. 942, 1521 Don, Robert 1209 Dellicour, Stephanie 237 Deye, Gregory 252 Diggs, Carter L. 1605 Dondji, Blaise 515, 1754 DellOca, Nicolas 1781, 1816 de Zeeuw, Janine 1655 Di Giussepe, Francesca 1497 Dondorp, Arjen 268, 904, 343, DeLorey, Mark 811, 1382, 846 Dezengrini-Slhessarenko, Renata Dimas, Hannatu J. 357 1101 Deloron, Philippe 987, 1557, 1797 1397 Dimatatac, Frederico 1062 Dong, Yuemei 1886 De los Santos, Maxy B. 545 Dhanaraj, Prabakaran 22 Dimopoulos, George 786, 1886 Dongol, Sabina 1641 de los Santos, Tala 481, 506 Dhanasekaran, Govindarajan 1378 DiNardo, Andrew R. 1126 Dongus, Stefan 1619 Del Piero, Fabio 1217 Dhar Chowdhury, Parnali 155, 801 Ding, Jianging 445 Donini, Cristina 673 del Portillo, Hernando A. 223, 233 Dhillon, Baljean 1101 Dinh, Ha S. 1501 Donkin, David A. 1462 Delshadi, Sarah 1055 Dhingra, Satish 1241 Dinis, João 1367, 1912 Donna, Denno M. 1880 Delves, Michael 287 Dia, Ibrahima 764, 1352 Dinis, Jorge 665 Donnelly, Christl 208, 209 Demas, Allison 18, 617 Dia, Seydou 962, 1552, 1798 Dinko, Bismarck 1865 Donnelly, Christl A. 2 DeMaso, Christina R. 624 Diabaté, Abdoulaye 119, 920, Dinman, Jonathan 166 Donnelly, Liam 796 de Matos Guedes, Herbert L. 1205 1373, 1625, 1817 Diop, Malal 122 Donnelly, Martin J. 760, 765, 768, Dembele, Adama 1539 Diakite, Mamadou A. 1495 Diouf, Ababacar 1799 773, 1189, 752, 776 Dembele, Ahmadou 619 Diakite, Moussa L. 1009, 1598 Diouf, Mamadou L. 1456 Doolan, Denise 609 Dembele, Benoit 1697, 1698 Diallo, A. 359 Diouf, Mame Birame 1250, 1251 Doolan, Denise L. 1567, 1604 Dembele, Massitan 1696, 1697, Diallo, Abdallah 491, 589, 1697, Di Pasquale, Aurelio 727 Dorigatti, Ilaria 195, 196 1698 1698 Direny, Abdel 1694 Doritchamou, Justin 987 De Montfort, Aymeric 835 Diallo, Abdoulaye 330, 658 DIRESA MINSA-Peru: Huanuco, Dorkenoo, Monique A. 1136 Deng, Bingbing 272 Diallo, Boubakar 1495 Junin, Ayacucho, San Martin, Dorn, Mauricio 548 Deng, Chang Sheng 949 Diallo, Diadier A. 900, 1520 Ucayali H. 453 Dorn, Patricia L. 1337 Deng, Haiyan 1495 Diallo, Hama 1561 Di Santi, Silvia M. 874, 901, 1540, Dornelas, Marina P. 1573, 1742 Dengela, Dereje 781, 1363 Diallo, Ibrahima 246, 1250, 1456 1545, 1573, 1742 Dorny, Pierre 458, 1196 Denholm, Ian 1242 Diallo, Nouhoum 323, 1493, 1527 Dittrich, Sabine 1046, 1214, 1852, Doroski, Victoria 292 De Niz, Mariana 223 Diamond, Michael S. 648, 1387 1853 Dorsey, Grant 24, 402, 893, 903, Dent, Arlene E. 1554, 1560 Diancourt, Laure 795 Diuk-Wasser, Maria A. 1219 935, 946, 1569, 1808, 1844, Deol, Arminder K. 1082, 1715 Diap, Graciela 1459 Divala, Titus H. 717, 1226 1864, 1903 de Oliveira, Alexandre M. 135 Diarra, Amidou 1590 Divis, Paul 1442 Doshi, Reena H. 85, 1430 De Oliveira, Camila I. 1203 Diarra, Bakary S. 1539 Dixit, Amruta 1613 Dosoo, David K. 1067, 1637 de Quadros, Ciro 1306 Diarra, Issa 380, 619, 1868 Djebbari, Habiba 709 dos Reis, Ediane M. Rodrigues. 135 de Quiroz-Castro, Maricel 739 Diarra, Seybou 25 Djegbe, Innocent 1192 dos Santos, Claudio R. A. 1813 Deran, Tong Chor M 1706 Diatta, Georges 99 Djenontin, Armel 118 dos Santos, Eduardo J. Melo. 1563 Deribe, Kebede 1690 Diawara, Aïssatou 642, 1719 Djimde, Abdoulaye 323, 324, 902, dos Santos, Gerusa B. 729 Derrick, Anne 882 Diawara, Halimatou 1509 619 dos Santos, Marcelo A. M. 1397 Derrick, Steven 230 Diawara, Lamine 1694 Djimde, Abdoulaye 1239, 1482, dos Santos, Marcia M. Martins. 135 Dery, Dominic B. 1637 Diaz, Carlos 1128 1491, 1492, 1493, 1527, 984 Dos Santos, Paulo Ricardo 551, 552 Desai, Meghna 237, 1478, 1503, Díaz, Francisco J. 793 Djogbenou, Luc 773, 776, 1192 Dossou, Ange D. 1655 1534, 1902 Diaz, James H. 507 Djossou, Laurette 1192 Dotson, Ellen 1819, 1908 Desai, Michael 907 Diaz, Kristians 51 Djuardi, Yenny 1895 Dougan, Gordon 1641, 1849, 1850 Desai, Sanjay 407, 1448 Diaz, Yamilka 206, 1423 Dlugosz, Lisa 1867 Douglas, Alexander D. 270 DeSalvo, David R. 204 Dibernando, Antonia 801 Dobaño, Carlota 338, 1567 Douglas, Nicholas M. 1905 Deseda, Carmen 577, 578 Diboulo, Eric 1011 Doblas-Reyes, Francisco 730 Douglass, Alyse 17 Deshotel, Michael B. 545 Dibusova, Lenka 1518 Dobson, Andrew 1543, 1820 Doumbia, Salif S. 1696 de Silva, Aravinda 575, 576, 1385, Dickinson-Copeland, Carmen M. Dodean, Rosie 291 Doumbia, Seydou 762, 1525 625, 626 226 Dodoo, Daniel 1067 Doumbo, Ogobara 668, 669, 902, de Silva, Aruna D. 822 Dicko, Alassane 25, 1509, 1520, Dodson, Brittany 1364 1009, 1561, 1598, 1599, 1868, deSilva, Aravinda M. 628 1539 Doggett, Joseph S. 1172 1592, 380, 796 de Souza, Dziedzom K. 1248, 594, Dicko, Ilo 1696, 1697, 1698 Doggett, Stephen 6 Doumbo, Ogobara K. 324, 389, 1114 Dicko, Yahia 25 Dogonyaro, Benjamin 1421 619, 962, 984, 1491, 1492, De Spiegeleere, Bart 1305 Dickson, Dorothy M. 583, 1288 Doker, Thomas J. 1041 1527, 1552, 1572, 1600, 1605, Despres, Philippe 12 DiClaro, II, Joseph W. 982, 1370 Dokladny, Karol 967 1796, 1798, 1866 Desruisseaux, Mahalia 216, 612 Diehl, Sean 583, 1288 Dokoto, Harishu 1136 Doumbo, Safiatou 405, 962, 1552, DeStefano, Joseph 1654 Dieng, Yemou 394 Dolan, Stephanie 877, 1528 1798, 1866, 1572 Destura, Raul V. 178 Dierckx, Suzan 266 Dolkart, Caitlin F. 886, 1907, 878, Doumtabe, Didier 962, 1552, 1798, Deubel, Vincent 1514 Diesburg, Steven 1318 1536 1866 Devaney, Eileen 486 Dietrich, Elizabeth A. 646 Douradinha, Bruno 609 astmh.org A-604 Abstract Author Index The number(s) following author name refers to the abstract number.

Dourng, Dany 1514 Dupke, Singye 921 Edison, Laura 1215, 1878 Engelbrecht, Christian 1375 Dow, Geoffrey S. 320 Duplantier, Jean-Marc 99 Edoh, Dominic 1092, 1093 Ennis, Francis A. 697 Dowd, Kimberly A. 624, 841 Dupuy, Lesley C. 13 Edorh, Patrick A. 1655 Enquselassie, Fikre 1690 Dowell, Floyd 121, 788 Duraisingh, Manoj 320, 415, 614, Edstein, Michael D. 249 Enyong, Peter 503, 1890 Downs, Philip 1259, 1076 1436, 616, 1450 Edwards, Chelsea L. 1437 Eom, Keeseon S. 1782 Doyle, Stephen R. 479 Durán-Arenas, Luis 1401 Edwards, Hannanh 337 Epis, Sara 1336 Dozie, Ikechukwu N. S. 260, 529 Durand, Patrick 99 Edwards, Leslie 1429 Epstein, Jon 1116 Drábek, Elliott F. 619, 1600 Durand, Salomon 277, 397, 1699 Edwards, Nick J. 270, 672, 671, Epstein, Judy 1005, 1610 Drabo, Maxime K. 1299 Durán-Rehbein, Gabriel A. 537 986 Erasmus, Jesse 159 Drake, Thomas 884 Durante, Amanda 921 Edwards, Rachel L. 317 Erhart, Annette 229, 274, 900, Drake, Tom 721 Durbin, Anna P. 181, 579, 822, 826 Edwards, Thomas E. 1038 959, 966, 1222, 1519 Drakeley, Chris 325, 348, 403, 655, Durham, David P. 519 Egea, Pascal F. 1943 Eriksson, Emily M. 385 700, 933, 946, 1109, 1521, Durnez, Lies 657, 926, 1033, 1514, Eghan, Kwesi 1077 Ermert, Volker 1497 1842, 139, 927, 41 1624, 1626 Egrot, Marc 703 Ernest, Samuel K. 238 Drame, Papa M. 1891 Durrant, Caroline 1811 Eguiluz, Maria 452 Ernst, Kacey 1736 Drammeh, Abdoulie 671 Duru, Valentine 1237 Ehounou, Genevieve 1855 Ersoy, Ilker 1453 Dranow, David M. 1038 Duthie, Malcolm 1228 Eigege, Abel 950 Escalante, Ananias A. 321, 379, Draper, Simon J. 672, 1556 Dutse, Abdulhamid I. 713 Eisele, Thomas P. 1470, 1507 971, 970 Drebot, Michael A. 155, 801 Dutta, Sheetij 389, 1867, 1605 Eisen, Lars 137 Escolano, Sylvie 251 Drevets, Douglas A. 1193, 1195 Duvall, Jeremy 286 Eisen, Rebecca J. 846 Esen, Meral 1608 Drewe, Julian 1109 Dweni, Celestine K. 1180 Eisenberg, Joseph 1761, 1764, Eshun, Miriam 848 Drews-Botsch, Carey D. 707 Dwivedi, Prabha 255, 1873 1530 Espetia, Susan 852 Drexler, Naomi 68, 661 Dwyer-Lindgren, Laura 1618 Ekawati, Lenny L. 327 Espina, Noel 1138 Druilhe, Pierre 1238 Dykes, Cherry L. 765 Ekomisa, Fabrice B. 1634 Espina Gomes, Luz 1383 Drumond, Betania 194 Dzabala, Nettie 717 Ekpo, Uwemedimo F. 1069 Espinosa, Diego 1606, 1607 Duan, Junhui 796 Dziedziech, Alexis Dziedziech 1176 Elaagib, Arwa 1706 Espinosa, Pablo 1268 Duangkhae, Parichat 818 Dzinjalamala, Fraction 258, 1474 Elakhal Naouar, Ines 1747 Espinoza, Angelica 803 Dube, Queen 678 Dzodzomenyo, Mawuli 570 Elder, John P. 153 Espinoza, Yrma 1134 Dube, Tina 581 Dzyublyk, Iryna 553 Elfitouri, Fatima 894 Espírito-Santo, Maria Cristina C. Dubhashi, Nagesh 910 Elfving, Kristina 927, 1040 1158 Dubray, Christine 1694 Elias, Sean C. 672, 1556 Essandoh, John 773 Duchateau, Magalie 1194 E Elizalde-Torrent, Aleix 223 Essondoh, John 776 Duchemin, Jean B. 1322 Elizondo, Douglas 191, 192 Estevez, Alejandra 1643 Eagle, Nathan 83 Duchon, Stéphane 122 Eller, Dirli E. 61 Estevez-Lao, Tania Y. 1818 Eagon, Scott C. 285 Ducusin, Maria Joyce U. 739 Elliott, Alison 403 Estivariz, Concepcion F. 1424 Eapen, Alex 367 Duda, Kirsten A. 360, 1422 Elliott, Karla P. 1135 Etard, Jean-François 1487, 1229, Eappen, Abraham 1594, 1620 Dudakova, Katarina 530 Elliott, Suzanne 675, 1488, 1489, 1855 Earle, Duncan 637 Dueger, Erica 112, 114 1490 Etchepare, Michel 703 Eastman, Richard T. 1900 Duez, Julien 1498 Ellis, Esther M. 1382, 1415 Ettwiller, Laurence 1896 Eaton, Will 1566 Duff, Kevin 905, 1532 Ellner, Jerrold J. 1124 Evans, Allan M. 244 Ebang Menye, Daniel 567, 1161 Duffield, Giles E. 748, 1016 Elmubarak, Wigdan 1706 Evans, Carlton A. W. 1278, 1279 Ebel, Gregory D. 648, 837, 845, Duffy, Craig 1442, 1445 Elnahas, Ayman 446 Evans, Christopher C. 1896 645 Duffy, Patrick E. 248, 366, 605, Elnaiem, Dia-Eldin A. 1743 Evans, Emily E. 1283, 1860 Ebels, Kelly 339 613, 1008, 1009, 1371, 1448, Elobied, Eyhab 437 Eversley, Tatiana 1325 Eberhard, Mark L. 1694, 1711 1539, 1593, 1598, 1599, 1621, Elphinstone, Robyn E. 1435, 1859 Ewer, Katie J. 671, 986, 1590 Ebihara, Hideki 1928 668, 669, 796, 1561, 1592 El-Sakkary, Nelly 1815 Ewing, Victoria 1252 Ebusu, Charles 1569 Duggal, Nisha K. 645 El-Sayed, Najib M. 1791 Existe, Alexander 978 Eccles-James, Iejoma 1569, 1864 Duggal, Priya 1266 Elwood, Dan 181, 579 Eyangoh, Sara 1855 Echazu, Adriana 514, 643 Dui, Le Thi 783 Elyazar, Iqbal R. F. 360 Ezeamama, Amara E. 714, 1121 Echeverri-Garcia, Diego F. 872 Dujardin, Jean-Claude 688 Embers, Monica E. 104 Ezeh, Ikenna O. 449, 450 Echezuria, Luis R. 821 Dukuzumuremyi, Janvier 358 Emeetai, Thomas 601 Ezeigwe, Nnenna 865, 882 Eckert, Erin 952, 1918 Dulley, Federico 718 Emegbuonye, Leslie 1907 Ezenduka, Ekene V. 74, 449, 450 Eckhoff, Grace 588 Duman-Scheel, Molly 686 Emerson, Paul M. 332, 1075 Eckhoff, Philip A. 46, 1542, 667, Du Mond, Jennifer 1526 Emery, Aidan 1810, 1812 1506 Dumonteil, Eric 1329, 1337, 1750, Emmanuel, Amlabu 1002 F Ecklu-Mensah, Gertrude 1717 1936 Emrich, Scott 686, 1341 Eckman, Brooke 1548 Fabiszewski de Aceituno, Anna Duncan, Elizabeth 1595 Encinas, Michele 391 Eden, John-Sebastian 852 1311, 1645 Duncan, Robert 1202 Endeladze, Marina 1304 Edgel, Kimberly 545, 608 Fabris, Clara 119 Dunne, David W. 59 Endsley, Janice J. 528 Edgerton, Sean V. 805 Fabris, Paolo 571 Duong, Dung T. K. 279 Endy, Timothy 1057, 1391 Edi, Ako Victorien C. 129 Facchinelli, Luca 1884 Duong, Tran T. 1519 Eng, Matthew W. 1345 Edi, Constant 776 Fadeyi, Ifey 255 Duong, Trinh 897 Eng, Samantha 146 Edi, Constant V. 773 Fagbohoun, Josias 134 Duparc, Stephan 675, 1486 Engel, Roberta 1343 astmh.org Abstract Author Index A-605

The number(s) following author name refers to the abstract number.

Fahle, Gary 270 Ferguson, Neil M. 195, 196, 1525 Fitzpatrick, Meagan C. 664 Francis, Filbert 304, 897 Failloux, Anna-Bella 1361, 1910 Feria, Teresa P. 1730 Flach, C 359 Franco, José R. 1740 Fair, Elizabeth 533 Fernandes, Guilhermina 1847 Flach, Clare 658 Franco-Muñoz, Carlos 541 Fair, Joseph 91 Fernandez, Facundo M. 255, 1873 Flanagan, Katie 671 Franekova, M. 867 Fairhurst, Rick M. 150, 298, 904, Fernandez, Mariano M. 821 Fleckenstein, Lawrence 1212, 1486 Frank, Molly F. 36, 37, 535, 715 1237, 1900, 410 Fernandez, Regina 807 Flecker, Robert R. 1199 Frantzreb, Charles 614 Fairlamb, Alan H. 287 Fernandez, Stefan 8, 168, 816, Fleming, Fiona 1151, 1160 Frasch, Alberto C. C. 1201 Fairley, Jessica K. 1681 863, 1272, 1274, 1429, 1770, Fletcher, Daniel A. 1208 Fraser, Claire M. 619, 1600 Faiz, M. Abul 1101 1774 Flordeliza, Nicole 319 Fraser, Jamie 471, 1051 Faizullabhoy, Adnan 1899 Fernandez-Arias, Cristina 610 Floreani, Annarosa 571 Fraser, Lisa 490 Fakiola, Michaela 691 Fernandez-Becerra, Carmen 223 Flores, Adriana E. 137 Frederick, Joseph 1908 Fakoli, Lawrence S. 982 Fernandez-Sesma, Ana 1839 Flores, Jorge 1102 Fredrick, Harrison 1879 Falendysz, Elizabeth A. 1427 Ferrari, Marilyn 1604 Flores Franco, Jorge L. 548 Freeman, Brandi D. 612 Falkard, Brie W. 434 Ferraz, Lorena d. Neres. 61 Flores Leon, Amilcar Alejandro 1104 Freeman, Matthew C. 596, 707, Fall, Mawo 1076, 1694 Ferreira, Jorge G. G. 858 Florey, Lia S. 704, 952, 1249, 1028, 602, 710, 1168, 1169 Familiar-Lopez, Itziar 24 Ferreira, José Jarbas B. 874 1289, 1617, 1623, 1632 Freiberg, Alexander N. 850 Fan, Erkang 1038 Ferreira, Karine S. 536 Flynn, Bailey 1285 Freire, Rosemeyre S. 1648 Fan, Hui 1724 Ferreira, Marcelo U. 656, 1224, Fobil, Julius 570 Freitas, Carolina R. C. 1090 Fan, Qi 941 1805 Fofana, Bakari 1493, 1527 Freitas, Vera L. Teixeira de. 718, Faneye, Adedayo 172 Ferreira, Paulo C. P. 858 Fofana, Bakary 324, 1492 1230 Fang, Qiang 382 Ferreira-da-Cruz, Maria de Fatima Fogg, Carole 268 Frempong, Eric H. 409 Fanthome, Amber 487, 501 390, 892 Foil, Lane D. 1370 Frempong, Kwadwo K. 1093 Farag, Tamer H. 582, 584 Ferro, Santiago 743 Fokou, Gilbert 659 French, Michael D. 1154, 1082 Faragher, Brian 23, 1856 Ferrufino, Lisbeth 548, 1102, 1930 Folefoc, Asongna 285, 891, 1674 Fresnay, Stephanie 1849, 1850 Faraj, Chafika 1370 Feschotte, Cedric 1177 Foley, Michael 286, 1585 Freyberger, Helen 1304 Fares Gusmao, Rafaelle 536 Festo, Charles 28, 255, 1578, 1579 Fomenjanahary, Verosoa 1056 Freyman, Jessica 734, 734, 734 Färnert, Anna 405, 964, 974 Fiandor, Jose M. 1932 Fongoro, Sahare 1495 Friant, Sagan 1760 Farrington, Lila 402, 1569 Fica, Alberto 1640 Fonkoua, Marie-Christine 1638 Fricke, Kai 605 Farris, Christina M. 111 Fidock, David 287, 675, 1237, Fonseca, Anna Maria 338 Fried, Michal 366, 605, 613, 1448, Faruque, Abu S. G. 425, 1174 1240, 1241 Fonseca, Benedito A. L. 1406 1539 Fasabi, Manuel M. 414 Fiekowsky, Elana 1551 Fonseca, Dina M. 1821 Friedman, Jennifer F. 56, 1727, 71, Fataki, Olivier A. 1547 Fielding, Burtram 158 Fonseca, Jairo A. 603 605 Fatunmbi, Bayo 256 Fields, Barry 819, 824, 1064, 1261, Fonseca-Ford, Maureen 723 Friedrich, Lindsey R. 622 Faulx, Dunia 481, 506 1382, 1045 Fontana, Mary 1565 Friedrich, Thomas C. 665 Faust, Aubrey 1805 Fievet, Nadine 1557, 1797 Fontenla, Santiago 1816 Fröberg, Gabrielle 929 Faust, Christina 1543 Figueiredo, Luiz T. M. 1406 Fontes, Raissa M. 1406 Fronas, Synne G. 1152 Favia, Guido 1336 Figueroa, Carlos A. 1420 Fontoura, Pablo S. 1224 Fryauff, David 883 Fawole, Olufunmilayo 523, 566 Figueroa Quintanilla, Dante A. 636 Fonville, Judith M. 828 Fu, Chi-Ling 32, 1792 Fawzi, Wafaie W. 722, 740 Filho, Olindo A. Martins. 61 Fonzi, Eugenio 1349 Fuchs, Jeremy 802 Fay, Michael 668, 669 Fillinger, Ulrike 3, 148, 1243 Fooks, Anthony 1419 Fujioka, Kenn 1376 Faye, Adama 1846 Fimmers, Rolf 1210 Forbush, Melissa 319 Fujiwara, Ricardo T. 1263 Faye, Babacar 254, 302, 394, 986 Finak, Greg 1800 Ford, Louise 483, 484, 495, 504, Fukuda, Mark 1494 Faye, Ousmane 330, 658, 764 Finan, Chris 1286 502 Fukumoto, Shinya 126 Fedak, Erin M. 1438 Fine, Ian 276 Fordyce, Sarah L. 1914 Fulakeza, Joseph M. 620 Feeney, Margaret E. 402, 1864, Fingeroth, Joyce 614 Fornace, Kimberly M. 1109 Fuller, Douglas 340 1565, 1569 Fink, James 606 Fornadel, Christen 781, 1363, 1551 Fullman, Nancy 1618 Feeser, Karla 1712 Finkelstein, Julia 1411 Forquer, Isaac 291, 673 Funakoshi, Ryota 1606 Feldman, Katherine 98 Finn, Tyler 580 Forshey, Brett 1413 Furin, Jennifer 55 Feldman, Michael 1044 Finnefrock, Adam C. 197 Forshey, Brett M. 807 Furini, Adriana A. C. 391, 393, 396 Feldmeier, Hermann 101, 102, Fiore, Jacqueline 953, 1522 Foster, Jeremy M. 1896 Fürst, Thomas 1871 1331 Fiorentino, Marion 1721 Foufopoulos, Johannes 1764 Furucho, Celia R. 1230 Felger, Ingrid 374, 955, 972, 1221, Firbas, Christa 12 Fournet, Florence 1403 Fusellier, Andrew 85 1550 Firdausi, Qadri 433 Fox, Chris 1606 Futami, Kyoko 1349 Felgner, Philip L. 389, 962, 1552, Firew, Heven Sime 591 Fox, LeAnne 496, 1694 1562, 1567, 1605, 1796, 329, Fischer, Katja 1218, 1828 Foxman, Betsy 1761 1223, 1510 Fischer, Kerstin 1785, 1826 Foy, Brian D. 755, 982 G Felix, Todd 837, 845 Fischer, Marc 839, 846 Fraga, Jorge 1095 Gabriel, Erin 668, 669, 1009 Felton, Amanda 880, 280 Fischer, Natalie 21 Fraga, Valeria D. 391, 393, 396 Gabriel, Sarah 625, 1196 Fenwick, Alan 1160 Fischer, Peter U. 65, 1707, 1785, Franca, Gabriel P. 340 Gabrieli, Paolo 1889, 1915 Ferdig, Michael T. 890, 898, 899, 1826 Français, Olivier 1498 Gacek, Rosana 690 1240, 1465, 908, 1897 Fishbauger, Matt 1829 Franca-Koh, Ana Claudia 1249, Gachara, George 558 Ferguson, Heather 77, 139 Fishbaugher, Matthew 1897 1249, 1289, 1617, 1623, 1028 Gadalla, Nahla B. 348 Ferguson, Neil 208, 209, 919 Fitzhenry, Robert 900 Franchard, Thierry 454 astmh.org A-606 Abstract Author Index The number(s) following author name refers to the abstract number.

Gaeddert, Mary 1124 Gardner, Christina L. 14 Gesase, Samwel 1227, 1483 Gobert, Geoffrey 1828, 1144 Gagova, Iveta 531 Gardner, Malcolm J. 329 Gething, Peter W. 271, 360, 944, Godfray, Charles 1 Gaké, Bouba 567, 1161 Garg, Seema 831 1422, 943, 945, 1929 Goes, Alfredo M. 61 Gakidou, Emmanuela 1618 Garimo, Issa 368 Geubbels, Eveline 727 Goethert, Heidi 1216 Galactionova, Katya 41, 1001, Garin, Benoit 1056 Ghafari, Caline 936 Goez-Rivillas, Yenny 103 1588, 1589 Garms, Rolf 593, 1693 Ghaffar, Atif 979 Gogtay, Nithya 611 Galaphaththi-Arachchige, Hashini Garn, Joshua V. 707 Ghai, Ria R. 665 Goh, Guat Kheng 965 N. 1794 Garske, Tini 208, 209, 1520 Ghani, Azra C. 43, 742, 1029, Goh, Kenneth C. 804, 808 Galappaththi, Hashini N. 1155 Gartrell, A. 1940 1525, 1843, 345, 919, 928, 999, Goh, Lucy M. L. 1215, 1878 Galappaththy, Gawrie N. L 864 Garver, Lindsey S. 1446 1520 Goheen, Morgan M. 1444, 1451 Galarce, Zaira H. 227 Garza-Hernández, Javier 1695 Ghansah, Anita 1239 Golassa, Lemu 1239 Galarion, Ma. Jowina H. 178 Garzoni, Luciana R. 536 Gharbi, Adel 1934 Gold, Dan 1943 Galarza, Ivonne 1405 Gass, Katherine 596 Ghawar, Wissem 1934 Goldberg, Daniels E. 1943 Galarza, Marco 1432 Gatkouth, Nyachare 319 Ghedin, Elodie 14, 488 Goldberg, Tony L. 92, 665, 1431, Galati, Fabio 439 Gatserelia, Lana 1304 Ghersi, Bruno M. 1755 842, 1760 Galdos Cardenas, Gerson 548, Gatton, Michelle L. 887 Ghimire, Anup 1679 Golden, Allison 481, 506 1102, 1930 Gaudart, Jean 658, 1491 Ghosh, Kashinath 547 Golding, Nick 360, 1422, 1740, Galera, Katia 1388 Gaul, Linda 1390 Ghosh, Partho 1724 1929 Galinski, Mary R. 218 Gaur, Deepak 1002 Giannini, Caterina 1140 Goldman, Ira 911 Galinsky, Kevin 1805 Gautam, Shalini 1751 Giardina, Steve 1606 Goldman-Yassen, Adam 615 Gallafrio, Christina N. 718 Gavino, Arquímedes 1278 Gibba, Balla 314 Goldstein, Harris 15 Gallardo, Emelda 1432 Gaye, Oumar 254, 302, 330, 394, Gibbons, Robert V. 697, 1391, Goldys, Anastasia 236 Gallego-Delgado, Julio 610 658, 359, 986, 1906 1770 Gomes, Cláudia 426 Gallegos, Marisa 1262 Gaye, Seynabou 246, 278 Gibbs, John 1385 Gomes, Edwin 910 Galloway, David 39 Gaynor, Bruce D. 1048, 1066 Gibot, Sebastien 838 Gomes, Juliana A. S. 536 Galson, Victor W. 732 Gazzinelli-Guimaraes, Ana Clara Gibson, Gabriella 1373 Gomes, Regis 1203 Galvani, Alison P. 519, 664 1263 Gibson, Greg 1802 Gómez, Giovan F. 1358 Gama, Elvis S. 555 Gazzinelli-Guimaraes, Pedro H. Gidado, Saheed 523, 853 Gomez, Maria Linares 294 Gamarro, Francisco 688 1263 Gidwani, Kamlesh 1751 Gómez-Pérez, Gloria P. 1602 Gamboa, Dionicia 388, 959, 966, Gbedande, Komi 1557, 1797 Gikunju, Stella 1382 Gomez-Puerta, Luis A. 451 1541, 1807, 1823, 1858 Gbolahan, Abass O. 566 Gil, Jose P. 324 Gomis, Jules 658, 1906 Gamo, Francisco Javier 250 Geall, Andrew J. 387 Gilbert, Amy 861 Gonahasa, Samuel 1460 Gamo-Benito, Francisco Javier 294 Geana, Mugur 812 Gilbert, Ian 287 Gonçalves, Bronner P. 325 Gan, Victor C. 1062 Geary, Timothy G. 498, 512 Gilbert, Sarah C. 270, 671, 1587 Gonçalves, Raquel M. 1224 Ganaba, Ramané 1196 Gebretsadik, Abeba 1690 Gilbreath, III, Thomas M. 1353 Gondwe, Esther N. 1558 Gandirilla, Omar 548 Gedefaw, Molla 1070 Gilchrist, Carol A. 1135 Gong, Bin 103 Ganesan, Anuradha 471, 1051 Gee, Christina 520 Gildengorin, Ginny 9, 177, 651, Gongadze, Nana 1304 Ganesh, Nandita 177 Geertruyden, Jean-Pierre van 1233, 1271, 1714 Gongora, Jennifher 1338 Ganga, Teresa 1153 1519 Gilder, Mary Ellen T. 1920 Gonin, Michelle L. C.. 1409 Gangopadhyay, Keshab 1303 Geier, Martin 1375 Gilders, Mary Ellen 940 Gonsu, Hortense K. 524 Ganley-Leal, Lisa 1124 Geiger, Stefan M. 1090 Gillan, Victoria 486 Gonzal, Analisa 1727 Ganter, Markus 1450 Gelaye, Woyneshet 332 Gillette, Michael 50 Gonzales, Michael A. 1941 Gantier, Jean-Charles 795 Gelderblom, Huub C. 1075, 1076 Gilliland, Jr., Theron 1396 Gonzales Hurtado, Patricia A. 1448 Gantin, Richard G. 481, 506 Geldhof, Peter 644 Gillrie, Mark R. 222 Gonzalez, Ana S. 1115 Gao, Qi 128, 958 Genco, Francesca 639 Gilman, Robert 329, 452, 564, 716, Gonzalez, Angie D. 971 Gao, Qin 1457 Gendrin, Mathilde 1335 852, 1778, 1930, 456, 51, 414, Gonzalez, Armando E. 451, 452 Gaona, Heather 291 Genton, Blaise 1065, 1767 457, 548, 636, 875, 1102, 1106, Gonzalez, Carmen 23 Gaona, María A. 1338 Gentsch, Jon 1643 1279, 1296, 1684 Gonzalez, Daniel 663 Garba, Amadou 1719 George, Dylan B. 1422 Gimnig, John 961, 1534, 1887 González, Eric 1672 Garba, Deen 1454 George, Gary 1016 Giordani, Maria Teresa 571, 1668 González, Grehete 857, 1416 Garber, Michael D. 741 George, Kristen 781, 1551 Giorgi, Emanuele 942, 1521 González, Guelsys 857, 1416 Garcia, Andres 1508 George, Miriam T. 994, 1585 Giorgi, Roch 1491 González, Iveth 334, 325, 461, Garcia, Hector H. 451, 452, 455, George, Sarah 182, 581, 1389 Giraldo-Calderon, Gloria I. 1341 868, 871, 1514 457 Geraldo, Juliana A. 1789 Girgis, Natasha 1941 González, John M. 537 Garcia, John 379 Gerardin, Jaline L. 46, 1506, 1542 Githeko, Andrew K. 120, 1020, Gonzalez, Kadir 1328 Garcia, Lineth 1095, 1231 Gerbasi, Vincent R. 604, 608 1887 Gonzalez, Karla N. 189 Garcia, Melissa N. 698, 1730 Gerber, Sue 85 Glanz, Joanna 605 González, Mariela N. 544 Garcia, Sylvie 1498 Gerena, Lucia 320 Glasner, Dustin 198, 1836 Gonzalez, Publio 663 Garcia-Blanco, Mariano 813 Gerloff, Dietlind 1000 Gleeson, James 1238 Gonzaléz, Rafael H. 1648 Garcia-Hernandez, Raquel 688 Gerrard, Anna 1083 Glesse, Nadine 1563 González, Raquel 338, 1478 Garcia-Rejon, Julian 137 Gerrard, John 606 Gmeiner, Markus 1608 Gonzalez, Sachy O. 1593 Garcia-Sastre, Adolfo 1839 Gerry, Stephen 1556 Gnadig, Nina F. 1237 Gonzalez, Sofia 1284 Garcia Tavares, Mara 1346 Gesase, Samuel 1467 Go, Chi-Jong 1062 Gonzalez Andrade, Pablo 1100 astmh.org Abstract Author Index A-607

The number(s) following author name refers to the abstract number.

Gonzalez-Figueroa, Frances 1402 Grasperge, Britton J. 1393 Guebey, Remy 1194 Gwandu, Conrad 670 Gonzalo, Alfonso 789 Grasperge, Brooke 1000 Guedri, Evelyne 1934 Gwanzura, Lovemore 151 Gonzalo-Rodriguez, J. 574, 1769 Gratz, Jean 585 Guelbeogo, Moussa W. 325 Gyan, Ben 1067 Good, Michael 1042 Graumans, Wouter 1018 Guelbeogo, Wamdaogo Moussa Gyan, Efua 1163 Good, Michael F. 606, 1003, 1006 Graupe, Bonita 653 117, 126, 1019 Gyapong, John 594, 1302 Goodhew, E. Brook 60, 1079, 1087 Graves, Justin C. 1907 Guelig, Dylan 1318 Gyapong, Margaret 296 Goodin, Doug 663 Graves, Patricia M. 950 Guerena, Fernando B. 464 Gyasi, Richard 1067 Goodman, Anna L. 672 Graves, Richard 1586 Guerin, Philippe 268, 1842 Gyeltshen, Sonam 921 Goodman, Catherine 28, 255, Graviss, Edward A. 1126 Guerra, Carlos A. 360 Gyorkos, Theresa W. 1269 1578, 1579, 1581, 1582, 1583 Gravitt, Patti E. 1684 Guerra, Eduardo 202 Goodman-Meza, David 458, 459, Gray, Darren 597, 1185 Guerra-Giraldez, Cristina 455, 457 1198, 1729 Gray, Darren J. 1144 Guerra Mendoza, Yolanda 666 H Goodson, Holly 73 Gray, David 287 Guerrant, Richard L. 1178, 1315, Ha, Kaitlin 967, 968 Gopalakrishnan, Anusha 417 Gray, Karen-Ann 958 1639, 1648 Ha, Kwon-Soo 993 Goraleski, Karen 1284 Gray, Meg 755 Guerreiro, João F. 1563 Haacke, Mark 20 Gorbach, Pamina M. 733, 737 Green, Amy 798 Guevara, Carolina 185, 803, 840, Habib, Abdulrazaq G. 713 Gorchakov, Rodion 698, 1730 Green, Angela M. 187, 806, 188 1420, 1928 Habluetzel, Annette 119 Gordon, Aubree 192 Green, Michael 1873 Gueye, Alioune B. 246, 1250, Habomugisha, Peace 593, 1693 Gordon, Catherine A. 1144 Green, Mike D. 255 1251, 1456 Hachizovu, Sebastian 1299 Gordon, Emile 1453 Green, Sharone 630, 697 Guezala Villavicencio, M. Claudia Hackman, Henry K. 1036 Gordon, Jeffrey I. 1654 Greenberg, Robert M. 1825 1757 Haddad, Danny 1076 Gordon, Scott W. 1375 Greene, Rebecca 682 Guggisberg, Ann M. 317 Hadi, Melinda 116 Gorena, Karla 983, 1832 Greenhill, Andrew R. 53 Guha, Shantanu 1476 Hadiwidjojo, Sri 608 Gosi, Panita 299, 912, 913, 1454 Greenhouse, Bryan 946, 947, 1565, Guiguemde, Robert T. 274 Hadjilaou, Alexandros 188 Gosinary, Fabiola 1055 1864, 1903 Guillermo-Cordero, Leonardo 1750 Haefeli, Walter 1492 Gosling, Roland D. 348, 1502, 342, Greenland, Katie 1083 Guimaraes, Ana F. 485, 487, 503 Haenle, Mark M. 443, 641 344, 1227, 1513, 654, 1500, Greenstein, Rebecca 1441 Guimarães, Luiz Henrique 1734 Haetrakul, Thanida 1137 1501, 1509 Greenwood, Brian 254, 348, 1067, Guindo, Merepen A. 668, 669, Hagerty-Hoff, Christopher 1165 Gosnell, William L. 1447 1520, 1917, 238, 1906 1009, 1598, 1599 Hagiwara, Mitika K. 1200 Gosse, Leslie 835 Greer, George 1015 Guindo, Merepen A. 796 Hahn, Andrew 1126 Gotia, Hanzel T. 1175 Gregoricus, Nicole 1643 Guiñez, Dannette 1640 Hahn, Micah B. 846 Goto, Hiro 1200, 1545, 1573, 1742 Gregory, Philip D. 1237 Guiraud, Issa 1854 Hahn, Sigrid 732 Goto, Yasuyuki 1228 Grenfell, Bryan 430 Guler, Jennifer 910 Haidara, Aboubacrine 1527 Gottardo, Raphael 1800 Gresh, Lionel 191, 1273, 1275 Gunasekera, Anusha 669, 1556, Haider, Najmul 1110 Gottdenker, Nicole L. 105 Grieco, John 797, 1324, 1191, 1610 Haigney, Mark 1481 Gottlieb, Keith 197 1369, 1735 Gunawardena, Sharmini 656 Haile, Ashley 682, 759 Gotuzzo, Eduardo 1129, 1284, Grieve, Eleanor 1809 Gundersen, Svein G. 1152, 1794 Hailemikael, Amha Kebede 591 1776, 1784, 1787 Griffin, Jamie 43, 345, 919, 928, Gundra, Uma Mahesh 1941 Hailu, Asrat 1690 Gough, Erik 1276 1520, 1525 Guodong, Lü 440, 442 Hailu, Elena 1286 Gould, Fred 1394 Griffin, Paul 675, 1488, 1489, 1490 Gupta, Charu 1105 Hajduk, S. L. 1940 Gounoue, Raceline 1893 Griffiths, Emily 1394 Gurarie, David 1149 Hakalima, Aves 925 Goupil, Brad A. 823 Griffiths, Yvonne 25 Gurary, Alexandra 629 Hakim, Lokman 200 Gouvras, Anouk 1810, 1812 Grigg, Matthew J. 22, 1109 Gurley, Emily S. 48, 1110, 1116, Hakimi, Mohamed-Ali 1943 Govella, Nico 1525 Grigg, Michael 547 1359, 1756, 1874, 162, 829 Hakizimana, Emmanuel 333, 1255, Govindan, Ramajayam 1193, 1195 Grignard, Lynn 325, 700 Gurwith, Marc 39 1362 Gowda, Charitha 1653 Grillet, María E. 820 Gushu, Montfort B. 1438 Haks, Marielle 992 Gowda, Kalpana 1604 Grimberg, Brian T. 604, 1455 Gutierrez, Andres 319 Halasa, Yara A. 203, 836, 1631 Gower, Charlotte 1184 Grinev, Andriyan 1383 Gutierrez, Gabriel 1606, 1607 Haldar, Bisakha 1433 Goyal, Kapil 1130 Grobusch, Martin 437 Gutierrez, Gamaliel 191 Haldar, Kasturi 304 Goyenechea, Angel 857, 1416 Grogl, Max 1934 Gutierrez, Juan B. 225 Halder, Amal K. 728, 1292 Gozze, Amanda B. 1224 Gromowski, Gregory D. 828 Gutierrez, Luz 1659 Hall, Aron 1643 Grabias, Bryan 272 Grossman, Marissa 1761 Gutierrez, Victoria 1432 Hall, Brantley 1344 Grabowski, Jeffrey M. 1276 Grove-Gaona, Megan 431 Gutiérrez-Meza, Juan Manuel 1133 Hall, Robert H. 39 Grabowski, Kate 1414 Grubaugh, Nathan D. 837, 982 Gutman, Julie R. 237, 1808, 1918 Haller, Aurelia 581, 1389 Graeter, Tilmann 443, 641 Gruener, Beate 443, 640, 641 Guy, Bruno 575, 576, 833 Hallet, Rachel 308 Graham, Andrea L. 1820 Grueninger, Heiner 1482 Guy, Elizabeth 1126 Halleux, Christine 476, 1063, 1716 Graham, Kirstie 1460 Grüring, Christof 1450 Guya, Bernard O. 263 Halliday, Alice 503 Graham, Stephen 1854 Gryseels, Charlotte 657, 1033, Guzman, Diamelis 202 Halliday, Jo E. B. 1758 Grandez-Castillo, G. 1769 1626 Guzman, Hilda 206, 1397 Halliday, Katherine 879, 1253 Grandez-Urbina, J. A. 574, 1769 Gu, Henry 486 Guzmán, Mitchel 1025, 1823, 1858 Hallyburton, Irene 287 Granjon, Laurent 99 Gu, Yaping 128 Guzmán, Rene C. 852 Halpenny, Carli M. 642 Grant, Edward 471, 1051 Guagliardo, Sarah Anne 787 Guzman, Yajaira 1405 Grant, Warwick N. 479 Gubler, Duane J. 199, 805 Gwadz, Robert W. 150, 1371 astmh.org A-608 Abstract Author Index The number(s) following author name refers to the abstract number.

Halsey, Eric S. 153, 733, 737, 807, Hardin, Rebecca 1764 Healy, Sara A. 668, 669, 1009, Herrera, Claudia P. 546 167, 1413, 1429 Harelimana, Jean de Dieu 358 1371, 1561, 1598, 1599 Herrera, Gianina 1128 Halweg, Sarah 73 Harenberg, Anke 835 Hearn, Pasco T. J. 211 Herrera, Julio 1784 Hamad, Ali 670 Harentsoaniaina, Rasamoelina 1056 Hedeen, David L. 844 Herrera, Raul 1008 Hamadani, Jena D. 1266 Harezlak, Jaroslaw 50 Hedeen, Meghan W. 844 Herrera, Samantha 1249, 1289, Hamainza, Busiku 701, 922, 924, Hariniaina, Elisoa 1055 Hedrick, Chris 278 1617, 1623 925, 963, 1504, 1507 Hariraju, Dinesh 585 Hedrick, Victoria 1276 Herrera, Sócrates 225, 334, 379, Hamapumbu, Harry 399, 938, Harkess, Graeme 1419 Heerwegh, Dirk 666 930, 1541, 1562, 1802 1496, 1529, 1629 Harman, Christopher 1559 Heffelfinger, James D. 162, 1874 Herrera, Víctor 815 Hamed, Kamal 1482 Harn, Donald 1144 Heffernan, Michael J. 1931 Herrera-Valdez, Marco 726 Hamel, Mary Harrabi, Rabiaa 1934 Hegde, Sonia 1116 Herrera-Varela, Manuela 3, 148 Hamel, Mary J. 86, 631, 666, 1503, Harrington, J. M. 1940 Heggen, Anne 1085 Herrick, Jesica A. 1891 1534 Harrington, Laura 756 Heilmann, Elizabeth 898, 899 Hertzmark, Ellen 740 Hamer, Davidson H. 637, 706, 728, Harris, Caroline 1619 Heimbaugh, Chelsea 1271 Herwaldt, Barbara L. 97 1292 Harris, Emma 106, 113 Heinen, Leticia B. S. 1397 Hes, Dominik 1516 Hamer, Diana 782 Harris, Eva 187, 188, 189, 190, Hein-Peters, Katarzyna 830 Hess, Jessica 1264 Hamer, Gabriel 842, 844 191, 192, 198, 651, 725, 726, Heise, Mark 9 Hester, James F. 1827 Hami-Adiamoh, Majidah 124 806, 1273, 1275, 1834, 1836, Heisey, Daniel A. R. 160, 1383 Hewson, Roger 211 Hamid-Adiamoh, Majidah 1352 1838 Helbok, Raimund 21 Heyderman, Robert S. 395, 1474 Hamidu, Buhari B. A. H. 510 Harris, Jason B. 34, 35, 36, 37, 432, Helegbe, Gideon K. 400, 985 Hickey, Patrick 1051 Hamilton, Elizabeth J. 930 433, 588, 1651 Helinski, Michelle 705 Hickey, Patrick W. 469, 1673 Hamilton, William L. 1899 Harris, Michael 282 Heller, Tom 1668 Hickman, Mark 252, 282, 283, Hammam, Olfat 1793 Harrison, Dustin J. 1766 Hemelaar, Simon 348 880, 280 Hammam, Omer 1229 Harrison, Lisa M. 519, 1526, 1717 Hemingway, Janet 767, 1188 Hidalgo Hernández, Bianca 1657 Hammar, Ulf 405 Hart, Kevin J. 975 Hemme, Ryan 811 Higa, Yukiko 1349 Hammond, Colleen 20 Hart, P. John 1824 Hemming, Elizabeth A. 909 Higazi, Tarig 1706 Hamory Hicks, Joan 64 Hart, Robert J. 979, 1830 Hemmings, Kay 767 Higgins, Sarah 1485 Hampson, Katie 77, 664 Hartinger, Stella M. 1295 Henao, Olga 1643 Higgins, Stephen 85 Hampton, Shay M. 1235 Hartl, Daniel L. 930, 1628, 1805, Henard, Calvin 1205 Higgs, Stephen 161, 205 Hamre, Karen E. S. 1857 1833 Henderson, Leslie L. 97 Hilaire, Isabelle J. 34 Han, Eun-Taek 864, 993 Harty, John T. 1571 Henein, Sandra R. 575 Hilaire, Johanne 37 Han, Jin-Hee 864 Harupa, Anke 1901 Henesse, Ian 278 Hildebrand, Vanessa 55 Han, Kay Thwe 1235 Harvey, Steven 1032, 1616 Heng, Somony 926, 1033, 1514, Hildenwall, Helena 271 Han, Soe-Soe 864 Hasan, Bilal 438 1626 Hildreth, Stephen W. 831 Hanafi, Hanafi H. 1370 Hasan, Herdiana 654 Heng, Sopheab 733 Hiliare, Isabelle J. 36 Hananiya, Anthonia M. 1441 Hasegawa, Tomoyuki 1591 Heng, Thay Kheng 912 Hill, Adrian 671, 986, 1590 Hanas, Jay S. 1193, 1195 Hashiguchi, Yoshihisa 1332 Henning, Tyler C. 100, 151 Hill, Adrian V. S. 270, 672, 1556, Hanbury, Blake R. 1132 Hashim, Kamal 1706 Hennink, Monique 573 1587, 1004, 1602 Hang, Jun 1851 Hashim, Ramadhan 1227 Henostroza, German 701, 963 Hill, Catherine A. 1276 Hangoma, Peter 1618 Hasim, Shamilah 1442 Henry, Andrew 1929 Hill, Danika L. 385 Hanieh, Sarah 53 Hasker, Epco 1305 Henry, Michael 121 Hill, Jenny 948 Hanisch, Benjamin R. 232 Hassan, Osama A. 437 Henry, Mirielle M. 1163 Hill, Terence E. 850 Hanna, Sheri L. 648 Hassan, Sumaia B. 1141, 1419 Hens, Niel 926, 1904 Hill, Vincent 1261 Hannaman, Drew 13, 1000, 1925 Hatano, Shinya 1591 Hensley, Lisa E. 662, 1924 Hillesland, Heidi 1038 Hannan, Abu 1756 Hathaway, Nicholas J. 1454 Henzler-Wildman, Katherine A. 995 Hillson, Rebecca 1587 Hanpithakphong, Warunee 1920 Hatwiinda, Sisa 701, 963 Heppner, D. Gray 1605 Hillyer, Julian F. 1818 Hansen, Diana S. 385, 1862 Hauser, W. Allen 1197 Herbein, Joel F. 1132 Hinderer, Jessica 613, 1593 Hansen, Elsa 415, 616 Haverstock, Ryan 1606 Herbert, De’Broski R. 1942 Hinjoy, Soawapak 1392, 1395 Hansen, Kristian S. 885, 1584 Havt, Alexandre 585, 1648 Heredia, Viviana 514, 643 Hinrichs, Dave 291 Hanson, Kara 1579, 1581, 1582, Hawkes, Michael 49, 350, 355, Herick, Jesica 1893 Hinson, Juanita 1374 1583 1485, 1859 Heritage, Rebecca 741 Hip, Phireak 108 Hanssen, Eric 606 Hawkins, Kenneth 339 Herman, Jonathan D. 676, 907 Hirayama, Kenji 33, 400, 985, 1074 Hao, Wen 440, 442 Hawksworth, Anthony 1429 Hermance, Meghan E. 212 Hiruy, Neway 332 Haq, Rouseli 1702 Hay, Christie 1205 Hermann, Laura 1485 Hiser, Michelle J. 1123 Haque, Ashraful 1437 Hay, Simon 1929 Hermsen, Cornelus C. 270 Hittner, James B. 1553 Haque, C. Emdad 155, 801 Hay, Simon I. 360, 832, 1422, 1740 Hernández, Bárbara 857 Hiwat, Helene 782 Haque, Farhana 1110, 1874 Hayden, Mary J. 846 Hernandez, Johanna 277 Hlaing, Tin Maung 1235 Haque, Rashidul 583, 585, 587, Hayford, Kyla T. 722 Hernandez, Libia Milena 745 Ho, May 222 891, 1132, 1135, 1174, 1266, Haynes, John D. 1867 Hernandez, Salvador 1050 Ho, Trung D. 1501 1288, 1426, 1654 Haystead, Timothy 813 Hernandez-Aguilar, Juan 1682 Hoang, Hang B. 279 Harahap, Alida 219 Healy, Jessica M. 839 Hernández Vasquez, Yolanda 544 Hobbs, Charlotte V. 248 Harayama, Rui 726 Healy, Sara 796, 1592 Herold, Jacqueline 1893 Hochberg, Natasha S. 1124, 1282 Harb, Omar S. 981 Herran, Oscar 815 Hochman, Sarah 15 astmh.org Abstract Author Index A-609

The number(s) following author name refers to the abstract number.

Hocker, James 1195 Hotez, Peter J. 517, 698, 1268, Huq, Sayeeda 1654 Inoue, Juliana 874, 901, 1540 Höde, Patrick 692 1730, 1931, 695, 1263, 1892 Hurd, Jacqueline M. 573 Inoue, Sandra 607, 1574 Hodel, Eva Maria 678 Hotwani, Aneeta 585 Hurth, Helene Verena 21 Insisiengmay, Bounnaloth 1782 Hodge, Dana M. 317 Houezo, Jean G. 1655 Husain, M. M. 1874 Introcaso, Camille E. 1215, 1878 Hodges, Theresa 1351 Houghton, Raymond 1934 Hussein, Abdullahi 1424 Invest, John 761 Hodgson, Abraham 883 Houpt, Eric R. 585, 1266, 1426, Hustedt, John 337, 1499 Inyama, Petrus U. 1363 Hodgson, Susanne 671, 1556, 587 Hutchinson, Eleanor 1809 Ioannidis, Lisa J. 1862 1602 Houweling, Tanja A. 67 Hutton, David W. 1630 Ipadeola, Banji 264 Hoerauf, Achim 30, 498, 1207, Howard, Elizabeth J. 1733 Huy, Nguyen T. 33, 400, 985 Ipadeola, Olabanji 265 1209, 1210, 1689, 1704, 1713 Howard, Hayford 1063, 1716 Huy, Rekol 108, 1334 Irani, Ayesha 690, 1173 Hoff, Nicole A. 85, 1430, 662, Howard, Randall F. 1008, 1595 Huynh, Bich-Tram 936 Iranzi, Gad 1362 1117 Howe, Matthew 813 Huynh, Trieu T. 462 Iriarte, Ivan 1405, 1407 Hoffman, Stephen 1609, 1800 Howell, Paul 681 Hwang, Jimee 1509 Iriarte, R. Ivan 1402 Hoffman, Stephen L. 606, 669, Howes, Rosalind E. 360 Hwang, Jusun 105 Irish, Seth 771, 1888 670, 876, 1515, 1556, 1561, Howie, Stephen 1139 Hyatt, Donna 580 Irungu, Lucy 356 1567, 1592, 1594, 1598, 1599, Hristov, Angelica D. 874, 901, 1540 Hyeroba, David 665, 1431 Irving, Helen 133 1600, 1602, 1608, 1610, 1620, Hsiang, Michelle 344 Hyman, James M. 843 Isa, Aisha N. 694 1005 Hsiao, Shih-Huai 724 Hyongvongsithy, Phouthasen 1853 Isaacson, Sinead C. 1279 Hoffmann, Ary A. 1322 Hsieh, Michael 32, 1792, 1793 Ishengoma, Deus S. 318, 304, 310, Hoffmann, Natalie 955, 972 Hsieh, Yi-Ju 1792 1227, 1239, 1467 Hofstede, Stefanie 1150 Htut, Ye 1235 I Ishino, Tomoko 973, 990 Hokke, Cornelis H. 59 Hu, Branda 576 Ishizuka, Andrew 1609 Iamsirithaworn, Sopon 1120, 1392, Holland, Martin 1877 Hu, Jinping 1349 Isingoma, Thomson 1693 1395, 1414 Holleran, John 1498 Hu, Renjie 105 Isiyaku, Sunday 68, 1069 Ianniello, Davide 1787 Hollingsworth, Deirdre 1082, 1723 Hu, Ricardo 1097 Islam, M. Ariful 1756 Iantorno, Stefano 547 Hollingsworth, T. D. 1525 Hu, Yan 520, 1267, 1724 Islam, M. Atiqul 1756 Ibarra-Cerdeña, Carlos N. 1339 Holloway, Kathleen A. 1875 Huang, Chiung-Yu 1552 Islam, Md. Saiful 1110, 1874 Ibe, Ogochukwu 1472 Holmen, Sigve D. 1155, 1794 Huang, Claire 581 Islam, S. 583 Ibeh, Victoria 988 Holmes, Edward C. 852, 1417 Huang, Fang 895 Ismail, Mashair Z. 1419 Ibikounlé, Moudachirou 1078 Holroyd, Nancy 488, 1811 Huang, Liusheng 1457, 1458 Ismail, Miriam D. 1225 Ibitokou, Samad 1557, 1797 Holston, James 726 Huang, Ming-Bo 226 Ismail Koleleni, Ismail 1287 Idampitiya, Damayanthi 1381 Homan, Tobias 727 Huang, Rui 958 Isoe, Jun 1909 Idika, Idika K. 450 Homs-Corbera, Antoni 223 Huang, Yan-Jang S. 161, 205 Isozumi, Rie 655 Idro, Richard 960 Honeycutt, Jared 1793 Huang, Yuzheng 1142 Issa, Issa H. 1015 Igbasi, Uche 1570 Hong, Nguyen V. 1519 Huang, Zhi 1929 Issiaka, Djibrilla 1539 Igboh, Ledor 1262 Hongwei, Pu 442 Huante, Matthew B. 528 Ito, Daisuke 998, 1591 Ige, Olusimbo K. 262 Honório, Nildimar 729 Huaringa, Maribel 1432 Ittiprasert, Wannaporn 1791 Ihantamalala, Felana 1056 Hontz, Robert D. 733, 737, 792, Huber, Curtis S. 1807 Ittiverakul, Mali 912, 1481 Ikeda, Allison K. 1452 1396, 1928 Huckvale, Thomas 1811 Ivers, Louise C. 34, 36, 37, 535, Ikegami, Tetsuro 850 Hooper, Jay W. 1925 Hudson, Mollie 533 715 Ikumapayi, Usman N. 1139 Hooper, P. J. 1076, 1697, 1698 Hue Kien, Duong Thi 783 Ivinson, Karen 666 Ilboudo, Hamidou 1100 Hopf-jannasch, Amber 1276 Huestis, Diana L. 1333 Iwalokun, Bamidele A. 362, 932 Ilias, Muhammad 1096 Hopkins, Heidi 871, 1514 Hue Tai, Luong Thi 783 Iwuchukwu, Nduka O. 1363 Ilnour, Mubarak 1229 Hopper, Douglas 1319 Huezo, Stephanie 291, 319 Iyer, Jayasree K. 1298 Imai, Natsuko 195, 196 Hoq, Mohammad Rubel 35, 1651 Huggins, John W. 464 Izulla, Preston 1528 Imali, Annette A. 1182 Horby, Peter 1768, 1929 Hughes, Angela 767 Immordino, Palmyra 93, 1675 Horna, Gertrudis 1659 Hughes, Laura 1101 Imnadze, Paata 1304 Horne, Kate M. 161, 205 Hughes, Molly 954 J Imoukhuede, Babatunde 986 Hornick, Jean-Luc 1765 Hughes, Philip 813 Imoukhuede, Egeruan 671, 720, Jack, Wanda 53 Hornston, Sureyya E. 1032 Hugo, Leon 1488 1590 Jackson, Belinda M. 1710 Horton, Daniel 1419 Huho, Bernadette 147 IMPACT Study Team 2 345, 928 Jackson, Nicholas 575, 576, 833 Horton, Katherine 112, 114 Hui, Tin-Yu 969 Impoinvil, Daniel 1908 Jacob, Benjamin 1700 Hoshi, Tomonori 1074 Hui, Wang 442 Imrie, Allison A. 629 Jacob, Christopher G. 895, 1235, Hosie, Heather 1595 Humphreys, Georgina 1842 Incani, Renzo N. 1150 1605 Hossain, Jahangir 1116 Humphries, Debbie 519, 600, 1526, Incardona, Sandra 461, 1807 Jacob, Melissa 1096 Hossain, Kamal 1110 1717, 1720 Inchauste, Lucia 1645 Jacob, Shevin T. 733, 737 Hossain, M. 583 Hun, Lewis 750 Indran, Sabarish V. 850 Jacobs, Emily 1663 Hossain, M. Jahangir 1139, 1756 Hung, Chris 1567 Ingabire, Chantal M. 333, 1255 Jacobs, Jan 261, 1806, 1854 Hossain, Md. K. 1874 Hung, Li-Yin 1942 Ingasia, Luiser 300, 306 Jacobs, Jonathan 166 Hossain, Muhammad Belal 1110 Hunsawong, Taweewun 168 Iñiguez, Volga 1311, 1645 Jacobs, Thomas 1863 Hossain, Shakhawat 155, 801 Hunsperger, Elizabeth 811, 1386, Inocêncio da Luz, Raquel 522, Jacobs Slifka, Kara 661 Hostetler, Jessica 410 1405, 1415 1157, 1786 Hunter, Gabrielle 1030 Jaeger, Stefan 1453 astmh.org A-610 Abstract Author Index The number(s) following author name refers to the abstract number.

Jaenisch, Thomas 806 Jerome, J. Gregory 37, 715 Josephine, Samaka 1065 Kakande, Celia 601 Jaffe, Olwen 1306 Jespersen, Jakobs S. 987 Joshi, Amritanshu 1202 Kakani, Evdoxia 681, 1889, 1915 Jagannathan, Prasanna 402, 1565, Jessica, Hernandez 319 Joshi, Sudhaunshu 888 Kakolwa, Mwaka 1478 1569, 1864, 1903 Jeun, Rebecca 695 Joshua, Rubona M. 1321, 1321 Kakumanu, Madhavi 109 Jahan, Assis 54 Jia, Peng 736 Journot, Valérie 251 Kakuru, Abel 26, 257, 1457 Jähnke, Richard 1873 Jia, Ping 1745 Joyce, Michelle 73 Kakuru, Mary 1569 Jahrling, Peter 1924 Jiang, Ju 108, 112, 114, 115, 1851 Juarez, Diana 167, 862 Kalanda, Gertrude 1922 Jahuira Arias, Martha H. 1106 Jiang, Nona M. 1266 Juarez, Marisa 514, 643, 695 Kalani, Rosalia 562 Jaidee, Anchalee 268 Jiang, Rays H. Y. 1450 Juin, Stanley 1300 Kalayanarooj, Siripen 630, 697, 816 Jain, Arti 1223, 1567, 1552, 1562 Jiang, Xioafang 1344 Juliano, Jonathan J. 377, 548, 913, Kalifa, Bojang 238 Jain, Jagrati 295 Jiao, Jin-An 1924 1454 Kalilani-Phiri, Linda 679, 1226, Jain, Komal 1766 Jiménez, Alfons 338 Juliano, Steven A. 785 1922 Jain, Surendra K. 1096 Jimenez, Guadalupe 1729 Juliao, Patricia 598 Kalimuthu, Kovendan 1656 Jalili, Zahraa A. 1059 Jimenez, Xyomara 1405 Julius Julian, Lutwana 1415 Kalinga, Akili 880, 1523, 1549, Jallow, Isatou K. 90 Jiménez-Cardoso, Enedina 1682 Juma, Dennis W. 353 280, 276 Jambou, Ronan 454, 1055, 1056, Jimma, Daddi 215 Juma, Elijah O. 116 Kalkhan, Mohammed A. 1303 1194, 1660 Jim On, Shelby 1934 Juma, Elizabeth A. 1556, 1919 Kalkoa, Morris 655 James, Anthony A. 684 Jin, Albert 1008 Juma, Jane 1261 Kallies, Axel 1862 James, Eric R. 1556, 1602, 1608, Jin, Hongfan 1606 Juma, Omar 670 Kalolella, Admirabilis 28, 255, 669, 1592, 1610 Jing, Li 440 Juma, Omary 743 1578, 1579 James, Kylie 1437 Jis, Mario 1144 Jung, Bong-Kwang 1782 Kaloleni, Ismail 351 Jameson, Samuel Jameson B. 546, Jo, Matthew 181, 579 Junhua, Wang 440, 442 Kalonji, Hercule 1856 792 Johanes, Boniface 255, 1578, 1579 Junior, Policarpo A. S. 1094 Kalsy, Anuj 588 Jamet, Helen P. 116 Johansson, Emily W. 271 Justin Babu, Josephin 1193 Kaltenbach, Tanja 443, 641 Jamil, Kh. Mahbuba 1426 Johansson, Michael 186, 650, Kalveram, Birte 850 Jamonneau, Vincent 1100 1611, 1380 Kama, Eugene L. 1544 Jangpatarapongsa, Kulachart 382 John, Chandy C. 232, 1857, 960, K Kamaka, Kassim 743 Janha, Omar 373, 376 1439, 1485, 1859 Kamal, Hany A. 1347 Kaadan, Abdulnasser 1089 Jankevics, Andris 688 John, Serene 1281 Kamanga, Aniset 136 Kaatera, Fred 1255 Janko, Mark 1454 Johnbull Ogboi, Sonny 265 Kamara, Anitta 243 Kabanywanyi, Abdunoor M. 1478 Janosczyk, Helene 831 Johns, Ben 731 Kamate, Bourama 669, 1598 Kabaria, Caroline W. 363 Janse, Chris J. 1004, 1594 Johns, Benjamin 1551 Kamath, Nitin 682 Kabasele, Freddy 1538 Jara, Jorge 1115, 1771, 1777 Johnson, Barbara W. 1064 Kamau, Edwin 300, 306, 1239 Kabatereine, Narcis 1160, 1184 Jara, Marlene 538 Johnson, Brian J. 1821 Kamau, Luna 760 Kabir, Furqan 585 Jaramillo, Liliana 361 Johnson, Kimberly 1239 Kambale, K. 1256 Kabir, Junaid 694 Jardim, Armando 512 Johnson, Mark 471, 1051 Kamgaing, Nelly N. 524 Kabir, Mamun 1135, 1174 Jarilla, Blanca 1727 Johnson, Reed 1924 Kamgno, Joseph 592, 1208, 1893 Kabongo, Madeleine 1157 Jarillo-Luna, Rosa Adriana 1133 Johnson, Roch C. 1655 Kamhawi, Shaden 1202 Kabongo, Mbuyi M. 1786 Jarman, Richard G. 816, 1272, John-Stewart, Grace C. 1848, 1880 Kamil, J. 1141 Kabore, Achille 1078 1391, 1429, 1770, 1851, 1414 Johnston, Emily 6 Kamiza, Steve 614 Kaboré, Bérenger 1854 Jarman, Rick 1774 Johnston, Kelly L. 483, 484, 485, Kamn’gona, Raphael M. 620 Kabula, Bilali 752, 768, 1189 Jaron, Peter 1261 495 Kampmann, Beate 671 Kaburu, Daniel 562 Jarusevicius, Jaqueline 1354 Johnston, Robert E. 817, 1385 Kampondeni, Samuel 20 Kabuye, Furaha 877 Jarvis, Adrienne 181, 579 Johnston, Sara C. 662 Kampondeni, Samuel D. 1438 Kabyemela, Edward 366 Jasim, Yousif 1141, 1141 Joice, Regina 614, 1225 Kamugisha, Erasmus 318 Kachur, S. Patrick 28, 255, 1578, Jasinskas, Algis 389, 1605, 1796 Jones, Chris 752 Kamuliwo, Mulakwa 767, 922, 1908 Jasseh, Momodou 1139 Jones, Claire 1849, 1850 924, 925, 1504 Kadarkarai, Murugan 1656 Jawara, Musa 124, 1352, 1355 Jones, David S. 668, 1009 Kamuyu, Gathoni 1556 Kading, Rebekah C. 861 Jayawardena, Priyankara 1381 Jones, Dean P. 218 Kamya, Moses 24, 866, 935, 1569, Kadri, Boubacar 1048 Jayaweera, V. P. D.. A.. 171 Jones, Kathryn M. 1931 1844, 1903, 242, 402, 893, 903, Kaewkhao, Karnrawee 301, 311 Jazuli, Farah 466 Jones, Lucy H. 1938 946, 1864 Kaewkungwal, Jaranit 1107, 1481 Jean, Samuel 1908 Jones, Malcolm K. 438 Kanakabandi, Kishore 1866 Kafkova, Jirina 531, 1518 Jean François, Trape 1822 Jones, Marcus 1798 Kanashvili, Marine 1304 Kagashe, Magreth 75 Jeffries, Cynthia 1096 Jones, Rebecca 25, 770 Kandyata, Alister 767 Kahindo, Aimée K. 1463, 1538 Jeffries, David 373 Jones, Sophie 325 Kaneko, Akira 655, 949 Kahn, Stuart 1934 Jemu, Samuel 1151 Jones López, Edward C. 1124 Kaneko, Satoshi 1074 Kain, Heather 17 Jenkins, Adam M. 683 Jongo, Said 670 Kang, Gagandeep 585 Kain, Kevin C. 49, 221, 722, 1435, Jenkins, Bethany J. 980 Jongsakul, Krisada 1494 Kangome-Ngwenya, Tokozile 1504 1859, 1485 Jenkins, Marion W. 708, 1762, 710 Joof, Fatou 1477 Kangwanrangsan, Niwat 1829 Kaindoa, Emanuel W. 1021, 1167 Jenks, Brenden P. 604 Jori, Giulio 119 Kanjala, Maxwell C. 717 Kajeechiwa, Ladda 328 Jenwithisuk, Rachaneeporn 217, Joseph, Gerard 1300 Kanjee, Usheer 1450 Kajubi, Richard 1457, 1458 1829 Joseph, Selina 670 Kanobana, Kirezi 522 Kakaire, Robert 714 Jeremy, Gilles R. L. 920 Joseph, Serene A. 1269 Kanoi, Bernard N. 997 astmh.org Abstract Author Index A-611

The number(s) following author name refers to the abstract number.

Kanokabana, Kirezi 1159 Katabarwa, Moses 593, 1693 Kelly, Jane X. 291, 673 Khoury, David 1437, 1489 Kanoute, Moussa B. 1539 Katakura, Ken 1332 Kelly, Kristina 1165 Khowawisetsut, Ladawan 183 Kanu, Musa S. 243 Kataliko, Kambale 1716 Kelly, Megan L. 317 Kiang, Richard K. 1771 Kanuka, Hirotaka 126 Katamba, Achillies 1577 Kelly-Hope, Louise A. 133, 1689, Kiattibutr, Kirakorn 941, 1537 Kanungo, Suman 423, 1433 Katana, Abraham 1478 1073, 1696, 1700, 1701, 1702, Kibendelwa, Tsongo 355 Kanyago, Christine 461 Katawa, Gnatoulma 30 1708 Kibiki, Gibson 348 Kanyi, Henry 60, 1079 Kateera, Fredrick 333, 358 Kelvin, Elizabeth A. 1197 Kibira, Simon Peter 871 Kanza, Eric 476, 1063, 1716 Katherman, Kyleray R. 236 Kemigisha, Elisabeth 1487 Kiconco, Sylvia 1457, 1458 Kapan, Durrell 798 Katile, Abdoulaye 669, 1561, 1592, Kempaiah, Prakasha 420, 967, 968, Kidima, Winifrida B. 228 Kapella, Bryan 866, 893 1598 1553 Kielian, Margaret 1387 Kaper, James 431 Kato, Cecilia Y. 107, 661 Kenangalam, Enny 1905 Kieu, Huong T. T. 279 Kapilananda, G M. G. 656 Kato, Hirotomo 1332 Kenawy, Mohamed A. 800 Kigondu, Sanyu 1014 Kapisi, James 24, 903, 1569 Kato, Nobutaka 286, 292 Kendjo, Eric 1473 Kigozi, Ruth 893, 935 Kapisy, Alain 454 Kato, Tomoyo 292 Kenefleck, Rupert 395 Kigozi, Simon Peter 946 Kapito-tembo, Atupele 888, 953, Kato, William 670 Kengne-Ouafo, Jonas Arnauld 503 Kiguli, James 268 1225, 1522 Katowa, Ben 399 Kennedy, Mark 975 Kihara, Jimmy 642 Kaplan, Aaron D, 1943 Katsuva, Jean Paul Makelele 350, Kennedy, Stephen H. 364 Kihombo, Aggrey 1631 Kapoor, Neera 132 355 Kenneth, John 1281 Kiknadze, Nino 1304 Kappe, Stefan, 17, 1018, 1829, Katurebe, Charles 866 Kent, Alida 1097 Kikuchi, Mihoko 400, 985 1897, 1568, 1901 Katureebe, Agaba 946, 1844, 1903 Kepha, Stella 508 Kilama, Maxwell 946 Karabou, Potochoziou K. 481, 506 Katureebe, Charles 893 Kephart, Josiah L. 1512 Kilian, Albert 1030, 1031, 1612, Karagiannis-Voules, Dimitrios- Katzelnick, Leah 190, 1273, 828 Kerbis, Julian 861 1845 Alexios 1069 Kaula, Henry 1580 Kerdahi, Khalil 1663 Killingbeck, Sarah 1834 Karamagi, Charles 870 Kaur, Harparkash 255, 341, 772 Kerkhof, Karen 657, 926 Kilpatrick, C. William 1337 Karani, George 1012 Kaur, Jasmeet 1049 Kern, Peter 437, 443, 446, 640, Kim, Charlie 1565, 1569, 1864 Karanja, Diana 57, 1795 Kaur, Upninder 1130 641 Kim, Chloe 875 Karanja, Joan 562 Kaushansky, Alexis 17, 1829 Kern, Petra 640 Kim, Deok R. 423, 1433 Karbwang, Juntra 985 Kavere, Emmy 57 Kersh, Gilbert J. 661 Kim, Eric K. 1462 Karchava, Marine 1304 Kavira, Nathalie 85 Kerubo, Emily 1879 Kim, Hani 1485 Karema, Corine 704, 1362 Kavishe, Reginald 310 Kessler, Albane 1932 Kim, Kami 15, 673 Karen, Ivinson 1488 Kawashima, Emiko 1349 Kessler, Maureen K. 100, 146 Kim, Ryung 615 Karhunen, Markku S. 360 Kaweme, Sydney 922, 924 Kessler, Robert A. 39 Kim, Saorin 1514, 1626 Karim, Shahid 1340 Kayan, Kinsley 1637 Kesteman, Thomas 42, 336, 703 Kim, Tae Yun 1783 Karim, Zachary 420, 967, 968, Kayembe, Serge M. 1463, 1538 Keswani, Tarun 386 Kim, Young Eun 595, 1870 1553 Kayentao, Kassoum 962, 1491, Keven, John 791, 1377 Kimani, Domtila 1556 Karim-Kos, Henrike E. 67 1552, 1798, 1866 Khainza, Annet 1693 Kimber, Michael 490, 492 Kariuki, Muthoni M. 1014 Kayiwa, John T. 213, 1773, 1415 Khalifa, Abdelrahman K. A. 437 Kimbi, Helen 918 Kariuki, Simon 19, 41, 86, 237, Kazadi, Théodore K. 460 Khalil, Syed M. 817 Kimura, Masatsugu 655, 949 631, 666, 911, 1503, 1534, Kazienga, Adama 900 Khalili, A. Hadi A. 444 Kindé-Gazard, Dorothee 1078 1902 Kazimoto, Teckla 1065 Khan, Ashraf I. 433 King, C. Richter 1604 Kariyawasam, K.k.g.d.u L. 542 Kazura, James W. 491, 791, 1212, Khan, Ashraful I. 35 King, Charles 9, 1714, 31, 55, 651, Karl, Stephan 1803 1377, 1554, 651, 1560 Khan, Kamran 1929 1086, 1149, 1271, 1719 Karmali, Anis 85 Kazwala, Rudovick R. 1758 Khan, M. Imran 1306 King, Christine A. 1281 Karron, Ruth A. 1119 Kearney, Brian 860 Khan, Mohammad I. 421 King, Christopher L. 31, 491, 604, Karunasagar, Indarani 1108 Kearney, Christopher 156, 1926 Khan, Salah U. 1116 1086, 1119, 1212, 1455, 1554, Karunaweera, Nadira D. 542, 656, Kebede, Amha 1690 Khan, Salah Uddin 1756 1585 1805 Kebela Ilunga, Benoît 1117 Khan, Shahid M. 1004, 1594 King, Nora E. 735 Karwa, Rakhi 80 Keck, Forrest 157 Khandelwal, Sanjay 16 Kingston, Hugh 268 Kasasa, Simon 599 Keegen, Brian P. 1931 Khang, La 839 Kinh, Nguyen Van 1768, 1837 Kasereka, Claude Masumbuko 350, Keenan, Jeremy D. 1048, 1066, Khantikul, Nardlada 315 Kiniboro, Benson 1531 355 1307 Kharabora, Oksana 1454 Kinunghi, Safari 1810, 1812, 123 Kasinathan, Ravi S. 1825 Keereecharoen, Lily 268 Khater, Emad I. M. 1347, 1348 Kinyari, Teresa 760 Kasonia, Kambale 476, 1063, 1716 Kegele, Josua 21 Khatun, Selina 1874 Kinyoki, Damaris K. 347 Kaspar, Naomi 351, 368, 1015, Kehn-Hall, Kylene 166 Khaw, Loke Tim 139 Kioko, Urbanus 237, 241 1287 Keift, Kristopher J. 210 Kheang, Soy Ty 322, 923, 1505 Kipanga, Purity N. M. 259 Kasper, Matthew 1417, 1428 Keiser, Jennifer 642, 1719 Khedher, Amina 1934 Kirawittaya, Tawatchai 697 Kasper, Matthew R. 744, 1755 Keiser, Philip H. H. 528 Khim, Nimol 1237, 1514 Kirby, Matthew 1888 Kass, Philip H. 141 Keita, Moussa 762 Kho, Kai-Ling 1342 Kirby, Matthew J. 705 Kassa, Tassew 87 Keita, Sekouba 1009 Khondowe, Shepherd 1299 Kirichareon, Naw Lily 328 Kassahum, Belay 866 Keitany, Gladys 1568 Khoo, Jing-Jing 1342 Kiricharoen, Suporn 364 Kassaye, Kebede Deribe 591 Kekre, Mihir 1899 Khoo, Saye 258, 1474 Kirkman, Laura 1176, 1441 Kassim, Kamaka 670 Kelly, Ben 689 Khor, Chee-Sieng 1342 Kirkness, Ewen F. 1798 Kasthuri, Raj S. 1444 Kelly, Gerard C. 279, 1500, 1501 Khoshnood, Kaveh 921 astmh.org A-612 Abstract Author Index The number(s) following author name refers to the abstract number.

Kirkpatrick, Beth D. 39, 583, 1426, Koles, Nancy 1747 Krisher, Lyndsay K. 916 Kyabayinze, Daniel 461, 871 579, 587, 822, 1288 Kolibab, M. 1517 Krishna, Sanjeev 21 Kyaw, Myat Phone 864, 1235 Kisalu, Neville K. 662 Kolibab, Martin 530 Krishnan, Sushma 1455 Kyaw, Ye Myat 1235 Kisinza, William 768, 774, 961, Kolla, Ravi V. 1275 Kroeger, Axel 200 Kyelem, Dominique 1697, 1698 1189, 1365, 1631 Kolling, Glynis L. 1178 Krogstad, Donald J. 1477, 1495 Kyle, Dennis 287, 320, 887, 890, Kiso, Karina 1230 Kolyada, Lena 1363 Krolewiecki, Alejandro 695 1465 Kissinger, Jessica C. 981, 1177 Kombila, Maryvonne 1473 Krolewiecki, Alejandro J. 514, 643 Kyomuhangi, Irene 705 Kiszewski, Anthony 915 Konan, Danielle 659, 1114 Krongthong, Thimasarn 864 Kitau, Jovin 770, 1247, 1365 Konate, Amadou 1598 Kronmann, Karl 471 Kitaura, Kazutaka 1840 Konaté, Lassana 330 Kroon, Erna 1383 L Kitron, Uriel 9, 153, 787, 842, Konate, Lassana 764, 1352 Kruczkiewicz, Andrew 1743 Labadie, Guillermo R. 281 1271, 844 Konate, Siaka 1697, 1698 Krystosik, Amy R. 1333 Laban, Natasha M. 369 Kittayapong, Pattamaporn 798 Kone, Abdoulaye K. 619, 1796 Kuanda, Seni 1403 LaBarre, Paul 339, 1318 Kittur, Nupur 57 Kone, Amadou 1561 Kublin, James G. 321 LaBeaud, A. Desiree 9, 651, 1271, Kityo, Robert 861 Kone, Aminatou 323, 324 Kubofcik, Joseph 589, 1277, 1696 1714, 31, 177, 1086 Kivi, Amétépé 1136 Kone, Bourahima 1561 Kuch, Ulrich 692, 1679 Lacerda, Marcus V. G. 233, 677, Kiware, Samson S. 1525, 1619 Kone, Mamady 1009 Kuchuloria, Tinatin 1304 1555 Kizza, Florence N. 1121 Kone, Younoussou 962, 1552, Kuesel, Annette C. 476, 1063, Lacerda, Marcus V.G. 340, 390 Kjetland, Eyrun F. 1152, 1155, 1794 1798 1716 Lackner, Peter 21 Klarmann, Ute 1207 Kone, Youssounou 1866 Kuhn, Andrea 1555 LaCon, Genevieve 153 Klarmann-Schulz, Ute 1210 Kong, Deok-Hoon 993 Kuhn, Jens 665 LaCrue, Alexis N. 887 Klei, Thomas 1892 Kong, Fanping 1753 Kuhn, Richard J. 1276 Lacsina, Joshua R. 516, 824 Kleinschmidt, Immo 760, 1245, Kong, Nareth 912 Kuipers, Irene 1861 Ladipo, Taiwo O. 365 1365, 1578 Kongere, James 949 Kulik, Margarete C. 67 Lafana, Alice 53 Klemm, Elizabeth 1641 Konnyebal, Godfrey P. 856 Kulkova, Nada 530, 531, 1516, Lafuente-Monasterio, Maria José Kleppa, Elisabeth 1155, 1794 Konradsen, Flemming 570 1518 250 Kligerman, Maxwell 88 Koonings, Jennifer 1513 Kulohoma, Benard 1731 Lage Rodrigues, Luis R. 730 Klimstra, William B. 14, 164, 214 Kopin, Alan S. 683 Kumagai, Takashi 33 Lagler, Heimo 1608 Klinge, Kari F. 1794 Koppinger, Kaitlin L. 515 Kumala, Justin 1073 Lago, Catherine 8, 1274 Klingler, Megan 1084 Koram, Kwadwo A. 468, 472, Kumar, Ashwani 910 Lago, Ednaldo L. 1734 Klion, Amy D. 1208, 1697, 1698, 1717, 1720, 883, 1035, 1067, Kumar, Geetha 991 Laguna-Torres, V. Alberto 807, 1696, 1893 1092, 1093, 1548 Kumar, Jessica A. 1212 1122, 1420 Kloprogge, Frank L. 1480 Korman, Lawrence 464 Kumar, Krishan 1008 Lai, Wen-Ter 724 Kluh, Susanne 1376 Korpe, Poonum 1174 Kumar, Nirbhay 235, 404, 417, Laidemitt, Martina R. 1180 Klungthong, Chonticha 8, 863, Kosek, Margaret 144, 585, 1646 991, 1000, 1586 Laing, Nicolas 93 1274, 1391, 1770 Koskey, Amber M. 1179 Kumar, Rajesh 1000, 1586 Lainhart, William 1823 Knappik, Michael 1853 Kothari, Neha 556 Kumar, Rajiv 1751 Lakshmanan, Viswanathan 996, Knight, Matty 1791 Kotloff, Karen L. 431, 582, 584, Kumar, Sanjai 16, 235, 272 1829 Knight, Philip 1521 1139 Kumar, Shiva 1449 Lakwo, Thomson Luroni 593, 1692, Knipes, Alaine K. 62, 68, 1084 Koudou, Benjamin 129, 130, 152 Kumar, Vinay 991 1712, 1693 Knobler, Stacey 54 Kouodjip Nono, Larissa 918 Kumar, Vipin 611 Lal, Sham 885, 1252 Knopp, Stefanie 1719 Koura, Ghislain K. 635 Kumari, Priti 1175 Lalani, Mirza 1294 Knowles, Sarah 1151 Kouriba, Bourema 380, 619, 1605, Kunene, Simon 45, 344 Lalani, Tahaniyat 471, 693, 1051 Knox, Geoffrey 1076 1796, 1868 Kuntawunginn, Worachet 912, Lalji, Shabbir 699 Kobayashi, Tamaki 369, 399, 1496 Kováč, Pavol 433, 588 1481, 1494, 1841 Lalloo, David 23, 258, 1474, 678, Kochar, Dhanpat K. 611 Kozakai, Yukiko 16, 272 Kunyu, Billy S. 1469, 1547, 1634 1252 Kochar, Sanjay K. 611 Kpawor, Mawolo 1716 Kuong, Khov 1721 Lam, Brandon 1566 Kochel, Tadeusz J. 807, 1413, 1928 Kraemer, Moritz 1929 Kurane, Ichiro 1840 Lam, Eugene 739 Kodama, Yukinobu 33, 985 Krajacich, Benjamin J. 982 Kuranova, Zuzana 867 Lam, Yukyan 1032 Kodio, Aly 1493 Kramer, Laura D. 198, 1364 Kurosaki, Tomoaki 985 Laman, Moses 1803 Koenker, Hannah 1030, 1032, Kramer, Vicki L. 839 Kurtis, Jonathan D. 56, 605, 1727 Lambert, Lynn 613, 1008 1612, 1616, 1845 Kratzer, Wolfgang 443, 641 Kurup, Samarchith P. 687 Lamberton, Poppy H. L. 1184, 1330 Koepfli, Cristian 1221 Krause, Lutz 609, 1218 Kurylo, Elizabeth 1076 Lameyre, Valérie 677, 1473 Koffi, Mathurin 1114 Krause, Peter J. 1219 Kuschner, Robert 1429 Lammie, Patrick 60, 1079, 491, Kohi, Yadon M. 1523 Krcmery, Vladimir 530, 531, 867, Kusi, Kwadwo A. 384 1087 Kohl, Alain 1910 1516, 1517, 1518 Kusuma, Andreas 219 Lamote, Francise 1839 Köhler, Carsten 21 Krebs, Bethany 842 Kutima, Lydiah H. K. 290 Lampah, Daniel 219 Koita, Fanta 1509 Kreishman-Deitrick, Mara 1934 Kvalsvig, Jane D. 1152 Lampah, Daniel A. 1905 Koita, Ousmane A. 1495, 1561 Kremer, Michael 64 Kwarteng, Alexander 1207, 1210, Lanar, David E. 1605 Kokaia, Nora 1304 Kremsner, Peter 21, 1602, 1608 1713 Lanaspa, Miguel 50, 470, 1280 Kokkonda, Sreekanth 1449 Krenz, Bonnie 878, 886 Kwiatkowski, Dominic 1239, 1445, Lanata, Claudio F. 1284 Kolarova, Iva 539 Kretchy, James-Paul 570 1899 Lanciatto, Robert 1064 Koleala, Tamarah 1803 Krisher, Jesse 916 Kwofie, Kofi D. 1092, 1093 astmh.org Abstract Author Index A-613

The number(s) following author name refers to the abstract number.

Landman, Keren Z. 1026 Le Blond, JeJennifer 1290 Lepore, Timothy 1216 Likicho, Lilian 601 Landouré, Aly 1719 Leboulleux, Didier 666 Le Quement,, Sebastian 286 Likwela, Joris Losimba 1533 Landry, Jasmine A. 1795 Lebwohl, Mark 1934 Lerdpanyangam, Kaewta 217 Lilay, abraham 215 Lang, Dennis 585 Le Clec’h, Winka 1810 ler Moo, Carit 268 Liles, W. Conrad 1485 Lang, Jean 576 Ledermann, Jeremy 861 Leroy, Odile 270, 986 Lillebø, Kristine 1155, 1794 Langer, Christine 606 Ledet, Grace 1586 Lerthiranwong, Maethira 1137 Lim, Caeul 415, 616 Langevin, Edith 576, 577, 578 Lee, Andrew H. 1240 le Rütte, Epke 1741 Lim, Chang-kweng 1840 Langham, Julia 1472 Lee, Anne C. C. 1312 Lescano, A. R. 1284 Lim, Pharath 298 Langshaw, Emma 1042 Lee, Bruce 279, 342, 1501 Lescano, Andres G. 184, 277, 545, Lima, Adeiana G. 1573, 1742 Langston, Anne 1632 Lee, Dongmin 1782 604, 1284, 1512, 1659, 1699, Lima, Aldo A. M. 585, 1315, 1648 Lanke, Kjerstin H. W. 1018 Lee, Gwenyth 1646 1737, 1738, 397 Lima, Danielle M. 543, 1406 Lankester, Felix 664 Lee, Jung Seok 1297, 1870 Leshem, Eyal 1300 Lima, Giselle F. M. C. 874, 901, Lansana, Sangare 1495 Lee, Kun-Lin 366, 796 Leslie, Denise 1197 1540 Lanteri, Charlotte 299, 912, 1481, Lee, Linda K. 825, 1678 Leslie, Glaister 315 Lima, Ila F. N. 585, 1648 913, 1454, 1841 Lee, Lydia 1674 Leslie, Toby 1809 Lima, Nathália F. 1224 Lanzaro, Gregory 1357 Lee, Marcus 287 Lesner, Tine H. 1584 Lima, Noélia L. 1315 Laoboonchai, Anintita 1841 Lee, Mieng-Chieh 941 Lessler, Justin 38, 40, 632, 829, Lima, Suelen R. F. 892 Lapierre, Didier 666 Lee, Ming-Chieh 1223, 1537, 1613 1392, 1414, 1650 Limbach, Keith 1603, 1604 Lapouble, Oscar 247 Lee, Moses 606, 1003, 1006 Leta, Gemechu Tadesse 591 Limin, Wang 442 Lapouble, Oscar M. 135 Lee, Patricia J. 1462 Leung, Daniel T. 35, 432, 433, 434, Limwagu, Alex J. 1023 Lara, Jenny 1771 Lee, Rachel M. 517 1651, 588 Lin, Aye T. 494 Lara, Victor 1528 Lee, Rosemary S. 1884 Leurent, Baptiste 869, 1809 Lin, Dan 1898 Larbi, John A. 510 Lee, Seong-Kyun 864 Levashina, Elena A. 1817 Lin, Daniel H. 995 Larocque, Regina C. 432, 433, 588 Lee, Sunhee 15 Levi, José E. 1540 Lin, Harold 1772 Larrauri, Paola 388 Lee, Susan Shin-Jung 513 Levine, Myron M. 39, 431, 582, Lin, Jessica 912, 913, 1454, 1841 Larremore, Daniel B. 1799 Lee, Wenn Chyau 1440 584, 1849, 1850, 1139 Lin, Renyong 438, 441, 447, 448 Larsen, David 922, 924, 925 Lee, Yeuk-Mui 60, 1079 Levine, Rebecca S. 844 Lin, Rone 1669 Larson, Heidi J. 720 Lee, Yoosook 787, 1357 Levitt, Brandt 813 Lin, Xiaoxu 1429 Larson, Peter S. 572, 1530, 1630, Lees, Rosemary S. 7, 1373 Levy, Jens 8, 1274, 1770 Linard, Catherine 363 1764 Lefèvre, Thierry 1373 Levy, Karen 1761, 1882 Lindblade, Kimberly 598, 1026, Larsson, Catherine 579 Legarda, Almudena 1602 Leyden, Jacinta 1126 1503, 1643 Larsson, Cathy 181 Legrand, Eric 1241 Li, Baiqing 382 Lindegardh, Niklas 26, 1920 Laserson, Kayla F. 582, 584, 631, Legrand, Fanny 1893 Li, Benwen 482 Lindh, Jenny 3, 148 1879 Legrand, Roger 835 Li, Jian 993 Lindh, Magnus 1040 Last, Anna 1877 Legros, Mathieu 896, 1394 Li, Jintao 1745 Lindner, Scott E. 413, 975, 1829, Lau, Colleen 521 Leguia, Mariana 167, 862, 1851 Li, Julin 128 1901 Lau, Louis 187 Lehman, Erik B. 1170 Li, Jun 438, 638, 1350 Lindow, Janet C. 822 Lau, Yee Ling 1440 Lehman, Leopold G. 918 Li, Peipei 941 Lindsay, Robbin 155, 801 Lauck, Michael 665, 1431 Lehmann, Tovi 1333 Li, Ping 686 Lindsay, Steve 3, 1243, 148, 946, Laufer, Miriam K. 321, 620, 717, Leite, Ruth M. 1230 Li, Qigui 252, 282, 283, 291 1844 888, 953, 1225, 1522, 1596, Leiva, Karina P. 397 Li, Shanping 962, 1552, 1798 Lindsey, Brianna 431 1808, 1226, 1559 Lembo, Tiziana 77, 664 Li, Tao 1391, 1594, 1620 Lindsey, Nicole 839, 846 Laurens, Laurens B. 984 Le Menach, Arnaud 344, 878, 886, Li, Xiangming 1606 Ling, Clare 328, 1670 Laurens, Matthew B. 270, 619, 1536, 1907 Li, Yuan 1791 Link, Andrew 608 717, 1602, 1605, 1796, 380 Leming, Matthew T. 748 Li, Yuesheng 1185 Linn, Anne 278 Laurenti, Marcia D. 1328 Lemnge, Martha 304, 310, 897, Li, Yuexin 291, 673 Linn, Patrick 278 Lauture, Daniel 1163 1467 Liang, Hong 1228 Linsuke, Sylvie 522, 1156, 1157, Lavazec, Catherine 1498 Lemnge, Martha M. 318, 474 Liang, Li 1567 1159, 1726, 1786 Lavigne, Aline 892 Lemoine, Jean F. 1908 Liang, Song 567, 1161 Liotta, Lance 1106, 1930 Law, Sha 1494 Lemos, Elenice M. 1090 Liang, Xiaowu 1567 Lipkin, Ian 1766 Lawrence, Elizabeth A. 1176 Lemus, Samuel 319 Liang, Yuejin 1205 Lipus, Adam 1311 Lawrence, J J. 637, 706 Lengeler, Christian 1026 Libke, Robert 1772 Lis, Agnieszka 1943 Lawrie, Alison 671, 1590, 672 Lenhart, Audrey 131, 135, 788 Lichtveld, Maureen 782 List, Justin 1121 Lawson, Daniel 1341 Lenk, Edeltraud J. 63 Licon, Meredith H. 546 Little, Eliza A. H. 794 Lawyer, Phil 547 Lentino, Miguel 971 Lieberman, Marya 73, 80, 1314 Little, Francesca 917 Layland, Laura 30 Leo, Yee-Sin 825, 1062, 1678 Liebman, Kelly 131, 788 Littrell, Megan 84, 905, 934, 1511, Lazo Chica, Javier Emilio 549, 550, Leon, Juan S. 741, 1311, 1645 Liendo, Iris 202 1532, 1533, 1580 551, 552 Leon, Kristoffer E. 1681 Lie-Tjauw, Samantha 1262 Liu, Canhui 478 Lazo-Chica, Javier E. 1666 Leonardo, Lydia 521 Lietman, Thomas M. 1048, 1066, Liu, Chengfang 1728 Le, Hung X. 1501 Leong, Cherng Shii 139 1307, 1877 Liu, Dong 1745 Le, Thien-Linh 1779 Leora Walter, Leora 1684 Lievens, Marc 666 Liu, Eugene 1552 Leach, Amanda 666 Lépine, Aurélia 1582 Lige, Bao 882 Liu, Fengchen 1877 Leal, Valéria S. Gomes. 1563 Le Pioufle, Bruno 1498 Lihoradova, Olga 850 Liu, Hui 895 astmh.org A-614 Abstract Author Index The number(s) following author name refers to the abstract number.

Liu, Jenny 915 Lopes, Stefanie C. P. 233 Lukens, Sarah 1705 Mace, Kimberly 1908 Liu, Jie 1426 López, Beatriz 598, 1643 Lukwa, Nzira 151 Macedo, Gleicy A. 1409 Liu, Meiting 1630 Lopez, Gerard 598 Lum, Koji 655 Macedo, Paula A. 839 Liu, Mingli 226, 418 Lopez, Maria R. 1643, 1665, 1777 Luna, Expedito J. A. 1230, 1876 Macete, Eusebio 338, 1478 Liu, Taiping 1564 López, María José 1410 Luna, Giannina 852 MacGillivray, Thomas 1101 Liu, Tong 1353 Lopez, N. Judith 1405, 1407 Lund, Troy 1439 Mach, Ondrej 1424 Liu, Xia 230 López-Cervantes, Malaquías 1401 Lundberg, Lindsay 166 Machado, Kim 677 Liu, Xue Q. 606 Lopez Moya, Mariel 1771 Lundie, Rachel J. 1938 Machado, Marta 901 Liu, Yang 1143 Lopez-Perez, Mary 225, 1562 Lunze, Karsten 637 Machado, Paulo 1206, 1749, 1734 Liu, Yaobao 128, 958 Lopez-Sifuentes, Victor 789 Luong, Hue Tai T. 784 Machado, Ricardo L. D. 391, 393, Liu, Yuanyuan 1058 Lopez-Urbina, Teresa 451 Lupidi, Giulio 119 396, 1563 Livengood, Jill A. 11 López-Varela, Elisa 470 Lupton, Emily J. 976 Macharia, Alex 1556 Li Wai Suen, Connie S. N. 385 Lopez-Verges, Sandra L. 1423 Luquero, Francisco J. 38 Macharia, Daniel 1260 Liyanagamage, Jayathra 171 López Yescas, Juan Guillermo 1388 Lusinde, Rose 699 Machevo, Sonia 338, 1280 Lizarazo, Erley F. 202, 820 Lora, Meredith 564, 1778 Lusingu, John P. A. 474, 310, 1483 Macias, Vanessa M. 684 Ljung, Annika 1040 Lord, Jennifer S. 1359 Lustigman, Sara 488, 1892 Maciel, Bruna 634, 1315 Llanos, Alejandro 959, 1858 Lorenz, Lena M. 774 Luteganya, Victor 811 MacInnis, Bronwyn 1239, 1445 Llanos-Cuentas, Alejandro 329, Lorenzana, Ivette 1410 Lutumba, Pascal 522, 1156, 1157, Mackenzie, Charles D. 498, 1207, 538, 966, 1025, 1095, 1510, Lorenzi-Pena, Olga D. 1402, 1405, 1159, 1726, 1786 1208, 1209 1541, 1823 696, 1379 Lutwama, Julius 861, 1773 Mackenzie, Grant A. 1139 Llewellyn, David 672 Lorimer, Don 1038 Lutwama, Julius J. 213 Mackenzie, Ian 23 Llewellyn, Martin S. 1231 Lorono-Pino, Maria A. 137 Luty, Adrian 1557, 1797 Mackroth, Maria S. 1863 Llewllyn, Stacey 597 Lotharius, Julie 675 Lutz, Jon E. 97, 1044, 1671 MacLean, Michael 1772 Lloyd, Alun 1394 Lotta, Ingrid A. 971 Luvira, Viravarn 1120 MacLennan, Calman A. 395 Lloyd, Jennifer K. 661 Loughlin, Anita M. 1282 Luxon, Bruce A. 1753 MacLeod, Bruce 727 Lmbert, Lynn E. 1593 Loughney, John 197 Luyendijk, Marianne 63 MacLeod, William B. 637 Lo, Ami Colle 394 Louis, Faustine 1361 Luz, Kleber Giovanni 577, 578 Maco, Vicente 1784, 1787 Lo, Aminata C. 302 Lourenco, Christopher 653 Lweno, Omar 670 Macom, John 1499 Lo, Eugenia 909, 937, 1223 Lourido, Sebastian 1943 Lwetoijera, Dickson W. 1013, 1619 Maculuve, Sonia 338, 1478 Lo, Michael 1064 Louya, Frédéric 65 Lwin, Khin M. 268 Madanitsa, Mwayiwawo 679, 377 Lo, Nathan C. 66 Lovchik, Janece 181, 579 Ly, Ann 1862 Maddur Ganesan, Suresh 1831 Loayza, Luis 277 LoVerde, Philip 1810, 1824 Ly, Antarou 174, 1403 Madebe, Rashid 318 Lobo, Neil F. 872 Lowe, Brett 1556 Ly, Kim 108 Madhivanan, Purnima 177 Lock, Michael 39 Lowe, Rachel 730 Ly, Nary 1766 Madi, Diego L. 391 Lodh, Nilanjan 1147, 1181 Loyola, Steev 1659 Ly, Po 1499 Madinga, Joule 1156, 1159 Loeffler, Felix 806 Lozano Beltran, Daniel 564, 716, Lycett, Gareth J. 1186 Madinga, Joules 1726 Logue, Kyle 5 716, 1778 Lye, David C. 825, 1062, 1678 Madison-Antenucci, Susan 1138 Lohfeld, Lynne 1504 Lozano-Fuentes, Saul 137, 779 Lyke, Kristen E. 270, 1602, 1605, Madoungou, Blondel 977 Lohy Das, Jesmin P. 1479 Lu, Feng 128, 993 1868, 1796 Madrid, Lola 50, 470, 1280 Lok, James B. 1264 Lü, Guodong 441 Lynch, Caroline 255, 310 Maestre, Amanda 408 Loke, P’ng 1941 Lu, Louise 1728 Lynch, Matthew 1030, 1032, 1616, Maestre, Ana 1839 Loker, Eric S. 1180 Lu, Miao 587, 1174 1031, 1612, 1845 Mafiana, Chiedu F. 1069 Lokko, Kojo 1845 Lu, Xin 584, 598, 601 Lynch, Michael 1918 Magalhães, Belisa 677 Lol, Carlos 131 Lubinda, Jailos 399, 938, 1529 Lynd, Amy 1186 Magalhães, Izanelda 247 Lombardini, Eric 1137, 1434 Luby, Stephen P. 162, 425, 829, Lyon, Caroline E. 39 Magalhaes, Ricardo Soares 521 Lombore, Bart 1212 1116, 1756, 1874 Lyon, Matt 94 Magalhães, Viviane 1749, 1749 Lon, Chanthap 299, 912, 913, Luca Di Tanna, Gian 376 Lysenko, Nikita 121 Mageni, Zawadi 774 1454, 1481, 1841 Lucantoni, Leonardo 119 Magesa, Stephen 768, 1189 Londono, Berlin L. 792 Lucas, Bradford 781, 1363 Magesa, Stephen M. 1247 Londoño-Barbosa, Diana 541 Lucas, Carmen 277 M Magill, Alan J. 1284 Londono-Renteria, Berlin L. 790 Lucas, John R. 761 Magiri, Charles 1556 Ma, Hai-Zhang 447, 448 Long, Carole 272, 1591, 1799 Lucchi, Naomi W. 911 Magistrado, Pamela 18, 617, 294, Ma, Jennie 587, 1174, 1288, 1266 Long, James H. 255 Luce-Fedrow, Alison 1220 676 Ma, Jinfeng 438 Long, Kanya C. 185 Luchini, A. 1106 Magnussen, Pascal 239, 254, 870, Mabey, David C. W.. 1048 Long, Kristin 9 Luciano, Jacinta 1847 885 Macaluso, J. 106 Long, Thulan 291 Luck, Ashley N. 1896 Magogo, Said 961 Macaluso, Kevin R. 104, 113, 1217, Longley, Rhea J. 1004 Lugemwa, Myers 893, 1460 Magubeia, Augusto 1087 106 Lonnroth, Knut 1278 Lui, James D. 520 Mag Uidhir, Fionn 992 Macareo, Louis 8 Loo, Jennifer D. 1258 Luis, Leopoldina 338 Maguina, Ciro 1851 MacCormick, Ian J. C. 1438 Loparev, Vladimir 1064 Luke, Catherine 579 Maguiña, Jorge L. 277 MacDonald, Andrew S. 1938 Lopera, Juan G. 1427 Luke, Thomas 1924 Maguire, Jason 471, 693, 1051 MacDonald, Nicholas 668, 1009, Lopes, Dinora 901 Lukens, Amanda K. 250, 292, 294, Magumba, Godfrey 705 1008, 1621 Lopes, Maria F. C. 1876 676, 1240 Magzamen, Sheryl 1196 astmh.org Abstract Author Index A-615

The number(s) following author name refers to the abstract number.

Mahajan, Babita 235, 272 Maloney, Kathleen M. 878, 886, Mapua, Salum A. 1010, 1021 Martins-Filho, Olindo A. 1090 Mahama, Emmanuel 1067 1536, 1907 Maradiaga, Johana 1337 Martins Gomes, Ivete 1658 Mahamar, Almahamoudou 1509, Malongo, Amos 1631 Maradova, Eliska 1517 Marube, Elizabeth 700 1539 Malonza, Isaac M. 1014 Marano, Nina 1382 Marzal, Alfonzo 227 Mahanty, Siddhartha 455, 457, 458 Malpede, Brian M. 995, 1898 Marapara, Jorge L. 144 Marzal, Miguel W. 455 Mahatovo, Jemima 939 Maltha, Jessica 1854 Marcenac, Perrine 1625, 1817 Masabho, Peter 121 Mahdi, Ramsan 1015 Malviya, Paritosh 1233 Marchesini, Paola 247 Masakhwe, Clement A. 855 Mahe, Gil 99 Maly, Dustin J. 1172 Marcos, Luis 1784, 1787 Mascarenhas, Anjali 910 Maher, Steven P. 297, 419, 976 Mamadou Ousmane, Ndiath 1822 Marcos, Pool 1432 Mascari, Thomas M. 1370 Mahfuz, Mustafa 634, 1654 Mambandu, Germain 476, 1063, Marcsisin, Sean R. 252, 283, 291 Mas-Coma, Santiago 1780 Mahmood, A. S. M. Sultan 1702 1716 Marfurt, Jutta 219 Mashauri, Fabian 123, 1883 Mahmudur Rahman, Mahmudur Mameli, Enzo 1915 Margolis, Harold S. 811, 824, 1405, Masiga, Dan 259, 158, 859 1110 Mamova, Alexandra 1517, 1518 1382, 1415 Masiye, Felix 1618 Mahopo, Cloupas 634 Mampuya, Ornella L. 1633 Marhone, Joseline P. 1163 Maskery, Brian 1297, 1870 Maiga, Bonaventure 25 Mamuchishvili, Nana 1304 Mariconti, Mara 439, 639 Masokano, Kabir 1907 Maiga, Hamidou 920 Mancha, Bulus 950 Marielle, Bouyou 1239 Mason, Carl J. 585 Maiga-Ascofare, Oumou 323, 1239 Mancini, Emiliano 1912 Mariezcurrena, Ainhoa 1915 Masse, Roseline 1566 Main, Bradley J. 1244 Mancuso, James 561 Marini, Roland D. 1305 Massey, Anna V. 1298 Maina, Alice 112, 114, 115, 1220 Mand, Sabine 1207, 1210 Marks, Morgan A. 1684 Massougbodji, Achille 251, 635, Maina, Mercy W. 80 Mandal, Tarun 1586 Markus, Miles B. 416 936, 1557, 1797 Maire, Nicolas 727 Mandala, Wilson L. 395 Marlow, Mariel A. 72, 1658 Massue, Dennis 774 Maitland, Kathryn 1861 Mandalakas, Anna M. 1126 Maro, Venance P. 1758, 1881 Massung, Robert F. 107 Maitland, Kirsty 486 Mandara, Celine 318, 1467 Marquart, Louise 1490 Masukidi, Malene B. 1469 Maiyaki, Musa B. 713 Mandike, Renata 699, 774, 1015, Marques, Jr., Ernesto T.A. 818, Matanock, Almea 598, 601 Majaliwa, Esther 93 1287, 1467 1399 Mathanga, Don 879, 888, 953, Majambere, Silas 1022, 1619 Mandike, Renatha 351 Marques dos santos, Rodrigo I. 212 1026, 1225, 1253, 1522, 1530, Majanja, Janet 559 Manesia, Javed M. 229 Marquette, Meghan 1620 1546 Majewski, Andrew C. 65, 1707 Manga, Isaac A. 278 Marrenjo, Dulcisária 1847 Mathe, Guidion 1535 Majji, Sai P. 607, 1574, 1601 Mangala, Sophia M. 1633 Marroquin, Lissette 812 Mathenge, Evan 760 Majumder, Eshaque 1101 Mangaza, Nono B. 1634 Marrs, Carl F. 1761 Mather, Frances J. 1495 Makanga, Michael 1052 Mangeni, Judith 370 Marsh, Andrew 731 Mather, Michael W. 407, 618, 673, Makepeace, Benjamin L. 1890 Mangesho, Peter 961, 1631 Marsh, Kennan 498, 675 980 Makharadze, Manana 1304 Mangham-Jefferies, Lindsay 884, Marsh, Kevin 989, 1556, 1602 Matheson, Alastair I. 824, 1260 Makhaza-Jere, Tikhala 678 1472 Marshall, Clara G. 738, 744 Mathew, Anuja 630 Makhviladze, Manana 1304 Mangham-Jeffries, Lindsay Marshall, John 1242, 1525 Mathieu, Els 62, 68, 1084 Maki, Elit 1212 Manhart, Lisa E. 1880 Marston, Barbara J. 1163 Mati, Vitor L. T. 1263 Maki, Jennifer 910 Manix Feintuch, Catherine 615 Marston, Denise A. 1419 Matinyan, Nick 1454 Makokha, Fredrick M. 1424 Manjurano, Alphaxard 123 Marte, Omely 796 Matos, Raissa F. 543 Makori, Euniah 700, 1521 Mann, Andrea 1579 Martens, Brian 890 Matovu, Enock 461 Makoutodé, Michel 1192 Mann, Jennifer 811 Mårtensson, Andreas 868, 927, Matovu, Fred 378 Makowski, Kai 801 Mann, Leah 500 929, 1040 Matowo, Johnson J. 752, 768, Malafonte, Rosely S. 1224 Manna, Byomkesh 1433 Marti, Matthias 320, 614, 930, 1247 Malania, Lile 1304 Manne-Goehler, Jen 1935 1225, 1436 Matowo, Nancy S. 1010, 1021 The Malaria Capacity Development Mann Flueckiger, Rebecca 1259 Martin, Estelle M. 170 Matsinhe, Graca 1535 Consortium Career Development Mann-Flueckiger, Rebecca 1075 Martin, James W. 464 Matta, Nubia E. 971 Groups 96 Manning, Jessica 299, 912, 913, Martin, Jason 580 Matte, Michael 873 MalariaCare 267 1481, 1841 Martin, Julio J. 1932 Matthews, Stephen A. 941 Malaviya, Paritosh 1856 Manoff, Susan 580 Martin, Kelcie 684 Mattioli, Mia 1058 Maldonado, Petraleigh 1385 Manoj, Anita 669, 1592, 1610 Martin, Nicholas 279, 1464, 1500, Mattocks, Melissa D. 817, 1385 Malecela, Mwele 304 Manore, Carrie A. 843 1501 Matusop, Asmad 1442 Malek, M. A. 425 Mans, Dennis 1097 Martine, Jackline L. M. 123, 1883 Matuta, Fabrice 1463 Malhotra, Indu 31, 1086, 1714 Manser, Tim 1264 Martinelli, Luzia 1230 Matute, Maria L. 1771 Malik, Naiela 255 Mantel, Nathalie 835 Martinez, Leonardo 1279 Mauck, Daniel E. 204, 1468 Malima, Robert 961 Mantel, Pierre-Yves 320, 1436 Martínez, Lily 110 Maude, Rapeephan R. 1060, 1310 Maliti, Deodatus F. 143 Mantilla, Jorge 840 Martinez, Melween 1385 Maude, Richard J. 343, 1060, 1101, Maljkovic-Berry, Irina 1429 Mantilla, Juan S. 971 Martinez, Nora 379, 1802 1310 Malk, Intithar 1685 Manu, Grace 1067 Martinez, Sandra 1054 Maulana, Pedro Manuel 1087 Malleret, Benoit 1440 Manzano, José Ignacio 688 Martínez, Sindy 1408 Mavere, Caroline 670, 743 Mallewa, Jane 258, 717, 1474 Manzano, Maria P. 1932 Martinez, Yanet 812 Maves, Ryan 471 Mallik, Buddhadeb 1621 Mao, Sivanna 298 Martínez-Puchol, Sandra 426 Mavuzi, Betricia 1538 Malloy, Halley 978 Mao, Sokny 657, 1626 Martinez-Vega, Pedro 1750 Mawili-Mboumba, Denise Patricia Malloy, Michael 1905 Maphumulo, Andile 1152 Martin-Herrou, Hadrien 122 977, 1473 Malone, David 134, 771 Mappin, Bonnie 271, 943, 944, 945 Martin-Jaular, Lorena 223 Mawindo, Patricia 1226 astmh.org A-616 Abstract Author Index The number(s) following author name refers to the abstract number.

Maxwell, Kilama 1844 McCracken, Michael K. 790, 823, Mehta, Saurabh 1281 Meurs, Lynn 1159 Maxwell, Kolawole 265, 1031 1393 Meier, Martin F. 1654 Meyer, Kari R. 1124 Mayanja, Harriet 1565 McCullough, Hazel E. 96, 1301 Meijgaarden, Krista E. van. 992 Meyers, Jacob I. 755, 982 Mayer, Benjamin 640 McCullough, Jeffrey 78 Meite, Aboulaye 1707 Meyers, Lauren A. 186, 664 Mayer, Leonard 1064 McCurdy, Gail 661 Mejia, Carolina 716, 1778 Meyers, Monica M. 1422 Mayer, Sandra V. 1397 McDermott, Adrian 1800 Mejia, Raul 569, 1411 Meymandi, Sheba K. 1050 Maynard, Kelley 1285 McDew-White, Marina 906, 908, Mejia, Rojelio 695, 1268 Mgamba, Janeth 1467 Mayor, Alfredo 338, 1478 1236 Mejía-Jaramillo, Ana M. 793 Mgando, Joseph P. 1013 Mayor, Pedro 1737, 1738 McDonald, Chloe E. R. 722, 1485 Mekaru, Sumiko 1929 Mgata, S. 280 Mayo-Smith, Leslie 36 McDonald, Circe E. 1711 Mekasha, Sindew 1692, 1712 Mgata, Saidi 880 Mayta, Holger 51, 457, 1296 McDonald, Emily A. 56, 1727 Mekonennen, Walelingn 509 Mgeni, Mohammed 670 Mayta Malpartida, Holger 456 McDonald, Robert W. 1681 Mekonnen, Seleshi K. M. 316 Mharakurwa, Sungano 313, 331, Mayxay, Mayfong 871 McDonald-Fleming, Renee 29 Mekuria, Asrat Hailu 591 889, 1629 Mazhar, Sabah N. 1061, 1685 McDonough, Erin 733 Melak, Berhanu 332 Mhashilkar, Amruta 478 Mazigo, Humphrey 743 McDonough, Erin A. 737 Melby, Peter C 690, 1746, 1748, Michael, Audrey 53 Mazinga, Charles 879, 1253 McElhinney, Lorraine M. 1419 1753 Michael, Charles A. 633 Mazitschek, Ralph 676, 907 McElroy, Peter 927 Melendez, Victor 252, 291 Michael, Daniel 877 Mazzalupo, Stacy 1909 McElvany, Ben 181, 579 Melendrez, Melanie C. 1391, 1429, Michael, Edwin 304, 1705, 1709 Mbabu, Murithi R. 660 McGaha, Tommy W. 1695 1774 Michaelides, Tula 1765 Mbacham, Wilfred 884 McGrath, Monica 585 Melnikov, Alexandre 1805 Michalikova, Lenka 530, 531, 1516, Mbacke, Sembene 1822 McGrath, Nuala 720 Melo, Gisely 340, 677 1518 Mbala, Placide K. 464 McGraw, Elizabeth A. 799 Menard, Didier 622, 657, 926, Michalski, Michelle L. 1896 Mbambara, Saidon 889 McGready, Rose 268, 364, 906, 1237, 1514, 1626 Michalski, Shelly 500 Mbanefo, Evaristus C. 33 908, 940, 1236, 1920 Mendell, Nicole L. 103 Michelini, Elisa 288 Mbanefo, Evaritus C. 985 McHardy, Stanton 1824 Mendes, Lilian d. Melo. 1563 Michelo, Charles 313 Mbarack, Zainabu 1065 McHenry, Amy M. 994 Mendes, Maria Tays 550 Michelow, Ian C. 605 Mbare, Oscar 1243 McIntyre, Tara 1569 Mendonca, Vitor R. R. 1555 Mickum, Megan L. 1795 Mbarek, Nasri 1934 McKay, Michael 368 Mendoza, A. Patricia 1738 Midzi, Nicholas 381, 1145 Mbété, Roger A. 1765 McKemey, Andrew R. 2 Menéndez, Clara 338, 1478 Mier-y-Teran, Luis 1414 Mbeye, Nyanyiwe M. 525, 702 Mckenney, Jennie L. 733 Meneses, Claudio 1202 Miguel, Camila 550, 551, 552, 549, Mbickmen, Steve 1208 McKibben, Maxim J. 31 Menezes, Maria José 1224 1666 Mbofana, Mariamo 1259 McLellan, Sandra L. 1179 Menezes, Taís 1749 Miguel, Edward 64 Mbogo, Charles 760 McLennan, John D. 95, 1291 Meng, Zaojing 1891 Mikautadze, Teona 1304 Mbogo, Loice W. 516 McLeskey, James T. 1165 Meng Chuor, Char 299, 1841 Mikita, Kei 1147 Mbondoukwé, Noé Patrick 977 McManus, Donald P. 638, 1185, Mengesha, A 527 Mikolajczak, Sebastian A. 1829, Mbonye, Anthony K. 870, 885, 1144, 1828, 438 Menon, Jayaram 22 1897 1252, 1536 McManus, Julie F. 1131 Menon, Sonia 1293 Mikolasova, Gertruda 867, 1516, Mbonye, Martin K. 275 McNulty, Ronan 252, 282 Mens, Petra 266, 1804, 358 1518, 531 Mborekere, Martini 1693 McNulty, Samantha N. 1785 Mensah, Athanasia V. I. 986 Milanoi, Sylvia 558 Mboua, Bitoungui 1855 McPhun, Virginia 1003, 1006 Mensah, David 1035, 1614 Mildenhall, Dallas 1873 Mboup, Souleymane 234, 1450, Mcqueen, Kelly 1313 Mensah, Kossi M. 1136 Miles, Michael A. 1231 1799 McQuilkin, Patricia 465 Mensah, Nii O. 267 Miles, Tim 1098 Mbugi, Patrick J. 1020 McShane, Helen 1800 Mensah, Sedzro 1035 Milesi, Pascal 1192 Mbuji, Peter 1467 McVicar, Daniel 630 Mensah-Brown, Henrietta 1443 Miley, Galen P. 673 Mbwasi, Ronald M. 1675 Md Idris, Zulkarnain 949 Menten, Joris 274, 1922 Millán Oñate, José 1562 Mbwili, Clara 925 Mduluza, Takafira 381, 1145 Menya, Diana 1162 Miller, Andre 1791 McArthur, Monica A. 1849, 1850 Mduma, Estomih R. 634 Mera y Sierra, Roberto l. 1780 Miller, Barry 861 McAuley, Alexander J. 161 Mead, Daniel G. 844 Mercereau-Puijalon, Odile 1237 Miller, Becky A. 150 McCall, Eileen 495 Mean, Phou 84 Merchant, Amina 1313 Miller, Christopher 1485 McCall, Philip J. 200, 763, 1883, Medeiros, Nayara I. 536 Mercier, Aurélien 795 Miller, Jessica 1568 1619 Medeji, Caroline 718 Merino, Kristen M. 991 Miller, John 922, 924, 1504, 1507, McCarthy, James S. 270, 597, 675, Medhin, Girmay 316 Meroni, Valeria 639 1511 1490, 1042, 1488, 1489 Medina, Alexis 1728 Meschino, Steve 1378 Miller, Mark 585 McCarthy, Kevin A. 667 Medina, Audrie A. 1746, 1748 Meseko, Clement 172, 1421 Miller, Melanie M. 520, 1267 McCarthy, William 1934 Medina, Freddy A. 1835 Meshnick, Steven R. 377, 548, 679, Miller, W. Allen 649 McCollum, Jeffrey 738 Medina, Juan F. 1835 1841 Miller, Woutrina 1762 McConville, Malcolm 1854 Medina, Norma H. 1876 Mesirov, Jill 50 Miller-Fellows, Sarah C. 55 McCormack, Shelley 26, 257 Mee, Peter T. 1322 Messenger, Louisa A. 1231, 548 Milligan, Gregg 802 McCormick, Benjamin J. J. 585 Meek, Sylvia R. 705, 1460, 1499 Messer, William 625, 628 Milligan, Paul 238, 302, 330, 359, McCornack, Jocelyn 515 Meeks, Janet 629 Messina, Jane P. 1422 658, 1520, 1906 McCoy, Andrea J. 737 Meester, Robert 1856 Methaneethorn, Janthima 1486 Millogo, Athanase 1196 McCoy, John Philip 1202 Meeyai, Aronrag 721 Metuge, Haelly Mejane 503 Milne, Kathryn H. 672 McCracken, John P. 1665, 1777 Meffre, Eric 1866 Metzger, Marco E. 105 Milner, Danny 20, 50, 614 astmh.org Abstract Author Index A-617

The number(s) following author name refers to the abstract number.

Min, Aung M. 364 Mohammed, Wahjib 1614 Moraes, Sandra L. 1573 Movila, Alexandru 610 Min, Aye 940 Mohammed Jabo, Aliyu 68 Morales, E. Angelo 1738, 1737 Moya-Alvarez, Violeta 1257, 1916 Min, Duk-Young 1782 Mohon, Abu N. 891, 1674 Morales, Javier 577, 578 Moyes, Catherine 1929 Min, Zhang 1465 Moi, Meng Ling 1840 Moran, David 1665 Moyle, Sarah 671, 672 Minagawa, Kogomi 1349 Moir, Juan C. 1643, 1777 Mordmüller, Benjamin 21, 1602, Moyo, Evance 1627 Minakawa, Noboru 1349 Moir, Susan 1866 1608 Mozo, Karen 427 Ming, Zhou 1891 Moiroux, Nicolas 122 Moreira, Rosa 811 Mpabanzi, Liliane 1156, 1159, Minja, Daniel 310 Moke, Fenny 582, 584, 1261 Moreno, Alberto 218, 225, 603 1726 Minja, Elihaika Mokuolu, Olugenga 238 Moreno, Brechla 663 Mpagala Kihomo, Robert 1631 Minja, Elihaika G. 75, 1167 Molina, Douglas 329, 1510, 1567 Moreno, Edwin 1128 Mpanya, Godéfroid 1533 Minta, Anna A. 962 Molina-Cruz, Alvaro 682, 759, Moreno, Elena 670 Mpina, Maximillian 670 Mintz, Eric D. 582, 584, 1163, 1008 Moreno, Marta 144, 149 Mrango, Zakayo 1483 1261, 1643 Molla, Yordanos B. M. 1290 Moreno, Yovany 512 Msamanga, Gernard I. 740 Miranda, Debora 1319 Mollel, Jackson 670 Moreno Leirana, Marta 1823 Msellem, Mwiniyi I. Miranda, Eduardo F. 453 Molnarova, Katarina 1516 Mores, Christopher N. 10, 165, Msellem, Mwinyi 927, 868, 929, Miranda, José C. 1203 Moloney, Grainne M. 347 790, 792, 823, 843, 1393 1040 Miranda, Maria Consuelo 577, 578 Molyneux, David 1700 Morgan, Juliette 1847 Msellemu, Daniel F. 354 Mirante, Clara 1153 Molyneux, Elizabeth 1285 Morgan, Marjorie 1331 Msipa, Patrick 1125 Mireji, Paul O. 259, 1323 Molyneux, Gemma 485, 489 Morgan, Oliver 1643 Mswanya, Christopher 276, 1523, Mireku, Michael O. 635, 1257 Molyneux, Malcolm E. 395, 614 Mori, Nicanor 1428 1549 Mirelman, Andrew J. 636 Molyneux, Sassy 1556 Morita, Masayuki 997 Mtali, Austin 879, 1253 Miri, Emmanuel 950 Mombo-Ngoma, Ghyslain 1211 Moritz, Robert L. 1901 Mtali, Sarah G. 75 Mishra, Nischay 1766 Momoh, Veronica 1576 Morkowski, Stan 1934 Mtoro, Ali 1483 Misiano, Paola 288 Monaghan, Andrew J. 846 Morris, Jamae 1261 Mtove, George 532, 563, 961 Misra, Kavita 877 Mondal, Dinesh 585, 1228 Morris, Sheldon 230 Mtumbuka, Esther 886 Misra, Pravas R. 708 Mondini, Adriano 827 Morris, Ulrika 927, 929 Mu, Jianbing 319 Missamou, François 65 Monge, Maria 1873 Morrisey, Joanne M. 980 Mu, Lina 48 Mitchell, Rebecca M. 1534 Mongkolsirichaikul, Duangrat 863 Morrison, Amy C. 153, 185, 733, Mubarik, Yusif 1210 Mitchell, Sara N. 681, 1915 Monje, Fred 1111 737, 792, 807, 1699, 1413 Mucci, Juan S. 1201 Mitre, Edward 1710, 1894 Monroe, April 1032 Morrison, Elaine 1747 Muche, Rainer 640 Mitreva, Makedonka 1785 Monroy, Carlota 1337 Morwabe, Alex 1879 Muchiri, Eric 9, 31, 1271, 651 Mitshiabu, Viviane 1633 Monroy Pérez, Eric 1657 Moschella, Samuel 1669 Muchoki, Terry 877 Miura, Kazutoyo 272, 997, 1591, Montague, Mark 1460 Moseley, Pope L. 967 Mudeppa, Devaraja 910, 1449 1799 Montalvo, Ana M. 1095 Moser, Janice M. 833 Muegge, Brian D. 1654 Miyashiro, Samantha I. 1200 Montaño, Nair 1095 Moser, Kara 1235, 1600 Muehlenbachs, Atis 1808 Miyingo-Kezimbira, Anne 213 Montano, Silvia M. 746, 1428 Moses, Hajara John 1317 Mueller, Ivo 53, 385, 609, 1221, Mizinduko, Mucho M. 1282 Montavon, Céline 592, 1208 Mosha, Dominic F. 1921 1531, 1550, 1803 Mkandawire, Felix A. 717 Montecino, Diego 1376 Mosha, Frank 961, 1631 Mueller, Yolanda K. 1229, 1855 Mkindi, Catherine 670 Monteiro, Daniela C. S. 543 Mosha, Franklin 752, 768, 1189, Mufamadi, Brenda 585 Mkoji, Gerald M. 1180 Monteiro, Wuelton M. 340 1247, 770, 771, 1365 Mugasa, Joseph 961, 1631 Mkopi, Abdallah 1015 Montgomery, Joel M. 184, 803, Mosha, Jacklin 1227 Mugayo, William 1693 Mkude, Sigsbert 886, 1467 819, 824, 1045, 1064, 1260, Mosha, Jacklin F. 348 Mugenyi, Levi 1904 Mkwanda, Square 1073, 1689 1869, 1902, 1261, 1382, 1415 Moshi, Irene R. 1167 Mugnier, Monica 1939 Mlacha, Yeromin P. 349 Montgomery, Susan P. 1071, 1643 Mosi, Lydia 659, 1114 Muhangi, Lawrence 403 Mlaganile, Tarsis 1065 Montoya, Magelda 190, 1273 Mosore, Mba T. 1092 Muheki Tukahebwa, Edridah 1184 Mlambo, Godfree 235 Montoya, Rosario 1278 Moss, Eli 292, 329, 907, 1628, Muhindo, Mary 24, 257, 903 Mmbando, Arnold S. 1013, 1021 Montresor, Antonio 1082 1241, 1799 Muhindo, Mary K. 1864 Mmbando, Bruno 304 Mony, Vidya 289 Moss, William 331, 1529, 369, Muia, Alfred 1556 Mnzava, Ruth 1631 Moo, Hilda 1920 399, 938, 956, 957, 1496 Muigg, Veronika 21 Moch, J. K. 1867 Moonah, Shannon N. 1266 Mossel, Eric 861 Muinde, Jackson 1086, 1714 Moe, Christine 573, 1262 Moonga, Hawela B. 701, 963 Mossoko, Mathias 1117 Muiruri, Samuel 651 Moebius, Jacqueline 1572 Moore, Brioni 1803 Mostaguir, Khaled 1679 Mukabana, Richard 727 Moechtar, Mischka 1450 Moore, Christopher C. 1283, 1860 Mott, Josh 562 Mukabana, Wolfgang R. 326, 356 Moehrle, Joerg 675, 1490 Moore, Emma A. 1136 Motta, Fernando C. 1775 Mukadi, Patrick 85 Mogasale, Vijayalaxmi V. 1870 Moore, Jason D. 774 Mouhamadou, Chouaibou S. 130, Mukaka, Mavuto 258, 678, 1474 Mogasale, Vittal 1297, 1870 Moore, Julie M. 224 152 Mukaka, Mavuto F. J. 620 Mogeni, Daniel O. 819, 1869 Moore, Laura B. 517 Moulds, JoAnn 1796 Mukele, Rodin 1157 Mogeni, Ondari D. 1045 Moore, Sarah J. 774 Mountford, Adrian P. 1937 Mukherjee, Angana 904 Mohamed, Abdinoor 1382 Moore, Sean M. 40, 1650 Moura, Sofia 1153 Mukherjee, Swati 624, 1387 Mohamed, Ally 1467 Moore, Sean R. 1648 Mourão, Marina M. 1789 Mukhopadhyay, Saikat 69 Mohamed, Gedi 1424 Moorhead, Andrew R. 1896 Moussa, Namountougou 775 Mukoko, Dunstan 1086, 1714 Mohamed, Hanan 1706 Moormann, Ann 1560 Moussa Djaouda, 567, 1161 Mukone Mudanga, George 101, Mohammed, Mohammed N. 1687 Moraes, Milena L. 1315 Moutairou, Kabirou 1557 102 astmh.org A-618 Abstract Author Index The number(s) following author name refers to the abstract number.

Mukonka, Victor 1708 Mushatt, David M. 1495 Mwinzi, Pauline N. 1071, 1182 Nartey, Alexander A. 296 Mukose, Aggrey 1577 Muskavitch, Marc A. T. 683 My, Phan Vu Tra 430 Nartey, Gloria A. 296 Mukunda, Faustin 522, 1157, 1726 Musoke, David 599, 1012 Myal, Yvonne 1745 Narum, David L. 668, 1008, 1009, Mukunda, Wilfried 1786 Musokotwane, Kebby 637 Mychaleckyj, Josyf C. 583, 1288 1621 Mukunzi, Silvanos 559 Musset, Lise 1241, 1805 Myers, Christopher A. 733, 737 Nasci, Roger S. 839, 846 Muleba, Mbanga 146, 956, 957 Musso, Didier 1923 Myers, Janette B. 1038 Nascimento, Eduardo J. M. 1399 Mulebeke, Ronald 1577 Mustafa, Ghulam 421 Myers, Nicholas 1314 Nascimento, Luis G. G. 1540 Mulembakani, Prime M. 662 Musuva, Rosemary 1071 Myers, Nick 80 Naser, Abu Mohd 162, 829 Mulenga, Modest 956, 957, 1922 Musuva, Rosemary M. 1072 Myers, Todd E. 111 Naser, Ibrahim 509 Muli, John M. 531, 1518 Mutabazi, Miriam 601 Myezwa, Helen 734 Nash, Scott D. 366 Müller, Ivo 955 Mutabingwa, Theonest 1922 Mylne, Adrian 1929 Nash, Theodore E. 455, 457, 1198 Müller, María L. 1115 Mutafungwa, Anold 351, 699, Myriam, Harry 1822 Nasir, Susann 866 Mulligan, Connie J. 978 1287 Mysore, Keshava 686 Nasrin, Dilruba 584, 1139 Mullins, Kristin E. 1851, 1852 Mutai, Beth 112 Mzilahowa, Themba 1073, 1188 Nasser, Julio 695 Müllner, Matthias 12 Mutambay, Cesar K. 464 Mzimbiri, Imam 664 Nassirou, Baido 1048 Mulogo, Edgar 873 Mutapi, Francisca 381, 1145 Mzungu, Elton 1086 Nataro, James P. 389, 431, 582, Mulrooney, Carol 286 Mutembo, Simon 637 584, 1639, 1648 Mulvaney, Shawn P. 733, 737 Mutesa, Leon 333, 1255 Nathavitharana, Ruvandhi 560 Mumba, D. N. 1256 Muthami, Lawrence 760 N Nation, Catherine S. 689, 1830 Mumba, Dieu-donné N. 460 Muthoka, Phillip 562 Nativel, Priscilla 1194 Nabasumba, Carolyn 268 Mumba, Dieudonne 1239, 1547 Muthoni, Rhodaly 1362 Naulikha, Jacqueline M. 516 Nabicassa, Meno 1877 Mumba, Peter 1035, 1622 Muthui, Michelle 1556 Naulikha, Jaqueline M. 1880 Nabukenya, Irene 1773 Mumbengegwi, Davis R. 1502 Mutuku, Francis M. 9, 1086, 1271, Naumenko, Anastasia N. 1913 Nackers, Fabienne 1229 Mumina, Ann 1103, 1229 1714 Nausch, Norman 1145 Nadjm, Behzad 532, 1768 Munawar, Kashif 1347, 1348 Mutuku, Martin W. 1180 Navaratnam, Visweswaran 244 Nag, Sidsel 310 Muné, Mayra 857, 1416 Muvunyi, Claude 333 Navarrete, Patricia 916 Nagarkatti, Rana 1105 Munene, Marianne 1556 Muwalo, Francis 717 Nawagi, Faith 526 Nagata, Melissa 629 Mungai, Peter 9, 1271, 1714 Muyembe, Jean-Jacques 662, 1430, Naw Nyo, Slight 328 Nagelkerke, Nico 1741 Mungai, Peter L. 31, 1086 464, 460, 1117 Nayak, Uma 583, 587, 1288 Nahar, Nazmun 1756 Mungwira, Randy G. 717 Muyembe-Tamfum, Jean-Jacques Nayakwadi Singer, Monica 1714 Nahum, Laila A. 1789 Munishi, Oresto 1287 85 Naylor, Caitlin 587 Nair, Shalini 908, 983, 1236, 1832 Munoz, Angel 1411 Muzia, Lucy 767 Nchimbi, Happy 1578 Naissant, Bernina 1176 Munoz, Benito 286 Mvuata, Marilyne L. 1469 Ndawula, Bbaale 871 Nakajima, Rie 389, 1605 Muñoz, Diana 1761 Mwabulanga, Adam 276, 880 Ndawula, Bbale 461 Nakajima-Sasaki, Rie 1796 Munoz, Elyse E. 413 Mwabulanga, J. 280 Ndege, Samson 83 Nakanwagi, Grace 1460 Muñoz, Fredy 598 Mwafongo, Winfred 351, 1287 Ndejjo, Rawlance 599 Nakasujja, Noeline 24 Munoz-Jordan, Jorge L. 811, 1390, Mwaipape, Osiah 699 Ndhlovu, Micky 1525 Nakayasu, Ernesto 1276 1405, 1835, 1064 Mwakapeje, Elibariki 351, 1287 Ndiath, Mamadou Ousmane 1187 Nakayiki, Teddie 861 Munsami, Mavis 1152 Mwakazanga, David 1856 Ndiaye, Daouda 234, 904, 1450, Nakhasi, Hira L. 542, 1202 Munthali, Clyton A. P. 678 Mwambulanga, Adam S. 1523 1477, 1694, 1799, 1833 Nakioba, Sarah 275 Munthali, Spy 731 Mwanaupanga, Mwajabu M. H. Ndiaye, Jean Louis 254, 394, 330, Nalikka, Betty 861 Munyakanage, Dunia 1362 1575 1906 Nalubega, Mayimuna 402, 1569 Mupere, Ezikiel 714 Mwancha-Kwasa, Magoma 461 Ndiaye, M. 359, 1906 Naluwu, Kate 402, 1569 Mupoyi, Sylvain 1157 Mwandama, Dyson 1026, 1546 Ndiaye, Magatte 254, 394, 986 Nalwoga, Angela 403 Muralidharan, Vasant 1946 Mwandawiro, Charles S. 508 Ndiaye, Yaya 1477 Namagambo, Barbara 1773 Muratova, Olga 668, 1621 Mwanga-Amumpaire, Juliet 1487 Ndiaye, Youssoupha 278 Namasivayam, Sivaranjani 1177 Murcia, Carlos 812 Mwangi, Paul W. 528 Ndibazza, Juliet 403 Namasopo, Sophie 49, 1485, 1859 Murdiyarso, Lydia 1543 Mwangungulu, Stephen P. 335, Ndiop, Medoune 246, 1250, 1251, Nambozi, Michael 1922 Murillo, Efrain E. A. 1050 1023 1456 Namirimu, Felistas N. 402 Murillo, Gabriela 812 Mwanziva, Charles 276, 881, 1549, Ndombi, Eric M. 57, 1795 Namwanje, Harriet 593, 1085 Murillo Solano, Claribel 1476 280, 880, 1523 Ndour, Cheikh T. 254 Nanayakkara, N. P. Dhammika 252, Muro, Florida J. 563 Mwapasa, Victor 258, 377, 679, Ndour, Moussa 246 674 Murphy, Jennifer 1261 1474, 1922 Ndour, Papa Alioune 1498 Nandy, Ranjan K. 423, 1433 Murphy, Laura L. 1296 Mwase, Enala 1708 N’Dri, Bedjou P. 152 Nankabirwa, Joaniter I. 242, 1460 Murphy, Sean C. 270 Mwebaza, Norah 1457, 1458 Ndyomugyenyi, Richard 885, 1252 Nankya, Annet M. 213 Murray, David C. 495 Mwenda, Reuben 879, 1253 Neafsey, Daniel E. 292, 329, 1241, Nankya, Felistas 1569, 1864 Murray, Kristy O. 698, 1730 Mwende, Faith 60, 1079 1628, 1799, 1833, 1805 Nanyiti, Aisha 378 Murray, Megan 37 Mwendwa, Mwenesi 351 Neal, Aaron T. 1900 Naples, Jean M. 1147, 1181 Murray-Kolb, Laura 634 Mwenechanya, M 20 Neal, Jillian 248 Naranjo, Nelson J. 140 Murshedkar, Tooba 1610 Mwenesi, Mwendwa 1287 Neatherlin, John 819, 824, 1064, Narayanan, Aarthi 157 Murugesan, Udayaprakash 407 Mweresa, Collins K. 326 1382 Narh, Charles 594, 659, 1114 Musapa, Mulenga 767 Mwesigwa, Julia 314, 376, 1544 Nebe, Obiageli J. 1069 Narh, Clement 296 Musenga, Junior 1633 Mwingira, Felista W. 374, 972 Nebie, Issa 325, 671 astmh.org Abstract Author Index A-619

The number(s) following author name refers to the abstract number.

Nebie, Roger 117 Nguyen, Hanh Tien T. 173 Nishat, Naoshin S. 35, 1651 Nosten, Francois 26, 268, 328, Needham, James 1934 Nguyen, Hien 279, 1464, 1500, Niu, Xinnan 608 364, 906, 940, 983, 1440, 1509, Negrao-Correa, Deborah 1263 1501 Nixon, Christian P. 605 1670, 1832, 908, 1236, 1920 Nelson, Christina 1044 Nguyen, Hoa L. 783 Nixon, Christina 605 Noulin, Florian 229 Nelson, Jacob T. 838 Nguyen, Hung B. T. 462 Nixon, Gemma L. 484 Nouvellet, Pierre 1232 Nelson, Martha 1417 Nguyen, Hung M. 1501 Njabi, Conica 918 Novak, Robert 1700 Nelson, Sara 605 Nguyen, Kieu T. T. 173 Njagi, Kiambo 760 Noviyanti, Rintis 219 Nene, Vishvaneth M. 1175 Nguyen, Linh 59 Njagi, Purity 877, 1528 Novoa, Italo 427 Nerurkar, Vivek R. 647 Nguyen, Nguyet Minh 783, 784 Njama-Meya, Denise 870 Novotny, Joseph 45 Nery, Susana V. 597 Nguyen, Phu H. 279 Njau, Ritha 1467 Novotny, Marian 539 Nestares, José 1784 Nguyen, Phuc T. H. 279 Njenga, Daniel 1556 Noyd, David 696, 1379 Nesterov-Mueller, Alexander 806 Nguyen, Quy Anh 1500 Njenga, Sammy 60, 1079, 508 Nozaki, Mamoru 973 Neto, José A. 1118 Nguyen, Thanh-Thanh H. 520 Njeru, Ian 660, 1424 Nsa, Henry 1363 Netto, Eduardo M. 1118 Nguyen, Thieu Q. 1501 Njie, Fanta 1544 Nsakashalo, Mutale 1708 Neves, Yuri 1118 Nguyen, Tran Bich Chau 814 Njih Tabah, Earnest 1855 Nsanzabana, Christian 27 Newbold, Chris I. 1900 Nguyen, Van T. T. 279 Njiné, Thomas 567, 1161, 1161 Nseka, Tommy M. 1538 Newell, Steven 1766 Nguyen, Vu 1008, 1621 Njiri, James 559 Ntadom, Godwin 865, 882, 988 Newman, Christina 842 Nguyen, Xa X. 279, 1464, 1500, Njogu, Julius 1532, 1580 Ntaro, Moses 873 Newman, Gale 226 1501 Njoku, Chinonyerem J. 529 Nteziyaremye, Julius 1861 Newport, Melanie J. 1290, 1690, Nguyen, Yen T. H. 279 Njuguna, Henry 562 Ntodom, Godwin 256 1286 Nguyen Van, Hong 1222 Njuguna, Patricia 1556 Ntoka, Adonis T. 1469 Newton, Charles 21 Nguyen Van, Van 1222 Nkamga, Vanessa 1638 Ntshalintshali, Nyasatu 45, 344 Newton, Paul 871, 1852, 1873 Ngwa, Alfred 314, 894 Nkanauena, Kondwani 953 Ntumngia, Francis B. 994, 1585 Newton, Paul N. 1046, 1214, 1853 Ngwenya-Kangombe, Tokozile 922, Nkanaunena, Kondwani 1522 Nu, Feiyang 1174 Newton, Samuel 1526 924 Nkemenang, Patrick 1855 Nuako, Godwin K. 1092 Neyra, Victor 144 Nhabomba, Augusto J. 383 Nkenfou, Celine N. 524 Nuckols, John T. 161, 205 N’falé, Sagnon 117 Nhacolo, Arsenio 338, 1478 Nkhama, Emmy 1525 Null, Claire 573 Ng, Caroline 675 Nhama, Abel 338 Nkhoma, Standwell C. 620, 983, Nundu, Sabin 1157, 1726 Ng, Marie 1618 Nhampossa, Tacilta 338, 470 1832 Nundu, Sylvain 1156 Ngadaya, Esther 1467 Nhu, Tran Do Hoang 430 Nkrumah, Francis 883 Nunes, Fernanda F. C. 1090 Ngalah, Bidii 300 Niangaly, Amadou 380, 619, 669, Nkurunungi, Gyaviira 403 Nunes, Jillian 319 Ngamboli, Jose 85 984, 1592, 1605, 1796 Nkuruziza, Emery 1908 Nuñez, Andrea 192 Ngan, Ta Thi Dieu 1768 Niangaly, Moussa 962, 1552, 1798 Nkwata, Allan K. 1121 Nunez, Jorge 1699 Ngandjio, Antoinette 1638 Niare, Karamoko 669, 1592 Nnadi, Donatus C. 529 Nuorti, Pekka J. 582 Ngangyung, Henrietta F. 1890 NiBuachalla, Cliodna 1489 Noack, Cassandra 673 Nutman, Thomas B. 29, 481, 491, Ngasala, Billy E. 269 Nic Fhogartaigh, Caoimhe 1046 Noblet, Raymond 1695 506, 589, 1208, 1213, 1277, Ngassa Mbenda, Huguette Gaelle Nichol, Stuart 861 Nobre, Flávio F. 1409 1696, 1697, 1698, 1891, 488, 307 Nickel, Ian 319 Noe, Amy R. 1606, 1607 1893 Ngatunga, Deogratias 318 Nicolete, Vanessa C. 1224 Nogueda-Torres, Benjamin 1682 Nuwaha, Fred 508 Ngeleka Mutolo, Daniel 1305 Nicoletti, Jacopo Giovanni 639, Nogueira, Ana Paula O. 1666 Nwakanma, Davis 124, 238, 373, Ng’eno, Eric 1260 1668, 439 Nogueira, Maurício L. 194 376, 932, 1352, 1477, 1806, Ngigi, Julius N. 934 Nicosia, Alfredo 672 Noh, John 458 261 Ngindu, Augustine M. N. 1014 Nielsen, Carrie 704 Nojoomi, Matthew 1285 Nwakwuo, Geoffery C. 260 Ngo, Thang D. 279, 1464, 1500, Nielsen, Morten 1201 Nola, Moïse 567, 1161 Nwankwo, Grace 264 1501 Nielsen, Morten A. 987 Nolan, Christina 291 Nwanta, John A. 74 Ngoc, Nguyen Minh 430 Nielsen, Rasmus 947 Nolan, Thomas J. 1264 Nwobi, Benjamin 68 Ngocho, James Samwel 1483 Nielsen, Robin 1610 Noland, Gregory S. 332, 950 Nwosuh, Ignatius C. 1421 Ngombe, Nadege K. 76 Nieves, Wildaliz 1942 Nold, Michael J. 1008 Nyabeyeu Nyabeyeu, Hervé 918 Ngomi, Nicholas 102 Nigo, Maurice M. 1716 Nolen, Leisha 477 Nyagol, Ronald 1258 Ngondi, Jeremiah 351, 699, 1015, Nikiema Ndong Ella, Rosalie 1473 Noor, Abdisalan M. 347, 363 Nyagonde, Nyagonde 897 1287, 368 Nikolay, Birgit 508 Norcross, Neil R. 287 Nyakarahuka, Luke 861 N’Goran, Eliézer K. 66, 1719 Nikolich, Mikeljon P. 1304 Nordin, Per 101 Nyaku, Mawuli 62, 68, 1084 Ngowo, Halfan S. 75 Niles, Jacquin C. 1831, 1944 Nordstrom, Karine 278 Nyambura, Janet 559 N’Guessan, Raphael 134, 769, 770 Nilles, Eric J. 1923 Norgan, Andrew P. 1140 Nyamuni, John 866 Ngufor, Corine 769, 770 Nilsen, Aaron 673 Noriega, Fernando 577, 578 Nyang, Haddy 314, 894 Ngufor, Corine A. 134 Nimol, Khim 657 Norris, Douglas E. 100, 146, 151, Nyangiri, Oscar A. 1902 Nguku, Patrick 357, 422, 523, 633, The NIMPE Advisory Groups 1501 1496, 1629 Nyarko, Alexander K. 472, 1093 853, 1652 Nin, Noch 1334 North, Ace 1 Nyathi, Emanuel 1639 Ngung’u, Joseph 461 Ning, Yue 1825 Norton, Diana C. 804, 808 Nyathirombo, Amos 476 Nguon, Chea 343, 904 Ninsiima, Hope 866 Norval, Suzanne 287 Nyehangane, Dan 1487 Nguyen, Anh Q. 279, 1464, 1501 Niranjankumar, Bhavana 177 Norwood, Jeanne 1934 Nyirenda, Osward M. 717, 888 Nguyen, Bich T. N. 462 Nisalak, Ananda 8, 630, 816, 1272, Nosamiefan, Iguosadolo 1799 Nyirenda, Tonney S. 395 Nguyen, Chau V. V. 462 1274 Nylen, Susanne 1751 astmh.org A-620 Abstract Author Index The number(s) following author name refers to the abstract number.

Nyothach, Elizabeth 1879 Odjo, Abibathou 769 Okolocha, Emmanuel C. 694 Omaswa, Freda 669, 1561, 1592, Nyunt, Myaing 1458, 1235 Odjo, Abibatou 134 Okombo, John 989 1599 Nzamba, Prycil 977 Odom, Audrey R. 317 Okoro, Chinyere 260 Omattage, Natalie S. 995 Nzelu, Chukwunonso O. 1332 Odongo, Wycliffe 41, 700, 1521 Okoronkwo, Chukwu 1069 Ombok, Maurice 760, 1887 Nzouankeu, Ariane A. N. 1638 Odoru, Abraham 883 Okoye, Patricia N. 133 Ome, Maria 53 Oduke, Jacqueline E. 326 Oksman, Anna 1946 O’Meara, Wendy 370, 882 Odundo, Elizabeth A. 1848, 1880 Okuboyejo, Titilope 256 Omedo, Martin O. 1071 O Odunga, Abuodha 666 Okui, Albert P. 1536 Omello, Martin 1856 Oduor, Clifford 1879 Okui, Peter 893 Omer, Rihab A. 446 Oakley, Miranda S. 235 Oduro, Abraham 1637, 1872 Okui, Scholastica A. I. 429 Omer, Saad B. 1777 Oattara, Amed 1605 Oeztuerk, Suemeyra 443, 641 Okumu, Fredros O. 75, 1010, 1013, Omololu-Aso, Joseph 1037 Obaid, K. Y. 1141 Offei Owusu, Irene 594 1021, 1023, 1167 Omololu-Aso, Oluwaseun O. 1037 Obala, Andrew A. 370 Ofili, Ikechuckwu A. 1570 Okumu, Moris 1693 Omondi, Angela 306 Obaldia, Nicanor I. 320, 951, 1436, Ofori, Joshua K. 1845 Okwera, Alphonse 1121 Omondi, Angela A. O. 305 930 Ofori, Michael 409 Olack, Beatrice 1869 Omondi, David 259 Obasanya, Joshua A. 1652 Ofosuhene, Mark 472 Oladejo, Abiodun J. 853 Omoniwa, Omowunmi F. 1907 Obenauer, Peter J. 1370 Ogala, William 256 Oladipo, Oladosu 882 Omore, Richard 582, 584, 1261 Obeng-Adjei, Nyamekye 398 Ogbuanu, Ikechukwu U. 633 Oladokun, Regina 262 Omulo, Sylvia 1220 Oberhammer, Lukas 21 Ogembo, Javier G. 614 Olaleye, Olufemi D. 175 Onah, Denchris N. 449 Oberhelman, Richard A. 51, 457, Ogola, Eric 660, 1220 Olamiju, Franca 1069 Onah, Denis N. 450 1279, 1296 Oguche, Stephen 256 Olander, William 1250 Onah, Dennis N. 74 Obi, Emmanuel 1030, 1031, 1612 Oguike, Mary 308 Olanga, Everline A. 356 Onapa, Ambrose 593, 1085 Obiakor, Harold T. 1448 Oguttu, David 593, 1693, 1184 Olaniran, Seyi B. 172 Onchiri, Frankline M. 516, 1848, Oboegbulem, Steve I. 74 Ogutu, Bernards 876 Olaosebikan, Rasaq R. 238 1880 Oboho, Ikwo 1300 Ogutu, Bernhards 306, 666, 1482, Olayinka, Adebola 365 Ondari, Daniel 1382 Obonyo, Nchafatso 1861 1483, 1556, 1602, 1919 Ole-Sangba, Marina 118 O’Neill, Maeghan 498 Obor, David 86 Ogutu, Michael 1071, 1146 Oliani, Sônia M. 393 O’Neill, Paul M. 484 Obrain, Benny 139 Ogwal, Alex 1536 Oliva, Clelia 1884 Oneko, Martina 631, 666, 1045 O’Brien, Kelsey 427 Ogwang, Caroline 1556 Oliveira, Carlo Jose Freire de 550, Onema, Willy 1533 O’Brien, Kieran S. 1307 Ogwel, Billy G. 1902 551, 552, 1666 Ong, Eugenia 180 Ocampo, Clara Beatriz 1403 O’Hara Ruiz, Marilyn 842 Oliveira, Claudia L. 1090 Ongaya, Violet A. 528 Ocampo, Karen 426 Ohashi, Kazunori 761 Oliveira, Danilo B. 858 Ong’echa, John M. 420, 967, 968, Ocana Gutierrez, Victor R. 1127 O’Hearn, Aileen 91 Oliveira, Guilherme 1789 1553 Ochako, Rhoune 877 Ohrt, Colin 276, 279, 342, 880, Oliveira, Jaquelline G. 858 Ongoiba, Aissata 405, 962, 1552, Ochiai, R. Leon 421, 1297, 1870 881, 1464, 1500, 1501, 1523, Oliveira, Maria Regina F. 340 1571, 1572, 1798, 1866 Ochieng, Benjamin 431, 584, 582 1549, 280 Oliveira, Natatia K. 892 Oniskey, Taylor 1942 Ochieng, Galex Orukan 1693 Ohta, Nobuo 33, 1092, 1093 Oliveira-Prado, Roberta 536 Onkanga, Isaac 58, 1071 Ochieng, John 1261 Ohuabunwo, Chima 633 Olkowski, Sandra 807 Onlamoon, Nattawat 183 Ochieng, Linus 1220 Ojaku, Alex 866 Olliaro, Piero L. 244, 245, 476, 642, Onmus-Leone, Fatma 1851 Ochieng, Melvin 819, 824, 1382 Ojo, Abiodun 265 1063, 1148, 1238, 1461, 1716, Onoja, Anyebe B. 175 Ochoa, Theresa J. 426 Ojo, Kayode K. 1038, 1172 1719, 200 Onoja, Bernard 172 Ocholla, Harold 19 Ojobor, Peter 1475 Olmeda, Raul 291 Onsrud, Mathias 1155, 1794 Ocholla, Stephen 851 Ojuawo, Ayodele 238 Olomi, Raimos 563 Onu, Adamu 265 Ocholla, Steven 559 Okado, Kiyoshi 126 Oloo, Florence 1919 Onwujekwe, Obinna 884, 1472 Ochomo, Eric O. 760, 1887 Okafor, Henrietta U. 256 Olotu, Olatunde 264 Onyango, Clayton 1045, 1064 Ockenhouse, Chris 1488 Okal, Michael N. G. 148 O’Loughlin, Samantha M. 4, 969 Onyango, Kevin Omondi 1483 Ockenhouse, Christian F. 1595 Okamoto, Emi 1102 Oloya, James 714 Onyango, Maria G. 1320 O’Connell, Elise M. 1213 Okamoto, Kenichi 1394 Olschner, Scott P. 860 Onyango, Wycliffe 1503 O’Connell, Kate 1532 Oke, Yetunde O. 264, 865 Olsen, Cara H. 1369 Onyona, Philip 911 O’Connell, Kathryn 84 Okebe, Joseph 314, 373, 376, 894 Olsen, Christian 1763 Oo, Htet Wai 864 O’Connell, Maile 647 Okech, Bernard A. 978, 1566 Olubayo, Luicer I. A. 411 Oo, Winny 937 O’Connor, David H. 665, 1431 OKech, Fredros O. 1022 Oluduro, Anthonia O. 1037 Ooi, Eng Eong 180, 199, 804, 808 O’Connor, Geraldine 630 Okeke, Iruka N. 424 Olugbade, Olukemi T. 365 Ooi, Winnie 1669 Odada, Peter S. 1560 Okeke, Joseph I. 1363 Olukosi, Adeola Y. 932 Oommen, Anna 1193, 1195 Odagiri, Mitsunori 708, 1762 Okell, Lucy 43, 345, 928, 1520 Olukosi, Yetunde A. 362 Oommen, Joyce 691 Oden, Maria 1285 Okezue, Godwin 853 Olupot Olupot, Peter 1861 Opanda, Silvanos 558 Odero, Christopher 1534, 666 Okhuysen, Pablo C. 431 Olusanya, Sholakunmi 365 Openshaw, John J. 1116 Odero, Norbert A. 86, 1503 Okiror, Samuel 1424 Oluwole, Akinola S. 1069 Opoka, Robert O. 24, 1857, 49, Odetoyin, Babatunde W. 424 Okitolonda, Emile 662, 1117, 1430 Olveda, Remigio M. 56, 1144, 71, 960, 1439, 1485, 1859 Odhiambo, Collins O. 1927 Okitolonda-Wemakoy, Emile 85 1727 Opoku, Millicent 1717 Odhiambo, Frank 631, 1879 Okitundu, Daniel L. 460 Omar, Ahmeddin 237, 949 Opoku, Nicholas O. 476, 1063, Odiere, Maurice 60, 1079, 1182 Okitundu, L. D. 1256 Omar, Rahila 927, 1040 1716 Odiwuor, Alfred 516 Okoh, Hilary I. 362, 932 Opole, Joseph O. 557 astmh.org Abstract Author Index A-621

The number(s) following author name refers to the abstract number.

Opot, Benjamin 558, 559 Otsuki, Hitoshi 419 Page, Anne-Laure 1283 Parra, Marcela 230 Ord, Rosalynn 310 Otsyula, Nekoye 666 Page-Sharp, Madhu 240 Parreira, Ricardo Cambraia 552 Ordoñez, Tania 916, 1411 Ottenhoff, Tom H. 992 Pagnoni, Franco 266 Parsons, Michele B. 582, 584, 1643 Ore, Marianela 545 Ottesen, Eric 1697, 1698, 1893 Pahlavan, Golbahar 1478 Partidos, Charalambos D. 11, 182, O’Reilly, Ciara E. 582, 584, 1261 Otto, Thomas D. 1899 Paige, Sarah B. 92, 1760 802 Oresto, Michael 351 Ouahabi, Souad 1370 Paine, Mark J. 765 Pasakia, Sonak 80 Oriero, Cheryll Eniyou 1352 Ouattara, Amed 324, 380, 984, Paing, May M. 995 Pasay, Cielo 1488 Oriero, Eniyou C. 261, 1806 1600, 1796 Pajuelo, Monica J. 51, 452, 1296, Pascale, Juan Miguel 663 Orikiriza, Patrick 1283, 1860 Ouch, Pidor 108 457, 564, 716, 1778 Pasos-Alquicira, Rafael 1329 Orillo, Beverly 838 Ouedraogo, Alphonse 1483, 1590 Pal, Subhamoy 834, 1396 Passos, Sara 1206, 1749 Orinda, George 1553 Ouédraogo, André Lin L. 325, 1525 Palafox, Benjamin 1582 Patankar, Swati 611 Orji, Bright C. 1317 Ouedraogo, Issa N. 1590 Palaniappan, Kannappan 1453 Patel, Bhumi 626 Orok, Bassey 932 Ouédraogo, Jean-Bosco 119, 274, Palevalut Scientific Committee 703 Patel, Jaymin C. 377, 598 Orok, John 1317 284 Pallansch, Mark 587 Patel, Jenish 1839 Oropesa, Suset I. 857, 1416 Ouédraogo, Jean-Bosco 1196 Palma, Noemí 426 Patel, Sandip K. 611 Orozco, Camilo 140 Ouédraogo, Robert K. 119 Palmer, Nicole 150 Patel, Saurabh D. 234 Orr, John M. 100 Ouedraogo, Sayouba 1856 Paltiel, A. David 664 Paternina, Luis E. 110 Orrego, Miguel A. 455 Ouedraogo, Smaila 346, 635, 1257, Palumbo, Anne 548 Paternina Gómez, Margaret 110 Orr-Gonzales, Sachy 613 1916 Pan, Michael 1285 Patrapuvich, Rapatbhorn 217, 976 Ortega-Lopez, Jaime 1750 Ouma, Collins 760, 1887 Pan, William K. 184, 803 Patrice, Combary 775 Ortiz, Lucía 1115 Ouma, David O. 158 Panchalingam, Sandra 431, 582, Patta, Shem 1382 Ortu, Giuseppina 518 Ouma, Fredrick R. 1182 584 Pattanapanyasat, Kovit 183 Oruko, Kelvin 1879 Ouma, Paul 148 Pandey, Alok Kumar 1002 Patterson, Amy E. 332, 950 Orumbie, Patrick 1475 Ouma, Peter 237, 1478 Pandey, Deb 692, 1679 Patterson, Noelle 1603, 1604 Orvis, Joshua 1175 Oumbouke, Welbeck A. 1192, Pandey, Manisha 1042 Patton, John 1264 Osabutey, Dickson 519 1246 Panella, Nicholas 861 Paudel, Vijaya 1679 Osada, Jorge 1659 Oundo, Joseph 431, 582, 584 Paniagua Contreras, Gloria Luz Paukovova, Zuzana 531 Osamu, Morinaga 1093 Ouologuem, Boucary 1598 1657 Paul, Moundipa F. 1130 Osarfo, Joseph 239 Ousterhout, Joep V. 717 Panigrahi, Pinaki 708 Paul, Repon C. 425 Osbert, Nicolas 706 Ovalle-Bracho, Clemencia 541, Pann, Sut-Thang 1481 Paul, Sinu 822 Oscar, Roland 1908 1099 Pan-ngum, Wirichada 1107 Pauley, Cindy 197 Osei, Isaac 1872 Overgaard, Hans J. 774, 1355 Panyanivong, Phonepasith 1214 Paulino, Marja 1050 Osei-Atweneboana, Mike Y. 510, Owada, Kei 521 Paolino, Kristopher 1610 Pauloi, Rossely 1700 1330, 1207 Owaga, Chrispin 700, 1521 Paolino, Kristopher M. 1925 Pauza, David 1559 Osei-Mensah, Jubin 1210 Owino, Emmanuel 666 Paone, Massimo 1740 Pavlin, Julie 1774 Osei-Poku, Jewelna 1248, 1335 Owiti, Preston 1879 Papa, Thomas 576 Pavlinac, Patricia 516, 1848, 1880 Osier, Faith H. A. 964, 1556 Owor, Nicholas 1773 Papageorghiou, Aris T. 364 Pavluck, Alex 1259 Osman, Ahmed A. 437 Owusu, Michael 847 Papaiakovou, Marina 1725 Paxton, Lynn A. 351, 1287 Osman, Omran 1743 Owusu-Agyei, Seth 1067, 1520, Papavasiliou, Nina 1939 Payne, Ruth O. 672 Osoga, Joseph O. 263 1637 Pappa, Vasiliki 289, 1441 Paz-Soldan, Valerie A. 153, 737, Osorio, E. Y. 1748 Oxborough, Richard M. 770, 771, Parapini, Silvia 288, 1461 733, 1295, 1296 Osorio, Elvia Y. 1746, 1753 1247, 1888 Parashar, Umesh 1300, 1643 Pearce, Richard 310 Osorio, Jorge E. 11, 163, 1427, Oyibo, Wellington A. 308, 882, Paredes, Adriana 455, 457 Peat, Elizabeth 1017 182, 581, 802, 1389 1570 Paredes, Gladys 514, 643 Pechacova, Daria 531 Osorio, Lyda 815 Oyieko, Janet 666 Paré Toé, Léa 1299 Peck, Robert 93 Osoro, Eric M. 660 Oyier, Lynette I. 989 Parham, Leda A. 1410 Peck, Roger 481, 506 Osoti, Victor 700 Ozaki, Masayo 950 Parikh, Sunil 26, 257, 921, 1457, Peery, Ashley 1344 Østerdal, Lars P. 1584 Oziemkowska, Maria 666 1458 Peeters, Koen 266 Osuna-Cabello, Maria 287 Ozwara, Hastings 290 Paris, Daniel H. 1214, 1852 Peeters Grietens, Koen 657, 1033, Otchere, Joseph 519, 1526, 1717 Park, Daniel J. 1240 1299, 1626 Otecko, Newton 1879 Park, Edward J. 469 Peixoto, Henry M. 340 Otero, William 320, 1436 P Park, Gregory S. 232 Pellé, Karell 50 Otiang, Elkanah 1220 Park, Jay 95 Pelleau, Stéphane 1241, 1805 Pablo, Jozelyn 389, 1605, 1796 Otieno, Allan 21, 666 Park, Jin Kyung 1870 Pelletreau, Sonia 1087 Pace, Cheryl 678, 1252 Otieno, Kephas 666, 911, 1478, Park, Jooyoung 317 Pemberton-Ross, Peter 1007 Pacheco, M. Andreina 379, 971, 1503, 1534 Parker, Daniel 941 Peña, Imanol 1932 970 Otieno, Michael F. 1553 Parker, Josephine 1883 Peña, José 189 Pacheco, Romina 388 Otieno, Peter 1503, 1534 Parker, Megan 1721 Pena, Mirna 1453 Pacheco-Yepez, Judith 1133 Otieno, Walter 666 Parker, Michael 1308 Peña, Víctor H. 793 Padilla, Deborah 1610 Otolorin, Emmanuel 1317 Parobek, Christian M. 377, 1454, Peñaranda, Katherin 388 Padilla, Norma 131 Ototo, Edna N. 120 1841 Peñataro Yori, Pablo 1646 Padrón, Alejandro 759 Ototo, Ednah N. 1020 Parra, Beatriz 815, 1403 Penilla, Patricia 137 Page, Andrea V. 466 O’Tousa, Joseph E. 748 Parra, Gabriel- Jaime 1232 Penilla-Navarro, Patricia 779 astmh.org A-622 Abstract Author Index The number(s) following author name refers to the abstract number.

Pennetier, Cedric 118, 122, 125, Phiri, Kamija 23, 525, 702 Plowe, Christopher V. 270, 321, Pozio, Edoardo 439 703 Phoenix, Inaia 850 324, 389, 619, 895, 984, 1235, Prabhakaran, Madhu 1800 Pennington, Luke F. 1792 Phok, Sochea 84, 905 1596, 1600, 1605, 1796, 1868, Prachumsri, Jetsumon S. 217, 1223, Penno, Erin 467, 473 Phommasack, Bounlay 1782 380 1488, 1537 Penny, Mary E. 600 Phommasone, Koukeo 871 Po, Ly 337 Praciano, Claudenia C. 543 Penny, Melissa A. 1001, 1007, 1588 Phonrat, Benjaluck 1120 Poer, Kathleen 767 Pradhan, Alana 614 Pensulo, Paul 888 Phuc, Hoang Le 430 Pogemiller, Hope 497 Pradhan, Anupam 890 Pepperl, Anathea 1165 Phuklia, Weerawat 1214, 1853 Poggensse, Gabrielle 1652, 422, Pradhan, Anupan 1465 Peprah, Dorothy 573 Phumratanaprapin, Weerapong 853 Prakash Babu, Senbagavalli 1893 Perdue, Christopher L. 744 1120 Poirot, Eugenie A. 1227, 1509 Pramundita Ramadani, Arba 1237 Perea, William 208, 209 Phung, Lam K. 173 Polanco, Myriam 1640 Prasanphanich, Nina S. 1795 Pereira, Ligia 910 Phyo, Aung Pyae 908, 1236 Polgreen, Lynda 1439 Prasetyo, Didot B. 1334 Perera, Rushika 1276 Phythian-Adams, Alexander T. 1938 Polhemus, Mark 916 Prata, Mara M. G. 1648 Perez, Jocelyn 1737, 1738 Piarroux, Renaud 1491 Pollard, Andrew J. 1849, 1850 Pratt, Drew 661 Pérez, Maria Angeles 191, 192 Piatt, Michael 750 Pollett, Simon 852, 1417 Prayoga, Pak 219 Perez, Omar G. 790 Pichardo-Rodriguez, R. 574, 1769 Polley, Spencer 325 Prendergast, Catriona T. 1937 Perez Padilla, Janice 1379 Pichler, Verena 1912 Polman, Katja 522, 1156, 1157, Pretzman, Viktoria L. 1165 Perez-Padilla, Janice 696, 1407, Pichyangkul, Sathit 1595 1159, 1721, 1726, 1786 Preux, Pierre-Marie 1196 1672 Pickering, Amy J. 709 Polo, Maria Rebollo 591, 1690 Prevots, Rebecca 366 Perez-Velez, Erika S. 545 Piedrahita, Estefani A. 1358 Pomaa, Mary A. 296 Price, Ric 219, 1842, 1905 Pérignon, Jean-Louis 1238 Pierce, Kristen 579 Pombi, Marco 1367, 1912 Price, Ric N. 22 Perignon, Marlene 1721 Pierce, Raymond J. 1789 Pompeu, Margarida M. Lima. 543 Price, Ric R. 958 Perkins, Alex 947, 1309 Pierce, Susan 398, 1453, 1866 Pompon, Julien 1817 Prichard, Roger 511, 642 Perkins, Douglas J. 420, 967, 968, Pierson, Theodore C. 624, 841, Ponce, Gustavo 137 Priest, Jeffrey 60, 1079, 1712, 1261 1553 1387 Ponce de Leon, Gabriel 1847 Prieur, Eric 1238 Perlman, Stanley 1924 Pigott, David 1422, 1929 Poncet, Antoine 1679 Pritham, Ellen 1177 Perrone, Alina E. 544 Pike, Andrew 786 Ponce-Terashima, Rafael 1179 Pritt, Bobbi S. 1140 Persampieri, Tania 1884 Pilishvili, Tamara 1258 Poncheri, Melanie 85 Proietti, Anna Barbara F. C. 1090 Pershing, April M. 618, 673 Pillai, Dylan R. 285, 891, 1674 Pondeville, Emilie 1910 Proietti, Carla 609 Petchampai, Natthida 113 Pillay, Pavitra 1794 Pondja, Maria do Rosario 1847 Protopopoff, Natacha 771, 772, Peter, Christopher R. 723 Pilotte, Nils 480, 1713, 1725 Pond-Tor, Sunthorn 56, 605, 1727 1245, 1365 Peters, Bjoern 822, 1275 Pimenta, Paulo F. P. 1357 Pone, Sheila M. 1409 Prouty, Michael 108, 1334 Peters, David H. 1875 Pimentel, Guillermo 112, 114, 737 Ponlawat, Alongkot 1391 Province, Michael A. 1654 Petersen, Christine 1732 Pinapati, Richard S. 1897 Ponnusamy, Loganathan 109 Prudhomme O’Meara, Wendy 83 Petersen, Ines 270, 986, 1590 Pinchoff, Jessie 136, 701, 701, 956, Pons, Maria Jesus 426 Psychas, Paul 1032, 1035, 1622, Petersen, Kyle 1284, 1659 957, 963, 1496, 1529 Poole-Smith, Katie 1386 1548 Peterson, A. T. 800, 1683 Pinchuk, IV, Iryna V. 1205 Popovici, Jean 622 Pu, Hongwei 438 Peterson, Jennifer K. 1820 Pindolia, Deepa K. 1508, 1536 Porcella, Steve 1555, 1866 Puerta Guardo, Henry 198, 1834, Peterson, Michael 1772 Pineda, Vanessa 1328 Porciani, Angélique 122, 125 1836 Peterson, Stefan S. 271 Pinggera, Leyla 21 Porco, Travis C. 1048, 1307, 1877 Pullan, Rachel L. 508, 1690, 1723 Petrarca, Vincenzo 1352, 1912 Pingnet, Jolien 606 Porta, Exequiel O. J. 281 Pulliam, Juliet R. C.. 1359 Petri, Jr., William A. 583, 423, 587, Pinheiro, Marcos Gabriel 1658 Porter, Chad K. 1647 Pullum, Thomas 704, 1632 1288, 1433, 1654, 1132, 1174, Pinheiro, Tauyne M. 194 Portugal, Silvia 1571, 1572, 1866 Puplampu, Naiki 1092 1266, 1426, 1135 Pinho, Flaviane A. 1200 Posey, Jamie 1064 Puta, Chilunga 637 Petronio, Joy Ann 178 Pinkerton, Relana 1639 Posey, Tanya 1376 Pyae Phyo, Aung 906 Petzold, Max 200, 271, 927, 1040 Piñón, Alexander 857, 1416 Posner, Jourdan 1447 Pyar, Khin Phyu 1235 Peyton, David H. 293 Pinsky, Benjamin A. 1058 Post, Rory J. 1330 Pybus, Brandon S. 252 Pfarr, Kenneth 1704 Pinto, Jesus 131 Postan, Miriam 544 Pfeil, Johannes 1254 Pinto, Joao 1352, 1367, 1912 Pota, Hirva 310 Pfeil, Sarah 9 Piola, Patrice 268, 336 Potchen, Michael J. 20, 1438 Q Pham, Duy 1642 Pion, Sébastien 592 Potet, Julien 1653 Qaadri, Kashef 1763 Pham, Giang T. 462 Pion, Sebastien D. S. 65, 1208 Pothin, Emilie 42 Qadri, Firdausi 35, 36, 432, 434, Pham, My-Lien 197 Pires, Leonidas M. 194 Potter, Brittney 283 583, 588, 1651, 1288 Pham, Phuong Thao 235 Pitanga, Thassila N. 1813 Potter, Lucas 1165 Qassimu, Ally 75 Pham, Tuan V. 279 Pitman, Phillip 464 Pou, Sovitj 673 Qorro, Grace P. 273 Pham Vinh, Thanh 1222 Pitt, C 1906 Poudel, Ram P. 1196 Quagraine, Josephine 1526, 1717 Phan, Qui T. 462 Pitzer, Virginia E. 1649 Poveda, Germán 361 Quang, Nguyen N. 249 Phann, Sut Thang 912 Pizarro, Juan C. 689, 1476 Póvoa, Marinete M. 135, 391, 393, Quartey, Alberta A. 1524 Philipp, Mario 1000 Plate, David 1136 396, 1563 Quartey, Joseph 1147, 1181 Philipsborn, Rebecca 1882 Platts-Mills, James A. 585, 1426 Powell, Michael D. 226 Quaye, Charles 659, 1114 Phillips, Margaret 675 Ploemen, Ivo H. J.. 1594 Powers, Ann 9, 1271 Quebrada Palacio, Luz P. 544 Phillips-Howard, Penelope 1879 Powery, Alece 105 Queiroz, Adriano 1734 Phiri, Faustina 767 Poyer, Stephen 877, 1532, 1533 astmh.org Abstract Author Index A-623

The number(s) following author name refers to the abstract number.

Quetz, Josiane S. 1648 Ramdas, Sahienshadebie 1097 Ravinetto, Raffaella M. 720, 1299, Renia, Laurent 233, 1440 Quevedo, Miguel 840 Ramiandrisoa, Sitraka 1194 1856, 1922 Rennoll-Bankert, Kristen E. 685 Quick, Robert 601 Ramirez, Grimaldo 1428 Raviprakash, Kanakatte 1924 Rentz, Jennifer Junghyon 380 Quicke, Kendra 750 Ramirez, Jose Luis 682, 759, 1886 Ravolanjarasoa, Leonora 939 Renyong, Lin 440, 442 Quillici, Marie-Laure 1638 Ramirez, Roberson 227, 1858 Rawiwan, Imerbsin 1434 Resch, Pamela 1604 Quinones, Mariam 759 Ramirez, Ronald 79, 277, 719 Ray, Debalina 1792 Resende, Paola C. 1775 Quintana, Fernando 916 Ramirez, Viviana 812 Ray, Sandipan 611 Retallack, Diane 1606 Quintela, Pedro H. 1648 Ramirez-Sierra, Maria Jesus 1329, Ray, William 515 Reuben, Hedrick 872 Quinten, Edwin 992 1750, 1936 Raychaudhuri, Syamal 1934 Reuter, Stephanie E. 244 Quintero, Juan J. 1358 Ramiro de Assis, Rafael 1810 Rayner, Julian 287, 410, 1899 Revollo, Rita 741, 1311, 1645 Quiroga, Rosario 1230 Ramos, Virginia 23 Razakandrainibe, Romy 454 Rey, Luiz Carlos 577, 578 Quispe, Antonio M. 1512 Ramos-Sanchez, Eduardo M. 874, Razsony, Tomas 1517 Reyburn, Hugh 532, 563, 869 Quispe, Renato 1129, 1776 1540, 1545, 1573 Razuri, Hugo R. 1755 Reyes, Lissette 598, 1643 Qureshi, Shahida M. 54, 585 Ramphul, Urvashi 682, 759 Read, Kevin 287 Reyes, Raquel 873 Rampling, Thomas 1556, 1590 Read, Lisa 252, 282, 283, 291 Reyes, Sharina 1610 Ramsan, Mahdi 351, 368, 1287 Reategui, Carmen 227 Reyes-Solis, Guadalupe 137 R Ramsauer, Katrin 12 Reaves, Erik 852 Reynales, Humberto 577, 578 Ramsey, Janine M. 1339 Rebar, Edward 1237 Reynolds, Joanna 1809 R, Daniel P. 907 Ranasinghe, Shiwanthi L. 1828 Rebick, Gabriel W. 715 Reynolds, Nathanael 746, 1661 Raballah, Evans 420, 967, 968, Rancier, Ramiro 459 Rebolledo, Mauricio 1173 Rezai, Ashley 1525 1553 Rand, Alison 403, 927 Rebolledo, Paulina A. 741, 1311, Rhee, Kyu 289 Rabone, Muriel 1812 Randall, Arlo 1552, 1567 1645 Rheinlander, Thilde 570 Rachmat, Agus 108, 1766 Randjelovic, Ana 1152 Rebollo, Maria 1708 Ribacke, Ulf 18, 617, 907, 1799, Radford, Kay 1064 Randremanana, Rindra 1055 Recalde, Cristina G. 569, 1411 676 Radley, David 580 Randriamanantena, Arthur 1056 Reda, Abeba 591 Ribeiro, Jose M. C. 488, 759 Rafael Cordova, Brisaida M. 636 Randrianarivelojosia, Milijaona 42, Reddy, K. Sony 1002 Ribeiro, José M. C. Rahal, Paula 194 336, 703, 939, 1239 Redekop, Ken 63 Ribeiro, Paula 1815 Rahantamalala, Anjanirina 454, Ranford-Cartwright, Lisa 1017, Reed, Steven G. 1008, 1228, 1595, Ribeiro-Rodrigues, Rodrigo 1124 1194 1900 1606 Ricchiuto, Arcangelo 1210 Raharinjanahary, Falinirina 454 Rang, Chandary 337, 1499 Reeves, Guy 1326 Ricci, Irene 1336 Raharinjatovo, Jacky 877 Rangel, Nonenipha 1603 Rehman, Andrea M. 238 Rice, Jennifer L. 1170 Raherinampinaina, Gisele 1055 Ranjalahy, Michel 1660 Rehman, Najeeb 954 Rich, Kirsty 495 Rahimy, Najeebullah 1120 Ransom, Janet 1934 Reich, Michael R. 1935 Richard, Stephanie A. 634 Rahman, Abu Hayat Md Waliur Ranson, Hilary 129, 130, 752, 767, Reich, Nicholas G. 1392, 1395 Richards, Allen L. 108, 111, 112, 1101 1019, 1186 Reich, Nick G. 816 114, 115, 1220, 1755, 1851, Rahman, M. Arifur 35, 433, 434 Rao, Mangala 1747 Reichard, Gregory A. 252 1852 Rahman, M. Ziaur 829, 1756 Rao, Ramakrishna U. 590 Reid, Caitlin 1743 Richards, Bethany R. 1337 Rahman, Mahmadur 1756 Rao, V. Bhargavi 345, 928 Reifman, Jaques 1597 Richards, Frank O. 950, 1693 Rahman, Mahmudur 162, 829, Rapoport, Laura B. 636 Reilly Ayala, Heather B. 236 Richards, Michelle J. 13 1874 Rasamoelina-Andriamanivo, Reiman, Jennifer M. 1003, 1006 Richards, Jr., Frank O. 332 Rahman, Mohammed Ziaur 162 Harentsoaniaina 1194 Reimer, Lisa 791, 1377, 1701, Richards-Kortum, Rebecca 1173, Rahman, Mujibur 1702 Rasgon, Jason 1364 1073, 1827 1285 Rahman, Ridwanur 1101 Rasheed, Murtaza 93 Reimer-McAtee, Melissa 564, 716, Richardson, Jason H. 1391 Rahman, Shamsur 1702 Rashid, Ramla 670 1778 Richardson, Mark 1934 Rahman, Taufiqur R. 1651 Rashu, Md. Rasheduzzaman 588 Reiner, Robert C. 652, 1413 Richardus, Jan Hendrik 63, 67, Rai, Madhukar 1751 Rashwan, Nour 511 Reinhart, Caroline 1228 1741 Rainwater-Lovett, Kaitlin 1272 Rasmussen, Stephanie 319 Reinherz, Ellis 1389 Richie, Thomas L. 670, 1005, 1574, Raj, Dipak K. 605 Rasmussen, Zeba A. 54 Reis, Crissiane C. 135 1601, 1602, 1603, 1604, 1608, Raja, Amber 1003, 1006 Rasmusson, Randall L. 1943 Reis, Eliana A. 1179 1610, 607 Rajahram, Giri S. 22 Raso, Fiona 415 Reis, Eliana A. G. 1813 Richman, Adam 1556, 1610, 1620, Rajamajhi, Bishnu 585 Raso, Giovanna 66, 1719 Reis, Mitermayer G. 1179, 1813 1594 Rajerison, Minoarisoa 1660 Rasoamampianina, Virginie 1194 Reis Castro, Luisa 1326 Richterova, Lenka 1017 Rajshekhar, Vedantam 1193 Rasoarimanana, Angeline 454 Reisen, William K. 141, 645, 839 Ricks, Philip 1035, 1622 Rakasz, Eva 182 Rasoloson, Dominique 939 Reiter, Karine 1008, 1621 Ricopa, Leonila 227 Rakotomanana, Domohina 1056 Ratanakorn, Parntep 1107 Reiter, Paul 795 Ricotta, Emily E. 1616 Rakotomanana, Fanjasoa 1056 Rath, Carolina 1204 Reithinger, Richard 1690 Ridde, Valéry 745, 1403 Rakotondrazaka, Mahenintsoa 454 Rathod, Pradipsinh 910, 1449 Rek, John 402, 1844 Riddle, Mark S. 1647 Ram, Pavani K. 48, 706, 711, 712, Ratka, Zachary A. 1462 Remarque, Edmond 966 Rieckmann, Karl 655 728, 1292 Ratsimandisa, Rova 877 Remes-Troche, Jose 577, 578 Riehle, Michael 750 Ramani, Enusa 1870 Ratsitorahina, Maherisoa 1056 Remoue, Franck 703 Rigat-Brugarolas, Luis 223 Rambaud Althaus, Clotilde 1767, Raucinova, Maria 1518 Ren, Ruilin 1579 Rigg, Chystrie 1328 1855 Rausch, Kelly 668, 1008, 1009 Renard, Emmanuelle 987 Riggs, Molly D. 1896 Rambeloson Zo, Jariseta 1765 Ravaoarisoa, Elisabeth 939 Renaud, François 99 Rijken, Marcus J. 364 astmh.org A-624 Abstract Author Index The number(s) following author name refers to the abstract number.

Rijnsburger, Rian 63 Rodrigues, Wellington Francisco Rosas-Aguirre, Angel 1541 Rulisa, Stephen 358, 1484 Riley, Christina M. 237 549, 550, 551, 552, 1666 Roschnik, Natalie 25, 879, 1253 Rund, Samual S. C. 1016 Riley, Lee W. 1658 Rodrigues-Gomes, Larissa 892 Rose, Andreas 1375 Rund, Samuel S. C. 748 Rim, Han-Jong 1782 Rodrigues-Neto, João Firmino 1744 Rose, Brenda 97 Runge-Ranzinger, Silvia 200 Rimarachin, Dolores 1699 Rodriguez, Ana 610, 1932 Rosen, Brian J. 1684 Rung-in, Siriwan 217 Rimoin, Anne W. 85, 464, 662, Rodriguez, Bibiana 1003 Rosen, Gail Emilia 1171 Rupérez, Maria 338, 1478 1117, 1430 Rodriguez, Hugo 329, 959, 1025, Rosenberg, Corey 779, 837 Rush, Amy H. 482 Rinaldi, Francesca 439, 571, 639 1510, 1823, 1858, 966 Rosenthal, Philip J. 893, 903, 946 Russell, Amy 580 Rinaldi, Gabriel 1781, 1814 Rodriguez, Idia V. 625 Rosowski, Emily E. 1943 Russell, Bruce 233, 1440 Riner, Diana K. 57, 1795 Rodriguez, Indra 1328 Ross, Allen G. 1144, 1185 Russell, Tanya L. 872 Ringwald, Pascal 895 Rodriguez, Ingrid B. 606 Ross, Amanda 1221 Russo, Francesco Paolo 571 Rios, Maria 160, 1383 Rodriguez, Pamela 388 Ross, Leila S. 250, 676 Rutebemberwa, Ellizeus 1577 Rios, Zonia 803 Rodriguez, Silvia 455, 457 Ross-Degnan, Dennis 1875 Rutishauser, Tobias 670 Rios Ribeiro, Steveen 1124 Rodriguez, Virginia 412 Rossi, Cynthia A. 860 Rutta, Acleus 304 Rippon, Emily J. 768, 1189 Rodríguez-Barraquer, Isabel 1272, Rossi, Patrizia 439 Rutvisuttinunt, Wiriya 849 Riscoe, Michael K. 673, 291 1400, 1414, 1903 Ross-Suits, Hannah 1170 Rwakimari, John 310 Risquez, Alejandro 821 Rodriguez de Armas, Lissette 1050 Rota, Paul 1064 Rwigi, Doreen 516 Risso, Cinzia 962 Rodriguez-Lainz, Alfonso 477 Roth, Alison E. 419 Rwiyereka, Angelique K. 1631 Ritmeijer, Koert 1229, 1743 Rodríguez Moctezuma, Raymundo Roth, Johanna 1804 Ryan, Edward T. 35, 36, 432, 433, Rivard, Robert G. 1304 1657 Roth, Alison 976 434, 588, 1651 Rivas, Ana M. G. 1390 Rodríguez-Morales, Kristel 1695 Rothenberg, Richard 1786 Ryan, Sadie J. 1411 Rivas, Enrique 577, 578 Rodríguez-Pérez, Mario A. 1695 Rothman, Alan L. 630, 697, 1391 Rychert, Jenna 36 Rivas, Kasey L. 1172 Roe, Kelsey O. 838 Rotimi, Charles 1286 Ryg-Cornejo, Victoria 1862 Rivera, Aidsa 696, 1379 Roederer, Mario 1800 Rotondo, Lisa 1076 Ryman, Kate D. 14, 164, 214, 818 Rivera, Andrea M. 455 Roesel, Sigrun 739 Roundy, Christopher 176 Rivera, Maribel 577, 578 Roestenberg, Meta 1602 Routh, Janell 1163 Rivera-Aguilar, Víctor 1133 Rogers, John H. 1580 Routray, Parimita 710 S Rivera-Hernandez, Tania 1042 Rogers, Matthew B. 14, 488 Roux, Guillaume 1055 Saad, Eduardo 247 Rivera-Sánchez, Aidsa 1672 Rogerson, Stephen J. 53, 1119, 679 Rowe, Alexander K. 1875 Saade, Camille A. 69 Rivero, Keytia 1411 Rogier, Christophe 42, 336, 703, Rowe, Casey 1489 Saal, Jessica 936 Riveros, Maribel 426 939 Rowe, J. Alexandra 1796 Saavedra, Marlon P. 149 Robb, Katherine 573, 1262 Rohloff, Peter 735 Rowe, Samantha Y. 1875 Saavedra-Rodriguez, Karla L. 137, Robert, Michael A. 165, 1885 Rohrbach, Petra 312 Rowland, Mark 134, 752, 769, 771, 779 Robert, Robert 258 Rojas, Ernesto 1095 772, 961, 1245, 1247, 1365, Sabat, Grzegorz 1790 Roberts, Chrissy 1877 Rojas, Nancy 1432 1631, 1888, 770 Sabbagh, Audrey 987 Roberts, Christine 580 Rojas Cabrera, Ernesto 1104 Rowland, Paul 250 Sabeti, Pardis 1805 Roberts, Kathryn W. 342 Rojas-Ferreyra, Percy A. 807 Rowlands, Christopher J. 1944 Sabin, Sabin 522 Roberts, Rachel 671, 986, 1590 Rojas-Peña, Monica 1802 Rowley, Carol A. 1438, 1452 Sabindy, Samira 1847 Roberts, Scott J. 387 Rollinson, David 1810, 1811, 1812 Rowton, Edgar D. 1369 Sabino, Ester C. 1090 Robinson, Annie 1055 Romain, Jean R. 978 Roy, Chad 11 Sabitu, Kabir 357, 422, 1652 Robinson, Leanne 609, 955, 1119, Romanha, Alvaro J. 1094 Roy, Sharon L. 598 Sabundayo, Beulah 181, 579 1531 Romano, Audrey 547 Royal, Scott R. 575, 625, 628 Sá Carvalho, Marilia 729, 730 Roca, Anna 1280 Romero, Candice 1417 Rozas, Marizabel 1428 Sacchi, Robert S. 458, 459 Roca Feltrer, Arantxa 315, 337, Romero, Claudia 815 Rozelle, Scott 1728 Sacci, John 1606 1499 Romig, Thomas 446 Rubach, Matthew P. 1881 Sacco, Dana 93 Roch, Dabire K. 775 Romo, Hannah E. 845 Ruben, Adam J. 1594 Sack, Brandon K. 1568 Rocha, Crisanta 192 Romo, Matthew L. 1197 Rubiano, Kelly 1562 Sack, David A. 38 Rocha, Manoel Otavio C. 536 Ronca, Raffaele 964 Ruck, Richard 1005, 1610 Sackey, William 1035 Rocha, Roberta D. R. 1090 Roncal, Norma 252, 282, 1462 Rudge, James 721 Sacko, Adama 762, 796 Roche, Claudine 1923 Rono, Josea 964, 974 Rudy, Lai a Fat 1097 Sacko, Moussa 25 Rochford, Rosemary 916 Roobsoong, Wanlapa 1829 Rueckle, Thomas 675 Sacks, David 547, 1751 Rocke, Tonie E. 1427 Rooth, Ingegerd 964, 974 Rueda, Jorge 1388 Sadiq, Aishatu A. 422, 1652 Rockett, Rebecca 270 Roper, Cally 310 Ruisenor-Escudero, Horacio 24 Sadumah, Ibrahim 1258 Roda, Aldo 288 Roque, Rosmery 857 Ruiton, Sila 1432 Saeij, Jeroen P. 1943 Roddick, Joanne 609 Roque-Barreira, Maria Cristina 549 Ruiz, Daniel 1411 Saenz, Patricia 455, 457 Rodó, Xavier 730 Rosado-Vallado, Miguel 1750, 1936 Ruiz, Fernando 389 Sae-Ung, Tipaporn 183 Rodpradit, Prinyada 863 Rosanas, Anna 900 Ruiz, Joaquim 426 Sagara, Issaka 668, 796, 902, 1009, Rodrigez, Idia 1385 Rosanas-Urgell, Anna 229, 959, Ruiz, Rosana 643 1371, 1491, 1492, 1493, 1527, Rodrigues, Amabélia 1367, 1912 966, 1222, 1519, 1531, 1803 Ruiz, Roxana 514 1598, 1599 Rodrigues, Janneth 759 Rosário, Virgílio 901 Ruizendaal, Esmee 266 Sage, Mike 1258 Rodrigues, Laura C. 1265 Rosas, Alicia 840 Ruiz-Ortega, Marta 610 Sagliba, Marianne J. 1727 Rodrigues, Nilton B. 1357 Rosas, Angel 959, 966, 1025 Ruiz-Roldán, Lidia 426 Sagnon, N’Fale 126, 1019 Rodrigues, Tamara S. 1648 Rosas, Reinaldo 1640 Rulisa, Alexis 333, 1255 astmh.org Abstract Author Index A-625

The number(s) following author name refers to the abstract number.

Saha, Amit 1651 Sanchez, Arianni R. 874, 1540, Sasaki, Hitoshi 33, 985 Schnabel, David 276, 880, 1523, Saha, Debasish 1139 1545, 1573 Sasaoka, Chisa 998 280 Sahar, Tajali 1002 Sánchez, Carlos 1410 Saterbak, Ann 1285 Schneeberger, Chandra 1261 Sahoo, Malaya K. 1058 Sánchez, Gerardo 1930 Satharath, Prom 912 Schneider, Angela J. 1732 Sahu, Priyadarshi 1762 Sanchez, Jose L. 744 Sather, D. Noah 17 Schneider, Kristan 321 Sahu, Rajnish 295, 674 Sanchez, Juan F. 277, 545 Sather, Mark 1495 Schneider, Matthew 1618 Sahu, Tejram 1593 Sanchez, Leny 548 Satofan, Samson 1212 Schoeller, George B. 789 Saidi, Alex 615 Sanchez, Maria Carmen A. 874, Sattabongkot, Jetsumon 382, 941, Schoepflin, Sonja 1221 Sai-gnam, Piyaporn 299 1540, 1545, 1573, 1742 976, 993, 998, 1591, 1829 Schoepp, Randal 91, 860 Saijo, Masayuki 1840 Sanchez-Martinez, Luis 959 Sauerwein, Robert W. 270, 992, Schofield, Louis 385 Saingam, Piyaporn 1774 Sanchez Mora, Bianca 319 1018, 1567, 1594, 1602, 1801 Schott, Whitney 600 Saito, Mayuko 1279 Sanchez-Vargas, Irma 648, 1276 Saulnier, Aure 575 Schousboe, Mette L. 310 Saiwaew, Somporn 220 Sande, John H. 879, 1253 Saunders, Catherine 86 Schrag, Stephanie 1258 Saiyasombat, Rungrat 649 Sanders, Kelly 654 Saunders, David 299, 912, 1481, Schreiber, Stuart 286, 292 Saizonou, Jacques 1192 Sandoval, Carlos A. 1265, 1268 1494, 1774, 913, 1454, 1841 Schriefer, Albert 1734 Saka, Yisa 1069 Sandvik, Leiv 1152 Saunders, Jessica 673 Schriewer, Alexander 708 Sakamoto, Hirokazu 998 Sanford, Christopher 1675 Sausen, Nicholas 960 Schultz Hansen, Kristian 870 Sakrejda, Krzysztof 816, 1392, Sang, David 1071 Sausser, Michele 580 Schulze zur Wiesch, Julian 1863 1395 Sang, Huldah 1081 Savariar, Vincent 1656 Schwartz, Ira B. 1414 Salama, Carlos 458 Sang, Rosemary 1927 Saverino, Elizabeth 1610 Schwarz, Erich M. 1267 Salamat, Maria Sonia 521 Sang, Rosemary C. 819 Saville, Melanie 576, 577, 578 Schwarz, Lara 1295 Salanti, Ali 987, 1557, 1797 Sangala, Jules 962, 1552, 1572, Savón, Clara E. 857, 1416 Schwem, Brian E. 178 Salawu, Mobolaji M. 1024 1798 Savransky, Tatyana 1446 Sciotti, Richard J. 252 Saldana, Azael 1328 Sangaré, Ibrahim 284 Sawa, Patrick 259 Scopel, Kézia K. G. 1224 Saldarriaga, Omar 690, 1746, Sangare, Lansana 1477 Sawadogo, Simon P. 119, 1373 Scott, Alan 331 1748, 1753 Sangare, Modibo 1697, 1698 Sawers, Larry 496 Scott, Charles 1934 Saldivia, Alejandra 1640 Sangare, Moussa B. 1697, 1698 Sayeed, Md. Abu 433 Scott, Thomas W. 153, 185, 787, Saleh, Amgad A. 1347, 1348 Sangenis, Luiz Henrique C. 536 Sazzad, Hossain M. S. 1116 807, 1391, 1413 Salgado, S. Rene 704 Sangoro, Peter 138 Scandale, Ivan 498, 1209 Screaton, Gavin 1837 Salih, Niven 1229 Sangweme, Davidson 235 Scaraffia, Patricia 750 Se, Youry 299, 1774 Salihu, Muhammad 265, 1576 Sania, Ayesha 740 Scaraffia, Patricia Y. 1909 Sea, Darapiseth 299 Salim, Amina 1556 Sani- Lamine, Mariam 1078 Scarpassa, Vera M. 1327 Sealy, Tara 861 Salinas, Jorge L. 218 Sanin, David 1937 Scarpino, Samuel V. 186 Searle, Kelly M. 938 Salinas, Nichole D. 995 Sankoh, Osman 736, 1872 Scavone, Lauren 157 Searson, Peter C. 875 Salje, Henrik 48, 162, 829, 1272, Sanogo, Koualy 1509 Schaad, Nicholas 1300 Seas, Carlos 1129, 1776 1392, 1395, 1414, 1756 Sanogo, Sintry 1009 Schaar, Steven 500 Sebayang, Boni 219 Sallau, Adamu K. 950 Sanou, Antoine 1019 Schäfer-Nielsen, Claus 1201 Sebina, Ismail 1437 Salman, Ahmed M. 1004 San Pedro, Alexandre 729 Schaffer DeRoo, Sarah 1596 Seckel, Laura 1759 Salman, Sami 475, 1059, 1677 Santamaria, Ana M. 930 Schaffner, Francis 795 Secor, W. Evan 60, 1079, 1795, 57, Salmon-Mulanovich, Gabriela 184, Santelli, Ana C. 135, 247 Schaffner, Stephen F. 1833 1071 803, 1295, 1755, 1757 Santiago, Araceli 1639 Schallig, Henk 266, 1097, 1804 Secundino, Nagila F. C. 1357 Sam, Baramey 298 Santiago, Gilberto 811, 1390, 1835 Scharf, Rebecca J. 1266 Sedegah, Martha 1005, 1603, 1610 Samake, Yacouba 669, 1598 Santiago, Luis M. 1835 Schats, Remko 992 Sedegah, Mary 389 Samalvides, Frine 1784 Santiago, Teresa 1385 Schaut, Robert 1732 Seder, Robert 1567, 1800, 1609 Samani, Hector L. N. 1720 Santillan-Romero, A. 1769 Schellenberg, David 28, 255, 345, Sedillo, Jennifer 231 Samb, Badara 764 Santolalla, Meddly L. 397 869, 928 See, Craig W. 1307 Sambo, Maganga 77 Santos, Erian d. 1563 Scherer, Christina 286, 286, 292 Seed, Kimberley D. 34 Sambol, Nancy 26, 257 Santos, Kathlyn 737 Scherrer, Alexandra U. 1719 Segata, Nicola 1625, 1817 Sambunny, Uk 657 Santos de Oliveira, Miguel 1873 Schiavetti, Benedetta 1305 Segbaya, Sylvester 1637 Samdi, Lazarus M. 1363 Santos Junior, Gilberto S. 1406 Schick, Laura 1566 Segura, Luis 1432 Sam El, But 1774 Sanuku, Nelly 1212 Schickel, Jean-Nicholas 1866 Segurado, Aluisio A. C. 901 Sameroff, Stephen 1766 Saphonn, Vonthanak 733 Schindler, Christine E. 1866 Sehgal, Kunal 611 Samie, Amidou 585, 1639 Sapparapu, Gopal 626 Schloegel, Jesse 994 Sehgal, Rakesh 1130 Samir, Parimal 608 Saraiva, Raúl G. 1886 Schloegel, Jesse L. 1585 Seif, Shekalaghe 1523 Samitier, Josep 223 Saraiva, Roberto M. 536 Schluter, W. William 739 Seifu, Enadale 509 Samol, Lorna 1531 Sardei, Daniela 1668 Schmaljohn, Connie S. 13, 1925 Seilie, Annette M. 270 Samon, Nou 1774 Sareth, Rith 1774 Schmid, Michael A. 189, 1838 Sekabira, Umaru 866 Sampaio, Vanderson 340 Sariol, Carlos A. 625, 628, 1385 Schmid, Scott 1064 Sekandi, Juliet 714, 1121 Samuel, Roshini 408 Sarpong, Doris 296 Schmidt, Elena 1083 Sekuloski, Silvana 1488, 1489, Samuels, Aaron 961, 1503, 1631 Sarr, Demba 224 Schmidt, Mark 1212 1490 Samy, Abdallah M. 800, 1683 Sarro, Yeya Dit S. 1561 Schmink, Susanna E. 1064 Selby, Richmond A. 1030, 1031, Sanabria, Miguel A. 790 Sartono, Erliyani 1895 Schmutzhard, Erich 21 1612, 1845 Sary, Men 150 Schmutzhard, Joachim 21, 23 Selling, Katarina E. 271 astmh.org A-626 Abstract Author Index The number(s) following author name refers to the abstract number.

Selvapandiyan, A 542 Shayo, Alex 309, 318 Sierra, Gloria M. 202, 820 Singh, Balwan 603 Selvaraj, Arokiyaraj 1656 Sheen, Patricia 452 Sierra-Davidson, Kailan 1800 Singh, Brajendra K. 1705, 1709 Semnani, Roshanak T. 1277 Shekalaghe, Seif 374, 670, 743, Sigaúque, Betuel 338 Singh, Naresh 231, 890, 976, 1465 Senarathna, SMDK Ganga 240 1602, 881 Sigle, Leah T. 1818 Singh, Neetu 1751 Senoga, Ronald 402 Shelat, Anang 1096 Siguas Salas, Mery 585 Singh, Om P. 132, 765 Senyonjo, Laura 1083 Shelite, Thomas R. 103 Sihuincha, Moises 1428, 1699 Singh, Om Prakash 1751 Sepulveda, Nuno 933 Shelly, Ellen 1736 Sikaala, Chadwick 136 Singh, Onkar M. P. 250 Serre, David 5, 621, 622, 983, Shen, Kui 759 Sikomyu, Esther 402, 1569 Singh, Raja B. 132 1805, 1827 Shepard, Donald S. 203, 836, 1631, Sikorskii, Alla 24 Singh, Shailesh 226 Sesar, Jillian 1724 961 Sikulu, Maggy 121, 1488 Singh, Toolika 691 Sessions, October M. 199, 804 Sheppard, Aaron D. 1016 Silal, Sheetal P. 917 Singhasivanon, Pratap 1107 Sette, Alessandro 609, 822, 1275 Sherbuk, Jacqueline 1102 Silber, Steven 498 Singleton, Jered 1318 Settgast, Ann 497 Sheth, Anandi N. 1681 Siles-Lucas, Maria del Mar 1816 Singleton, Joseph 661 Sevene, Esperanza 338, 1478 Shetty, Nandini 1046 Silharova, Barbora 867 Sinha, Anuradha 423, 1433 Severens, Johan L. 63 Shewale, Jitesh 431 Silkey, Mariabeth 955 Sinnis, Photini 1607, 1901 Severson, David W. 686, 1343, Shi, Baoxin 638 Siloka, Griffith 653 Sinyangwe, S 20 1345, 1913 Shi, Lirong 1901 Silterra, Jacob 50 Siqueira, Andre M. 233, 340, 677 Sewe, Maquins 86 Shi, Ya Ping 911, 1503, 1534 Silumbe, Kafula 1507 Siqueira, Marilda M. 1775 Sewe, Maquis 631 Shiao, Shin-Hong 751, 754 Silva, Antônio V. A. 1648 Siregar, Josephine 1543 Seyde, Karl 888 Shidi, Calixite 1430 Silva, Joana C. 619, 1175, 1600 Sirichaisinthop, Jeeraphat 941, Seydel, Karl 20, 615, 1225, 1452, Shields, Timothy M. 956, 957, Silva, Juliana A. 1734 1223, 1537 1522, 614, 620, 1438 1512, 1529, 938 Silva, Lia M. B. 1230 Sirima, Sodiomon 671, 1483, 1590 Seynabou, Diedhiou M. 1822 Shiff, Clive J. 369, 1147, 1181, Silva, Luciano K. 1179, 1813 Siriwardena, H.v.y. D. 542 Seynabou, Sougoufara 1822 1512 Silva, Maria E. 1420, 1755 Sirohi, Devika 1276 Seyoum, Aklilu 781, 1363, 1622 Shikanai-Yasuda, Maria A. 718, Silva, Marita 185, 852, 1417 Siryan, Eman S. 1419 Shackleford, David 673 1230 Silva, Mercy 916 Sissako, Aliou 1495 Shadomy, Sean V. 1110 Shikani, Henry 216 Silva, Rubens A. 1230 Sissinto-Savi de Tovè, Yolande 1078 Shaee, Amro 1688 Shilling, Andrew J. 1695 Silva, Sheila C. Vicente. 718 Sissoko, Kourane 1009 Shafi, Oumer 1690 Shimp, Jr., Richard 1008 Silva Jr., Claudeir D. 1399 Sissoko, Mahamadou S. 669, 1592, Shafritz, Lonna 1765 Shin, Dongyoung 1366 Silva-Nunes, Mônica 1224 1598, 1599 Shagari, Shehu 700, 1521 Shin, Jang-Sik 1486 Silvas, Jesus 1928 Sissoko, Mahamdou S. 1561 Shah, Harishchandra 1691 Shindruk, Averyl 515 Silveira, Cassio 1230 Sisya, Tamika J. 620 Shah, Jui 1249, 1289, 1617, 1623 Shirai, Kenji 1840 Silver, Taryn 1163 Siv, Sovannaroth 337, 657, 1514 Shah, Kamal 1285 Shivastava, Arpit 1762 Sim, B. Kim Lee 670, 995, 1592, Siwo, Geoffrey 890, 898, 1465, Shah, Monica 1026 Shone, Alison 502 1594, 1602, 1608, 1610, 1556, 899 Shah, Nirmish 1451 Shongo, Robert 1117 1620, 669, 1600, 1609, 1800 Skerrett, Erica 1285 Shah, Vishal 1102 Shono, Yoshinori 761 Sima, Michal 539 Skinner, Jeff 1552, 1798 Shahum, Andrea 530 Shott, Joseph P. 1371, 1598, 1599 Simao, Julia 1873 Sladeckova, Veronika 530, 1518 Shaik, Rhiaz B. 910 Shotwell, Sandra 293 Simarro, Pere P. 1740 Slater, Hannah C. 43, 1843 Shakely, Deler 927, 929, 1040 Shoyama, T 1093 Simasathien, Sriluck 1770 Slatko, Barton E. 1896 Shakoor, Sadia 585 Shuaibu, Mohammed N. 400, 985 Simbauranga, Rehema H. 70 Sloan, Lynne M. 1140 Shaman, Jeffrey 794 Shuker, Samer 1677 Sime, Heven 1690 Sloots, Theo 270 Shanker, Lauren M. 105 Shukullari, Ada 795 Simmons, Cameron 173, 814, Slotman, Michel A. 1351, 1355, Shanks, G. Dennis 372, 249 Siame, Mwiche P. 313, 889 1384, 1837, 783, 784 1356 Shapiro, Jesse 34 Siba, Peter 5, 53, 609, 791, 1221, Simms, Benjamin T. 1526 Slutsker, Laurence 631, 666, 1478, Sharakhov, Igor V. 1190, 1911, 1531, 1550, 1803, 385, 491, Simo, Pierrette 1638 1908 1913, 1344 1119, 1212, 1377, 1554 Simoes, Rejane C. 1357 Sluydts, Vincent 657, 926, 1033, Sharakhova, Maria V. 1911, 1913, Sibley, Carol 287, 1842 Simon, Cleophas 77 1514, 1624, 1626 1344 Sibley, Samuel D. 665, 1431 Simon, Gregory G. 1077 Small, Scott T. 491, 1827 Sharma, Ambika 910 Siciliano, Giulia 288 Simon, Jakub K. 39 Smartt, Chelsea T. 1366 Sharma, Amit S. 1057 Sicuri, Elisa 595 Simons, Mark 852, 1417, 1755 Smedley, James 1856 Sharma, Ankur 613 Siddique, Abdullah 1132, 1174 Simons, Noah D. 665 Smircich, Pablo 1816 Sharma, Archana 1002 Sidibe, Bakari 1527 Simon-Salvador, Jocelyn 1682 Smit, Cornelis H. 59 Sharma, Atashi 1190, 1911 Sidibe, Bouran 1492 Simpson, Julie A. 1905 Smith, Amanda P. 818 Sharma, Namita 1057 Sidibe, Youssoufa 1539 Simpson, Steven 1663 Smith, Brian 1390 Sharma, Raman 502, 504 Sidik, Saima M. 1943 Simubali, Limonty 1629 Smith, Bryan 252, 320 Sharma, Sanjib K. 692, 1679 Sidney, John 609, 822, 1275 Sinden, Robert 287, 284, 1029 Smith, Darci R. 837 Sharma, Sumana 410 Sidoti, Laura 1780 Siner, Angela 616 Smith, Darvin Scott 1728 Sharp, Tyler M. 696, 811, 1379 Sie, Ali 1011 Singa, Benson O. 516, 1848, 1880 Smith, David 652, 1929, 946, 947, Shashikumar, Soumya 607, 1574 Siebelink-Stoter, Rianne 1018 Singer, Alexandra 1610 1508, 1903 Shaw, Leah B. 1414 Siedner, Mark 873, 1283, 1860 Singer, Giselle 1934 Smith, Derek J. 828 Shaw, W. Robert 1817 Siegl, Peter 287 Singh, Amritpal 1057 Smith, Emily 608, 1603 Shaw Saliba, Kathryn 415 Sieke, Scott 837 Singh, Balbir 616, 1442 Smith, Hilary A. 680 astmh.org Abstract Author Index A-627

The number(s) following author name refers to the abstract number.

Smith, Joe 222 Sotto-Calle, Veronica 959 Stanistreet, Debbi 1258 Strein, Andrea K. 1603 Smith, Joshua D. 100 Soubigou, Guillaume 1660 Stanton, Michelle C. 1689, 1700, Stresman, Gillian 700, 1521 Smith, Larry R. 1604 Soukou, Koffi Bhonna 118 1708 Stricker, Kirstin 1482 Smith, Nicholas 1337 Soulama, Issiaka 1550, 1590 Staples, J. Erin 650 Strode, Clare 767 Smith, Roger S. 899 Souleymane, Doucoure 1822 Starr-Spires, Linda 831 Stromdahl, Ellen Y. 98 Smith, Samuel J. 243 Soumaoro, Lamine 589, 1696, States, Sarah L. 1219 Strub, Nathalie 1483 Smith, Stanton Q. 1462 1697, 1698 Stauber, Christine 1786 Stryker, Gabrielle A. 1754 Smith, Thomas 42, 727, 1007, Soumare, Harouna 1509 St. Clair, Kristina 1673 Stuckey, Erin M. 41, 1511 1221, 41, 914, 955, 1001, 1588 Sousa, Carla A. 1367 Steeg, Christiane 1863 Sturdevant, Daniel E. 1866 Smrekova, Eva 1517 Sousa, Giovane R. 536 Steel, Cathy 1213 Sturrock, Hugh J. W. 342, 344, Snead, Andrew T. 817 Sousa, Jason C. 252, 283 Steen, Keith 752, 768, 776, 1189 348, 1502 Snounou, Georges 1440 Sousa-Figueiredo, José Carlos 1719 Steeves, Tanner K. 1219 St. Victor, Yves 978 Snow, Robert W. 363, 948 Soussou, Efoué 1136 Stefaniak, Maureen E. 1604 Su, Qi 619 So, Mary 1481 South, Adam 1889 Steindel, Mário 72 Suangtho, Paphanij 1392, 1395 So, Peter T. 1944 Souza, Katia 1383 Steinhardt, Laura 1026, 1908 Suarez, Julianne 1337 Soares, Alberto M. 1315 Souza-Neto, Jayme A. 1354 Steketee, Richard W. 637, 1511 Suárez, Milagros 538 Soares, Irene S. 233, 966 Souza-Santos, Reinaldo 729 Stenger, David A. 737 Suaza, Juan 361 Soares, Maria J. 811 Sovann, Ly 1774 Stephens, Henry A. F. 630 Subramanaim, Kris 1441 Soares Magalhaes, Ricardo 518 Sovannaroth, Siv 150, 905, 1499, Stephenson, Rob 332 Subramaniam, Krishanthi 760 Soarin, Kim 657 1624 Stepniewska, Kasia 268, 1842 Subramanian, Sathish 1654 Soboslay, Peter 481, 506 Sow, Doudou 394 Sterk, Esther 1103, 1425 Suchdev, Parminder 1311, 1645 Sobuz, Shihab U. 1266 Sowunmi, Akintunde 253, 256 Sterling, Charles 1736 Sucupira, Izis M. C. 135 Sochantha, Tho 926, 1033, 1514, Spaccapelo, Roberta 1884 Sternberg, Paul W. 1267 Sudathip, Prayuth 315, 1513 1624, 1626 Specht, Sabine 498, 1207, 1209, Stete, Katarina 1719 Sudo, Moe 990 Sodiomon, Sirima, Ouagadougou, 1210, 1713 Steven, Andrew 485 Sudoi, Raymond 1532 Burkina Faso B. 1484 Speer, Brittany 813 Stevens, Eric 481, 506 Sudoi, Raymond K. 934 Sodjinou, Joel 1551 Speich, Benjamin 642 Stevens, Lori 1337 Sufyan, Ahmed 422 Soe, Myat Thu 864 Spenassatto, Carine 1354 Stevens, Michael P. 1165 Sugiarto, Paulus 1905 Soebiyanto, Radina P. 1771 Spence-Lewis, Infanta M. N. 1047 Stevens, Yvonne 1934 Sugiharto, Victor A. 1369 Sognikin, Koffi S. 1136 Spencer, Alexandra J. 1004 Stevenson, Jennifer 41, 700, 1496, Sukapirom, Kasama 183 Sogoba, Nafomon 762 Spencer, Christopher S. 747 1521, 1629 Sukhbaatar, Munkzhul 629 Soh, Eugene 1481 Speybroeck, Niko 966, 1159, 1222, Stevenson, Mary M. 512 Sukupolvi-Petty, Soila 1387 Sohn, Woon-Mok 1782 959 Stewart, Lindsay 1443 Sulaiman, Irshad M. 1663 Soisson, Lorraine 1605 Spiegelman, Donna 740 Stewart, Matthew 1610 Sulaiman, Syuhaida 1342 Sok, Sopheak 1856 Spithill, Terry 606, 1003 Stewart, V. A. 1369 Sulca, Juan 185 Sok, Touch 1766 Spratt, Heidi 1753 Stewart, Zachary 1888 Sule, Doyinsola A. 436 Sokhna, C. 1906 Spring, Michele 912, 913, 1481, Stewart-Ibarra, Anna M. 201, 569, Suleiman, Anthony 422 Sokhna, Cheikh 302, 658 1494, 1841 1411, 916 Sullivan, David 369, 875 Sokleng, Sun 1856 Sravanam, Praveen 1015 Stewart Schicker, Rebekah 332 Sullivan, Eddie 1924 Sokny, Mao 1624 Sreng, Sokunthea 298 Stickles, Allison 673 Sullivan, Richard T. 1565 Solmo, Chelsea 1536 Sridhar, Revathi 965 Stienstra, Ymkje 1655 Sultana, Tania 588 Solomon, Anthony 1259 Srikanth, Rapole 611 Stiles, Jonathan K. 226 Sumaye, Roberty D. 1023 Soloniando, Sandrine 1055, 1056 Srikiatkhachorn, Anon 8, 630, 697, Stillwaggon, Eileen 496 Summers, Phillip J. 535 Soloviov, Sergii 553 1274 Stinchcomb, Dan T. 11, 182, 581, Sun, Cheng 1177 Soltis, Bryony 464 Srinath, Meghna 1311 1389, 802 Sun, Chengqun 14, 164 Soma, Dieudonné D. 777 Srinivasan, Prakash 1008 Stine, O Colin 431 Sun, David 661 Sombo, Marie-Thérèse A. S. 460 Srivastava, Sanjeeva 611 St. Jean, Denise 1219 Sun, Longhua 686 Somethy, Sok 912 Sriwicha, Sabaithip 299 St. Jean, Yvan 1908 Sun, Tiffany 387 Sommethy, Sok 1481 Sriwichai, Patchara 145 St. Laurent, Brandyce 150, 1371 Sundac, Lana 606 Somony, Heng 657 Sriwichai, Sabaithip 1481, 1494 Stockamp, Nathan 1772 Sundar, Shyam 691, 1233, 1751 Somsakchaicharoen, Raweewan Ssebuliba, Joshua 1457, 1458 Stoddard, Steven 153, 1413, 787 Sundrakes, Siratchana 299 328 Ssekitooleko, James 705 Stoler, Justin 1404 Suninthaboon, Panya 168 Sondén, Klara 405 Ssempebwa, John C. 599 Stolk, Wilma 67, 1208, 1741, 63 Suntarattiwong, Piyarat 1770 Songolo, Peter 1708 Ssewanyana, Isaac 1565, 1903 Stoller, Nicole E. 1048, 1066, 1307 Suon, Seila 150, 298 Soong, Lynn 103, 1205 Staedke, Sarah 242, 1844, 1903, Stone, Chris 1080 Supali, Taniawati 1895 Sopha, Chantha 298 1904, 243, 893, 946 Stone, William J. R. 1018 Supaprom, Chonthida 108 Sopoh, Ghislain E. 1655 Stager, Charles 1126 Storme, Casey K. 1595 Sup-Yeom, Joon 1495 Sorichetta, Alessandro 1508 Stahl, Stacey 1824 Stothard, J.Russell 1708 Surat, Guezin 1608 Sorvillo, Brian 1376 Stamler, Jonathan S. 1435 St. Pierre, Timothy G. 1803 Susapu, Melinda 1701 Sosa, Nestor 663 Stanek, Danielle 1390 Strachan, David P. 1265 Sutamihardja, Awalludin 279, 876, Sotelo, M. 1769 Stanhope, Kaitlyn 1311 Straimer, Judith 1237 880, 881, 280 Soti, David 760 Stanisic, Danielle I. 606, 1006, Strauss, Kathleen A. 270 Sutcliffe, James 1026 Soto-Becerra, Percy 1128 1119, 1531 Streat, Elizabeth 866, 871 Sutherat, Maleerat 1120 astmh.org A-628 Abstract Author Index The number(s) following author name refers to the abstract number.

Sutherland, Claudette S. 1080 Tami, Adriana 202, 820, 1150 Tediosi, Fabrizio 595, 1001, 1080, Thiptharakun, Supan 364 Sutherland, Colin 302, 308, 323, Tamoka, Tamiwe 1808 1588, 1589 Tho, Sochantha 657 325, 1493, 1527, 348, 1865 Tan, Asako 890, 1240, 1465 Tefera, Mesfin 87, 509 Thoma, George 1453 Sutherland, Laura J. 31 Tan, Hwee Cheng 180, 199, 808 Teissier, Anita 1923 Thomas, Alaina C. 738, 1494 Suvada, Jose 867 Tan, Jingwen 1312 Teixeira-Carvalho, Andrea 536, Thomas, Brent 1708 Suwanarusk, Rossarin 1440 Tan, Kim-Kee 1342 1090, 1383 Thomas, Dana L. 1270, 1390 Suwanpakdee, Sarin 1107 Tan, Yan 50 Tejada, Magaly 746 Thomas, Phaedra J. 231 Suzaki, Yuriko 1840 Tang, Douglas 1481 Tejada Santiesteban, Milagros G. Thomas, Stephen J. 630, 697, Suzuki, Mitsuko 409, 1092, 1093 Tang, Hanqi 14 636 1272, 1391, 1429 Suzuki, Ryuji 1840 Tang, Jianxia 128 Teja-isavadharm, Paktiya 299, 1481 Thomas, Tom 613 Suzuki, Takashi 1248 Tang, Linhua 895 Tejeda, Gustavo 200 Thompson, Corinne 430 Swai, Johnson K. 1022 Tang, Minh L. 661 Tekete, Mamadou M. 1492 Thompson, Rachel 877 Swales, Danielle 166 Tangnararatchakit, Kanchana 183 Tekola-Ayele, Fasil 1070, 1286 Thompson, Ricardo 1087 Swamidoss, Isabel 255, 788, 1873 Tangy, Frederic 12 Tekwani, Babu L. 295, 674, 1096 Thompson, Trevor A. 1495 Swanepoel, Robert 1927 Tanih, Nicoline 1639 Telford, Sam 1216 Thomsen, Edward 767 Swanstrom, Jesica A. 628 Taniuchi, Mami 587, 1266, 1426 Tellen, Karina 270 Thomsen, Edward K. 1212 Swe, Pearl M. 1218 Tannenbaum, Brynne 580 Temanni, Ramzi 1791 Thomson, Madeleine 1743 Swearingen, Kristian E. 1901 Tanner, Marcel 374, 670, 1221 Temba, Herilinda J. 463 Thomson, Nick 1877 Sweeney, Alexandra 1743 Tanowitz, Herbert B. 1729 Tembine, Intimbeye 1009 Thomson, Rebecca 255, 1578, Sy, Ngayo 1694 Tanya, Vincent N. 1890 Temu, Lucky 276, 880, 1523, 1549, 1579 Sy, Ousmane 330, 359, 658 Tappero, Jordan 26, 257, 1569, 280 Thongkukiatkul, Amporn 1591 Sykes, S. E. 1940 1864 Teng, Crystal 9, 1271, 651 Thongsripong, Panpim 798, 1830 Sylla, Khadime 394 Taramelli, Donatella 288, 1461 Teng, Jessica E. 34, 36, 37, 715 Thorne, Peter S. 1164 Sylla, Lakamy 796 Tarazona-La Torre, J. O. 574, 1769 Tenu, Filemoni 961, 1631 Thornton, Andrew 598 Sylvain, Pechangou N. 1130 Tarleton, Rick 687 Teo, Teck Hui 392 Thornton, Haley 109 Szabo, Flora 39 Tarnagda, Zékiba 1196 Teramoto, Isao 949 Thoryk, Elizabeth 1378 Szempruch, Anthony 1940 Tarning, Joel 26, 301, 311, 1480, Terashima, Angelica 1784, 1787 Thuma, Philip E. 313, 331, 938, Sztein, Marcelo B. 1849, 1850, 1920 ter Kuile, Feiko 19, 525, 679, 237, 1496, 399 1868 Tartaglino, Lilian 690 377, 702, 948 Thumar, Bhagvanji 1119 Szucs, Thomas D. 830 Tashi, Sonam 921 Terlouw, Dianne J. 23, 678, 1252, Thuo, Wangechi 1160 Szumlas, Daniel 1622 Tate, Jacqueline E. 582 942 Thurber, Mary I. 665 Tatem, Andrew J. 83, 736, 946, Ternier, Ralph 37 Thurman, Kathleen 1064 653, 1508, 1929, 344, 1611 Terrazas-Nuñez, E. 574, 1769 Thwing, Julie 246 T Tauber, Erich 12 Terry, Frances 1713 Thwing, Julie I. 1250, 1251, 1846 Taukobong, Nomathemba P. 734 Tesfaye, Berhanu 509 Tibenderana, Becky 1283 Ta-aksorn, Winita 1481 Tavárez, Mariana 1407 Tesh, Robert B. 206, 1397, 1423 Tibenderana, James 1460 Tabernero, Patricia 1873 Tavera, Gloria 1554 Teshima, Hayato 761 Tibery, Cecilia 181, 579 Tachibana, Mayumi 990 Tawfeek, Haifa I. 1688 Teshome, Birke 87 Ticona, Eduardo 1428 Tadei, Wanderli P. 1357 Tawfik, Harvey 1498 Tessema, Zegeye H. T. 527 Tiendrebeogo, Justin 1011 Tadesse, Zerihun 332 Tawfik, Nahla O. M. 1068, 1687 Tetteh, Kevin K. 989 Tietje, Kathleen 339 Tagbor, Harry K. 239, 1053, 1922 Tay, Sun-Tee 1342 Tettevi, Edward J. 510 Tilahun, Hiwot 1856 Tahina, Vonimbola 1056 Taylor, Alexander B. 1824 Teuscher, Franka 887 Tilley, Drake H. 427, 435, 852, Tahir, Rehman 421 Taylor, Cameron 704, 1034, 1249, Tewari, Kavita 1610 1043, 1662, 1170, 746, 1428 Tahita, Marc C. 274, 900, 1854 1289, 1617, 1623 Thaisomboonsuk, Butsaya 8, 168, Timinao, Lincoln 1221, 1550 Tairou, Fassiath 330 Taylor, David W. 1890 863, 1272, 1274 Timmerman, Ans 1774 Tairou, Fassiatou 359, 658 Taylor, Diane W. 228 Thangamani, Saravanan 212 Timoshevskiy, Vladimir A. 1913 Takahashi, Richard 839 Taylor, Mark 255, 487, 502, 503, Thanh, Nguyen X. 249 Tina, Lucas O. O. 666 Takala-Harrison, Shannon 321, 888, 504, 1578, 1579, 483, 484, 485, Thanh, Pham V. 1519 Tindana, Paulina 1239 895, 984, 1235, 1600, 1605, 489, 495, 501 Thapa, Chabilal 1679 Tine, Roger C. K. 254, 394 1796, 389 Taylor, Myra 1152, 1794 Thatte, Urmila 611 Ting, Jie 1772 Takasaki, Tomohiko 1840 Taylor, Robert W. J. 1679 Theander, Thor G. 474, 1557, 1797 Ting, Li-Min 673 Takashima, Eizo 997, 998, 1591 Taylor, Steve M. 377, 1451, 679 Thein, Tun-Linn 825, 1062, 1678 Ting, Nelson 665 Takeo, Satoru 998 Taylor, Terrie E. 20, 321, 614, 620, Theisen, Michael 394 Tinoco, Yeny 1417 Takken, Willem 727, 1351 717, 953, 1225, 1438, 1522, Thera, Mahamadou A. 380, 389, Tinto, Halidou 274, 900, 1299, Talaat, Kawsar 668 615, 888, 1226, 1452 619, 984, 1600, 1605, 1796 1854, 1922 Talledo Rodríguez, Joselyne O. 456 Taylor-Salmon, Emma 682 Thera, Philippe 25 Tiono, Alfred 671, 1550, 1590 Tallo, Veronica 56, 71 Tchalim, Mawèkè 1136 Thesing, Phillip 1226 Tipismana, Martin 1428 Talukdar, Arunansu 611 Tchaparian, Eskouhie 257 Thi, Long Vo 783 Tirta, Yusrifar 219 Tam, Mifong 512 Tchatchueng, Jules B. 1208 Thiele, Elizabeth A. 1692, 1712 Tisch, Daniel J. 791, 1827 Tamariz, Jesus 1659 Tchum, Kofi 1637 Thielecke, Marlene 101, 102 Tisdale, Michele 693 Tamarozzi, Francesca 439, 571, 639 Teal, Allen 1138 Thiga, Jacqueline 112 Tiyo, Gelila D. 554 Tambo, Munyaradzi 1502 Techasaensiri, Chonnamet 183 Thinraow, Suradet 328 Tobby, Roselyn 1119 Tamfum-Muyembe, J. J. 1256 Thiongo, Mary 1856 Todd, Jim 563 astmh.org Abstract Author Index A-629

The number(s) following author name refers to the abstract number.

Todd, Suzanne R. 661 Toutous Trellu, Laurence 1855 Trinies, Victoria 602, 1168, 1169 Tyagi, Rajeev 1238 Toé, Laurent 1695 Tovar, Marco 1278 Tripathi, Abhai K. 235, 1886 Tyndall, Lily M. 1882 Tofail, Fahmida 1266 Towers, Catherine E. 1883 Tripet, Fréderic 920 Tyner, Stuart 1774 Togo, Amadou 1493, 1527 Towers, David 1883 Tripura, Rupam 904 Tyrer, Hayley E. 485, 487, 503 Tohme, Rania A. 1300 Towner, Jonathan 861 Trobaugh, Derek W. 14 Toi, Cheryl 6 Townes, Lindsay R. 1546 Trollfors, Birger 1040 Tokarz, Rafal 1766 Townsend, Jeffrey P. 519 Trongnitpatt, Namtip 1841 U Tokunaga, Naohito 973 Townsend, R. R. 1785 Troupin, Andrea 176 Ubalee, Ratawan 912, 1841 Tolia, Niraj H. 317, 995, 1455, Townsley, Elizabeth 630 Troye-Blomberg, Marita 655, 964, Ubol, Sukathida 168 1585, 1898 Townson, Simon 1209 1557, 1797 Uddin, M. I. 583 Tolliver, Marcee 109 Tozan, Yesim 1254, 1615 Troyo, Adriana 778 Uddin, Md. Jashim 1426 Tolo, Youssouf 380, 619, 984 Tracey, Alan 488 Trueba, Gabriel 1761 Uddin, Muhammad I. 1651 Tolouei Semnani, Roshanak 29 Traina, Mahmoud I. 1050 Truman, Richard 477 Uddin, Taher 35, 433, 434 Tolstoy, Nikolai S. 1170 Tran, Duong T. 279, 1464, 1500, Truong, Hieu M. 279, 1500 Udenze, Kenneth O. 1465 Tomaino, Francesca 480 1500, 1501 Truong, Nguyen Tan 783 Udhayakumar, Venkatachalam 911, Tomashek, Kay M. 696, 811, 1379, Tran, Man 197 Truscott, James E. 1715, 1722 1807 1382, 1386, 1402, 1415, 1672, Tran, Tuan M. 1552, 1572, 1798, Try, Vorleak 298 Udhayashankar, Kanagasabai 465 1390 962 Tsai, Annabel 1934 Udomsangpetch, Rachanee 382 Tomaz, Franciele M. B. 391, 393, Tran, Vanessa 221 Tsai, Hung-Chin 513 Ugot, Iyam 1031 396 Tran, ViLinh 218 Tsai, Jih-Jin 826 Ujuju, Chinazo N. 934 Tomchaney, Michael 686 Tran Thanh, Duong 1222 Tsai, Wen-Yang 826 Uk, Sambunny 1033, 1626 Tomczyk, Sara 1070 Tran Vu Thieu, Nga 1641 Tsegay, Girmay 1070 Ukaga, Chinyere N. 260 Tompkins, Adrian M. 1497 Traore, Abdrahamane 1798 Tsegaye, Mesfin M. 179 Ul Alam, Mahbub 728, 1292 Ton, Thanh 1428 Traore, Aliou 1493, 1527 Tsertsvadze, Tengiz 1304 Umar, Abba 264, 265, 1576 Tong, Carlos G. 144, 149, 1823 Traoré, Boubacar 405, 962, 1552, Tshala-Katumbay, Desiré 460, 460, Umesumbu, Solange E. 1463, Tonga, Calvin 918 1571, 1572, 1598, 1798, 1866 1256 1469, 1538, 1547, 1634 Tongren, J. Eric 704 Traore, Diahara 25 Tshangela, Akhona N. 1644 Umlauf, René 871 Tonkin, Daniel R. 817 Traore, Djibril 1009 Tshefu, Antoinette 1533 Undurraga, Eduardo A. 203, 836 Torchinsky, Miriam B. 1670 Traore, Karim 380, 619 Tsuboi, Takafumi 973, 990, 993, Ung, Sam An 1766 Torii, Motomi 973, 990, 1591 Traore, Maminata 1922 997, 998, 1591 Unger, Holger W. 53, 1119 Tornieporth, Nadia 577, 578 Traore, Modibo 25 Tsuji, Moriya 1606 Unicomb, Leanne 728, 1292 Torondel, Belen 710 Traore, Moussa 1539 Tu, Ying 1096 Unnasch, Thomas R. 481, 1695, Torr, Steve 148 Traore, Omar B. 1492, 1527 Tu, Zhijian 1344 478, 1693, 1700 Torrero, Marina N. 1710 Traore, Sekou 1509 Tubert-Bitter, Pascale 251 Unni, Susmita 515 Torres, Amanda R. A.. 1200 Traore, Sekou F. 1371, 1696, 1697, Tuikue Ndam, Nicaise 987, 1557, Unosson, Klara 655 Torres, Brenda 1386 1698 1797 Unshur, Ahmed 1424 Torres, Eliana 959 Traore, Sekou F. 796 Tukahebwa, Edridah Muheki 593, Unwin, Vera T. 483 Torres, Giselle 1835 Traore, Sekou Fantamady 762 1085, 1160, 1693 Upfill-Brown, Alexander M. 46 Torres, Jaime R. 821 Traore, Sitan 762 Tukesiga, Ephraim 593, 1693 Uppal, Karan 218 Torres, Juan P. 1672 Traore, Souleymane 1598 Tulantched, Steve 1633 Upton, Leanna M. 1029 Torres, Katherine 1807 Trapaidze, Nino 1304 Tullo, Gregory 1799 Upton, Richard N. 244 Torres, Katherine J. 388 Trape, Jean-François 99, 1187 Tumbo, Anneth 670 Urassa, Honorati 77 Torres, Melissa 1713 Traub, Rebecca 597 Tumukunde, Alex 665 Urassa, Willy S. 722, 740 Torres, Michael 167, 862 Travassos, Mark A. 389, 1605, Tun, Nay Win 940 Uribe, Sandra 361 Torres, Rolando 206 1796, 380, 1596, 619, 1600 Tunes, Luiza G. 1094 Urnov, Fyodor 1237 Torres, Sonia M. 329 Travassos, Travassos A. 984 Tung, Nguyen H. 1092, 1093 Usman, Aishat B. 523, 566 Torres-Estrada, José 1695 Travi, Bruno 690, 1748, 1753 Tungu, Patrick 1189, 1888 Usuf, Effua-Abigail 666 Torres-Velasquez, Brenda 1382, Tree, Maya O. 210 Turab, Ali 634 Utami, Retno 219 1672 Trehan, Indi 1668 Turabelidze, George 661 Uth, Sophal 337, 1499 Torrico, Faustino 564, 716, 1778 Trentadue, Jordan 500 Turell, Michael J. 154, 789, 1369, Uthaimongkol N, Nichapat 1774 Torrico Rojas, Mary Cruz 1104 Trentini, Bruno 72 1374 Uto, T 1093 Tort, Jose F. 1781, 1816 Tretina, Kyle 1175 Turkson, Anthony J. 1614 Utrianen, D 20 Torto, Baldwyn 148 Triana, Omar 1820 Turner, Gareth 1434 Utzinger, Jürg 66, 1069, 1719 Tortolero, Mery 202 Triana, Paula 202 Turner, Hugo C. 1722 Uysal, Selver D. 1615 Tosh, Donna 1494 Triana-Chávez, Omar 793 Turner, Joseph D. 484, 487, 502, Uzochukwu, Benjamin 1472 Tosi, Michael 493 Triantafilo, Vjera 1640 503, 504 Uzonna, Jude 1745 Totino, Paulo Renato R. 390 Trianty, Leily 219 Turyaguma, Patrick 1085 Touch, Sok 721, 1774 Tribble, David 471, 693, 1051 Tusting, Lucy S. 1844 Tougher, Sarah 1579, 1581, 1583 Trieu, Angela 609 Tuxun, Tuerhongjiang 447, 448 Tougoue, Jean Jacques 1078 Trilisinska, Jana 1517 Twesigye, Roger 268 V Tounkara, Moctar 1539 Trimarsanto, Hidayat 958 Twumasi, Mieks 1067 Vaca, Maritza 1265, 1268 Touré, Mahamoudou B. 1525 Trindade, Pamella C. A. 391, 393, Twum-Danso, Kingsley 1036 Vaca, Sergio 1657 Toure, Sekou 1492, 1527 396 Ty, Maureen C. 610 astmh.org A-630 Abstract Author Index The number(s) following author name refers to the abstract number.

Vafa-Homann, Manijeh 405, 974 Van Overmeir, Chantal 900 Vestergaard, Lasse S. 897 Vos, Martijn W. 1018 Vaidya, Akhil B. 618, 673, 980 Vanraes, Joelle 1483 Vi, Lu Lan 430 Voss, Kelsey 157 Vaillant, Michel 476, 1063, 1716, Van Roey, Karel 657, 1624, 1626 Viana, Giselle M. R. 1563 Vounatsou, Penelope 1069 1719 van Schaijk, Ben C. L.. 1594 Viana, Susana A. S. 1224 Vu, Dung K. A. 279 Val, Fernando F. A. 340 van Seventer, Emily E. 1277 Vianna, Priscila 1563 Vu, Tuan Trung 814 Vala, Anifa 338, 1478 Van Sprundel, Marc 1157 Vianou, Bertin 1557, 1797 Vu, Tuyet T. A. 279 Valanparambil, Rajesh M. 512 Van Voorhis, Wesley C. 1038, 1172 Viboud, Cecile 54 Vuitton, Dominique A. 440, 442 Valderrama, William 840 Van Vugt, Michele 1255 Vicente, José L. 1367, 1912 Vujcic, Jelena 711, 712 Valderrama Cumbrera, Anayansi van Woensel, Job 1861 Vicuña, Yosselin 1268 Vujcikova, Julia 530 1346, 663 van Wyk, Albert 255 Vidadala, Rama S. R. 1172 Vulule, John 582, 760, 1261, 1560, Valdés, Odalys 857, 1416 Varan, Aiden K. 723 Viebig, Nicola 671, 986, 1590 1879, 1887 Valdez, Melissa 481, 506 Vareta, Jimmy A. 620 Vieira, Paula 1263 Vuong, Chau 283 Valdovinos, Alexis 319 Varga, Laszlo 11 Vigdorovich, Vladimir 17 Vuyyuru, Raja 1264 Valea, Innocent 1922 Vargas, Elsy 812 Vigninou, Estelle 769 Vythilingam, Indra 139 Valecha, Neena 910 Vargas, Paola 514, 643, 695 Vila-Sanjurjo, Anton 167, 862 Valencia, Braulio M. 538, 1095 Vargas-Calla, Ana M. 451 Vilcarromero, Stalin 807 Valencia Ayala, Edward 1930 Vargas-Zambrano, Juan C. 537 Vilchez, Samuel 586 W Valencia-Hernández, Carlos A. 1234 Varma, Santosh G. 611 Villadiego, Kelly 1408 Waddington, Claire S. 1849, 1850 Valente, Vanessa A. 536 Vasconcelos, Pedro F. C. 650 Villalobos, Fabricio 1339 Wadegu, Meshack 559 Valenzuela, Jesus 1202 Vasilakis, Nikos 206, 1397, 1928 Villamor, Eduardo 815 Waggoner, Jesse J. 1058 Valerie, Exler 387 Vasireddy, Vamsi 905, 934, 1532, Villani, Fernanda N. A. 858 Waghabi, Mariana C. 536 Valerio, Laura 1884 1533, 1580 Villanueva-Lizama, Liliana 1750, Wagman, Joseph 1191 Valero, Natalia R. 532 Vasques, Thais Perez 550 1936 Wagstaff, Simon 489 Valerrama, William D. 1757 Vasquez, Bruna Perez 550 Villar, Luis Angel 577, 578, 745, Wahala, Wahala 1385 Valian, Adams 68 Vasquez, Carlos 1610 815 Waiboci, Lilian W. 819, 1382 Valim, Clarissa 50, 614, 1477, 1522 Vasquez, Karla 1761 Villasante, Eileen 608, 1005, 1574, Waisberg, Michael 1453 Valle, Denis 375 Vasquez-Valderrama, Alex 807 1604, 1610, 1601, 1603 Waitumbi, John 112, 263 Valle, Giancarlo A. 1129, 1776 Vasudevan, Prabhakaran 1193, Villasis, Elizabeth 388, 1807 Waleckx, Etienne 1329 Vallejo, Andres F. 334, 1802, 379 1195 Villegas, Leopoldo 1872 Wali, Sadiq S. 713 Vallur, Aarthy C. 1228 Vaughan, Ashley M. 1018, 1829, Villinger, Jandouwe 158, 259, 859 Walke, Henry 1110 Van, Hai 337 1897, 1901, 1568 Vincent, Naomi 791 Walke, Jayashri 910 Vanachayangkul, Pattaraporn 912, Vaughan, Jefferson A. 1374 Vincenti, María F. 202, 820 Walker, Allison T. 584 1481 Vazeille, Marie 1361 Vinekar, Kavita 1300 Walker, David H. 103 Vanaerschot, Manu 688 Vazquez-Prokopec, Gonzalo M. Vinetz, Joseph M. 144, 149, 149, Walker, Don 675 VanArendonk, Laura G. 1654 153, 787 227, 329, 414, 1025, 1224, Walker, Edward 5, 842, 1368, 791, Van Beneden, Chris 1215, 1878 Vedantam, Rajshekhar 1195 1510, 1823, 1823, 1823, 388, 931, 1377, 1534, 1887 VanBlargan, Laura A. 841 Vedovello, Danila 194 1807, 1858 Walker, Larry A. 252, 295, 674, Van den Abbeele, Jan 229 Vedvick, Tom Vedvick 1606 Vinh Nguyen, Chau Van 783 1096 Van Den Borne, Bart 333 Veintimilla, Marcia 916 Vinje, Jan 1643 Walker, Lucy 395 Van Den Eede, Peter 1519 Vekemans, Johan 666 Virnig, Beth 1459 Walker, Martin 1330 Vanden Eng, Jodi 1026, 1847 Velasco-Salas, Zoraida I. 202, 820 Visser, Leo G. 992 Walker, Neff 731 Van der Auwera, Gert 1095 Velo, Enkelejda 795 Viswanathan, Anusha 493 Walker, Patrick 919, 1520, 1843 Van Der Elst, Josiane 1305 Venepally, Pratap 1798 Vital-Brazil, Juliana M. 1058 Walker, Peter J. 1322 van der Geest, Sjaak 1097 Venkatachalam, Indumathi 965 Vittinghoff, Eric 1227 Walker, Thor 1285 van der Werf, Tjip S. 1655 Venkatesh, Apoorva 611 Vittor, Amy Y. 663 Walldorf, Jenny A. 953, 1225, 1522 van de Vegte-Bolmer, Marga G. Vennervald, Birgitte 403, 1794 Vizcaino, Lucrecia 131, 788 Walling-Maiga, Amelia 324 1018 Venter, Marietjie 1927 Vlaminck, Johnny 644 Wallqvist, Anders 1595, 1597 van Diepen, Angela 59 Ventocilla, Julio A. 397, 1737, 1738 Voinescu, Bianca 1517 Walson, Judd L. 516, 824, 1260, van Doorn, Rogier 1768 Ventocilla, Julio A. Vojtasova, Lenka 867 1848, 1880 van Eijk, Anna Maria 23, 948 Ventrone, Cassandra 181, 579 Volcik, Kelly 431 Walther, Michael 669, 1561, 1592 van Eijk, Annemieke 679 Venturelli, Claudio 1375 Volf, Petr 539 Walyambillah, Waudo 290 Van Geertruyden, Jean-Pierre 274, Vera Hernandez, Marcos 1582 Volkman, Sarah 18, 320, 331, 617, Wamala, Samuel 402, 1569 275, 522, 900, 1157, 261, 734, Vera-Maloof, Farah 779 904, 1805, 19, 234, 930, 1240, Wamboga, Charles 461 1806, 318, 1786 Verbel, Daniel 110 1241, 1477, 1628, 1799, 1833 Wami, Welcome M. 1145 van Gemert, Geert-Jan 1018 Vercellotti, Gregory M. 232 Voller, Katja 1419 Wamola, Newton 1045 Van Kempen, Luuk 1255 Vercruysse, Jozef 644 Volney, Béatrice 1241 Wamoyi, Joyce 1466 Van Kerkhove, Maria 208, 209 Verduguez, Aleida 1095 Volta, Bibiana 544 Wampfler, Rahel 955, 1550 Vanlandingham, Dana L. 161, 205 Verfaillie, Catherine M. 229 von Fricken, Michael E. 1566 Wandera, Bonnie 242, 1460 van Liere, Genevieve 1150 Verhoeve, Victoria I. 113, 106 von Horn, Charlotte 30 Wanderley, Dalva M. V. 1230 van Loen, Harry 1856 Verity, Robert 999 von Seidlein, Lorenz 22 Wang, Amy 1235 Vann, W. F. 588 Verkerke, Hans P. 1132 Vontas, John 1186 Wang, Annie 1764 Van Ngoc, Tran 1384 Verma, Saguna 838 Voorhees, Matthew 91, 1418, 860 Wang, Bo 864, 993 Vannier, Edouard G. 1219 Vernick, Kenneth D. 1910 Voronin, Denis 501 astmh.org Abstract Author Index A-631

The number(s) following author name refers to the abstract number.

Wang, Chunling 190 Webb, Lauren 1938 White, Clinton 1173 Wilson, Michael D. 1248, 1330, Wang, Eryu 663 Webster, Bonnie 1810 White, Gregory 839 1335, 1717, 1720 Wang, Hui 438, 638 Webster, Bonnie L. 1812 White, John 910, 1449 Wilson, Michael David 594 Wang, Liman 197 Webster, Jayne 1466 White, Karen 673 Wilson, Nana O. 226, 1694 Wang, Michael Zhuo 1091 Webster, Joanne P. 1184, 1810, White, Laura F. 1282 Wilson, Tony 1860 Wang, Molin 722 1812, 1811 White, Laura J. 817, 1385 Win, Tun Tun 1920 Wang, Qian 1931 Wee, Siok-Bi 98 White, Lisa J. 343, 917, 1107 Winchell, Jonas 1064 Wang, Ruobing 329, 1568 Weeks, Andrew R. 1322 White, Michael 47, 964, 1242 Wingfield, Tom 79, 719, 1278 Wang, Shengyue 638 Weerasooriya, Mirani V. 590 White, Nicholas 268, 904, 1873, Winskill, Peter 2, 1330 Wang, Tsun 754 Weetman, David 752, 773, 776, 1920, 301, 311, 328, 343, 940 Winter, Alan 486 Wang, Wei-Kung 826 1367, 1912 Whitehead, Stephen 173, 579, 822, Winter, Rebecca 704, 1028, 1249, Wang, Weiming 128 Wegbreit, Jennifer 1500 826, 1378, 181, 828 1289, 1632 Wang, Xiaohong 1350 Wegbreit, Jennifer A. 342, 1464 Whitehorn, James 783, 784 Winter, Rolf 291, 673 Wang, Xiaoming 1353 Weger-Lucarelli, James D. 163 Whittlin, Sergio 287 Winters, Anna M. 136, 925, 1504, Wang, Xuelei 31 Wei, Junfei 1892 Whitton, Jane M. 1151 922, 924 Wang, Ying 371, 1537 Weier, Christina 1704 Wichaidit, Wit 728, 1292 Winters, Benjamin 136 Wangai, Hannah 563 Weigl, Bernhard 1318 Wickremasinghe, Renu 655 Winterstein, Eric 515 Wangnapi, Regina 53 Weil, Ana A. 432 Widdowson, Marc-Alain 1771 Winzeler, Elizabeth 287, 1805, 292, Wangrawa, Dimitri W. 117 Weil, Gary J. 65, 482, 590, 1707, Widen, Steven 206, 1397 294 Wani, Tasaduq H. 542 1785, 1212 Widman, Doug 625, 627, 628 Wirth, Dyann F. 18, 234, 250, 292, Wanja, Elizabeth 263 Weill, Mylène 1192 Wiegand, Roger 50, 294, 676 294, 614, 656, 676, 904, 907, Wanjala, Christine L. 120, 1020 Weina, Peter 1434 Wiegand, Ryan 60, 1079, 1908, 930, 1240, 1241, 1477, 1628, Wanji, Samuel 503, 592, 1208, Weinbaum, Bradley 1860 1795 1799, 1833, 50, 320, 331, 415, 1704, 1890 Weinstein, Corey S. 404 Wieringa, Frank 1721 617, 1436, 1805 Wansaicheong, Gervais 1062 Weinstein, Philip 6 Wierzba, Thomas F. 1297, 1870 Wirtz, Robert 788 Wanyoike, Salome K. 660 Weir, William 486 Wigant, Wilson 1640 Wirtz, Veronika J. 1935 Wanzira, Humphrey 26, 257, 946 Weirzba, Thomas F. 423, 1433 Wijayalath, Wathsala 607, 1574, Wise, Emma 1419 Wanzirah, Humphrey 1844 Weiskopf, Daniela 822, 1275 1601 Wiseman, Virginia 884, 1472, 1582 Ward, Abigail 878, 886, 1907 Weiss, Christopher M. 164 Wijewickrama, Ananda D. 171, Witek-McManus, Stefan 879, 1253 Ward, Danielle 1892 Weiss, Daniel J. 943, 944, 945, 1381 Witkowski, Benoit 1237 Ward, Stephen A. 484, 502, 483, 1929 Wiladphaingern, Jacher 328 Wiwa, Owens 1907 495, 504 Weiss, Louis M. 1176 Wilcox, Bruce 798 W/Kidan, Samuel 509 Wardrop, Nicola 1290 Weissenberger, Giulia 1664 Wilding, Craig S. 768 Woc-Colburn, Laila 698 Ware, JeanAnne 1198 Weitzel, Thomas 1640 Wilkins, Patricia P. 458 Woda, Marcia 630 Warit, Saradee 168 Wekesa, Wakoli 1376 Willey, Barbara 1579 Wojcik, Genevieve L. 1266 Warraich, Gohar 954 Weldon, William 587 William, Timothy 22, 1109 Wolbers, Marcel 173, 783 Warrell, David A. 1679 Wellems, Thomas E. 319, 1805 William, Yavo 1239 Woldehanna, Sara 1759 Warren, Cirle A. 1178 Wen, Hao 438, 441, 447, 448, 638 Williams, Craig 6 Wolf, Katherine 1627 Warren, William 833 Wenbao, Zhang 440, 442 Williams, Gail 1144, 1185 Wolkon, Adam 1847 Warrenfeltz, Susanne 981 Wendel, Silvano 1545, 1742 Williams, Jack L. 1446 Won, Kimberly 60, 1079, 1182 Wassmer, Samuel C. 610 Weng, Shih-Che 753 Williams, Jacob 1622 Wondji, Charles S. 133, 764, 1188 Wastling, Jonathan M. 1890 Weng, Tzu-Ping 724 Williams, John 1872 Wong, Jacklyn 787 Waswa, Sadic 861 Wenger, Edward A. 46, 1542, 667, Williams, Maya 1429, 1924 Wong, Joshua G. 825, 1678 Watanabe, Noriko 655 1506, 1535, 1613 Williams, Priscilla L. 860 Wong, Meng Li 139 Watanaveeradej, Veerachai 1770 Weny, Geoffrey 665, 1431 Williams, Steven A. 480, 1725, Wong, Mimi 581 Waterhouse, David 502 Wepnje, Godlove 918 1713 Wong, Paolo A. 1128 Waterhouse, Robert M. 683 Weppelmann, Thomas A. 1166, Williams, Thomas N. 1556, 1902 Wong, Wesley 930, 1628, 1833 Waterman, Stephen 477, 723 1566 Williams, Zachary 500 Wongarunkochakorn, Saowaluk Waters, Norman C. 887 Were, Patrick S. 290 Williams-Newkirk, Amanda J. 105 1494 Watila, Ismaila 256 Were, Phelix 80 Williamson, John 582, 631, 1478 Wong-Madden, Sharon 1561 Watkins, Simon C. 818 Were, Vincent 1534 Williamson, Jonathan 79, 719 Wongsombat, Chayaphat 406 Watson, Alan M. 14, 214 Wertheim, Heiman 1768, 1837 Willilo, Ritha 351 Wongsrichanalai, Chansuda 693 Watson, Nora L. 56 Wesolowski, Amy 83 Willilo, Ritha A. 1015 Wongstitwilairoong, Tippa 1774 Watts, Douglas M. 647, 1928 Wesson, Dawn M. 546, 792, 843, Willis, Paul 287 Wood, Thomas G. 206, 1397 Waybright, Nicole 166 1372 Willis, Rebecca 1259 Woodall, Patricia A. 1703 Wearing, Helen J. 10, 165 West, Nicole 921 Wills, Bridget 173, 462, 783, 1837 Woodrow, Charles J. 268, 904 Weaver, Abigail A. 73, 80, 1314 West, Phillipa 1245, 1365 Wilschut, Jan C. 202 Woods, Edward 1083 Weaver, Anne M. 48 West, Sheila K. 1048, 1066 Wilson, Alan 59 Woolhouse, Mark 1145 Weaver, Marcia R. 275 Westby, Katie 785 Wilson, Danny W. 385 Worley, Sam 93 Weaver, Scott 663 Weurtz, Stefan 708 Wilson, Leslie 1772 Woukeu, Arouna 1856 Weaver, Scott C. 11, 206 Whalen, Christopher C. 714, 1121 Wilson, Mark L. 572, 1225, 1530, Wozniak, Edward J. 1730 Webale, Mark 1553 Wheatley, Adam 1800 1546, 1630 Wright, Alexandra 772, 1365 Webb, Claire 93, 1675 Wheeler, Brian 1908 Wilson, Michael 519, 1526 Wright, David W. 505 astmh.org A-632 Abstract Author Index The number(s) following author name refers to the abstract number.

Wright, Gavin J. 410 Yan, Hong-Bin 1814 Zheng, Aihua 1387 Wright, Richard 1083 Yan, Li 26 Z Zheng, Hong 235, 272 Wu, Haipeng 1630 Yanagi, Tetsuo 400, 985 Zaatour, Amor 1934 Zheng, Qiang 252 Wu, Haiwei 56 Yang, Amy 230 Zabala, Julian R. 140 Zhong, Daibin 758, 1353 Wu, Hua 1924 Yang, Chao-Fu 207 Zachem, John 1044 Zhong, Lina 1048 Wu, Lin-Kun 724 Yang, Henglin 895 Zahm, Jacob A. 995 Zhou, Guofa 371, 937, 941, 1020, Wu, Lindsey 742 Yang, Sihyung 1091 Zaidi, Anita K. 585 1353, 1537 Wu, Shuenn-Jue 1396 Yang, Tsong-Han 207 Zaidi, Irfan 669, 1561 Zhou, Huayun 128, 958 Wu, Xiao-Jun 1790 Yang, Zhaoqing 371 Zaidi, Syed Sohail Zahoor 54 Zhou, Taoli 1564 Wu, Yimin 668, 796, 1009, 1371 Yannow, Stephanie 1003 Zakrzewski, Martha 1218 Zhou, Tingyi 197 Wubie, Moges 1070 Yanow, Stephanie K. 408 Zaky, Weam I. 480 Zhou, Xiao-Nong 1719 Wu-Freeman, Ying 588 Yao, Phil 1752 Zaman, K. 583 Zhou, Xiaonong 895 Wunderlich, Juliane 312 Yap, Alan 1945 Zamanian, Mostafa 492 Zhou, Yan 638 Wurapa, Eyako 276, 1523 Yaro, Jean Baptiste 671, 1483, Zambon, Maria 1046 Zhou, Yiyang 1926 Wurapa, K. 280 1590 Zambrano, Betzana 578 Zhou, Zhaoxia 1048 The WWARN AL Dose Impact Study Yasnot, Maria Fernanda 412 Zambrano, Maria L. 105 Zhou, Zhiyong 1534 Group 27 Yatsunenko, Tanya 1654 Zamsaeur, Nicole 563 Zhu, Daming 668, 1009 Wyatt, Paul 1098, 1933 Yazdanbakhsh, Maria 1895 Zamudio, Carlos 1129, 1776 Zhu, Guoding 128 Wyka, Katarzyna 1197 Ye, Maurice 1011 Zani, Carlos L. 1094 Zimic, Mirko 51, 1296 Ye, Yazoume 1028, 1034, 1249, Zapol, Warren 1487 Zimic, Mirko, for the Cysticercosis 1289, 1579, 1617, 1623, 1872 Zariquiey, Carlos M. 1738 Working Group in Peru 452 X Yeboah-Antwi, Kojo 637, 706 Zarroug, Isam 1706 Zimicki, Susan 1759, 1765 Yeboah-Manu, Dorothy 1335 Zimmerman, Peter 5, 1805, 491, Xayavong, Maniphet V. 97, 98 Zavala, Fidel 1606, 1607 Yee, Angela 1567 621, 791, 983, 1377, 1455, Xaymounvong, Khounkham 1759 Zayed, Alia 114 Yeka, Adoke 1577 1827 Xia, Dong 1890 Zébazé Togouet, Serge Hubert 567, Yen, Minmin 34 Zinoecker, Severin 1866 Xia, Hui 382 1161 Yeo, Tsin W. 22, 1905 Zinszer, Kate 935, 1293 Xiao, Lihua 1261 Zegers de Beyl, Celine S. 1845 Yerbanga, Serge R. 119, 284 Zogo, Barnabas 118 Xiao, Wenyuan 1177 Zeh, Clement 1879 Yeung, Shunmay 255 Zornetzer, Heather 725 Xiao, Yunfeng 441 Zeidner, Nordin S. 1110 Yewhalaw, Delenasaw 568 Zorowka, Patrick 21 Xiaomei, Lu 440, 442 Zeitlin, Jennifer 936 Yeya, Sarro 1598 Zorrilla, Diana 746 Xie, Lisa H. 282, 283, 291 Zeituni, Amir E. 1799 Yi, Chenda 108 Zottig, Victor E. 252, 1462 Xiong, Xu 635, 1733 Zelaya, Carla E. 1650 Yimamnuaychok, Narathatai 217, Zou, Cathy 608 Xu, Guang 103 Zeli, E 20 1829 Zubieta-Zavala, Adriana 1401 Xu, Jiabao 758 Zelman, Brittany 915 Yin, Kuo 197 Zuchi, Nayara 1397 Xu, Peng 433, 588 Zelner, Jon 430 Yin, Xiaoping 714 zu Erbach-Schoenberg, Elisabeth Xu, Shulin 419 Zeng, Qiang 282, 283 Yingyuen, Kritsanai 299 653 Xu, Sui 128 Zenklusen, Isabelle 670 Yip, Fuyuen 1258 Zulu, Siphosenkofi G. 1152 Xu, Wei Ping 1040, 868, 927, 929 Zevallos, Eric 94 Yman, Victor 964, 974 Zulu, Siphosenkosi G. 1794 Xu, Weixin 1914 Zevallos, Karine 1278 Yoboue, Charlene A. 130 Zumbua, Graciete 1153 Xu, Wenyue 1564 Zhan, Bin 1892 Yokobe, Lindsay 481, 506 Zunt, Joseph R. 1428 Xuya, Marvin 1777 Zhang, Chao 128 Yong, Tai-Soon 1782, 1783 Zhang, Chuanshan 438 Zwang, Julien 245, 1148 Yoo, Eun-Hye 48 Zhang, Jin-Hui 447, 448 Zwet, Erik v. 992 Y Yoon, In-Kyu 8, 168, 630, 697, Zhang, Jing 282, 283 816, 863, 1272, 1274, 1391, Zhang, Karen 1720 Yacoub, Sophie 1837 1429, 1770 Zhang, Lei 1237 Yactayo, Sergio 208, 209 Yoon, Nara 1149 Zhang, Lixin 1761 Yadav, Prashant 1630 Yoon, Steven S. 893 Zhang, Min 898 Yadav, Rajpal 118 Yori, Pablo P. 585 Zhang, Ping 282, 283 Yahathugoda, Thishan C. 590 Yoshino, Timothy P. 1790 Zhang, Qunyuan 1654 Ya Jagne, Jankey 671 Yost, Stacy 831 Zhang, Rou 1440 Yakasai, Ahmad M. 713 Young, Ryan M. 1695 Zhang, Summer 180 Yakob, Laith 521 Yount, Boyd L. 625, 628 Zhang, Summer L. 199 Yakpa, Kossi 1136 Yousif, Maitham G. 1667 Zhang, Wenbao 438, 638, 638 Yakubu, Habib 573 Yu, Hannah 1016 Zhang, Xue 438 Yamaoka, Shoji 1092, 1093 Yu, Sun N. 1048, 1066, 1307 Zhang, Zhi-qiang 197 Yamasaki, Tsutomu 1591 Yu, Weiwei 1360 Zhang, Zhongsheng 1038 Yaméogo, Bienvenue K. 284 Yu, Yanan 588 Zhang, Zhuangzhi 638 Yamo, Emmanuel 866 Yuan, Melissa 1285 Zhao, Chenghao 1564 Yan, Guiyun 371, 758, 909, 937, Yui, Katsuyuki 985 Zhao, Jin-Ming 447, 448 941, 1020, 1223, 1353, 1537, Yukich, Josh 1470, 1507 Zhao, Jincun 1924 1613, 1887, 120 Yukich, Rudy K. 1507 Zhen, Huajun 638 Yun, Heather C. 471 astmh.org