376 but different for the human-baited tent, with a majority (41%) of An. by plastic containers and small pools along the way. The main specie vagus. Eight of the nine morphological species represented in this study collected was Ae. aegypti followed by Culex quinquefasciatus. Anopheles were captured on each of the three hosts, suggesting a plasticity in mosquitoes were not found in the highway area. Tire repair shops were host attraction behavior. Multiple host feeding and flexibility in feeding the places with the highest number of infected tires; we also found behavior could have important implications for malaria control. abandoned tires infected with mosquito larva. 1250 1252 ASPECTS OF ECOLOGY OF POTENTIAL RIFT VALLEY FEVER A COMPARATIVE EVALUATION OF SIX DIFFERENT MALARIA VIRUS MOSQUITO VECTORS, KHARTOUM STATE, SUDAN VECTOR COLLECTION METHODS IN LOW-LYING MALARIA ENDEMIC REGIONS OF WESTERN KENYA Deena M. Abdelgadir Federal Ministry of Health Sudan, Khartoum, Sudan Nabie Bayoh1, George Olang1, Nico Govella2, Gerry Killeen2, John Gimnig3 Rift valley fever epidemics are disruptive and expensive to local and 1Kenya Medical Research Institute, Kisumu, Kenya, 2Ifakara Health regional economies. After a devastating outbreak of Rift Valley Fever Institute, Dar es Salaam, United Republic of Tanzania, 3Centers for Disease in khartoum state, Sudan 2007; ecological baseline surveys were Control and Prevention, Atlanta, GA, United States conducted in Khartoum State, Sudan, during the rainy season (end of July to the beginning of September) 2008 in order to identify mosquito Outdoor biting and other forms of behavioral adaptation by malaria species present and evaluate their emergence and survivorship. Larval vectors to domestic insecticide-based control measures may compromise identification of species of Culicine and Anopheline mosquitoes present the sensitivity of conventional sampling tools operating indoors such as in Khartoum State taken from five study sites represents Khartoum state light traps or indoor resting catches, thus preventing effective surveillance indicated that Anopheles arabiensis is the only species of the Anopheline and management of vector populations. We evaluated six different vector mosquitoes found. Three species of culicine mosquitoes were found: collection methods to optimize a protocol for operational sampling of Culex quinquefasciatus, Cx univittatus and Cx arbeeni, Species of Aedes malaria vectors robust to variations in vector behavior, notably variations were found in irrigated schemes at one study site and was absent from associated with the presence of important malaria control methods. Over the other four study sites, these species were Ae. vittatus and Ae. vexans, 30 days, we replicated a Latin square design 10 times at sites in 4 districts whose presence was recorded after the onset of the rainy season.the in western Kenya: Kisumu, Bondo, Nyando and Rachuonyo. Each site same breeding site was first occupied by Ae.vittatus then Ae. vexans, consisted of 3 locally representative houses through which the six different with an interval of habitat drying. Daily emergent adults Culicine and sets of trapping methods were rotated every 3 nights in a random order Anopheline mosquitoes present were taken from randomly selected of three possible arrangements: 1) Indoor human landing catch (HLC) and breeding sites in the five study sites, population measurements were outdoor HLC, 2) CDC Light trap placed beside an occupied insecticide- performed. The absolute number of emergent adults was obtained by treated net indoors combined with Ifakara tent traps outdoors, and 3) collecting mosquitoes under net-traps covering the breeding sites. Records Window traps to catch exiting mosquitoes combined with both pot and were taken each day for seven constitutive days, synchronized emergence box formats of resting traps placed both indoors and outdoors. At each of males and females was observed at all the study sites, showing an site, a fourth house was selected for pyrethrum spray catch (PSC). The overall marked predominance of females in emergence trap catches. Adult top collection methods with their corresponding number of Anopheles survival rate was the most important factor determining the stability of the per collection effort were PSC (10.5), HLC indoor (3.0), Light trap (3.0) population and total egg production. Females that become infected when HLC Outdoor (2.8) and Ifakara tent traps (2.7). Resting Boxes and Pots taking a blood meal must survive throughout the incubation period of the positioned both indoors and outdoors caught less than 1 Anopheles per pathogen. Under controlled laboratory environment, effect of food types collection effort. HLC outdoor collected the highest amount of Culex at (sucrose 10%, sucrose 10% and blood diet) on longevity of adult female 77.4 per collection effort. Irrespective of the intensity or type of insecticide mosquitoes was conducted, sugar-fed and blood-fed mosquitoes exhibited based vector control method in place and of biting behavior of the local very high percentage of surviving rates beyond the 15 days (incubation malaria vectors, we conclude that pyrethrum spray catch is the most period for RVFV),However these have varied among the five study areas. sensitive method for vector collection in low lying malaria endemic regions Also results indicated prolonged survival of sugar-fed female mosquitoes of western Kenya. more than blood and sugar fed females, this served to increase survivorship of females until they find the appropriate host. 1253 1251 HETEROGENEOUS FEEDING PATTERNS OF AEDES AEGYPTI IN HOUSEHOLDS IN IQUITOS, PERU MAIN MOSQUITO BREEDING SITES FOR AEDES AEGYPTI IN 1 2 1 THE PAN-AMERICAN HIGHWAY: CUCUTA-PAMPLONA AREA Kelly Liebman , Helvio Astete , Steven T. Stoddard , Moises 3 4 5 1 (NORTE DE SANTANDER - COLOMBIA) IN 2010 Sihuincha , Eric Halsey , Tadeusz J. Kochel , Amy Morrison , Thomas W. Scott1 1 2 Berlin L. Londono-Renteria , Flaminio Londono , Jenny Carolina 1University of California, Davis, Davis, CA, United States, 2Navy Medical 3 1 Cardenas , Lucio Daniel Cardenas Research Unit-6, Iquitos, Peru, 3Hospital Apoyo Iquitos, Iquitos, Peru, 4Navy 1Universidad de Pamplona, Pamplona, Colombia, 2Politecnico Jaime Isaza Medical Research Unit-6, Lima, Peru, 5Navy Medical Research Center, Silver Cadavid, Bello, Colombia, 3Hospital Los Patios-Norte de Santander, Los Spring, MD, United States Patios, Colombia Heterogeneous biting by female mosquitoes can significantly alter Aedes aegypti is the principal dengue vector in Colombia where dengue transmission of mosquito-borne pathogens. Previous studies show Aedes transmission is limited by the presence of the vector; unfortunately in this aegypti, the primary vector of dengue viruses (DENVs), more frequently country, the presence of Ae. aegypti has been documented up to 2200 bite individuals with higher body mass index (BMI). Because BMI increases m.a.s.l. Norte de Santander is the second most endemic area for dengue with age, we expect positive linear relationship with age and biting. in the country. Previous studies have associated travel and transport as Studies show, however, that young adults receive more bites than older key factors in the spread of diseases and vectors. With this pilot study, we adults. Factors such as sex, mosquito exposure time and previous DENV investigated the main breeding sites and mosquito larva species on the infection should, therefore, be used to analyze heterogeneous feeding highway from Cucuta (325 m.a.s.l) to Pamplona (2342 m.a.s.l) in 75km patterns. Between October 2009 and November 2010, 2,035 interviews distance. We found that tires where the main breeding site followed with 280 participants were conducted in 19 households in Iquitos, Peru. www.astmh.org 377 Interviews focused on anthropomorphic characteristics and time spent in 1255 houses. In the week following interviews adult mosquitoes were collected twice daily, yielding 1,878 engorged and partially engorged mosquitoes. A COMPUTER SYSTEM FOR FORECASTING WEST NILE VIRUS Engorged abdomens were excised and participant DNA was obtained by RISK USING EARTH OBSERVATION DATA cheek swab. All DNA was extracted using Qiagen extraction columns. 1 1 1 Human DNA was amplified at 10 microsatellite loci, and allelic profiles Michael C. Wimberly , Ting-Wu Chuang , Aashis Lamsal , 1 1 2 identified using capillary electrophoresis. A computer program matched Alemayehu Midekisa , Geoffrey M. Henebry , Gabriel Senay , 1 1 participant profiles to mosquito blood meals. To date, 99 of 115 identified Michael B. Hildreth , Yi Liu blood meal profiles have been matched to participants. 29 young adults 1South Dakota State University, Brookings, SD, United States, 2USGS Earth (ages 15 - 35) received 50 bites (1.72 bites/person). 14 children (<15) and Resources Observation and Science Center, Sioux Falls, SD, United States 23 older adults (>35) received 154 and 34 bites, respectively (1.07 and Although there have been many calls to expand the use of earth 1.48 bites/person). In one household of 12 residents ranging in age from observation technologies in the health sciences, there are few examples 5 to 70 years with BMI of 13 to 32 kg/m2, 2 young adults ages 27 and 31 of operational systems with demonstrated impacts on public health. Our with BMI’s of 23 and 21.2 contributed to 46% of the 26 identified blood research objective was to bridge the gap between remote sensing and meals, consistent with the idea that young adults are bitten most often, public health by developing decision support systems to provide health and indicating that age better predicts biting frequency than BMI. Analysis scientists and practitioners with access to environmental information of the remaining 1,763 mosquitoes and interview data will be completed for surveillance and forecasting of mosquito-borne diseases. Specific in the next 4 months. Results will be used to model virus transmission and objectives were to automate the processing of remote sensing data to compare various vaccine delivery strategies. to generate environmental metrics, analyze the predictive capabilities of these metrics using retrospective datasets of human disease cases, 1254 and develop a web-based system for visualization and analysis of the WHOLE GENOME SEQUENCING OF ANOPHELES resulting products. The system was programmed using JAVA for user interface development and overall system control. Spatial analyses were PUNCTULATUS SIBLING SPECIES OF PAPUA NEW GUINEA carried out using Python scripts to call ArcGIS geoprocessing functions. David Serre1, Kyle Logue2, Lisa Reimer2, Ernest Chan1, Cara PostgreSQL was used for the storage and manipulation of the resulting Halldin2, Peter Siba3, Peter A. Zimmerman2 data summaries. We implemented a prototype of the system to forecast 1Cleveland Clinic, Cleveland, OH, United States, 2Case Western Reserve outbreaks of West Nile virus in the northern Great Plains. Environmental University, Cleveland, OH, United States, 3Papua New Guinea Institute of variables included MODIS land surface temperature (LST) and vegetation Medical Research, Goroka, Papua New Guinea indices (e.g., NDVI, EVI) derived from the MODIS nadir BRDF-adjusted reflectance product. We also used these data to compute actual The Anopheles punctulatus (AP) group in Papua New Guinea and evapotranspiration (ETa) using the simplified surface energy balance Southwest Pacific consists of at least 13 sibling species that include the method. Statistical analysis using generalized additive models (GAMs) vectors of malaria and lymphatic filariasis. Understanding the population revealed non-linear associations between interannual variability in WNV organization of the mosquitoes as well as the molecular basis for incidence and interannual deviations of cumulative LST, NDVI, and ETa the phenotypic variability related to vector competence or control is throughout the spring and early summer. There was an early-season complicated by limited data on the genetic diversity of these mosquitoes. influence of the timing of spring onset (captured by NDVI) as well as a We present here data generated by whole genome sequencing from late spring/summer influence of accumulated moisture and temperature individual AP mosquitoes and show that this approach provides extensive (captured by LST and ETa). Forecasts are currently being disseminated via catalogues of genetic polymorphisms and can significantly contribute to a web atlas (http://globalmonitoring.sdstate.edu/eastweb) and will be better understand the biology of these mosquitoes. We extracted DNA validated using surveillance data from the 2011 WNV season. from individual mosquitoes, and after determination of the species status by species-specific PCR-based assay, sheared the DNA molecules into 250- 1256 300 bp fragments and prepared libraries for two Anopheles punctulatus mosquitoes, one An. farauti 1, one An. farauti 2 and one An. koliensis. DYNAMICS OF ANOPHELES GAMBIAE POPULATIONS IN THE We sequenced each library on individual lanes of an Illumina GAIIx (paired- SAHEL: NEW PATTERNS AND NEW PUZZLES AWAIT NEW end 51 bp) or HiSeq 2000 (paired-end 100 bp). Overall, less than 1.5% of UNDERSTANDING the reads generated could be mapped to the An. gambiae (AG) reference 1 2 2 genome sequence suggesting that the sequence divergence between AP Tovi Lehmann , Adama Dao , Abdoulaye Adamou , Yaya 2 2 2 2 and AG is too great for the latter to serve as a useful reference sequence. Kassogue , Alpha S. Yaro , Moussa Diallo , Adama I. Traore , 2 1 We therefore reconstructed large chromosomal segments (“contigs”) Seydou Timbiné , Diana L. Huestis using solely the sequence information contained in the reads. Using this 1National Institute of Allergy and Infectious Diseases/National Institutes procedure we successfully assembled the entire mitochondrial genome of Health, Rockville MD, United States, 2Malaria Research and Training sequence for each of the five mosquitoes which confirmed the deep Center/University of Bamako, Bamako, Mali divergence between AP and AG but also revealed deep divergences Malaria remains a top public health priority across Sub-Saharan Africa, among the AP sibling species. In addition, we assembled 50-60% of each where it is transmitted primarily by Anopheles gambiae s.s. and An. genome into fragments larger than 1,000 bp and identified more than arabiensis. Populations of these species exploit diverse environments 40,000 DNA polymorphisms that can now be used in association studies including dry savannahs and semi-desert areas, where surface waters for traits related to insecticide resistance, preference to human blood meal required for larval development are absent for large parts of the year. or capacity to transmit malaria and filariasis. How mosquitoes survive the long dry season has been debated without resolution for over 60 years. Although recent studies provide evidence for aestivation (extended survival throughout the 4-7 month-long dry season) of M form An. gambiae, the role of long-distance migration from areas with year-round breeding remains unclear. Here, we analyze the dynamics of the members of the An. gambiae complex in the Sahelian village Thierola (Mali), focusing on the dry season and its preceding and subsequent transition periods, over a period of three years (2008-2011). The dry season mosquito populations were characterized by low overall density (<0.05 mosquito/house), and were predominantly composed of www.astmh.org 378 the M form (>95%), with the remainder being An. arabiensis. Males were 1258 found throughout the dry season, both indoors and in swarms, albeit in very low numbers. Interestingly, the dry-season dynamics were not stable: MOSQUITO COMMUNITIES AND VECTOR-ASSOCIATED in early April, ~2 months before the first rain, density surged up to three MICROBIOMES SAMPLED ACROSS A HABITAT GRADIENT OF orders of magnitude and receded to typical dry-season density within THAILAND days. This surge was observed in both 2010 and 2011 and consisted only 1 2 3 of the M form. Five to seven days after the first rains (early June), before Panpim Thongsripong , Amy Henry , Pattamaporn Kittayapong , 4 5 1 a new generation of adults could be produced, the M form surged again Durrell Kapan , Bruce Wilcox , Shannon Bennett over one order of magnitude, and continued to increase gradually at an 1Department of Tropical Medicine, Medical Microbiology and average rate of 50%/week, for several weeks. Unlike the M form, the S Pharmacology, University of Hawaii at Manoa, Honolulu, HI, United States, form and An. arabiensis remained virtually zero for over four weeks after 2Microbiology Department, University of Hawaii at Manoa, Honolulu, the first rains; thus it is unlikely that they aestivated but would emerge HI, United States, 3Biology Department, Mahidol University, Bangkok, only ~5 weeks after all larval sites filled. These results suggest that both Thailand, 4Center for Conservation Research and Training, University of the S form and An. arabiensis persist in the Sahel primarily by migration Hawaii at Manoa, Honolulu, HI, United States, 5Office of Public Health whilst the M form aestivate. Final analysis and implications for malaria Studies, University of Hawaii at Manoa, Honolulu, HI, United States control will be presented. Changes in biodiversity have the potential to affect the risk of infectious diseases in plants and animals, including humans, since infectious 1257 disease distribution is largely dependent inter-specific interactions. In DIRECT AND INDIRECT COSTS OF PLASMODIUM INFECTION particular, mosquito-borne diseases are well-suited to study how changes ON MOSQUITO REPRODUCTIVE SUCCESS in interacting species, namely mosquitoes, their hosts, and associated microorganisms in changing habitats may affect infectious disease Rodrigo J. González, Guha Dharmarajan, Theodore W. James, risk. Current knowledge of mosquitoes and their associated microbial Tovi Lehmann communities in natural habitats is, however, limited. Here we explored Lab of Malaria and Vector Research, National Institute of Allergy and the composition and diversity of mosquitoes and mosquito-associated Infectious Diseases, National Institutes of Health, Rockville, MD, United microbes in relation to habitats ranging from forest to urban areas in States the central plain of Nakhon Nayok province, Thailand. During the rainy season in 2008, adult mosquito collections from 24 sites using CDC light Infection with malaria parasites reduces the immediate reproductive traps, BG sentinel traps, Mosquito Magnet traps, and CDC backpack success of mosquitoes, but the life-long effects, as well as their interaction aspirators yielded a total of 62,511 identifiable female mosquitoes of 54 with stress, are not well known. Additionally, the negative effects of confirmed taxa. Female mosquito abundance was highest in the rice field infection may be exacerbated by the nutritional cost of feeding on anemic habitat and lowest in the forest habitat with 27,041 (43.26%) and 4,840 blood. We evaluated the effect of Plasmodium gallinaceum infection on (7.74%) mosquitoes collected, respectively. The diversity of mosquito reproductive success of stressed and unstressed Aedes albopictus, fed communities was characterized using a variety of diversity measurements on either infected or uninfected chicken blood. Each of these treatment including statistical sampling approaches to extrapolate species richness. combinations were subdivided into three subgroups that were either fed: In general, the rural habitat was the most diverse while the least diverse (i) directly on an infected (or uninfected) chicken (Live); (ii) membrane-fed habitat varied depending on the indices used. The Vishnui subgroup of on fresh blood from the same chicken (MemFRESH); (iii) membrane-fed on Culex species was the most common taxon found overall and also the the same blood incubated at 4°C for 12 h (rendering infectious blood most common in the fragmented forest, rice field, rural, and suburban non-infectious; MemUNINF). The mosquitoes were subsequently fed two habitats, while Uranotaenia sp. was the most common taxon in the forest more times on uninfected blood from the same chicken. The egg batch habitat and Cx. quinquefasciatus was the most common species in urban size (EBS) of individual mosquitoes was determined 7 d after each feed. settings. Aedes aegypti and Ae. albopictus were most abundant in urban Preliminary analyses revealed that EBS was lower in infected vs. uninfected and rural area respectively. To explore the diversity and composition of and stressed vs. unstressed mosquitoes. However, the interaction between vector-associated microbiomes, the microbiota from three vector species stress and infection was not significant. Likewise, there was no significant Cx. quinquefasciatus, Ae. aegypti, and Ae. albopictus from different interaction between infection and feeding type (i.e. Live, MemFRESH, and habitat types were studied using 454 pyrosequencing of ribosomal RNA. MemUNINF), indicating that the fitness costs of being fed on an infected Patterns of microbiota community assembly in mosquitoes by habitat chicken were similar in both infected-infectious (MemFRESH) and infected- type and vector species using both alpha- and beta-diversity analyses non-infectious (MemUNINF) blood. We also found that the negative effects will be discussed. Our results are particularly relevant for understanding of infection and stress on EBS were not restricted to the first oviposition the dynamics of mosquito vectors and their associated microbiomes in cycle, but rather that these factors could lead to a dramatic decline in landscapes of Thailand. the lifelong reproductive success of individuals. Our results highlight both the life-long and indirect (i.e. due to anemic blood) fitness costs of 1259 Plasmodium infection to both stressed and unstressed mosquitoes. Such costs are important from an ecological and epidemiological perspective, LINKING OVIPOSITION-SITE CHOICE TO OFFSPRING FITNESS as they could affect evolution of resistance/tolerance mechanisms, and in IN AEDES AEGYPTI: CONSEQUENCES FOR TARGETED LARVAL turn affect mosquito population dynamics and vector potential. CONTROL OF DENGUE VECTORS Jacklyn Wong1, Yui Yin Chu1, Imaan Baseer1, Steven T. Stoddard1, Helvio Astete2, Amy C. Morrison1, Thomas W. Scott1 1University of California, Davis, Davis, CA, United States, 2Naval Medical Research Center Unit-6, Lima, Peru Maternal oviposition-site choice and its repercussions for offspring fitness are known to influence population dynamics of insects. Using four experimental container treatments (size [large vs. small] x water management [manually filled vs. unmanaged]), we tested the hypothesis that wild Aedes aegypti in Iquitos, Peru choose egg-laying sites to maximize offspring survival and growth. Among 80 containers located

www.astmh.org 379 in 20 houses, females consistently laid more eggs in large vs. small 1261 containers (β = 9.17, p < 0.001), and in unmanaged vs. manually filled containers (β = 5.33, p < 0.001). There was poor correlation, however, USING MOSQUITO SURVEILLANCE DATA TO PREDICT between oviposition preference and two components of mosquito fitness, HUMAN WEST NILE VIRUS TRANSMISSION RISK pupation probability and adult size. Probability of pupation was higher 1 2 for mosquitoes developing in small, unmanaged containers than any A. Marm Kilpatrick , W. John Pape other container type (β = 3.4, p < 0.001). Adult body size decreased for 1University of California Santa Cruz, Santa Cruz, CA, United States, individuals developing in large containers (females: β = -0.19, p < 0.001; 2Colorado Department of Public Health and Environment, Denver, CO, males: β = -0.11, p = 0.002) and unmanaged containers (females: β = United States -0.17, p < 0.001; males: = - 0.11, p < 0.001). Our data suggest that β West Nile virus (WNV) has become endemically established across the the majority of Ae. aegypti eggs are laid in non-optimal sites, such that Americas with enzootic activity and significant human illness. Despite this, selective oviposition behavior contributes to population regulation by funds for surveillance and control are limited and decreasing. Predicting limiting the production and size of adults. Targeted larval control strategies the risk of human infection to initiate timely preventative measures is removing the most productive containers may have the unintended the primary goal of public health mosquito surveillance. Many arbovirus effect of encouraging females to spread their eggs more evenly among response plans outlines recommended public health and mosquito remaining containers. By tracking egg-laying patterns of individual control actions based on levels of virus activity determined by statewide females inside a semi-field enclosure, we found that the probability of mosquito surveillance. However, studies linking mosquito surveillance data any container receiving eggs increased when preferred container were to the spatio-temporal risk of human WNV infection, have rarely been removed (but the total number of containers remained constant) ( = β attempted. We quantified the links between mosquito surveillance data 1.36, p < 0.001). We suspect that in Iquitos, and possibly other locations, and the spatio-temporal patterns of 3,827 human WNV cases reported selective oviposition behavior by Ae. aegypti, along with a potential switch in Colorado from 2003-2007. Mosquito data were strongly predictive from clustering eggs to spreading them out, will render targeted larval for spatio-temporal variation in human WNV infections several weeks control less effective than anticipated. in advance in a statewide analysis, and with temporal variation within a county. Correlative and predictive relationships were strengthened by 1260 using pooled estimates of prevalence from across the state in estimating TOWARDS A CONSERVED cis-REGULATORY MODULE of risk early and late in the season when few mosquitoes were trapped at the local scale. However, we found that when current year prevalence WITH CROSS-STRAIN/SPECIES APPLICATION FOR DRIVING data was not available, as could occur with reduced or eliminated ANTI-PATHOGEN EFFECTOR TRANSGENES: COMPARATIVE surveillance budgets, no meaningful predictions of human risk could be TRANSCRIPTOMICS TO DISCOVER EARLY BLOODMEAL- made to determine appropriate public health response. Overall, our results RESPONSIVE, CIS-REGULATORY SEQUENCES FROM demonstrate that mosquito surveillance provides valuable predictive data MOSQUITO MIDGUT RNA-Seq about the risk of human infection which can be used to trigger emergency Augustine Dunn, Osvaldo Marinotti, Xiaohui Xie, Anthony James response actons and allocate limited public health and mosquito control resources. University of California, Irvine, Irvine, CA, United States Empirical definition of active cis-regulatory elements (CRE) through 1262 classical “promoter bashing” is difficult in mosquitoes due to the time and effort required to produce transgenic mosquito strains. Bioinformatic DOCUMENTING THE POTENTIAL INTRODUCTION OF methods combined with existing biological knowledge and quality mRNA DENGUE VIRUS INTO KEY WEST, FLORIDA THROUGH abundance data should allow the inference of active CRE combinations, AIRLINE AND CRUISE SHIP PASSENGERS FROM DENGUE- cis-regulatory modules (CRM), without requiring construction of ENDEMIC LOCATIONS transgenic mosquito strains. The ecdysone (20E) response cascade is Ali M. Messenger, Dana A. Focks, Kelli L. Barr conserved throughout insects and has been shown to drive changes in mRNA abundance following the ingestion of a bloodmeal. This supports University of Florida, Gainesville, FL, United States the hypothesis that it should be possible to deduce a conserved CRM For the first time in decades, sporadic cases of locally-acquired dengue by studying bloodmeal-regulated transcript abundance across multiple were reported in Key West in 2009 and again 2010. Current hypotheses mosquito species. 20E has had multiple early-response factors described regarding this continuance include vertical transmission, the establishment previously including the ecdysone receptor (EcR), its binding partner of an endemic state with undetected transmission between years, and ultraspiracle (USP), and the 20E-inducible gene E74. Other laboratories multiple introductions via visitors from endemic countries during both have shown that levels of 20E early-response factor isoforms vary in a years. Regarding the third hypothesis, country- and year-specific dengue time- and tissue-specific fashion in response to pulses of 20E following a incidence data (PAHO) and the numbers of airline passengers originating bloodmeal. This allows one hormone to regulate diverse cellular responses. in dengue-endemic countries in this hemisphere with a final destination of Tissue-specific, time-course RNA-Seq data with high temporal resolution Key West were used to estimate the relative the magnitude of potentially (2 hours) will be used to compare 20E early-response factor isoform viremic passenger-days experienced per year. These estimates suggest mRNA expression levels across evolutionarily distant species (Anopheles multiple introductions per year are not uncommon and that potential gambiae, Aedes aegypti, and Culex quinquefasciatus) to infer transcripts introductions in 2009 and 2010 were higher than in 2007 and 2008 that display probable time-lagged induction by 20E and harbor known as a result of an increase in air travel and major dengue activity in the 20E early-response factor motifs. This transcript set will leverage a Caribbean and Central America. Both years were El Niño years that combined comparative-genomics and expression-profile based CRE/CRM historically are associated with elevated temperatures and higher dengue discovery strategy to reveal putative CRMs expected to provide a better activity in the region. A similar analysis of potential introductions via the understanding of 20E-regulated transcript regulation in the midgut. The cruise ship industry will also be presented. discovered CRMs will serve as the basis for validation of a set of conserved CREs that may be combined to drive robust anti-dengue effector transcription in the midguts of Ae. Aegypti mosquitoes directly following the ingestion of each bloodmeal.

www.astmh.org 380 1263 1265 CRYPTIC BREEDING: A POTENTIAL CAUSE OF LOCAL DENGUE INTEGRATION OF A SAFE WATER SYSTEM WITH ANTENATAL TRANSMISSION IN KEY WEST, FLORIDA SERVICES, MACHINGA DISTRICT, MALAWI, 2010-2011 Kelli L. Barr1, Dana A. Focks1, Ali M. Messenger1, Andrea Leal2 Janell Routh1, Anagha Loharikar1, Elly Chemey2, Martin 3 2 4 3 1University of Florida, Gainesville, FL, United States, 2Florida Keys Mosquito Msukwa , Aulive Msoma , Kate Sabot , Annie Michaelis , Robert 1 Control District, Key West, FL, United States Quick 1Centers for Disease Control and Prevention, Atlanta, GA, United States, June 2009 marked the beginning of a 2-year outbreak of locally-acquired 2Clinton Health Access Initiative, Machinga, Malawi, 3Clinton Health dengue in Key West, Florida. Despite increased control efforts by mosquito Access Initiative, Lilongwe, Malawi, 4Clinton Health Access Initiative, control and local residents, the number of dengue cases in 2010 nearly Boston, MA, United States doubled that of 2009. Surveillance on the abundance of immatures was inconsistent with magnitude of the adult population of Aedes aegypti. Antenatal clinic (ANC) visits provide an opportunity to integrate additional Similar disconnects(?) between immature and adult abundance in other interventions to improve maternal and neonatal health and motivate dengue-endemic regions have been the result of cryptic breeding which pregnant women to attend ANC services. In Malawi, although 93% of occurs when mosquitoes reproduce in locations that escape control women attend at least one ANC visit, 57% deliver in health facilities, efforts. The majority of homes in Key West were built prior to municipal and 7% have postnatal checks. To reduce the risk of diarrhea, a leading utilities and stored water in cisterns and disposed of waste through septic cause of childhood mortality, we integrated free hygiene kits (safe water systems. Cisterns and unused septic tanks are several cubic meter in storage containers, water treatment solution [WaterGuard], soap, and oral size and most are not easily accessible. Though historical maps exist, the rehydration salts) with ANC services. To receive the hygiene kit, women true number of cisterns and septic tanks is unknown thus complicating had to have a spouse/partner present; HIV testing was also offered to the control efforts. Presented here are the combined efforts of the University couple. At subsequent ANC visits, up to 3 refills of WaterGuard and soap of Florida and Monroe County Mosquito Control to identify and eliminate were provided. We surveyed 106 women receiving ANC care at baseline cryptic breeding sources for Ae. aegypti in Key West. before program implementation and at follow-up 12 months later to assess water treatment; test drinking water for residual chlorine; observe 1264 hand-washing; and determine ANC service utilization. From baseline to follow-up, there was an increase in the percentage of women who had THE QUALITY OF DRINKING WATER IN COMMUNITIES ever used WaterGuard (38% vs. 100%, p<0.001), knew how to use it ALONG THE MARANON RIVER IN THE PERUVIAN AMAZON correctly (23% vs. 81%, p<0.001), were observed to have a bottle in their home (3% vs. 77%, p<0.001), had residual chlorine in their stored water Yayi Guo, Kellogg J. Schwab (0 vs. 71%, p<0.001), and were able to demonstrate proper handwashing Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United technique (21% vs. 65% p<0.001). At follow-up, 89% of respondents States had ≥3 ANC visits, 90% delivered at a health facility, 99% were tested Water is one of the world’s most critical resources, however international for HIV, 99% of partners were tested for HIV, and 98% had disclosed water quality surveillance and monitoring is often not implemented, their status to their partner. Women in this program showed statistically obscuring associations and etiologies of potentially related illnesses. We significant increases in water treatment and hygiene practices, and high conducted an evidence-based approach to understand the sources and utilization of ANC services and HIV testing. This evaluation suggests that types of water contaminants as well as the overall safety of available integration of hygiene kits, refills, and HIV testing during ANC is feasible, drinking water in Peru. A comprehensive, portable, water quality can serve as an incentive to increase use of health services, and may help assessment toolbox was used to quantify key microbial (total coliforms, motivate changes in health behavior. E.coli and enterococci) and chemical (metals, anions and pesticides) contaminants. This assessment system was applied in the field to evaluate 1266 the drinking water of 20 rural villages bordering the Maranon River in the Peruvian Amazon. In total, 32 households, 32 drinking water IMPACT OF COMPLEXITY OF HANDWASHING INSTRUCTIONS sources, and 2 water treatment systems were assessed. All household ON ADHERENCE IN A LOW INCOME SETTING, DHAKA, drinking water samples and 93% of source water samples contained BANGLADESH, 2010 moderate to high levels of E.coli contamination. Water treatment systems Dawn D. Sagerman1, Fosiul A. Nizame2, Md Nuruzzaman2, varied in contaminant removal, ranging from 2.03 logs to 4.15 logs of Jihnhee Yu1, Stephen P. Luby3, Pavani K. Ram1 measured bacterial removal. Multiple water samples contained chemical 1University at Buffalo, Buffalo, NY, United States, 2International Centre contaminants in excess of WHO guideline levels including phosphate for Diarrhoeal Disease Research, Bangladesh, Dhaka, Bangladesh, (anion); aluminum, iron, and manganese (metals); and lindane (pesticide). 3International Centre for Diarrhoeal Disease Research, Bangladesh (Dhaka, Current international water quality screening and evaluation efforts are Bangladesh) and Centers for Disease Control and Prevention, Atlanta, GA, not adequate to address the burdens caused by the adverse health effects United States of waterborne contaminants, thereby demonstrating the need for portable water quality screening. In the Peruvian Amazon, results comparing Handwashing reduces diarrhea risk in young children. Interventions source water and household contamination suggest recontamination to improve handwashing usually include instructions on how and during transport. Analysis of chemical pollutants revealed a need for when to wash hands. These instructions vary in complexity, with some water treatment to address metal contaminants. Treatment system results recommending multiple steps including duration of lathering and indicated that standardized treatment measures are required. The use scrubbing various aspects of the hands. To assess whether complex of our water quality assessment toolbox provided more comprehensive handwashing instructions result in reduced adherence, we conducted a detection and analysis of waterborne threats to the public. These data randomized trial in a low-income area of Dhaka, Bangladesh. Mothers can help local governments and non-governmental organizations to select of young children were randomly assigned to one of three sets of appropriate treatment solutions. handwashing instructions: simple, moderate, or complex. Simple instructions were to wet, lather, and rinse hands; moderate instructions included simple instructions and additional steps to scrub palms, scrub backs, and dry hands by waving them in the air; complex instructions included moderate instructions and additional steps to scrub between fingers, scrub under nails, and lather for 20 seconds. The field worker www.astmh.org 381 taught the participant the randomly assigned set of instructions, without importance of promoting affordable means of increasing handwashing. mention of the other two sets. Immediately, two days, and two weeks Interventions that emphasize soap availability and the efficacy of after the teaching, participants were asked to demonstrate handwashing inexpensive soap may be effective in increasing handwashing behavior at to the field worker. Adherence was defined as demonstration of all of critical times. the instruction steps prescribed for the assigned treatment arm. We enrolled 244 participants (simple n=85, moderate n=75, complex n=84). 1268 Compared with the simple group, in which 100% adhered to prescribed instructions at all post-intervention assessments, the more complex IMPACT OF INTENSIVE HANDWASHING PROMOTION groups had lower adherence at two weeks (moderate 43%, p<.0001; ON HOUSEHOLD TRANSMISSION OF INFLUENZA IN A complex 31%, p<.0001). Adherence to air-drying hands was low at LOW INCOME SETTING: PRELIMINARY RESULTS OF A immediate, Day 2 and Week 2 assessments (moderate: 49%, 39%, and RANDOMIZED CONTROLLED CLINICAL TRIAL 47%; complex: 57%, 46%, and 38%). Exclusion of the air drying step 1 2 2 from the outcome yielded adherence rates of 99%, 91% and 88% for Margaret A. DiVita , Kaniz Khatun-e-Jannat , Manoshi Islam , 1 1 2 the moderate group and 81%, 69% and 71% for the complex group. Emily Cercone , Kimberly Rook , Badrul Munir Sohel , Makdum 2 3 2 In a low-income community in Dhaka, highly complex instructions for Ahmed , Eduardo Azziz-Baumgartner , W. Abdullah Brooks , 1 2 1 handwashing resulted in decreased adherence. Future research should Jihnhee Yu , Stephen P. Luby , Pavani K. Ram investigate whether adherence to the highly complex set results in greater 1State University of New York at Buffalo, Buffalo, NY, United States, hand decontamination than adherence to the simple or moderate set 2International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, of instructions. When developing materials to promote handwashing Bangladesh, 3Centers for Disease Control and Prevention, Atlanta, GA, behavior, handwashing promotion programs should consider the United States complexity of the overall set of instructions, as well as the microbiological Although handwashing with soap decreases the risk of all-cause impact and feasibility of adherence to specific instructions, such as air respiratory illness, there is little published empirical evidence for drying. the efficacy of handwashing with soap for prevention of influenza transmission in resource-poor settings. We tested the impact of 1267 handwashing promotion on the risk of household transmission of CONSISTENT SOAP AVAILABILITY CORRELATES WITH influenza, influenza-like-illness (ILI), and fever in rural Bangladesh. In 2009 and 2010, we identified index case patients (ICPs), individuals USE OF INEXPENSIVE SOAP PRODUCTS AND IMPROVED who developed ILI within the previous two days and were the only HANDWASHING BEHAVIOR IN LOW-INCOME HOUSEHOLDS symptomatic person in their household. ILI was defined as fever in IN DHAKA, BANGLADESH children <5 years old and fever with cough or sore throat in persons > 5 Meghana Gadgil1, M. Abu Yushuf Sharker2, Leanne Unicomb2, years old. Households were randomized to intervention or control). The Stephen Luby2, Pavani Ram3 intervention group received handwashing stations with soap and daily handwashing motivation at critical times for pathogen transmission, such 1Stanford University, Stanford, CA, United States, 2International Centre as after coughing or sneezing. We conducted daily surveillance and tested for Diarrheal Disease Research, Bangladesh, Dhaka, Bangladesh, 3SUNY household members with fever for influenza viruses by polymerase chain Buffalo School of Medicine and Biomedical Sciences, Buffalo, NY, United reaction. Secondary attack ratios (SAR) were calculated for influenza, States ILI, and fever in each arm. We used logistic regression with generalized Handwashing (HW) with soap reduces diarrhea in children < 5 years of estimating equations to estimate the significance of the SAR comparison age in low-income countries. Understanding characteristics of households while controlling for clustering by household. Among 274 ICPs enrolled, with increased HW could inform interventions to increase this behavior. 33 (12%) had laboratory-confirmed influenza infections. The SARs for Amongst low-income households in Dhaka, Bangladesh, we studied influenza among household contacts of ICPs with confirmed influenza consistent soap availability as a possible indicator of increased HW. virus infection were 7.5% in the control arm (10/133) and 11.0% in the Households were selected randomly from controls of a case-control study intervention arm (11/100) (p = 0.362). The SAR for ILI among household on hygiene and respiratory illness; all had ≥ 1 child ≤ 5 yrs. We visited 220 contacts of all ICPs was 11.9% in the control arm (146/1,226) and 14.2% households 8 times over 4 weeks in Feb-Mar 2010. Respondents were in the intervention arm (186/1,314) (p = 0.232). SARs for fever were interviewed about soap availability. Fieldworkers observed the presence of 12.1% and 15.0%, respectively, in the control and intervention groups soap and water at HW stations, the cleanliness of respondents’ palms and (p = 0.113). When an intensive handwashing intervention was initiated administered a validated 14-question tool on the strength of handwashing after illness onset in a household member, we found no protective effect habits. We used data from structured observations, conducted several against influenza virus infections. Handwashing behavior may not have months prior, to estimate HW behavior at critical times. We used logistic changed rapidly enough to match the pace of influenza virus transmission regression to adjust for socioeconomic status, and compared households between household members. Courtesy bias among intervention that had soap for HW available at 100% of visits to households that households, who received daily motivation as well as hardware to facilitate did not. Soap for HW was available in 1513 (88%) of 1716 visits to handwashing, may have led to greater reporting of respiratory symptoms. 220 households. In 110 households (50%), soap was available at every Future efforts should consider whether handwashing behavior can be visit. Compared to those with inconsistent soap availability, households changed quickly after illness onset in order to blunt household influenza with consistent soap availability at each visit were more likely to be in transmission. the highest socio-economic status quintile (determined by principal component analysis on household assets) (OR 1.9; 95% CI 1.4- 2.4), more 1269 likely to have soap present at the HW station (ORadj 1.6; 95% CI 1.3- 2.0), CONSUMER INPUT TO DESIGN AND DEVELOPMENT OF A more likely to wash hands with soap at critical times (ORadj 1.4; 95% CI 1.1 -1.7) and more likely to identify cheaper detergent soap rather than NOVEL HOUSEHOLD WATER TREATMENT DEVICE bar soap as the main HW product (OR 2.2; 95% CI 1.6- 2.9). Consistent adj Jeffrey F. Williams, Michael Bridges, Nevada Ruehlen, Shannon availability of soap for handwashing was not associated with scores for the Schelinder, Jose I. Santiago, Lori Trimpe key components of habit. Households that had soap available in the home during each visit were also more likely to keep soap where it was needed HaloSource Incorporated, Bothell, WA, United States for washing hands, and to wash hands more frequently at times relevant We collected consumer preference data in urban, periurban, and for hand contamination and pathogen transmission. Reliance on less rural areas in India and Indonesia to use in design and development expensive soap may facilitate consistent soap availability, underscoring the of a novel POU device for use in Asia. The end product incorporates a www.astmh.org 382 drinking water disinfection medium (registered by USEPA-#72083-3, 1271 2009). Consumer exposures ranged from 1-month in-home use of functional prototypes, to use-pattern questionnaires, and from household EFFICACY VERSUS EFFECTIVENESS OF WATER placement of life-size cut-outs of proposed designs to 3- dimensional CHLORINATION IN RURAL COASTAL ECUADOR models based on these designs. Householders showed a preference for 1 2 1 gravity feed device configurations that: could accommodate ~ 10 L of Karen Levy , Larissa Anderson , Katharine Robb , Joseph 2 source water; allow for collection of filtered, disinfected product water Eisenberg after no more than a few hours; ensured collection of clear, uncolored 1Emory University, Atlanta, GA, United States, 2University of Michigan, Ann water with no detectable taste, taint, untoward mouth-feel or odor on Arbor, MI, United States immediate consumption or after storage; offered ease of use in cleaning Chlorination can provide a low-cost method of treating drinking waters of upper chamber filtration elements; ensured high convenience in secure and is known to be efficacious for reducing bacterial loads, but actual replacement of the water treatment train (prefilter/filter/adsorption media/ effectiveness under household conditions may not reduce microbial disinfecting cartridge) after a useful life of no less than several months’ contamination to the same extent as under lab conditions. In a previous daily use (i.e., > 1000L); required minimal assembly at start-up; provided study we found no significant differences in log reductions in drinking for ready access to product water via a faucet/outlet with reliable, drip- water of households that reported chlorination of their water in rural free function; and (critically important) had a ‘modern’ and attractive coastal Ecuador. We present the results of a follow-up study at the same appearance, enhancing the household working and living environment. field site in which we observed and quantified chlorination procedures at From in-home observations we determined that: construction needed to the household instead of relying on self-reported chlorination. We also be robust, include auto- shut-off at the end-of-life, plus a visual indicator tested source waters and water from control containers stored under of approaching termination, and include an option for ‘dialing in’ varying protected conditions outside of the household. We collected three sets of efficacy levels (up to US-EPA 6/4/3). HaloPure Waterbird emerged from samples (source water, water stored in the home, and water stored under this process, a gravity-feed purifier capable of 6/4/3 log reduction, auto control conditions) from 145 households: 67 that did not chlorinate, 42 shut-off at 1500L (± 20%), and leak-free cam-lock cartridge placement. that used locally available chlorine according to local practices, and 35 Imperceptible halogen residual provides for continued protection of that used chlorine dosed to recommended standards. Covariates included product water, in the device or upon transfer. Listening to the “voice of physicochemical data and household level indicators. The efficacy of the consumer” can lead to enhanced product design aimed at household chlorine treatment in our field laboratory-matched control samples was water treatment device development. higher than the effectiveness in corresponding household samples, which is most likely the result of recontamination in the household during 1270 storage. Recontamination of water in containers in the household over USE AND ACCEPTABILITY OF A POINT-OF-USE WATER a 24-hour storage period was observed between pairs of household and matched control samples for both E. coli and total coliform concentrations, FILTRATION DEVICE IN HIV-1 INFECTED ART NAÏVE KENYAN with mean log differences ranging from 0.4017-0.6147 (p < 0.0001). ADULTS 63.8% of samples had greater microbial contamination in household Patricia B. Pavlinac1, Linda Chaba2, Benson Singa2, Jacqueline M. samples than in their matched control. The reduced effectiveness can Naulikha2, Naomi Kimani2, Laura Sangaré1, Ben K. Piper1, Grace also be explained by other factors such as source water turbidity, socio- John-Stewart1, Judd L. Walson1 economic status, unsafe water storage behaviors. Negligible disparities were found between the two chlorine treatment groups, suggesting 1University of Washington, Seattle, WA, United States, 2Kenya Medical that dosing practices did not greatly modify the relationship between Research Institute, Nairobi, Kenya chlorination and log reduction in contamination. Household effectiveness Among HIV-infected adults and children in Africa, diarrheal disease of chlorine treatment was significantly reduced over laboratory efficacy. remains a major cause of morbidity and mortality. WHO recommendations This research provides important new insight about the relationship suggest HIV-infected individuals should treat drinking water at the point- between household storage practices and chlorination under village of-use. While simple and effective water filtration devices are available, conditions. limited data exist regarding the use and acceptance of these devices in this population. We enrolled ART naïve HIV-positive adults into a two- 1272 year cohort study in western Kenya. Individuals were visited in their home at least once to assess acceptability and use of a study water filtration ROTAVIRUS OUTBREAK AMONG CHILDREN IN DAY CARE device. Of 417 participants enrolled and subsequently visited, most were CENTER, ZAPORIZHZHYA, UKRAINE female (81%), married (64%), had at least a primary school education Tetyana Chumachenko1, Volodymyr Khomutov2, Lyudmyla (72%), and had CD4 cell counts above 350 cells/µl (76%). At enrollment, 2 2 participants reported the most common sources of drinking water to be Giba , Olena Kochetkova shared pipe or tank source (45%) followed by well water (25%) and river 1Kharkiv National Medical University, Kharkiv, Ukraine, 2Zaporizhzhya City or stream (25%). Among participants with a functioning device, more SES, Zaporizhzhya, Ukraine than half (57%) reported using the water filtration device in all of the last In the city of Zaporizhzhya the incidence of acute gastroenterocolitis (GI) 5 instances of obtaining water to drink (always) and 25% reported using has recently increased. The proportion GI illness due to rotavirus infection the device at least 3 of the last 5 times. Only 3% reported never using (RI) has increased from 21.6% in 2008 to 40.6% in 2009. Among children the device. We found household monthly income greater than 5,000 incidence of RI has increased 2.5 times. We investigated an outbreak of Kenyan Shillings (~$57 US) to be associated with always using the device GI in a daycare center (DCC). Samples of drinking water, food and human (OR: 2.12 (95%CI 1.23, 3.65)). A trend towards an association between specimens were examined bacteriologically for intestinal pathogens. Ill increasing age and always using the device was also observed (OR per persons, contacts and water were tested for rotavirus antigen by ELISA 5 year increase: 1.11 (95%CI 0.98, 1.30)). While 38% of participants and dipstick testing. During a two week period in April, 17 cases of RI reported drinking water outside of the home within the last 24 hours, were reported. Cases were identified in 8 of 11 classes. In a class which most (77%) reported filtering their drinking water. Almost all participants attended only three hours per day there were no cases. Through testing found the device very acceptable, with 97% willing to purchase the water we identified 11 carriers (1 caregiver from class 1 and 10 children). The filtration device if their current device were to break. Providing simple highest incidence of RI (4 patients and 7 carriers) was observed in class 1 point-of-use water filtration devices to HIV infected adults may be an which consists of children under 3 years. The primary case was identified inexpensive and practical intervention to improve water quality and reduce in this class. Rotavirus was found in these 4 children. Due to a staffing the risk of diarrheal disease among this high-risk population. shortage care-givers served food to the children against normal sanitary www.astmh.org 383 regulations. Using a retrospective cohort study design we established for human-specific contamination. B. fragilis is a strict anaerobic bacteria that the route of transmission was beet salad (RR=3,5; CI 1,07-11,36). and is known to be one of the predominant microbial flora in human The salad was served from a single bowl and distributed to children gut. For this, an on-site wastewater treatment system was used as a by class. Children from the two classes in which there were no cases testing ground. Water samples were collected from various points: septic of RI received the salad first. The first 4 cases of RI were not identified tank effluents; after bioreactor; and after chlorine dioxide treatment. until laboratory testing was performed. The study established that the The level of B. fragilis were tested with IMS/ATP using B. fragilis-specific caregiver was infected at DCC. Transmission is believed to have occurred antibody attached magnetic beads and qPCR targeting gyr B gene. The via asymptomatic carriers. Caregivers serving food to the other classes B.fragilis (Bf) levels measured by IMS/ATP showed 1.5 log reductions are thought to have transmitted disease to them. No cases of RI occurred after bioreactor, and 2.0 reductions after ClO2 treatment, respectively, in classes that received salad before class 1 was served. This study when compared with the original levels in the septic tank. The Bf levels demonstrates the necessity of strict adherence to the sanitary and hygiene determined by qPCR showed 1.6 log reduction after bioreactor and 2.3 regulation in DCC’s and the ongoing problem of RI. log reduction after ClO2 treatment. The Bf levels measured by IMS/ATP and qPCR correlated well (y=0.8834x+0.8791, R = 0.998). In summary, IMS/ 1273 ATP rapidly determined the levels of Bf in an on-site wastewater treatment system with sensitivity and specificity. Thus, it can provide near real-time SPATIO-TEMPORAL PATTERNS OF DIARRHEAL DISEASE (1.5 hr) results of the on-site wastewater treatment efficiency prior to CAN REVEAL TRANSMISSION PATHWAYS IN AN EMERGING its release into the environment. This is the first study that the new IMS/ URBAN REGION OF ECUADOR ATP assay targeting Bf was applied for determining on-site wastewater treatment efficiency. This assay can be applied for a broad range of rapid 1 1 2 Darlene Bhavnani , Jason E. Goldstick , William Cevallos , detection of human-specific fecal contamination in water where fecal 1 Joseph N. Eisenberg contamination is suspected. 1University of Michigan, Ann Arbor, MI, United States, 2Universidad San Francisco de Quito, Quito, Ecuador 1275 Diarrheal disease is caused by a variety of pathogens that exploit multiple FURTHER INSIGHTS INTO THE PHYSIOLOGICAL transmission pathways. The patterns of diarrheal disease in space and time may reveal which transmission pathways are dominant; e.g., direct MECHANISMS THAT UNDERLIE TSETSE’S BENEFICIAL person-to-person spread produces temporary clusters of cases; whereas SYMBIOSES environmental pathways result in constant clusters around environmental Brian L. Weiss, Michele Maltz, Serap Aksoy sources. We explored these spatial and temporal distributions of diarrhea Yale University, New Haven, CT, United States in Borbón, a small urban region of northwestern Ecuador. The relationship between these patterns and household and neighborhood WASH Bacterial symbioses are ubiquitous in nature, yet to date few studies have characteristics was also estimated. We conducted a series of six nested been performed to determine the physiological mechanisms that underlie case control studies between December 2008 and May 2009. Surveys these relationships. Insects represent a group of advanced multi-cellular were carried out monthly to collect data on WASH factors. The river as organisms that harbor well-documented symbiotic associations. One such well as all houses and outdoor latrines were mapped using GPS. We insect, the tsetse fly (Glossina spp.), harbors 2 maternally-transmitted employed spatial point pattern analyses assuming an inhomogeneous bacterial symbionts, mutualistic Wigglesworthia and commensal Poisson process. We used the K-function to measure clustering and the Sodalis, that are intimately involved in maintaining the overall fitness ratio of intensity between cases and controls to estimate spatial variation of their host. In this study we examine the functional mechanisms that of risk by month. Generalized linear and generalized additive models were underlie these symbioses by producing tsetse flies that lack all of their used to estimate the association between WASH factors and household endogenous microbiota. The resulting aposymbiotic offspring are highly diarrhea. We found both spatial and temporal variation of diarrhea in susceptible to infection with normally non-pathogenic E. coli, and this Borbón. The spatial variation was associated with different risk factors immuno-compromised phenotype is characterized by the absence of each month; the exception to this finding was living with children under phagocytic hemocytes and the irregular expression of immunity-related five, which was found to be a consistent risk factor. For example, early genes. When hemocytes collected from wild-type tsetse are transplanted in the rainy season (December and January), use of an unimproved into aposymbiotic flies, the recipient individuals regain their refractory sanitation facility was significantly associated with diarrhea. In the middle phenotype. We also supplement the diet of pregnant aposymbiotic of the season, significant WASH effects were absent. Towards the end females with Wigglesworthia and Sodalis in an attempt to compliment the of the rainy season (May) better household hygiene was significantly fitness of their offspring. Our observations provide further insights into protective for diarrhea. These results provide insight on where and when the evolutionary adaptations that anchor the steadfast relationship shared improvements to WASH factors may protect from diarrheal disease, between tsetse and its symbiotic microbes. highlighting the importance of indirect transmission through contaminated latrines in the dry season. 1276 1274 PUNIQUE VIRUS, A NOVEL PHLEBOVIRUS, RELATED TO SANDFLY FEVER NAPLES VIRUS, ISOLATED FROM SANDFLIES RAPID VIABLE DETECTION OF HUMAN-ORIGINATED FECAL COLLECTED IN TUNISIA AND ITS POTENTIAL IMPACT ON CONTAMINATION USING IMS/ATP AND qPCR TARGETING PUBLIC HEALTH BACTEROIDES FRAGILIS Khalil Dachraoui1, Laurence Bichaud2, Sonia Sakhira1, Ifhem Jiyoung Lee, Perfect S. Agidi, Karen Mancl Chelbi1, Gregory Moureau2, Saifedine Cherni1, Mohamed Derbali1, Ohio State University, Columbus, OH, United States Remi Charrel2, Xavier de Lamballerie2, Elyes Zhioua1 1Institut Pasteur de Tunis, Tunis, Tunisia, 2Medical School of Marseille, Human-originated fecal contamination of our drinking water source and Marseille, France recreational water is a continuous public health concern around the world. Timely and cost-effective ways in detecting contaminants in water is very Sand flies are widely distributed around the Mediterranean. Therefore, important for protecting human exposure to possible presence of potential human populations in this area are exposed to sandfly-transmitted enteric infectious agents. This study aimed to determine the effectiveness diseases, including those caused by phleboviruses. While there is of a rapid detection method, immunomagnetic separation coupled with substantial data in countries located in the northern part of the ATP bioluminescence (IMS/ATP) and qPCR targeting Bacteroides fragilis www.astmh.org 384 Mediterranean basin, few data are available for North Africa. Sand flies 1278 were collected from the site of Utique, a well-known site of visceral leishmaniasis in northern Tunisia, during the summers of 2008, 2009 and FIELD EVALUATION OF A WICKING ASSAY FOR THE RAPID 2010. In 2008 and 2009 sand flies were captured and pooled by sex and DETECTION OF RIFT VALLEY FEVER VIRAL ANTIGENS IN species. A vast majority of sand flies belong to Phlebotomus perniciosus. MOSQUITOES (DIPTERA: CULICIDAE) Thus species identification was abandoned in 2010 and sand flies were 1 1 1 pooled by sex. Sand flies were tested for the presence of phleboviruses by Elizabeth W. Wanja , Zahra F. Parker , Tobin Rowland , Michael 2 3 3 4 PCR. Viral RNA corresponding to a novel virus closely related to Sandfly J. Turell , Kirti Dave , Sonia Dave , Rosemary Sang fever Naples virus (SFNV) was detected in pools of sand flies collected in 1Walter Reed Army Institute of Research, Silver Spring, MD, United States, 2008 and 2009. Virus isolation in Vero cells was achieved. Genetic and 2U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, phylogenetic characterisation based on sequences in the three genomic MD, United States, 3VecTOR Test Systems, Inc., Thousand Oaks, CA, segments showed that it was a novel virus distinct from other recognized United States, 4Kenya Medical Research Institute, Nairobi, Kenya members of the species. This novel virus was provisionally named Punique Rift Valley fever virus (RVFV) causes outbreaks of severe disease in virus. Viral sequences in the polymerase gene corresponding to another domestic ungulates as well as humans in Africa. There is a concern phlebovirus closely related to but distinct from Sandfly fever Sicilian virus that outbreaks of RVF may continue and that this virus may spread (SFSV) were obtained from positive pools collected in 2008 and 2010. into regions where it had not previously been detected. Surveillance Isolation of this virus temporarily named Utique Virus remained to be and rapid detection are critical to the initiation of an effective disease achieved. The public health impact of those two new viruses remained to control program. Here we report on the field evaluation in Kenya of the be determined. VectorTest® RVFV antigen assay, modeled on the VecTest® assay for West Nile virus. The dipsticks provided results in less than 20 min, were 1277 easy to use, and did not require a laboratory with containment facilities. ANTIBODY RESPONSES OF GUINEA PIGS TO SALIVARY Although none of the field-collected mosquitoes were infected with RVFV, ANTIGENS OF TRIATOMA INFESTANS FOR THE the dipstick provided a clear positive result with pools of field-collected mosquitoes spiked with a single positive, irradiated (to inactivate an DEVELOPMENT OF TRIATOMINE EXPOSURE MARKERS infectious virus) mosquito. Similarly, the dipstick was able to detect virus Veronika Dorňáková, Alexandra Schwarz from pools of mosquitoes captured during the RVFV outbreak in 2007. Institute of Parasitology, Biology Centre, ASCR, v.v.i., Ceske Budejovice, The RVFV dipstick assay was highly specific with only a single weak false Czech Republic positive out of 266 pools tested (specificity >99.6%). The RVFV assay can provide a rapid, safe, easy to use preliminary test to alert public health Antibody responses to salivary antigens of the most effective vector personnel to the presence of RVFV in mosquitoes in a given area. Results of Chagas disease, Triatoma infestans, offer the potential to develop from this assay will allow for more rapid medical threat assessments and exposure markers for detecting the presence of small numbers of the focusing of vector control measures on high-risk areas. triatomines, especially after vector control measures have been implemented. Previous studies have detected a salivary apyrase as a main 1279 candidate exposure marker using guinea pig sera, but this protein is frequently found in the saliva of different haemathophagous insects and IDENTIFICATION OF A NEW GROUP OF LACTATION- thus not triatomine specific. Futhermore, antibody responses to saliva of ASSOCIATED PROTEINS IN THE TSETSE FLY, GLOSSINA different developmental stages were not considered, although the immune MORSITANS MORSITANS responses may vary if using nymphal or adult saliva. Therefore in this study, guinea pigs were exposed weekly to 5 nymphs or adults of different Veronika Michalkova, Joshua Benoit, Geoffrey Attardo, Serap T. infestans strains from Chile, Argentina and Bolivia over a period of 11 Aksoy weeks and they were bled 5 days after each exposure. IgG responses Yale University, New Haven, CT, United States of guinea pigs to nymphal and adult saliva were detected 11 days after Tsetse females generate a single larva during each gonotrophic cycle. All the first exposure and both responses differed significantly. Saliva of nutrients for larval development are provided by the mother in the form nymphs and adults revealed complex protein profiles that uncovered of lactation products generated by the milk gland. The nutrients within differences not only between the T. infestans strains but also between the milk are primarily composed of equal amount of lipids and proteins. the developmental stages. The most prominent bands in all strains were Four proteins have been associated with tsetse lactation products, milk of 85, 72, 44 and 25 kDa. Although the saliva of nymphs was richer in gland proteins 1-3 (gmmmgp1-3) and transferrin. However, little is its protein composition than the adult saliva, more salivary proteins of known about other protein components of tsetse milk. In this study, adults (n=10) were recognized by guinea pig sera than nymphal proteins we performed an Illumina based transcriptome analysis of differential (n=6) during the long-term study. Four antigens (85, 79, 72 and 44 kDa) gene expression in pregnant flies compared to those post parturition to were recognized by all guinea pig sera. Candidate exposure markers, identify lactation-specific genes. This analysis revealed 11 transcripts that such as a truncated pallidipin-like salivary protein (gi|148469123), were are upregulated during pregnancy including the previously identified characterized, identified and synthesized as recombinant protein forms. gmmmgp1-3 and transferrin. Seven new MGPs (gmmmgp 4-10) were The immunogenicity of these antigens was evaluated by sera of guinea identified in this analysis. These proteins appear to be related as the amino pigs from the laboratory and field studies. acid composition of these proteins is similar to gmmmg2-3. Genomic analysis of gmmmgp2-10 revealed that they are located on the same genomic loci. Analysis of the predicted upstream regulatory regions for gmmmgp4-10 found conserved binding sites previously identified in the regulatory regions for gmmmgp1-3. Search for homologus sequences to these genes has only revealed a single uncharacterized sequence from the flesh fly, Sarcophaga crassipalpis. The predicted amino acid sequences for gmmmgp2-10 contain a high percentage of hydrophobic amino acids and a conserved secretory signal peptide, however they lack characterized functional domains. Expression patterns of gmmmgp2-10 are female specific and localized to the milk gland tissue. Temporal analysis of transcript levels for these genes is similar to the other genes

www.astmh.org 385 associated with lactation. This expression pattern results in increased completed a self-administered survey answering questions about their transcript levels in correlation with larvigenesis followed by immediate ease of use, setup, dismantling, and comfort. Results of this acceptability decline after parturition (birth). Knockdown of gmmmgp7 utilizing siRNA study will be presented in the context of military force health protection. injections resulted in a significant reduction of fecundity. The discovery of gmmmgp4-10 reveals a family of genes essential for viviparity and novel in 1283 form and function. IMMUNOGENIC AND BIOCHEMICAL PROPERTIES OF 1280 IXOLARIS, A TICK SALIVARY TISSUE FACTOR PATHWAY INHIBITOR RETENTION OF DUPLICATED LONG WAVELENGTH OPSIN 1 2 3 GENES IN THE GENOMES OF THE MOSQUITO VECTORS Ivo Francischetti , Patrick Brown , Robson Monteiro , Jose 1 4 2 AEDES AEGYPTI, ANOPHELES GAMBIAE AND CULEX Ribeiro , David Narum , Peter Schuck QUINQUEFASCIATUS 1Laboratory of Malaria and Vector Research, Rockville, MD, United States, 2National Institutes of Health, Bethesda, MD, United States, 3UFRJ, Rio de Gloria I. Giraldo-Calderon, Michael J. Zanis, Catherine A. Hill Janeiro, Brazil, 4National Institutes of Health, Rockville, MD, United States Purdue University, West Lafayette, IN, United States Ixolaris is a potent Tissue Factor inhibitor from tick saliva. It binds to Understanding the role of mosquito vision in mating, host detection Factor X(a) and the binary complex Ixolaris/FX(a) interacts with FVIIa/TF and oviposition, may help to improve or develop new control strategies thus blocking the coagulation cascade. Ixolaris has been sucessfully tested to reduce the incidence of vector-borne diseases. Here we report the as an antithrombotic in rats, and also displays anti-cancer properties in first molecular analysis of light receptors (opsins) from three mosquito a glioblastoma model. Because Ixolaris displays therapeutic potential, vectors - the yellow fever mosquito, Aedes aegypti, the malaria understanding its immunogenic and biochemical properties is of interest. mosquito, Anopheles gambiae, and the southern house mosquito, Culex Here we demonstrate that ixolaris elutes as approximately 18 kDa protein quinquefasciatus. Opsins are receptors that interact with photons to according to gel-filtration chromatography. Light scattering plot and initiate visual processes. Typically, insects have three classes of opsins ultracentrifugation experiments also indicate that Ixolaris is a monomeric that are stimulated by ultraviolet, short, and long (LW) wavelengths. We protein of approximately 18 Kda. Since the predicted mol mass for used expression data to improve the 10 A. aegypti and 11 A. gambiae Ixolari is 15.5 kDa, the discrepancy is attributed to glycosilation. This published opsins gene models, and we report the first manual annotation contention has been confirmed by a smear observed by SDS-PAGE and of 13 opsin genes from C. quinquefasciatus. Opsin transcripts were mass spectrometry analysis of Ixolaris. Elisa also demonstrate that Ixolaris confirmed using published expression data and Reverse Transcriptase-PCR. is non-immunogenic in rabbits and in mice. Taken together, these results Phylogenetic analyses predicted six putative LW opsins in A. aegypti, six provide further support for the potential therapeutic use of Ixolaris in a in A. gambiae and eight in C. quinquefasciatus, suggesting an expansion number of conditions with abnormal expression of Tissue Factor, including of these genes in the Culicidae relative to other insect taxa. Time of thrombosis, cancer, sepsis and malaria. divergence suggests the mosquito LW opsins originated from several duplication events between 167 to 1 million years ago (MYA), and that 15 1284 LW genes may have originated following a duplication event that occurred approximately 126 MYA. LW opsins share approximately 100% and 60% MOLECULAR MECHANISMS OF WOLBACHIA-MEDIATED amino acid similarity within and between mosquito taxa, which raises VIRAL INTERFERENCE intriguing questions regarding the retention of these genes in the three Xiaoling Pan, Zhiyong Xi mosquito genomes. Seven amino acids were identified under positive selection in the N and C termini, and one in a third trans-membrane Michigan State University, East Lansing, MI, United States domain suggestive of opsin spectral tuning. Conserved nucleotide Dengue is one of the most important arboviral diseases currently sequence in 6 out of 38 ortholog pairs and in 8 out of 14 paralog threatening human populations, with over 50 million cases in tropical and pairs of the non coding regions, up- and or down-stream, of the LW subtropical regions each year. No treatment or vaccine is currently available opsins is indicative of coordinated gene regulation. We discuss potential for dengue fever. Recently, the endosymbiotic bacterium Wolbachia has mechanisms, including positive selection and differential gene regulation, been proposed to be used as a tool to reduce mosquito vectorial capacity for the conservation of LW opsins in these mosquitoes. for dengue viruses through population replacement. Our previous studies showed Wolbachia alone can induce resistance to dengue virus in Aedes 1281 aegypti, which was associated with a boosted basal immunity in the Wolbachia-infected mosquito. To understand the molecular mechanisms FIELD USER ACCEPTABILITY EVALUATION OF A NOVEL, SELF- underlying Wolbachia-mediated viral interference, we examined SUPPORTING, LONG-LASTING INSECTICIDAL NET (LLIN) Wolbachia-induced physiological changes in mosquito by comparison of John Paul Benante, Gabriela Zollner, Jason Richardson genome-wide transcriptome between Wolbachia-infected and -uninfected Ae. aegypti. Experiments were also conducted to compare full scale Walter Reed Army Institute of Research, Silver Spring, MD, United States physiological responses of the two groups of mosquitoes to dengue Insect bed nets provide protection against arthropod-borne disease virus infection. We found that the Toll signal pathway was prominently pathogens such as malaria, dengue, and leishmaniasis. United States activated by Wolbachia in response to dengue virus infection. Interestingly, Army service members currently have a choice between two types of bed the genes related to redox stress response systems and mitochondria were nets to use in field environments; however, both have various limitations strickly regulated by the Wolbachia in Ae. aegypti. Further studies were that preclude effective long-term use by non-mobile forces. Therefore, also conducted to investigate how the Toll signal pathway was activated the US Army was faced with a challenge to develop an improved bed net by Wolbachia in Ae. Aegypti. As the effector genes of Toll signal pathway, that does not have any of the limitations associated with these existing the defensins and cecropins genes induced by Wolbachia were confirmed bed nets. The Walter Reed Army Institute of Research has partnered with to play roles in control of dengue infection. Our studies provide evidence Tritons Systems, Inc. to develop a novel, self-supporting, long lasting, to support that Wolbachia induces resistance to dengue virus in Ae. insecticide-impregnated net (LLIN). The purpose of this study was to aegypti through modulation of host immunity. We discuss the results in evaluate the new bed net in comparison with the existing Standard and relation to develop Wolbachia-based control strategies for population Self-Supporting Low-Profile bed nets using an acceptability threshold of replacement. 70%. Upon completion of a large scale field training exercise in which these bed nets were used over the course of several nights, soldiers www.astmh.org 386 1285 among island populations and between island and mainland sites. We observed a positive correlation between geographic and genetic distance, THE INFLUENCE OF HABITAT ON THE GENETIC STRUCTURE which suggests that open water does not necessarily act as a barrier to OF GLOSSINA FUSCIPES FUSCIPES IN UGANDA AND tsetse dispersal. Genetic data also suggest that males disperse farther than IMPLICATIONS FOR VECTOR CONTROL females and that populations are stable over wet and dry seasonal cycles. We will discuss the results in light of other recent genetic studies and Chaz Hyseni, Adalgisa Caccone compare them to previous ones based on ecological data. Yale University, Department of Ecology and Evolutionary Biology, New Haven, CT, United States 1287 Human and animal forms of African trypanosomiasis represent a burden THE CELL BIOLOGY OF CANDIDATUS RICKETTSIA ANDEANAE to the public health and economy of many African countries. For effective trypanosomiasis management, controlling its vector, the tsetse fly (Diptera: Alison Luce-Fedrow1, Chelsea Wright2, Holly Gaff2, Daniel Glossinidae), is necessary, but long-term success in vector control efforts Sonenshine2, Wayne Hynes2, Allen L. Richards1 requires a better understanding of tsetse dispersal and breeding ecology. 1Naval Medical Research Center, Silver Spring, MD, United States, 2Old We have collected genetic data over several years in Uganda for a major Dominion University, Norfolk, VA, United States trypanosomiasis vector, Glossina fuscipes fuscipes (G.f.f.). This genetic information coupled with publicly available environmental data (climate, Candidatus Rickettsiae andeanae is an incompletely characterized spotted hydrology, land cover) was used to assess habitat selection and dispersal fever group rickettsia (SFGR), first detected in Amblyomma maculatum and breeding capacity of tsetse in Uganda. Connection networks between and Ixodes boliviensis ticks collected in 2002 from northern Peru during a G.f.f. sampling localities were constructed and a modified inverse febrile outbreak investigation. Phylogenetic analysis of the 17-kDA, gltA, distance weighting method was used on these networks to interpolate ompB, ompA, and sca4 genes demonstrated alignment with SFGR, but the a ‘landscape’ of genetic variation in Uganda. Genetic variation captured molecular isolates were not found to be identical to any rickettsial agent in this way was used with environmental data to carry out environmental listed in GenBank, and Candidatus R. andeanae was deemed a novel niche modeling in Maxent v. 3.3.3. The inferred distribution of G.f.f. rickettsial agent. Despite molecular characterization of the Candidatus represents the flow of genetic information on the environmental substrate R. andeanae, the successful in vitro cultivation of this bacterium has of Uganda. We used circuit theory methods implemented in the program remained a challenge. We recently used one half of the Candidatus R. Circuitscape v. 3.5.4 to model genetic connectivity of the environmental andeanae-positive A. maculatum tick collected in Portsmouth, VA to landscape in Uganda and estimate environmental resistance to dispersal successfully infect cultures of Vero, DH82, and S2 cells. Infections were between G.f.f. populations. The environmental ‘friction’ estimates were confirmed using quantitative real-time PCR (qPCR) assays, acridine orange used to explore local genetic structuring of tsetse flies via spatially explicit staining, and DNA sequencing of gltA, ompB, and sca4 fragments. principal components analysis (sPCA) with the ‘adegenet’ R package. Current investigation of the cell biology by electron microscopy of Environmentally explicit modeling of gene flow provides information about Candidatus R. andeanae shows that the coccobacillus is approximately 0.3 the influence of the environment on genetic variation and connectedness. um long and 0.2 um wide, it has a double cell membrane similar to other Environmental-genetic inferences about habitat selection and dispersal SFGR, but it is has only been observed growing in the cytoplasm and not in tsetse could substantially improve vector control by helping to identify in the nucleus of the three cell lines assessed. Nuclear extraction studies areas to be targeted for control and minimizing the probability of re- are ongoing to more specifically determine if this agent replicates within infestation from neighboring areas. the nucleus. The studies described herein will more fully characterize this newly discovered rickettsia, which has now been established in culture for 1286 the first time in our laboratory. FINE-SCALE GENETIC DIFFERENTIATION OF GLOSSINA 1288 FUSCIPES FUSCIPES IN THE LAKE VICTORIA BASIN AND THE ROLE OF BIOFILM FORMATION IN COLONIZATION AND IMPLICATIONS FOR VECTOR CONTROL TRANSMISSION OF THE COMMENSAL SYMBIONT SODALIS Agapitus B. Kato1, Chaz Hyseni1, Loyce M. Okedi2, Charles GLOSSINIDIUS WITHIN THE TSETSE FLY Masembe3, Johnson Ouma4, Serap Aksoy1, Adalgisa Caccone1 Michele Maltz, Brian Weiss, Serap Aksoy 1Yale University, New Haven, CT, United States, 2National Livestock Resources Research Institute, Tororo, Uganda, 3Makerere University, Yale University, New Haven, CT, United States College of Natural Sciences, School of Biological Sciences, Department Awareness of diversity and abundance of beneficial microbes has greatly of Biology, Kampala, Uganda, 4Trypanosomiasis Research Centre, Kenya increased with the advancement of molecular technologies. Recently, the Agricultural Research Institute, Nairobi, Kenya influence of beneficial microbes in onset or prevention of disease has been The primary vector of Human African Trypanosomiasis (HAT) in Uganda is shown, indicating an opportunity for harnessing these microbes for control Glossina fuscipes fuscipes (G.f.f.). Little information is available on genetic of disease. Sodalis glossinidius is a gram-negative commensal symbiont differentiation and population dynamics of G.f.f. in the Lake Victoria of the tsetse fly, the sole vector of the African trypanosome. Sodalis is basin. We screened for genetic diversity among tsetse populations both harbored throughout the fly both intra- and extracellularly, primarily in on mainland and island sites in southern Uganda. The aim of this work the midgut tissue in close proximity of the trypanosome and is maternally- is to provide empirical data to support short-term vector control efforts transmitted to offspring. The proximity and the ability to genetically and inform long-term monitoring with the ultimate goal of creating tsetse manipulate Sodalis makes it a great candidate for paratransgenesis; i.e., free zones. We used genetic data from 19 microsatellite loci and the expression of antitrypanosomal compounds in the Sodalis within tsetse’s mitochondrial cytochrome oxidase gene (530bp) to estimate population midgut. One essential aspect of paratransgenesis is understanding sizes and levels and patterns of genetic differentiation and gene flow transmission biology of Sodalis and recolonization efficiency of genetically within and among 13 tsetse populations in the Lake Victoria basin. We modified Sodalis in tsetse lines. Biofilms are dense populations of microbes also used mark-release-recapture data to estimate population sizes and that adhere to surfaces and each other secreting extracellular polymers. movement patterns and related these to genetic inferences. Temporal Only a few studies have shown the role of biofilm formation in vector born collections from the same sites were used to evaluate seasonal fluctuations disease; i.e., Yersinia pestis within the flea gut. The ability of Sodalis to (dry vs. wet) of tsetse demography. Both nuclear and mitochondrial produce a biofilm was investigated using a classical microtiter plate biofilm markers suggest the existence of past and current genetic exchange assay and was shown to produce a biofilm under in vitro conditions. In this study we assessed genes important for biofilm formation in the fly gut www.astmh.org 387 colonization process, transmission to progeny and trypanosome infection major persistent reservoir of infection for these pathogens in the surveyed rates. Our studies provide enhancement of paratransgenic methodology communities. Associations of disease distribution with demographic, by understanding the role of biofilm formation in both recolonization geographic, and meteorological data were performed to define and of Sodalis and trypanosome infections, which will guide us in applying map the prevalence and indirect estimates of incidence as the basis for paratransgenesis in the future. assessing actual disease burden. 1289 1291 NEW VECTOR CONTROL MATERIALS FROM THE ARMED INVESTIGATION OF A SUSPECTED OUTBREAK OF ACUTE FORCES PEST MANAGEMENT BOARD FEBRILE ILLNESS IN MALINDI, KENYA IN DECEMBER 2010 Graham B. White1, Douglas A. Burkett2, Daniel A. Strickman3 Eyako K. Wurapa, Jeremy Kambi, Sam Lumbaso, David Oluoch, 1University of Florida, Gainesville, FL, United States, 2Armed Forces Pest M. Abdirizak, G. Batonjo Management Board, Forest Glen, MD, United States, 3U.S. Department of GEIS Kenya, DPO, AE, United States Agriculture, Agricultural Research Service, Beltsville, MD, United States Acute febrile illness (AFI) refers to sudden illnesses with fever. It is a The Deployed War-Fighter Protection research program (DWFP) is an common clinical presentation in Kenya where its aetiology remains initiative to develop and validate novel methods to protect United States unknown. Information on the prevalence and causes of AFI in Kenya is military deployed abroad from threats posed by disease-carrying insects. limited. Walter Reed Project’s (WRP) AFI surveillance site in Malindi noticed Starting in 2004 and administered by the Armed Forces Pest Management a 3 fold surge in AFI cases from Sept 2010 to Nov 2010 (Sept #10, Oct #5, Board the program is funded at $5M per year. The DWFP research Nov #15 cases). On average 2 cases are enrolled monthly. This prompted portfolio is concentrated in 3 specific areas: novel insecticide chemistries/ an investigation. Of note cases were from the same locality. Aliquots of formulations, application technology, and personal protective systems. malaria negative blood by RDT were sent to WRP reference lab for PCR Program consists of a noncompetitive funding process for USDA ARS- and ELISA for Malaria, Salmonellosis, Brucellosis, Leptospirosis, Aborviruses based research, and a competitive grants process open to non-USDA ARS and nasopharyngeal swabs tested for Flu. Study team: WRP and MOH. scientists (PIs from academia, industry, and military entomologists: 55 Study Area: Malindi District Hospital and Kisumu-dogo area. Investigation projects funded). Up to $3 million per year is given to USDA ARS National Period: 15 - 18 Dec. Case size: All 21 cases. A standard questionnaire Program 104, dealing with Veterinary, Medical, and Urban Entomology. was administered to all. All lab records from the period were reviewed. Ultimate objective is to find industry partners and get useful products into Data from the questionnaires was entered into an EPI-INFO database the market/military stock system. Presentation highlights DWFP products and analyzed. Majority, males (57.1%, n=12). The median age 30 yr. with examples of equipment, insecticides, and ~300 refereed publications. Most below 20 yr (42.9%, n=9). Most from Kisumundogo (23.8%, n=5). Majority presented with headache (42.9%, n=9), joint aches (28.6%, n=6) 1290 and myalgias (19%, n=4). 42.9% of cases classified as having AFI actually had undiagnosed malaria. 42.9% were malaria positive on PCR. All cases COMPREHENSIVE EPIDEMIOLOGICAL RESEARCH EFFORT ON were negative for viruses by PCR, Elisas and cell culture. Importantly, the FEBRILE ILLNESSES AND HEMOGLOBINOPATHIES ALONG aetiology of fever remained unknown in 57.1% of cases. Malaria RDT’s THE BANGLADESH-MYANMAR BORDER are not sensitive enough in low malaria transmission areas and when parasitemia is low. A negative RDT may not be enough to rule out malaria 1 1 1 Paul Swoboda , Peter Starzengruber , Hans-Peter Fuehrer , Julia in regions of low malaria endemnisity. There is clinical and lab evidence 1 1 1 1 Matt , Kamala Thriemer , Benedikt Ley , Markus Fally , Rashidul of low parasitimia having being cleared in malaria immune patients as no 2 2 1 Haque , Wasif A. Khan , Harald Noedl PCR positive was given antimalarias but on repeat PCR all were negative 1Institute of Specific Prophylaxis and Tropical Medicine, Medical University for malaria. This could be explained by self treatment but all denied it. A of Vienna, Vienna, Austria, 2International Centre for Diarrhoeal Disease comprehensive study to discover both common and uncommon pathogen Research, Dhaka, Bangladesh causes of acute febrile illnesses is needed. PCR may be a complement In order to estimate the burden of febrile illnesses in the border region to RDT and Microscopy in low malaria endemic areas. Continue vector of Bangladesh toward Myanmar a comprehensive prevalence study on control. Malaria naive persons should continue to be offered prophylaxis febrile illnesses and hemoglobinopathies was conducted in an area with or preventive measures. suspected high endemicity of tropical infectious diseases. Little is known about the prevalence of febrile illnesses in the Chittagong Hill Tracts, 1292 the southernmost region of Bangladesh bordering Maynmar and India, EFFICACY, SAFETY AND PK OF ARTEMETHER- an area with limited access to medical care due to inaccessible terrain and lack of infrastructure. Samples were collected from patients enrolled LUMEFANTRINE DISPERSIBLE TABLET IN THE TREATMENT during two separate cross-sectional studies in the years 2007 to 2010 OF ACUTE UNCOMPLICATED PLASMODIUM FALCIPARUM covering the same rural communities in rainy and dry season to assess MALARIA IN INFANTS <5 KG BODY WEIGHT seasonal trends. In a parallel ongoing hospital-based fever survey data M. Meremikwu1, M.J. Alao2, A. Gbadoẻ3, A. Tshefu4, G. Lefèvre5, of febrile participants from the catchment area of the Bandarban Sadar V. Walter5, M. Cousin5, K. Hamed6, B. Ogutu7 Hospital were collected. Out of a total population of 2123 enrolled in the 1Institute of Tropical Disease Research and Prevention, University of studies 671 acute febrile patients were diagnosed for the most common Calabar Teaching Hospital, Nigeria, 2Paediatrics and Clinical Genetics, infectious diseases: malaria (RDT, microscopy and PCR), typhoid fever, Cotonou, Benin, 3Chef de l’Unité d’Infectiologie et d’Onco-hématologie, leptosirosis, dengue (serological assays) and influenza (RDT and PCR) as CHU-Tokoin de Lomé, Togo, 4Kinshasa School of Public Health, University well as hemoglobinopathies. The collected data allow for an estimation of Kinshasa, The Democratic Republic of the Congo, 5Novartis Pharma AG, of long term trends in the epidemiology of the investigated diseases as Basel, Switzerland, 6Novartis Pharmaceuticals Corporation, East Hanover, well as short-term variations such as seasonal fluctuations and emergence NJ, United States, 7Centre for Clinical Research-Kenya Medical Research of rare conditions. Falciparum malaria remains the major health threat Institute, Nairobi, Kenya with a cross-sectional prevalence of 40.9% (CI: 35.4 - 46.7%) during monsoon months (May - October). However numbers vary significantly WHO recommends artemisinin-based combination therapy (ACT) as with the season and show an overall declining trend over the years. A high first-line therapy for infants with uncomplicated Plasmodium falciparum number of seropositive cases for leptospirosis (n= 194, 28.9%; SD: 25.6 malaria who have body weight (BW) ≥5kg. However, no ACTs are - 32.5%) and typhoid fever (n=203, 30.3%; SD: 26.9 - 33.8%) indicate a indicated in infants <5kg. Poor safety profile of current standard of care, www.astmh.org 388 quinine, limits its usage. Coartem (20mg artemether-120mg lumefantrine, 1294 AL), with an available pediatric formulation, has the largest clinical trial and postmarketing safety experience in infants ≥5kg to-date. This open- INVESTIGATION OF A CLUSTER OF DEATHS ATTRIBUTABLE label, single-arm, multicenter study in Sub-Saharan Africa will enroll TO MALARIA IN RURAL SENEGAL inpatient neonates and infants of <5kg BW with a confirmed diagnosis 1 2 2 of uncomplicated P. falciparum malaria in two sequential cohorts of 15 Mame Birame Diouf , Robert Perry , Julie I. Thwing , Moussa 1 infants each: term age >28 days (cohort 1) and term age ≤28 days (cohort Thior 2) to minimize any theoretical risk. A joint data monitoring committee 1National Malaria Control Program, Dakar, Senegal, 2Centers for Disease will review efficacy, safety, and pharmacokinetic (PK) data from cohort 1 Control and Prevention, Atlanta, GA, United States and recommend whether to proceed to cohort 2, with or without dose Since 2005, malaria control interventions, including insecticide-treated adjustment. The primary objectives are to evaluate the efficacy and safety nets, rapid diagnostic tests, and artemisinin-combination therapies have of AL dispersible tablet administered as 1 tablet bid over 3 days (to adjust been scaled up in Senegal, resulting in a large decrease in the malaria- if required), and to determine plasma levels of artemether, its active associated morbidity and mortality. However, in Touba district, deaths metabolite dihydroartemisinin, and lumefantrine. Exclusion criteria include attributed to malaria from September - November (transmission season) severe malaria, signs and symptoms of a critical condition, hepatic or renal increased by 58% in 2009 compared to 2008, while admissions for abnormality, and major neurological malformation. Neurodevelopment malaria decreased 19%. One health center reported 80% (38/47) of the status follow-up of patients is planned until day 42 and at 3, 6 and 12 deaths. The National Malaria Control Program led an investigation of months. Primary endpoint is PCR-corrected parasitological cure at day 7. these deaths, consisting of interviews with families and care providers and Secondary endpoints include reduction in parasite density at 24 hours; retrospective chart reviews. Charts were reviewed for 38 malaria deaths, PK assessments; PCR-corrected parasitological cure at days 14, 28, and all confirmed by rapid diagnostic testing. The median age was 5 years 42; time to parasite, fever and gametocyte clearance; and safety and (39% < 5 years and 37% 5-9 years) and 59% were male. Only 17% (6) tolerability assessments. Appropriate use of antipyretics and quinine as sought care within 24 hours of symptom onset, with a median of 3 days. a rescue medication will be permitted. Protocol approval will be sought Anemia was laboratory-confirmed in 37% (14) and diagnosed clinically from ethics committees in Switzerland, and in each participating country. in 26% (10); mean hemoglobin was 3.9 g/dL in those tested, two of Written informed consent will be sought from all parents/guardians. Study whom received a blood transfusion. Rapid blood glucose was performed results are expected in 2014. in 18% (7) and complete blood counts in 37% (14). Of the 12 patients with elevated leukocytes, 8 received an antibiotic. Of 37 patients for 1293 whom treatment was documented, 65% received a correct medication EVALUATING THE READINESS OF OUTPATIENT HEALTH regimen, 26% had dosing errors, and 5% died prior to starting quinine. The majority (92%) of patients died the day of admission or the following FACILITIES TO MANAGE MALARIA CASES IN BENIN day. Recommendations included expanding the home-based management Abdou S. Gueye1, Alexander K. Rowe1, Bruno Aholoukpe2, Emile program, reinforcing preventive community based interventions, educating Bongo3, Simplice Takoubo3, Michele Seibou3, Milton Amayun3 the community on danger signs and accessibility of treatment, and staff 1Centers for Disease Control and Prevention, Atlanta, GA, United States, training to improve referral practices, performance and documentation 2National Malaria Control Program, Cotonou, Benin, 3United States Agency of history and physical exams, complete blood counts and blood glucose, for International Development, Cotonou, Benin correct dosing of antimalarials, and differential diagnosis of fever. Additional studies would be needed to determine if the delays in seeking In 2008, the government of Benin and its partners began implementing care and deficits in care were associated with the deaths. A standardized a new national malaria case-management policy in 787 public health tool for investigation of deaths will help improve case management and facilities (HFs) that recommended the use of artemisinin-based the response to clusters of deaths. combination therapy (ACT). We evaluated the readiness of outpatient HFs to manage malaria cases in Benin about one year later. In late 2009, 1295 we conducted a nationally representative cross-sectional survey of a stratified random sample of 60 HFs. Surveyors observed consultations EFFECTS OF GASTROENTERITIS EPISODES ON MAINTENANCE and interviewed and re-examined patients seeking care for any illness OF POLIO VACCINE TITERS IN CHILDREN THREE YEARS AND and pregnant women seeking antenatal care. In addition, health workers UNDER IN RURAL COASTAL KENYA (HWs) were interviewed, and HFs were assessed to determine the 1 2 1 availability of drugs and equipment. Results were weighted. Altogether, A. Desiree LaBeaud , Peter Mungai , Ginny Gildengorin , 3 3 3 57 HFs, 113 HWs, and 448 patients were included in the analysis. All HFs Elisabeth McKibben , Maxim McKibben , Christopher L. King , 3 had a thermometer, 70.8% (95% confidence interval [CI]: 59.3-82.3%) Indu Malhotra had a scale for weighing children, and 66.3% (95% CI: 56.6-80.1%) of 1Children’s Hospital Oakland Research Institute, Oakland, CA, United HFs had a booklet or chart with ACT algorithms. Although all hospitals States, 2Ministry of Health, Msambweni, Kenya, 3Case Western Reserve could perform malaria testing, only 40.8% of non-hospitals could University, Cleveland, OH, United States perform testing. In the three months before the survey, 46.7% (14/30) of Evidence shows that infants with concurrent gastroenteritis (GE) are hospitals and 33.3% (9/27) of health centers had stock-outs of all types of less likely to respond to oral polio vaccination than those without artemether-lumefantrine blister packs (i.e., none in stock) for at least three gastroenteritis. Our objective was to determine whether further days. Adherence to the testing policy (i.e., test all patients with a febrile episodes of gastroenteritis in the first three years of life had an effect illness, and do not test patients without a febrile illness) was 52.9% (95% on maintenance of polio titer. Children enrolled in a birth cohort in rural CI: 48.3-57.5%) among all 448 patients, 24.7% (95% CI: 18.2-31.2%) coastal Kenya received four trivalent polio vaccinations before 6 months of among 170 patients <5 years old, and 70.1% (95% CI: 64.7-75.5%) life. Sera were then drawn at 6 month intervals until age 36 months and among 278 patients ≥5 years old. Nearly all of the 79 patients who polio titers were measured using poliovirus IgG ELISA kits. GE episodes tested positive were given an antimalarial (98.2%; 95% CI: 95.3-100%). were documented during scheduled follow-up visits and at any time However, 22.1% (17/77) of patients who tested negative were given an of illness during the 3 year period. Student’s t-test was performed to antimalarial. HF readiness varied. HWs prescribed antimalarials when tests compare those with and without GE at each time point. Of 545 children in were positive. However, ACT stock-outs were common, HWs did not the study, 159 had 246 episodes of gastroenteritis in the first three years follow testing guidelines for children <5 years old, and they sometimes of life. GE episodes were more likely to occur between 6 and 18 months prescribed antimalarials even when tests were negative. of life. The range of GE episodes per child was 0-4. Polio titers did not significantly differ between children with and without GE from 6 to 36 www.astmh.org 389 months of age. Although concurrent gastroenteritis may hamper immune age 5 mortality is 9.8%. 87% of the population is anemic based on WHO response to oral polio vaccine, further episodes of gastroenteritis after the standards. Access to Medical Care and treatment remains difficult for this vaccination series do not appear to alter the maintenance of polio titers. population We are significantly limited by lack of accurate ages, mobility of the population and changing demographics. Malaria is very prevalent 1296 in this rural area of Sierra Leone. Primary treatment and prevention has had some impact on mortality rates compared to prior studies, however CLINICAL IMPLICATIONS OF ADHERENCE TO WHO there still remains a significant disease burden in this area, with significant GUIDELINES FOR THE MANAGEMENT OF THE FEBRILE PHASE morbidity and mortality. OF DENGUE Luis A. Villar1, Beatriz Parra2, Doris Salgado3, Janeth Florez1, Irene 1298 Bosch4 ARTEMETHER, DIHYDROARTEMISININ AND LUMEFANTRINE 1Centro de Investigaciones Epidemiologicas UIS, Bucaramanga, Colombia, DO NOT INDUCE IN VITRO DRUG METABOLIZING ENZYMES 2Departamento de Microbiologia, Universidad del Valle, Cali, Colombia, AND METABOLISM OF ORAL CONTRACEPTIVES 3Universidad Surcolombiana, Neiva, Colombia, 4One Gustave L. Levy, Department of Microbiology, New York, NY, United States Hilmar Schiller, Judith Streckfuss, Bertrand-Luc Birlinger Novartis Pharma AG, Basel, Switzerland According to WHO guidelines the use of acetaminophen is indicated during the febrile phase of dengue and aspirin or non-steroidal anti- The goal of this study was to evaluate in vitro the components of inflammatory agents (NSAIDs) should be avoided as these drugs may Coartem/Riamet (artemether and lumefantrine) and the active metabolite aggravate gastritis or bleeding. However, there is little clinical evidence to dihydroartemisinin (DHA) for their potential to induce drug-metabolizing support this recommendation. We conducted a prospective cohort study CYP enzymes and the metabolism of oral contraceptives. The experiments in an endemic area in Colombia to evaluate the potential association were conducted according to the FDA drug drug interaction guidance. The between noncompliance with this guideline and the risk of developing assessment was done in vitro in cryopreserved primary human hepatocytes severe dengue. Acute febrile outpatients (less than 96 hours of onset) of at least three individual donors. Induction of mRNA, relative to the with dengue (confirmed by viral isolation, RT-PCR or a shift from vehicle control, was determined by real-time PCR and evaluation of negative to positive IgM test) were followed daily until the seventh day changes in cytochrome P450 (CYP) enzyme activities were assessed after of disease. Subjects were excluded based on the following: diabetes, 48-h induction periods by LC/MS/MS analysis of CYP-selective probe AIDS, hematologic disorders, cancer or cardiac disease and the presence substrate metabolism. Metabolism of the oral contraceptives was tested of a major bleed, albumin < 3gr/dL, effusions or shock at presentation. by HPLC analysis. Human hepatocytes were incubated with the three Inappropriate Initial Treatment (IIT) was considered when the patient test substances up to concentrations which exceeded their therapeutic reported having taken NSAIDs, aspirin or dipyrone. Data collected concentrations by a factor of 10. Ethinyl estradiol and levonorgestrel included signs and symptoms, and daily microhematocrit determinations were selected as active ingredients of oral contraceptives and were tested to recognize hemoconcentration. A complicated case was defined by the at their therapeutic concentrations of 1 nM and 20 nM, respectively. following: a platelet count ≤100.000/mm3; any spontaneous hemorrhagic Rifampicin at 0.1, 1, and 20 μM, and phenobarbital at 1000 μM were manifestation (or one positive tourniquet test); and evidence of plasma used as positive controls for induction of genes regulated by PXR and/ leakage (i.e. pleural effusion, ascitis, hypoalbuminemia or a variation or CAR like CYP2B6, CYP2C, and CYP3A; β-napthoflavone at 10 μM of hematocrit greater than 10%). Of 596 patients, 97% appropriately was included as positive control for AhR-mediated induction of genes received acetaminophen but 54% also received IIT. 63.2% (n=98) of cases like CYP1A. Artemether, DHA, and lumefantrine were determined not receiving IIT were complicated compared with 36.8% (n=57) complicated to be inducers of CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, or cases in the 271 subjects treated only with acetaminophen [OR crude: CYP3A enzyme activity in hepatocytes or CYP1A1, CYP1A2, CYP2B6, 1.62 ; 95% CI: 1.96- 6.39; OR : 1.51; IC95% (1.03-2.2) adjusted by age CYP2C8, CYP2C9, CYP2C19, CYP3A4, or CYP3A5 mRNA. Metabolism of and sex]. In conclusion, adherence to WHO guidelines during the febrile ethinyl estradiol and levonorgestrel was determined not to be induced by phase of dengue is important to reduce the risk of complications. This artemether, DHA, and lumefantrine. As per FDA criteria, these conclusions study is registered with Colciencias (Departamento Administrativo de were based upon the levels of mRNA or activity at least less than 2-fold, Ciencia, Tecnología e Innovación de Colombia), number: 110245921561. with respect to the vehicle control, and/or less than 40% of the maximal positive control induction response, indicative of a non-inducer in vitro. 1297 1299 MALARIA PREVALENCE AND MORTALITY IN RURAL SIERRA LEONE EFFECT OF DIARRHEA ON GROWTH IN INFANTS IN URBAN SLUM OF SOUTH INDIA John J. Flaherty 1 1 1 NorthShore University Health Systems, Evanston, IL, United States Deepthi Kattula , Prabhu Sivarathinasamy , Rajiv Sarkar , Sitara S. Ajjampur1, Jayaprakash Muliyil1, Honorine Ward2, Gagandeep Malaria is a leading cause of morbidity and mortality in rural areas of Kang1 Sierra Leone. Mortality from malaria is as high as 28% in the under age 5 1Christian Medical College,Vellore, Vellore Tamil Nadu, India, 2Tufts Medical group of pediatric patients. The Village Medical Project provides medical Center, Boston, MA, United States care and treatment to women and children in several villages in Gorama Chiefdom, Kono District of rural Sierra Leone The purpose of this study Diarrheal diseases are the second leading cause of morbidity and mortality is to document the prevalence of malaria infection, anemia and crude in children less than 5 years of age. The vicious cycle of diarrhea and mortality. The project has been working for 3 years to ascertain the malnutrition has long been recognized, and it has been shown that early success of primary treatment and prevention of malaria in a rural area of childhood diarrhea has long term effects on growth and development. To Sierra Leone. Adult and Pediatric Patients were tested for Plasmodium study the association of diarrhea in early infancy with growth faltering at falciparum malaria and hemoglobin. Patients were selected from each one year of age, longitudinal data (N = 897) from 3 birth cohort studies village based on prior census data and followed for a 2 year period, from conducted between 2002-2011 in an urban slum of South India were 2008-2010. Primary prevention with bed nets were provided for children analyzed. Children were followed biweekly/weekly to assess diarrhea and under 5 years of age. 1043-1463 patients were seen annually over a 2 other morbidities. Monthly anthropometric (height/ weight) measurements year period from 2008-2010. Overall, malaria prevalence varied from 67- were obtained. We assessed the effect of diarrhea, on acute (WHZ < 97%. The overall crude mortality rate from 2008-2010 was 8%. Under - 2 SD) and chronic (HAZ < - 2 SD) malnutrition, using the WHO Child www.astmh.org 390 Growth Standards 2006. The median number of diarrheal episodes 1301 among children in the cohort was 2 (1-3). At 1 yr, 33.9% of infants had chronic malnutrition and 26.9% had acute malnutrition. Three or more PRELIMINARY RESULTS OF A HOSPITAL-BASED LABORATORY episodes of severe diarrhea was significantly associated with chronic SURVEILLANCE FOR INFECTIOUS ETIOLOGIES OF (OR=2.45, P=0.02) and acute malnutrition (OR=2.8, P<0.01). Other UNDIFFERENTIATED FEBRILE ILLNESSES IN GEORGIA factors associated with chronic malnutrition were living in a mud house, 1 1 2 an indicator of lower socioeconomic status (OR=1.8, P<0.01), presence of Tinatin Kuchuloria , Tamar Akhvlediani , Manana Makhviladze , 3 4 5 an older sibling (OR=1.6, P<0.01). Duration of exclusive breasting, more Marina Endeladze , Tengiz Tsertsvadze , Maiko Chokheli , Paata 6 7 7 than primary schooling as highest education in the family and being a Imnadze , Margaret Farell , Mohamed Abdel Maksoud , Guillermo 7 8 8 9 girl offered protection of 22% (P<0.01), 44% (P<0.001), 30% (P<0.01) Pimentel , Danielle Clark , Christian Bautista , Robert G. Rivard , 9 7 respectively. As expected, severity of diarrhea and poverty are associated Matthew J. Hepburn , Brent House with acute and chronic malnutrition, with exclusive breastfeeding and 1I. Javakhishvili Tbilisi State University, Tbilisi, Georgia; Technology higher education being protective. Lower rates of malnutrition were noted Management Company, Tbilisi, Georgia, 2V. Bochorishvili Anti-Sepsis in girls, an unexpected finding. Center, Tbilisi, Georgia, 3Infectious Pathology, AIDS and Clinical Immunology Research Center, Tbilisi, Georgia, 4Infectious Pathology, AIDS 1300 and Clinical Immunology Research Center, Tbilisi, Georgia; I. Javakhishvili Tbilisi State University, Tbilisi, Georgia, 5National Center for Disease Control U.S. MILITARY FORCE HEALTH PROTECTION POLICIES MAY and Public Health, Tbilisi, Georgia, 6National Center for Disease Control IMPACT PEDIATRIC MALARIA PROPHYLAXIS PRESCRIBING and Public Health, Tbilisi, Georgia; I. Javakhishvili Tbilisi State University, PATTERNS Tbilisi, Georgia, 7Global Disease Detection and Response Program, U.S. Naval Medical Research Unit No. 3, Cairo, Egypt, 8Walter Reed Army Patrick W. Hickey, Ruth Gardner, Rodney Coldren Institute of Research, Silver Spring, MD, United States, 9U.S. Army Medical Uniformed Services University of the Health Sciences, Bethesda, MD, Research Institute of Infectious Diseases, Fort Detrick, MD, United States United States Since 2008, laboratory-based sentinel surveillance for undifferentiated To date, there have been no large scale systematic surveys of antimalarial febrile illness (UFI) has been ongoing in six hospitals to establish the prescribing practices in the United States. Although pediatric patients frequency of nine infectious causative agents of febrile illness in Georgia. are at higher risk of severe disease due to malaria than adults, there is Hospitalized patients ≥ 4 years of age with fever ≥38°C for ≥48 hours a relative scarcity of information on the prevention of malaria among were asked to voluntarily participate. Blood culture and serologic testing pediatric travelers versus adult travelers. This study consists of a systematic (ELISA) were conducted for Leptospira spp., Brucella spp., West Nile virus search of the military health system electronic medical record system for (WNV), Crimean-Congo hemorrhagic fever (CCHF) virus, Coxiella burnetii, all prescriptions of chloroquine (CQ), mefloquine (MQ), and atovaquone- tick-borne encephalitis virus (TBEV), hantavirus, Salmonella Typhi and proguanil (AP) to military family members 8 years of age and under in the Rickettsia typhi. A total of 478 subjects have been enrolled in the study. years 2005-2010. Prescribing patterns were assessed for changes over Of these, 71% were outpatients and 53% were males with the mean time to identify if Department of the Army and Department of Defense age of 36 years. Fever of unknown origin was the preliminary diagnosis policies, published in 2009, limiting the use of mefloquine in deployed in 88% of patients. Patients also reported having fatigue (90%), rigors forces coincided with changes in prescribing patterns for young children. (87%), sweats (82%), pain in joints (48%), and sleep disturbances (40%). A total of 3404 prescriptions were written for these medications during Acute and convalescent samples from 403 patients (n=473) were initially the study period. In total, CQ, AP, and MQ, respectively accounted for tested by IgM ELISA. Sixty-nine patients were seropositive for hantavirus 7%, 43%, and 50% of all prescriptions. Overall prescription volume (16%), 52 for Leptospira spp. (13%), 17 for Coxiella burnetii (4%), 16 increased from a low of 507 prescriptions (60% MQ) in 2005 to a high of for TBEV (4%), and 3 for WNV (0.7%). Additionally, 33 patients were 726 (39% MQ) in 2010 (p <0.001). While the total volume of antimalarial seropositive for Brucella spp. (8%), 3 patients for S. Typhi (0.7%), and 8% prescriptions rose, this change was reflected almost entirely by an increase (34) of patients showed positivity by IgG ELISA for R. typhi. Highest cross- in the usage of AP. In 2010, in contrast to prior years, 44% of all AP reactivity was observed for hantavirus and Coxiella burnetii, in 13(2.8%) prescriptions were for amounts in excess of a 30 day supply, compared to samples. Preliminary laboratory results indicate a high prevalence of 37% for earlier prescriptions (p=0.015). This trend of progressively more antibodies against hantavirus, leptospirosis, brucellosis and rickettsioses prescriptions for AP in absolute and relative terms exists over the entire among febrile patients in Georgia. This hospital surveillance for UFI has study period (p = 0.003). This study documents that military physicians significantly enhanced laboratory capacity for the detection of specific providing pediatric travel services now prescribe less MQ relative to AP. infectious etiologies as well as established a valuable network of clinical This occurs even in settings where the duration of travel has led many sites that can be used for future syndromic surveillance studies. Confirmed experts to recommend MQ as the drug of choice. The timing of these laboratory results will allow the Georgian public health authorities to make changes suggests that military force health protection policy, as well as better informed decisions regarding screening and prevention strategies. patient/family and provider awareness regarding adverse effects associated with MQ may be impacting prescribing practices for these medications. 1302 FACTORS INFLUENCING HIGH RATES OF CATCH UP GROWTH AFTER EARLY CHILDHOOD STUNTING IN CHILDREN OF URBAN SLUMS OF SOUTHERN INDIA: A COHORT STUDY Amos Lal, Prasanna Samuel, Victoria Jiang, Deepthi Kattula, Jayaprakash Muliyil, Gagandeep Kang Christian Medical College, Vellore, India Malnutrition and stunting in early childhood is a major public health problem in less developed countries. A lack of long term cohorts leads to a paucity of data on factors that influence catch up growth in children not enrolled in supplementary feeding programs. Our study in an urban slum in southern India investigated catch up in growth in children after early stunting. The study group was a birth cohort of 452 children,

www.astmh.org 391 followed intensively for three years, but at 7-8 years, 273 children were case was cultured from CSF. These preliminary study results suggest the contacted in 2010. Data was collected using a structured questionnaire presence of a wide-spectrum of pathogens among patients with suspected and anthropometry. For analysis, the cohort was divided into categories meningitis and encephalitis. This surveillance study serves as a model of children who were ever stunted at 12, 24 and 36 months and those for enhancing patient care through understanding disease prevalence, who were never stunted. Of available children, 189/273 (69.2%) were building laboratory diagnostic capacity and designing future syndromic ever stunted, but more than 80% of the 189 showed catch up growth by surveillance projects in Georgia. 2010. The mothers of the ever stunted group were younger by 1.4 years (p = 0.009), shorter (p = 0.009) and weighed less (p = 0.02) than mothers 1304 of never stunted children. Another variable that predisposed to stunting was household debt (Crude OR 1.82, 95 % CI 1.07-3.08). Ever stunted CLINICAL MANAGEMENT OF DENGUE: A PHYSICIAN children were divided into 2 groups, persistently stunted (33, 17.5%) EDUCATION PROGRAM TO IMPROVE CLINICAL OUTCOMES, and children with catch up growth (156, 82.3%) at the current follow PUERTO RICO up. In univariate analyses, factors associated with catch up growth were having <3 children, use of sunflower oil, use of a ration card, schooling Janice Perez-Padilla, Rosa Rodriguez-Acosta, Christopher of child in an unaided private school and using liquefied petroleum gas Gregory, Eunice Soto-Gomez, Hilda Seda, Carmen Perez-Guerra, as cooking fuel. After multivariate logistic regression analysis, the factor D. Fermin Argüello, Kay M. Tomashek independently associated with catch up growth was use of a ration card Centers for Disease Control and Prevention, San Juan, PR, United States (Adjusted OR 3.16, 95% CI 1.01-9.76). Our study shows remarkably high In 2007-08, the Centers for Disease Control and Prevention (CDC) rate of catch up growth, which was associated with use of a ration card Dengue Branch and the Puerto Rico Department of Health (PRDH) issued by the public distribution system, indicating that there is potential conducted a survey to assess physician’s knowledge of dengue and for governmental interventions to decrease malnutrition in poor urban clinical management practices. The survey identified limited knowledge communities. of warning signs for severe dengue and early signs of shock and non- standard treatment practices including use of corticosteroids and non- 1303 isotonic crystalloid solutions. A review of fatal dengue cases in 2007 corroborated these findings. In 2008-09, CDC Dengue Branch developed INFECTIOUS ETIOLOGIES OF ACUTE MENINGITIS AND and pilot tested a physician training course to address the deficiencies ENCEPHALITIS IN GEORGIA identified by the survey and fatal case review. Focus groups and Tamar Akhvlediani1, Margaret Farrell2, Christian Bautista3, interviews were conducted with attendees of the pilot course to evaluate Tinatin Kuchuloria1, Tengiz Tsertsvadze4, Roman Shakarishvili5, instructional process and content, and the course was revised accordingly. Nana Tatishvili6, Rusudan Chlikadze7, Annemarie Wasley8, The course was approved by CDC and accredited by the Accreditation Matthew Hepburn9, Guillermo Pimentel2, Robert Rivard9, Brent Council for Continuing Medical Education for 4 CME credits in February House2 2009. The Secretary of Health of Puerto Rico mandated that physicians take the training as a prerequisite for re-certification by 2013. To fully 1TMC/U.S. Army Medical Research Institute for Infectious Diseases implement the case management course, 52 physicians were selected, Clinical Research Unit; I. Javakhishvili Tbilisi State University, Tbilisi, trained and certified as Master Trainers. From February 21, 2009 to Georgia, 2U.S. Naval Medical Research Unit No. 3, Cairo, Egypt, 3Walter December 31, 2010, 55 courses attended by 8,301 of the 12,929 licensed Reed Army Institute of Research, Silver Spring, MD, United States, 4I. physicians in Puerto Rico were conducted. Most physicians (6,294, 76%) Javakhishvili Tbilisi State University; Infectious Pathology, AIDS and Clinical were trained between September 1 and October 31, 2010 in response Immunology Research Center, Tbilisi, Georgia, 5P. Sarajishvili Institute to another mandate from the Secretary of Health that all primary care of Clinical Neurology and Neurosurgery; I. Javakhishvili Tbilisi State physicians be trained immediately due to the increased number of dengue University, Tbilisi, Georgia, 6Neurology Department of the Iashvili Children’s fatalities. An evaluation of the impact of training on clinical practices Hospital, Tbilisi, Georgia, 7National Center for Disease Control and Public will be conducted in the Fall of 2011. Findings from this evaluation Health, Tbilisi, Georgia, 8World Health Organization Regional Office for will be used to redirect continuing training efforts and to develop an Europe, Copenhagen, Denmark, 9U.S. Army Medical Research Institute of online dengue clinical management course. Lessons learned from the Infectious Diseases, Frederick, MD, United States implementation of this training initiative will be shared with dengue In Georgia, there are diverse etiologies of meningitis and encephalitis, endemic countries planning to train physicians on the clinical management including vaccine preventable agents such as mumps virus, varicella of dengue. zoster virus (VZV), Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae type B (Hib), and others viral agents (e.g. Epstein- 1305 Barr virus (EBV), tick-borne encephalitis virus (TBEV) and West Nile virus (WNV)). Prevalence information regarding these infections in Georgia is PERFORMANCE OF STABILIZATION TUBES FOR EXTENDING limited. In October 2010, a hospital-based surveillance study was initiated TIME TO ANALYSIS OF COMPLETE BLOOD COUNTS FROM to determine the incidence of infectious etiologies of acute meningitis TRIAL PARTICIPANTS AT A RURAL FIELD SITE IN MALI and encephalitis; enhance laboratory capacity for the diagnosis of central 1 2 2 nervous system (CNS) infections; determine antimicrobial susceptibility Joseph P. Shott , Merepen A. Guindo , Sekouba Keita , Sintry 2 2 2 3 profiles; and describe the risk factors and clinical presentations associated Sanogo , Alassane Dicko , Issaka Sagara , Michael P. Fay , Patrick 4 4 with etiologic agents of CNS infections. Patients with suspected meningitis E. Duffy , Ruth D. Ellis and encephalitis were enrolled from three hospitals in Tbilisi. Cerebral 1Clinical Monitoring Research Program, SAIC-Frederick, Inc., spinal fluid (CSF) and acute and convalescent sera were collected for Bethesda, MD, United States, 2Faculty of Medicine, Pharmacy, and bacterial culture and RT-PCR testing for HSV types 1 and 2, mumps virus, Odontostomatology, University of Bamako, Bamako, Mali, 3Biostatistics enteroviruses, VZV, S. pneumoniae, Hib, and N. meningitidis. The diagnosis Research Branch, Division of Clinical Research, National Institute of of WNV, TBEV, and EBV was conducted via commercial ELISA assays. As Allergy and Infectious Diseases, National Institutes of Health, Bethesda, of 21 March 2011, 66 patients have been enrolled (23 adults and 43 MD, United States, 4Laboratory of Malaria Immunology and Vaccinology, children) and 61 CSF samples tested. Initial laboratory results indicate the Division of Intramural Research, National Institute of Allergy and Infectious frequency of HSV-1, enteroviruses and VZV to be 43%, 38% and 2%, Diseases, National Institutes of Health, Rockville, MD, United States respectively. For both TBEV and WNV, the frequency was determined to be Stabilization tubes (ST, Beckton Dickinson, Franklin Lakes, NJ) extend 7%. Nine samples were positive for TBEV and seven samples were positive the pre-analytical stability of whole-blood (WB) specimens taken for for EBV in 131 pairs of acute and convalescent sera. One S. pneumoniae CD4 measurements by utilizing a proprietary supplement targeted at www.astmh.org 392 cell surface membranes and clusters of differentiation (CD) on white 1307 blood cells (WBC). We hypothesized that this membrane-protective effect might confer specimen stability by acting on other membrane- “LOOKING FOR GOLD, FINDING MALARIA” THE 2010 bound cellular components. ST were assessed using twenty (n=20) WB MALARIA SURVEILLANCE OF THE SURINAME GOLD MINING specimens collected during a malaria vaccine clinical trial in Mali, West MALARIA CONTROL PROGRAM Africa. WB specimens were collected into ST and EDTA Vacutainer tubes for comparison, and complete blood counts (CBC) were conducted at day Hedley Cairo 0 and then every 24-hrs for 7 days thereafter. All measurements of WBC Ministry of Health, Paramaribo, Suriname parameters deteriorated (> 10% erroneous or missing values) after 24 Malaria is endemic in the forested interior of Suriname. Since 2006 malaria hours post-collection, while all red blood cell (RBC) parameters remained cases have declined tremendously with dispersed foci remaining in the largely unchanged through 6 days post-collection. Data analysis revealed gold mining areas. Currently the majority of malaria infections occur that ST do not provide stability of WB after collection in our setting. among persons (ca. 15,000) engaged in small-scale gold mining and Further investigations validating and implementing novel technologies in related activities. Because there were no formal health services in these the field are greatly needed to ensure quality specimens are analyzed in remote areas, a Global Fund supported malaria program was initiated in clinical research. 2009 to fill the gap. This control program builds further on a surveillance system established in 2006 as a pilot under the Medical Mission Malaria 1306 Program. The surveillance system gathers weekly information from the EVALUATING BLOOD CULTURES IN GUATEMALA AFTER Tourtonne diagnostic and treatment facility in the Capital city Paramaribo IMPLEMENTATION OF A DEDICATED PHLEBOTOMY TEAM and from a network of 18 home-based diagnostic and treatment facilities (Malaria Service Deliverers) in the gold mining areas. Malaria cases are Kim A. Lindblade1, Jennifer Gray2, Wences Arvelo1, Lissette diagnosed by blood film or rapid diagnostic tests. A descriptive analysis Reyes3, Aleida Roldan2, John McCracken2, Amarilis Motta3, of preliminary surveillance data of 2010 will be presented and compared Jennifer Verani1 with data from the previous year. The system recorded 1548 cases of 1Centers for Disease Control and Prevention, Atlanta, GA, United States, malaria in 2010 among gold miners; 1388 (90%) confirmed by microscopy 2Universidad del Valle de Guatemala, Guatemala, Guatemala, 3Ministerio and 160 RDT cases. This number represents a decrease of 31.5% from de Salud Publica y Asistencia Social, Guatemala, Guatemala the 2259 cases reported for 2009. Plasmodium falciparum 39%, P. vivax 52%, mixed P. falciparum plus P.vivax (7%) and P. malariae (2%) were Blood cultures (BCs) are important diagnostic and surveillance tools the species identified. Among the 1548 cases 961 (62%) were classified to identify invasive bacterial diseases. In January 2008, we established as imported from neighboring countries and 26 (2%) were of unknown automated BCs at a rural hospital in Guatemala and provided frequent origin. 40 cases were reported in pregnant women of which 8 were P. trainings, job aids and all supplies and materials. After the first year vivax relapse. The Annual Blood Examination Rate (ABER) was 52.83, of implementation, we found poor adherence to protocols and high Slide Positivity Rate (SPR) was 17.51 and the Annual Parasitic Index (API) contamination rates. In August 2009, we implemented a dedicated, calculated from autochthonous cases was 30 in 2010 compared to ABER round-the-clock phlebotomy team. To determine whether this intervention 48.34, SPR 22.45 and API 39 in 2009. In comparison to 2009 a notable decreased contamination rates and improved pathogen isolation, we decrease in the number of malaria cases from gold mining areas was conducted segmented regression analysis of a monthly time series of BC reported in 2010. Conveying the importance of adhering to appropriate outcomes from the laboratory database. A blood culture was defined preventive measures for malaria to the population at risk is mandatory for as one or two blood culture bottles filled with blood taken from the the decrease in malaria cases to be sustainable. same site. We collected data on 2,140 BC prior to intervention (January 2008-July 2009) and 1,525 blood cultures after the intervention was 1308 fully implemented (October 2009 to September 2010). There was an increase in the median number of BC per month among children ADULT REFERENCE VALUES FOR COMMONLY USED <10 years old (41 per month pre-intervention vs. 58 per month post- BIOCHEMICAL AND HEMATOLOGICAL TESTS IN CENTRAL intervention, p=0.05) but not among persons aged 10 years and older GHANA (63 per month pre-intervention vs. 68 post-intervention, p=0.69). Among 858 BC in children <10 years old during the pre-intervention period, David K. Dosoo1, Evans Kwara1, Kingsley Osei-Kwakye1, Seeba 14% of cultures were contaminated and 7% produced a pathogen, Amenga-Etego1, Philip Bilson1, Ruth Owusu1, Stephen Apanga1, compared to 10% contaminated and 4% with a pathogen among 695 Kwaku Poku Asante1, Emmanuel Mahama1, Kingsley Kayan1, cultures post-intervention. Among persons aged 10 years and older, 3% Dennis Adu-Gyasi1, Elizabeth Awini2, Elizabeth Awini2, Kofi of 1282 cultures taken pre-intervention were contaminated and 8% Tchum1, Kwadwo Koram3, Seth Owusu-Agyei1 yielded a pathogen, compared with 1% contaminated and 4% yielding a 1Kintampo Health Research Centre, Kintampo, Ghana, 2Dodowa Health pathogen of 830 cultures taken post-intervention. Segmented regression Research Center, Dodowa, Ghana, 3Noguchi Memorial Institute for analysis showed no impact of the intervention on the contamination Medical Research, Accra, Ghana rates among children <10 years old (β=-15.3, p=0.13) or persons aged Laboratory results and clinical examinations, provide useful information 10 years and older (β=-2.5, p=0.45). Similarly, there was no effect of the in screening, diagnosing and monitoring of diseases. Interpretation of intervention on pathogen isolation rates among children <10 years old laboratory results depend on reference values obtained from apparently (β=-2.2, p=0.62) or persons aged 10 years and older (β=-3.6, p=0.55). healthy individuals from the population they are intended to serve. The The results from this evaluation suggest that contamination rates among reference values obtained from healthy residents of the communities young children are unacceptably high and may be preventing isolation of used for clinical studies will help in determining eligibility and assessing pathogens. In addition to strengthening efforts to reduce contamination the safety of those participating in these studies. This study was aimed during venipuncture, particularly of young children, a review of laboratory at establishing gender-specific haematological and biochemical reference protocols and procedures may identify further opportunities to reduce values for healthy adults in central Ghana. A total of 625 adults between contamination and identify meaningful pathogens. the ages of 18 and 60 were enrolled within Kintampo and its environs. The medians, 2.5th and 97.5th percentiles were determined for five haematology and five biochemistry parameters commonly considered during screening/enrolment and follow up monitoring of individuals who usually participate in clinical trials and also for health management.

www.astmh.org 393 The Clinical Laboratory and Standards Institute (CLSI) guidelines for 1310 defining reference values were used. Values established in this study were compared with those derived in the developed countries. The percentage ETIOLOGY OF UNCOMPLICATED FEBRILE ILLNESS AMONG of our healthy population which had out of range values based on the CHILDREN 2 - 59 MONTHS OLD ATTENDING A PRIMARY data from the United States and the United Kingdom were determined. HEALTH CARE CENTRE IN ZANZIBAR The red blood cell (RBC)) parameters (haemoglobin, haematocrit and 1 1 2 RBC count), total leucocyte and platelet counts and urea values were Kristina Elfving , Deler Shakely , Kimberley Baltzell , Annika 3 4 5 6 significantly lower compared to values derived in the developed countries. Ljung , Mwinyi I. Msellem , Birger Trollfors , Max Petzold , 4 1 7 Higher values were, however, obtained in our study for parameters such as Abdullah S. Ali , Anders Björkman , Magnus Lindh , Andreas 8 Alanine aminotransferase, aspartate aminotransferase and total bilirubin. Mårtensson Up to 53% and 75% of the haematology and biochemistry values, 1Malaria Research, Department of Medicine, Karolinska Institutet, respectively from our healthy population would have been declared as Stockholm, Sweden, 2Department of Physiological Nursing, University of abnormal results if data for the developed countries were to be used. The California San Francisco, San Fransisco, CA, United States, 3Department results from this survey support the need to establish reference values of Infectious Medicine, University of Gothenburg, Gothenburg, Sweden, using individuals from the population it intends to serve. This will help 4Zanzibar Malaria Control Programme (ZMCP), Zanzibar, United Republic of reduce the inappropriate exclusion of potential clinical trial participants Tanzania, 5Department of Paediatrics, The Queen Silvia Children’s Hospital, based on reference values derived in the developed countries. University of Gothenburg, Gothenburg, Sweden, 6Nordic School of Public Health, Gothenburg, Sweden, 7Clinical Virology, Department of Infectious 1309 Medicine, University of Gothenburg, Gothenburg, Sweden, 8Division of Global Health, Department of Public Health Sciences, Karolinska Institutet, CLINICAL OBSERVATIONS OF HUMAN MONKEYPOX Stockholm, Sweden INFECTIONS IN THE DEMOCRATIC REPUBLIC OF THE CONGO A better knowledge of non-malarial fevers is a critical component for James Martin1, Mark Withers1, John Huggins1, Jean-Jacques improved case management of childhood fevers in the new context of Muyembe2, Mbala-Kingebeni Placide2, Cesar L. Cesar2, Bryony low malaria transmission in Zanzibar. We therefore conducted a health Soltis1, Fernando B. Guerena1, Lawrence Korman1, Phillip R. facility based prospective cohort study of the relative frequencies of Pittman1 infectious disease etiologies of uncomplicated febrile illness in North A 1U.S. Army Medical Research Institute for Infectious Diseases, Frederick, District, Zanzibar. We enrolled 650 children aged 2 to 59 months old MD, United States, 2Institut National de Recherche Biomedicale, Kinshasa, presenting to a primary health care center with confirmed fever (axillary The Democratic Republic of the Congo temperature ≥37,5°C) or history of fever within the preceding 24 hours. Patients were clinically managed according to the newly adapted Zanzibar We describe the results of an observational study of the clinical natural Integrated Management of childhood illness (IMCI) guidelines, which history of human monkeypox (MPX) infections at the remote L’Hôpital include a Rapid Diagnostic Test (RDT) for malaria. Naso-pharyngeal Général de Référence de Kole in the rainforest of the Congo River basin and rectal swabs were collected for PCR-detection of respiratory tract of the Democratic Republic of the Congo (DRC). The cardinal observations infection and diarrhoeal etiologies. Further, full blood count, malaria from 244 subjects enrolled in the study are summarized. All subjects who microscopy, C-reactive protein, chest X-rays, urine cultures with antibiotic were positive by pan-orthopox MBG PCR -- utilizing an onsite quantitative susceptibility testing, rapid detection of pneumococcal antigen in urine real-time assay (LightCycler) -- were also positive by a MPX-specific MGB and of Group A Beta-Hemolytic Streptococci from throat swabs were PCR assay, suggesting that MPX may be the only poxvirus circulating in analyzed. Preliminary results from a sub-sample of 292 patients of whom the area. Sequencing of MPX DNA from one subject’s scab showed only 41% had measured fever show the following distribution of clinical 17 nucleoside changes from the MPX Zaire 79 strain (collected in 1979) diagnoses (a patient can have >1 diagnosis): 52% pneumonia, 50% which was circulating during the WHO clinical characterization studies upper respiratory tract infection, 22% diarrhea, 8% tonsillitis, 4% skin of 25 years ago when the case-fatality rate (CFR) was about 10%. This infection, 3% ear infection and 2% dysentery. After laboratory testing no is the same isolate USAMRIID has used to develop non-human primate RDT or microscopy confirmed malaria infection was found. Streptococcus MPX models for drug and vaccine evaluation. The spectrum of disease A rapid test was positive in 8% of all patients. Chest x-ray was performed severity in our study was broad as evidenced by lesion counts ranging in patients classified as pneumonia according to IMCI (cough and/or 7 from 2 to 8,617, viremia (by PCR) ranging from undetectable to 6.3 x 10 difficult breathing and increased respiratory rate). Radiological pneumonia genomes ml/blood, and clinical status ranging from very mild to critically consolidation was verified in 5% of those with pneumonia as initial ill. The CFR to date is only 0.9% in subjects aggressively treated with diagnosis. Urine culture from non-diarrheal patients showed significant antibiotics, antimalarials, antiparasitics, anti-inflammatory drugs, and IV growth in 8%. These preliminary results indicate that a majority of the fluid. A strong correlation appears to exist between maximum lesion count fever episodes were related to respiratory tract infections and diarrhea. and maximum viral load. Low albumin and total protein levels, as well as However, most IMCI classified pneumonias could not be confirmed with elevated liver enzyme and alkaline phosphatase levels, were seen in nearly chest x-ray. Further PCR analyses will provide insight into the viral and all cases. In one case, in which viremia was detected before the onset of bacterial etiologies of these febrile episodes. clinical illness, the maximum viral load occurred before the appearance of lesions and coincident with the onset of symptoms. Fetal demise due to maternal transmission of MPX infection occurred in three of four cases of pregnancy. A high percentage of cases involved transmission within households. The severity of disease within families varied widely without discernable pattern. This observational study is expected to lead to future hypothesis driven studies.

www.astmh.org 394 1311 The patient presented with left-sided empyema, which was drained with a chest tube under water seal. Sputum was positive for acid-fast bacilli, PRE-TRAVEL VACCINATIONS, PRESCRIPTIONS AND and anti-TB therapy and antibiotics were initiated. After two weeks, a COUNSELING FOR MEDICAL MISSIONARIES AND persistent air leak remained and chest radiography showed failure of RESEARCHERS lung reexpansion. The chest tube was trimmed to 4 centimeters and the 5th finger of a plastic glove with both ends cut was attached to the 1 1 1 Natasha S. Hochberg , Hari S. Iyer , Davidson H. Hamer , William end and lubricated with Vaseline, which allowed for one-way exit of air 1 2 3 4 B. MacLeod , Mary E. Wilson , Lin H. Chen , Adolf W. Karchmer , and fluid. The tube was left in place for three months to create a chest 5 6 7 Christine M. Benoit , Winnie W. Ooi , Laura Kogelman , Elizabeth window, after which the tube was removed, leaving an epithelialized 5 D. Barnett passage between the pleural space and external environment. Scant 1Boston University School of Public Health, Boston, MA, United States, fluid continued draining from the valve and the chest window during the 2Harvard School of Public Health, Boston, MA, United States, 3Mount treatment course but no other complications were noted. Bacteriological Auburn Hospital, Cambridge, MA, United States, 4Beth Israel Deaconess cure was confirmed by negative control sputum at two and five months. Medical Center, Boston, MA, United States, 5Boston Medical Center, After 12 months the window had closed spontaneously, and the lung had Boston, MA, United States, 6Lahey Clinic Medical Center, Burlington, MA, completely reexpanded. Radiographs illustrate the entire clinical course. United States, 7Tufts Medical Center, Boston, MA, United States The method described here using a widely available resource−a non-sterile plastic glove−to make a modified one-way valve was successfully used Overseas volunteers and researchers face unique risks related to their for the treatment of tuberculous bronchopleural fistula and persistent travel purpose and duration. We sought to characterize these travelers and pneumothorax. Ongoing drainage of chronic empyema and formation identify whether they received appropriate vaccinations, prescriptions, and of a chest window is thus possible without advanced thoracic surgical counseling on travel-related issues. Boston Area Travel Medicine Network intervention. The glove drain is worth considering in resource-limited is a research collaboration of five travel clinics in the greater Boston settings for this challenging complication of pulmonary TB. area that sees ~7,500 travelers per year. We evaluated characteristics of travelers who reported their reason for travel as “missionary/volunteer” or “researcher/student.” Between March 2008-July 2010, 15,440 travelers 1313 were seen in BATMN clinics. Of these, 1451 (9.4%) were missionaries/ THE BURDEN OF CHRONIC HEPATITIS B IN IMMIGRANTS IN volunteers, 1216 (7.9%) researchers/students, and 65 (0.4%) reported QUEBEC, CANADA: A POPULATION BASED STUDY both reasons. The median age of all 2,732 was 24 years (range 8-85), and 907 (33.2%) were male. The median travel duration was 21 days Christina Greenaway1, Sonya Cnossen2, Carmine Rossi2, Kevin (range 1-1,096). Among 4,035 destinations, the most common were Schwartzman3, Christina Holcroft2, Marina Klein4 Haiti (308; 7.6%), India (228; 5.7%), Kenya (195; 4.8%), China (190; 1Jewish General Hospital, Montreal, QC, Canada, 2Lady Davis Institute 4.7%), and Tanzania (176; 4.4%). Documentation was available for up for Medical Research, Montreal, QC, Canada, 3Montreal Chest Institute, to date vaccination status, evidence of immunity, or vaccine receipt at the Montreal, QC, Canada, 4McGill University Health Centre, Montreal, QC, clinic visit for 1,466 (53.7%) for Td/Tdap, 1,835 (67.2%) for hepatitis B, Canada 2,323 (85.0%) for hepatitis A, 1,088 (39.8%) for influenza, and 199/286 (69.6%) for meningococcus (among persons going to at-risk countries). Immigrants have higher mortality from chronic hepatitis B (HBV) and Commonly prescribed medications included ciprofloxacin (1599; 58.5%), hepatocellular carcinoma as compared to those born in Canada. Despite azithromycin (590; 21.6%) and levofloxacin (50; 1.8%) for traveler’s this disparity there are no screening programs to detect chronic HBV, diarrhea and atovaquone/proguanil (850; 49.0%), mefloquine (174; and HBV vaccine is not routinely given to immigrants after arrival in 10.0%) and doxycycline (120; 6.9%) for those 1732 persons traveling to Canada. This is because there is no population based data describing the chloroquine-resistant malaria risk countries. HIV post-exposure prophylaxis burden of chronic hepatitis B in immigrants. To fill this gap we created was prescribed for 1,235/1,598 (77.3%) travelers, and 385/1,943 (19.8%) a cohort of all cases of hepatitis B reported from 1991-2008 in Quebec had documentation of tuberculin skin testing. Blood-borne pathogen through linking administrative databases. We linked the MADO (Quebec counseling was documented for 1,235/1,599 (77.2%), evacuation Reportable Disease database), the MICC (Quebec Landed Immigrant insurance counseling for 1,243/1,878 (66.2%), and rabies or animal bite database) and the RAMQ (Quebec provincial health insurance and counseling for 215/1,940 (11.1%). Missionaries, volunteers, researchers physician billing database). For incidence rate estimates, denominators for and students make up less than 20% of BATMN travelers. Although it is immigrants were obtained from the MICC database (N=757,650 newly likely that not all of these travelers had direct patient contact overseas, arrived immigrants from 1991-2008); for non-immigrants, denominators there are still critical gaps in the vaccinations and counseling they receive. used 1991, 1996, 2001, and 2006 Quebec census data (immigrants removed). Rates and rate ratios and 95% CI were calculated using the 1312 Poisson distribution. 13,889 cases of chronic hepatitis B were reported during the study period. Non-immigrant cases were older (mean age 43.4 SUCCESSFUL USE OF MODIFIED HEIMLICH VALVE vs 33.4 p <0.01) and were more likely male (69% vs 51%, p <0.01). The USING PLASTIC GLOVE FOR MANAGING TUBERCULOUS rate of chronic hepatitis B overall was 10 fold higher in immigrants as BRONCHOPLEURAL FISTULA compared to non-immigrants [rate ratio; 95% CI = 10.0 (9.7-10.30) and rates/100,000 person years (PY) were 73.9 vs 7.4]. Rates were highest Alexis Cambanis1, Finly Zachariah1, Mary Electa Lybarfe2 for immigrants from East Asia/Pacific [rate/100,000 PY 95% CI = 280 1PIH-UCI Family Medicine Residency Tropical Medicine Program, Whittier, (268-293)], Sub-Saharan Africa [280 (262-298], Eastern Europe [86 (77- CA, United States, 2Tuberculosis Unit, St. Elizabeth General Hospital, 94)]; they ranged from 38-42/100,000 PY for immigrants from South Shisong, Kumbo, Northwest Province, Cameroon Asia, the Middle East/North Africa and Latin America. Immigrants are at increased risk for chronic hepatitis B and its associated sequelae, including Sequellae of pulmonary tuberculosis (TB) include pleural effusion, potential transmission to close contacts. Immigrants would therefore likely empyema, and bronchopleural fistula. After thoracostomy and appropriate benefit from screening for chronic hepatitis B and verification of hepatitis medical therapy, failure of lung reexpansion may signify bronchopleural B immune status, so that appropriate treatment of chronic infection and fistula due to underlying pulmonary destruction, which often results in vaccination of susceptible contacts can be provided. recurrence of empyema, sepsis and death. We present a 23-year old HIV-negative male in Cameroon with smear-positive TB complicated by empyema and bronchopleural fistula who was successfully managed using a modified Heimlich valve made from the finger of a plastic glove.

www.astmh.org 395 1314 with filariasis. A course of Doxycycline was initiated for the management of both acute Lyme arthritis and chronic filariasis. Antifilarial therapy CORRELATION OF MALARIA RAPID DIAGNOSTIC TESTING with Doxcycline was directed toward the symbiotic bacteria, Wolbachia WITH CLINICAL-BASED ALGORITHM IN A RURAL VILLAGE IN associated with microfilaria. While it is possible our patient’s positive Lyme UGANDA serology reflected cross reacting antigens to filaria; his clinical presentation was consistent with Lyme associated arthritis. This case is unique in that 1 2 1 Jessica A. Kumar , John Kalule , Carlos Elguero , Elizabeth antimicrobial management of one endemic infection was useful in the 1 1 Dufort , Ellis Tobin management of a geographically separate pathogen. 1Albany Medical Center, Albany, NY, United States, 2Engeye Health Clinic, Ddegeya Village, Uganda 1316 Uganda has the world’s highest malaria incidence and mortality. The STRATEGIES TO PREVENT MEASLES, MUMPS AND RUBELLA Engeye Clinic was created in 2006 as a U.S./Ugandan non-governmental AMONG NEWLY ARRIVED ADULT IMMIGRANTS AND organization based in the Ddegeya Village. In this resource poor setting REFUGEES IN CANADA: A COST-EFFECTIVENESS ANALYSIS lacking microscopes and trained technicians, rapid diagnostic testing (RDT) was initiated to confirm clinically suspected malaria. Issues facing this Louis-Patrick Haraoui1, Kevin Schwartzman2, Chris Greenaway3 community include little to no use of mosquito nets, failure to complete 1Department of Infectious Diseases and Medical Microbiology, McGill treatment and/or use of paracetamol substituted for malarial treatment University, Montréal, QC, Canada, 2Respiratory Division, Montreal by village merchants. The purpose of this study was to evaluate the Chest Institute, McGill University, Montréal, QC, Canada, 3Centre for implementation of a clinical algorithm in a resource-limited setting for Clinical Epidemiology and Community Studies of the Lady Davis Institute the diagnosis of malaria compared to RDT. Over a two week period in for Medical Research, Jewish General Hospital and Department of February 2010, 344 patients were assessed by the on-site clinician using Microbiology and Infectious Diseases, Jewish General Hospital, McGill a clinical algorithm for the diagnosis of malaria. This included fever, chills, University, Montréal, QC, Canada sweats, headaches, muscle or abdominal pains, nausea and vomiting for greater than 48 hours with no other obvious cause. RDT was performed Adult immigrants are an unrecognized group at risk for measles, mumps on patients meeting clinical criteria for malaria by obtaining whole blood and rubella (MMR). They have been over-represented in rubella outbreaks samples using immunographic testing. Treatment for suspected malarial and most cases of congenital rubella (CRS) in Canada have occurred in cases was initiated with artemether/lumefantrine based on weight and infants born to foreign-born mothers. Despite this gap, MMR vaccine pregnancy/lactation status. 117 patients met clinical criteria for malaria is not routinely given to adult immigrants after arrival in Canada. We diagnosis. All clinically diagnosed cases were positive when confirmed performed a cost-effectiveness analysis to define the optimal strategy with RDT for the detection of parasite specific antigens for P. falciparum. to prevent MMR in this population. We constructed a decision analysis The prevalence of malaria as a cause of presenting symptoms was 34% (Markov modeling) to compare the cost-effectiveness of four preventive in this cohort. This clinical algorithm was found to be highly specific. The strategies using MMR vaccine, compared to no intervention (status quo). specificity and the positive predictive value of the clinical algorithm was Interventions tested were: 1 and 2) Vaccinating all adult immigrants with 100% when compared to the RDT in this cohort. In patients without either 1 or 2 doses of MMR vaccine without prior serotesting, or 3 and clinical criteria for a diagnosis of malaria there were no positive RDT 4) serotesting for MMR then vaccinating those found to be susceptible results. It was determined that the clinical algorithm could be used by rural to one or more of the diseases with either 1 or 2 doses of MMR vaccine. health care workers to accurately diagnose malaria as misdiagnosis leads A hypothetical cohort of 250,000 newly arrived immigrants 18 years to a delay in treatment causing an increased mortality and unnecessary of age or older was modeled over a 20-year period. The expected prescription of malarial medications and increased drug resistance. number of cases of MMR, associated complications and costs (health care system perspective) for each of the strategies were calculated with 1315 3% discounting for costs and clinical outcomes. Vaccinating all adult immigrants with 2 doses of MMR vaccine without prior serotesting saved LYME DISEASE AND FILARIASIS - A WOLBACHIA 41 million Canadian dollars and avoided 6,362 measles cases, 9 measles CONNECTION: A CASE REPORT deaths, 5,051 mumps cases, 8,181 rubella cases, 1 rubella death and 38 cases of CRS over a 20-year period, as compared to no intervention. Jessica A. Kumar, Graham Atkins, Ellis Tobin, Richard Blinkhorn Comparing both interventions involving vaccination without serotesting, Albany Medical Center, Albany, NY, United States the 2-dose strategy cost an additional $1.69 per person (total of $422,500) compared to the 1-dose strategy but prevented an additional Health care providers must consider neglected tropical and regional 273 measles cases, 405 mumps cases, 273 rubella cases and 1 CRS. Both endemic diseases. Lyme disease, commonly found in North America strategies that involved serotesting before vaccination were cost-saving requires a diagnosis of erythema migrans with confirmatory serology. compared to no intervention. However, both serotesting strategies were Lymphatic filariasis, endemic to Africa, is a neglected chronic disease that more costly and less effective than the vaccination-only strategies. All new can be easily overlooked in immigrants coming to North America. We adult immigrants should be given two doses of MMR vaccine without prior present a case of a 21 year old Liberian male complaining of one week serotesting: this strategy is cost-saving, prevents individual morbidity and of right knee pain and swelling during the summer. He had a history mortality, and decreases the potential for outbreaks. of malaria and filariasis. He immigrated to Albany, New York eighteen months ago as a refugee from civil war and was taking isoniazid and pyridoxine for a positive tuberculin test. He had no known tick exposure or 1317 trauma. He was afebrile and hemodynamically stable complaining of right DETERMINANTS OF ANEMIA AMONG YOUNG CHILDREN IN knee discomfort, warmth, swelling and decreased range of motion. His WESTERN KENYA white blood cell count was 4.7 with 52% neutrophils, 26% lymphocytes, 9% eosinophils and 12% monocytes. C-reactive protein (101 mg/L) Eric M. Foote1, Kevin M. Sullivan1, Laird Ruth2, Cliff Ochieng3, erythrocyte sedimentation rate (73 mm/hr) and IgE level (4123 U/ml) were Parminder S. Suchdev1 elevated. Knee radiograph showed joint effusion. Synovial fluid aspiration 1Emory University, Atlanta, GA, United States, 2Centers for Disease Control contained 116.5 tho/cmm white cells (95% neutrophils) but gram and Prevention, Atlanta, GA, United States, 3Safe Water and AIDS Project, stain and culture were negative. Lyme PCR from synovial fluid was not Kisumu, Kenya submitted for analysis. Lyme C6 peptide was positive as was confirmatory Western blot which demonstrated IgG bands 18, 23, 30, 31, 39, 41, 58 Anemia among children in developing countries is often attributed to and 93. Antifilarial IgG4 was positive indicative of past or chronic infection iron deficiency caused by low dietary iron intake. However, the causes www.astmh.org 396 of anemia are multi-factorial and interlinked. In sub-Saharan Africa, 1319 sickle cell disease (SCD), α-thalassemia, and infections are widespread and are known risk factors for anemia. Data on multiple risk factors for MID-UPPER-ARM CIRCUMFERENCE IS A USEFUL TOOL FOR anemia are needed to design more effective prevention and treatment ASSESSING NUTRITIONAL STATUS IN YOUNG CHILDREN programs. We conducted a cross-sectional cluster survey of 841 children WITH DIARRHEAL DEHYDRATION aged 6-35 months in 60 randomly selected villages in Nyando District, 1 2 2 3 western Kenya. Anemia prevalence (hemoglobin 8.3 mg/L) 75%, vitamin Dilruba Nasrin , Md Shahnewaz , A. S. Faruque , Dipika Sur , 4 4 1 1 A deficiency (retinol binding protein (RBP) 10 mg/L) 30%, reported Adeyemi Mitchell , Debasish Saha , Tamer Farag , Myron Levine , 1 fever in the last 24 hours 27%, stunting (height-for-age z-score<-2) Karen Kotloff 30%, wasting (weight-for-height z-score<-2) 3%, sickle cell trait 17%, 1University of Maryland Baltimore, Baltimore, MD, United States, SCD 2%, heterozygous α-thalassemia genotype 38% and homozygous 2International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, α-thalassemia genotype 9%. In bivariate analysis, anemia was associated Bangladesh, 3NICED, Kolkata, India, 4MRC, Basse, Gambia with iron deficiency, vitamin A deficiency, malaria, inflammation, fever, Undernutrition is implicated in over half of all child deaths related to stunting, wasting, homozygous and heterozygous α-thalassemia diarrhea. Cohort studies which assess the relationship between diarrheal genotypes, age<30 months, male sex, and low socioeconomic status (SES) disease and nutritional status typically use weight-based anthropomorphic (p<0.05). In linear regression, accounting for cluster design, the best fit measures. However, such measures are not useful to study the acute model included TfR, RBP, malaria, CRP, SCD, homozygous α-thalassemia nutritional changes induced by diarrhea because children with diarrhea genotype, male sex, age<30 months (R2= 0.59, p<0.0001). Age <30 are often dehydrated. Mid-upper-arm circumference (MUAC) is a cheap, months, homozygous α-thalassemia genotype, and CRP modified the simple tool often considered to be a better indicator of acute malnutrition relationship between iron deficiency and hemoglobin. Fever, height-for- than weight. MUAC measures musculature, a proxy for protein stores. If age z-score, height-for-weight z-score, and low SES were not significantly truly unaffected by interstitial water content, or dehydration, MUAC could associated with hemoglobin when included in the best fit model and did be used to assess the acute nutritional changes induced by diarrhea. In not confound the relationship between TfR and hemoglobin. Interventions February 2011, we initiated a case control study in Mirzapur (Bangladesh), designed to prevent anemia should utilize an integrated approach, Kolkata (India), and Basse (The Gambia) to determine whether diarrheal ensuring optimal iron intake while also addressing malaria and other dehydration affects MUAC. A case, enrolled at a clinic, was defined as a infections. child 0-59 months old with acute (< 7 days) diarrhea (≥3 abnormally loose stools in the previous 24 hours) who had ≥1 of the following: sunken eyes, 1318 skin tenting, dysentery, IV rehydration, or hospitalization, and underwent COMPLICATIONS OF MONKEYPOX INFECTIONS IN HUMANS rehydration therapy. A healthy control matched for age and gender was enrolled at home within 3 days of case enrollment. Weight and MUAC Placide K. Mbala1, John Huggins2, Jean-Jacques Muyembe1, were measured for each case and his/her matched control at enrolment Cesar K. Cesar1, Mark Withers2, Bryony Soltis2, James Martin2, and again 4 hours later. To date, 66 cases and 66 controls have been Fernando B. Guerena2, Phillip R. Pittman2 enrolled. Cases had substantial weight gain within 4 hours, which was 1Institut National de Recherche Biomedicale, Kinshasa, The Democratic significantly more than the controls within 4 hours [mean kg 0.29 (0.16, Republic of the Congo, 2U.S. Army Medical Research Institute for 0.42) vs. 0.10 (0.02, 0.19) p=0.02]. In contrast, the change in MUAC Infectious Diseases, Frederick, MD, United States before and after rehydration in cases was comparable to that in controls measured at two time points 4 hours apart [mean cm 0.03 (-0.01, 0.07) We report on the complications of monkeypox infections in humans vs 0.01 (-0.01, 0.04) p>0.09]. These preliminary results suggest that as observed during a 4 year (2007-2011) study at the remote L’Hôpital rehydration therapy results in weight gain but does not affect MUAC, thus Général de Référence de Kole in the rainforest of the Congo River basin providing a useful tool for epidemiologic studies. of the Democratic Republic of the Congo (DRC). The study was conducted jointly by the Institute National de Recherche Biomedicale (INRB) and the 1320 US Army Medical Research Institute of Infectious Diseases (USAMRIID). Human monkeypox infections were first identified during the final stages EMERGING BIOMARKERS FOR THE DIAGNOSIS OF SEVERE of smallpox eradication when laboratory testing determined that some NEONATAL INFECTIONS APPLICABLE TO LOW RESOURCE cases clinically presenting as smallpox were actually caused by monkeypox SETTINGS virus. The present study was conducted at one of the two previous WHO MPX study sites (1981-1986) staffed by the same Spanish medical order Thor A. Wagner1, Courtney A. Gravett2, Janna C. Patterson1, of Catholic sisters who have continued the treatment of MPX patients Sara A. Healy1, Viju Soma1, Michael G. Gravett3, Craig E. Rubens2 after others in the country stopped hospitalizing such cases. A total of 1Seattle Children’s Hospital and University of Washington, Seattle, WA, 244 patients were consented and enrolled into our study. Generally, United States, 2Global Alliance to Prevent Prematurity and Stillbirth, patients presented with fever, chills, sore throat, pox lesions, and general Seattle Children’s, Seattle, WA, United States, 3Global Alliance to Prevent malaise, fatigue. The number of pox lesions ranged from two lesions to Prematurity and Stillbirth, Seattle Children’s and University of Washington, more than eight thousands lesions. Complications included death, coma Seattle, WA, United States and other neurologic manifestations, co-infections, infected wounds, secondary dermatitis, miscarriages, keratitis, staphyloma, and caseation Severe neonatal infections are one of the most significant causes of of eye lesions. The case fatality rate was 0.9% in our study. Case histories pediatric mortality, resulting in more than 500,000 deaths each year, with appropriate graphics, including photographs will be presented, when of which 99% occur in low-resource settings. Compared to clinical appropriate, to demonstrate findings. algorithms, new point-of-care diagnostics that could distinguish neonates with or without severe infections may have potential to substantially improve the global management of severe neonatal infections. This review sought to characterize promising biomarkers for the diagnosis of severe neonatal infections. Biomarkers extensively reviewed elsewhere (procalcitonin, C-reactive protein, tumor necrosis factor-α, interferon-γ, and interleukin-6 and 8) were not re-reviewed. Hundreds of other biomarkers have been associated with “sepsis”; this review focused exclusively on biomarkers with published performance data for the diagnosis of severe neonatal infections. We identified infant diagnostic

www.astmh.org 397 performance data on 21 biomarkers: seven acute phase reactants (Serum 1322 Amyloid A(SAA), LPS Binding Protein(LBP), Inter-α Inhibitor Proteins(Iα- Ip), Antithrombin, Soluble E-Selectin, Fibronectin); five pro-inflammatory QUASISPECIES VARIANT ANALYSIS OF A 2010 DENGUE 3 cytokines (Interleukin-1α, Interleukin-1β, Interleukin-12p70, Interleukin-18, VIRUS FROM KAMPHAENG PHET, THAILAND Granulocyte Colony Stimulating Factor(G-CSF)); two anti-inflammatory 1 1 cytokines (Interleukin-10, Interleukin-1 Receptor Antagonist(IL-1RA)); five Melanie C. Melendrez , Piyawan Chinnawirotpisan , Alongkot 2 1 1 chemokines (Growth Related Oncogene α, Interferon-γ-Inducible Protein Ponlawat , Chonticha Klungthong , Stephen J. Thomas , Robert V. 1 3 4 1 10(IP-10), Monokine Induced by Interferon-γ, Regulated upon Activation Gibbons , Alan L. Rothman , Tim P. Endy , In-Kyu Yoon , Thomas 5 2 1 Normal T cells Expressed and Secreted, Monocyte Chemoattractant 1); W. Scott , Jason H. Richardson , Richard G. Jarman one soluble cell surface marker (soluble intercellular adhesion molecule-1); 1Armed Forces Research Institute of Medical Sciences, Department of and one molecule involved in triglyceride metabolism (apolipoprotein Virology, Bangkok, Thailand, 2Armed Forces Research Institute of Medical CII(apoC2)). Seven soluble biomarkers (G-CSF, IL-RA, IP-10, SAA, LBP, Sciences, Department of Entomology, Bangkok, Thailand, 3Institute for IαIp, apoC2), compatible with point-of-care immunodiagnostics (defined Immunology and Informatics, Department of Cell and Molecular Biology, as a concentration > 1ng/ml), emerged as promising candidates, with University of Rhode Island, Providence, RI, United States, 4Division sensitivity and specificity generally > 90% (range 33 to 100%). These of Infectious Disease, State University of New York, Upstate Medical biomarkers seem particularly attractive for future prospective studies of University, Syracuse, NY, United States, 5University of California, Davis, CA, diagnostics for severe neonatal infections. United States All four serotypes of dengue viruses exist as quasispecies. Quasispecies 1321 are described as a spectrum of variants (‘candidate genomes’), genetically OUTCOME OF FOUR PREGNANCIES IN CONGOLESE WOMEN linked through mutation, creating an interactive population where selection acts on the population rather than the individual variant. We WITH MONKEYPOX INFECTION explored an assertion of quasispecies theory that the fitness (ability to Mark R. Withers1, Placide M. Kingebeni2, Jean-Jacques T. infect and cause disease) of a particular viral sequence is determined Muyembe2, James Martin1, Therese Riu-Rovira2, John Huggins1, more by its freedom to mutate than by its ability to replicate. A Fernando B. Guerena1, Phillip R. Pittman1 quasispecies from dengue virus serotype 3 (DENV3) was cloned from a 1U.S. Army Medical Research Institute for Infectious Diseases, Frederick, single mosquito collected within a cluster of human dengue infections MD, United States, 2Institut National de Recherche Biomedicale, Kinshasa, (100 meter radius) in Kamphaeng Phet, Thailand, in 2010, to understand The Democratic Republic of the Congo diversity and mutational effects apparent in the population. Sequences were combined with other published DENV3 sequences and maximum The outcomes of four pregnancies in women with clinically apparent, likelihood phylogenetic analysis revealed quasispecies populations PCR-confirmed, community-acquired monkeypox (MPX) virus infections removed from the baseline ‘consensus sequence’ diversity of human are described. During 2007 to 2011 we studied the clinical features of DENV3 circulating in Thailand. Mutational analysis showed a high human MPX infections in Kole, the Democratic Republic of the Congo. proportion of nonsynonymous mutations and 2.8% of the population was 244 subjects were enrolled of which four were pregnant. The outcomes evolving faster per site than average despite overall low diversity. Forty- of these four pregnancies along with the maternal pox lesion counts and four percent of the sequences were under positive selection while 19% the PCR-confirmed viremia were carefully documented. This is the first were under purifying selection. Quasispecies analysis identified amino acid report of intrauterine demise due to complications of human monkeypox. substitutions that have been reported to lead to phenotypic changes in In Case 1, MPX viremia rose rapidly and abruptly upon cessation of fetal viral like particle assembly, prM/E protein production, glycosylation and/or movement at the 18th week of gestation, some 24 days after onset of antibody binding ability. Other uncharacterized amino acid substitutions rash. Marked fetal hepatomegaly and peritoneal effusion (hydrops fetalis) identified are predicted to be deleterious. The diversity of the quasispecies were seen at necropsy. In Case 2 a spontaneous miscarriage occurred suggests there are variants with altered abilities to infect and disperse with in a subject without significant viremia or remaining lesions at the 6th overall diversity being constrained in the mosquito. An altered ability to week of gestation, 22 days after rash onset. The third spontaneous infect or disperse will potentially affect how the population responds to miscarriage occurred at 7 weeks of gestation and the 10th study day in a selective pressures such as innate immunity and vaccine implementation. mother with over 1,000 lesions and viremia exceeding 105 genomes/mL of blood. (No pathology examination was performed and no determination 1323 of viral load was made on the aborted material for Cases 2 and 3.) The fourth pregnant subject was enrolled for observation as a healthy family VALIDATION OF DENGUE SEVERITY PREDICTIVE member of an index MPX case. She was then noted to be about 14 weeks ALGORITHMS DERIVED FROM PRIMARY CARE AND pregnant. On her second study day she was noted to have MPX lesions on HOSPITALIZED CASES IN AN ADULT SECONDARY CARE her genitals. Her lesion count never exceeded 20 and she had a low level COHORT viremia by PCR. At her study day 75 follow-up visit (24 weeks gestation), the fetus was alive. Although the sample size is small, we observed a very Victor C. Gan, Chi Jong Go, Tun L. Thein, Yee Sin Leo, David C. high abortion rate in cases of maternal MPX infections, but death is not Lye inevitable. Tan Tock Seng Hospital, Singapore, Singapore Dengue is the most prevalent arthropod-borne infection worldwide. In well-resourced centers where diagnosis can be rapidly established, the next crucial step is to triage for appropriate care. Singapore has primarily adult dengue disease and recent epidemics have led to development of predictors to guide admission to secondary care. We validate three algorithms in a prospective cohort of 137 laboratory confirmed adult cases referred to a hospital-based dengue clinic. Cases that have already fulfilled severity criteria at presentation are excluded from analysis. First, the decision tree classifier developed from a febrile (<=72hrs) primary care cohort with laboratory-confirmed dengue fever, as reported previously: a cycle threshold of real time reverse-transcriptase polymerase chain reaction <=20.9 with positive dengue IgG at presentation, or platelet

www.astmh.org 398 count of <108 000/mm3. Reported sensitivity (Sn) was 78.2% and 1325 specificity (Sp) 80.2% in predicting a platelet nadir of 50 000/mm3. In our cohort, Sn/Sp=88.9%/66.7% in an identically defined subgroup GENETIC DIVERSITY OF DENGUE VIRUS IMPACTS TO THE (n=30), with no significant difference between previously published and DETECTION SENSITIVITY OF RT-PCR BASED METHOD, ONE our Sn/Sp. Second, comparison was made with a decision tree developed CAUTION FOR METHOD DEVELOPMENT AND QUALITY from a retrospective hospitalized cohort to predict dengue hemorrhagic CONTROL ASSESSMENT fever (DHF) (Lee et al Trop Med Int Health. 2009 Sep;14(9):1154-9). Reported Sn/Sp=100%/46% using any of a history of bleeding, serum Chonticha Klungthong1, Piyawan Chinnawirotpisan1, urea >4 mmol/L, or serum protein <=67g/L. In our cohort (n=115), Sn Wudtichai Manasatienkij1, Prinyada Rodpradit1, Ananda Nisalak1, was significantly lower at 85.7% but difference in Sp at 49.4% was Siripen Kalayanarooj2, Robert V. Gibbons1, Richard G. Jarman1 not significant. Last, the predictive equation for DHF using history of 1Armed Forces Research Institute of Medical Sciences, Department of bleeding, serum urea, serum protein and lymphocyte proportion from Virology, Bangkok, Thailand, 2Queen Sirikit National Institute of Child the same cohort (Lee et al, J Clin Virol. 2008 May;42(1):34-9) had Sn/ Health, Bangkok, Thailand Sp=97.6%/60.3%. Our Sp was significantly lower at 32.2% but Sn was Dengue virus (DENV), transmitted by Aedes mosquitoes, causes the not significantly different at 100%. While our cohort was more severe disease in 50-100 million people per year in tropical and subtropical than the hospitalized training cohort and the primary care cohort (24% vs regions worldwide. Four DENV serotypes (DENV-1 to -4) can cause 4% vs 2.6% DHF), it is reassuring that sensitivities remain high. Given the infections ranging from asymptomatic or mild febrile illness to severe wide spectrum of dengue disease and varying presentations in different hemorrhagic disease. Various RT-PCR techniques have been developed populations, a thorough exploration of the utility of prognostic algorithms for rapidly detecting and typing DENV. The rapid diagnosis allows early taking into account population and clinical factors such as time to initiation of patient care and specific preventive health measures. RT-PCR presentation will be required to safely triage dengue patients. We showed is also a useful diagnosis tool for surveillance studies. Our laboratory has that the utility of predictors may vary even within the same country used the modified Lanciotti’s conventional RT-PCR method as one of the depending on source of patients. diagnostic tests for DENV since 1994. The method was evaluated and classified as an optimal method for DENV diagnosis and surveillance by 1324 an international External Quality Control Assessment (EQA). However, RAPID DIAGNOSIS OF DENGUE IN A HOSPITAL-BASED the analysis of PCR results of 13,532 dengue confirmed cases by ELISA COHORT tested over 11 years (2000-2010) showed 17-42% negative PCR results. Among these negative PCR results, 36% were from acute sera from Victor C. Gan, Frederico Dimatatac, Tun-Linn Thein, David C. Lye, patients within 0-4 days of illness onset, typically a viremic period with a Yee-Sin Leo high percentage of virus isolation. We tested 300 samples from patients Tan Tock Seng Hospital, Singapore, Singapore with dengue confirmed by ELISA and with negative conventional PCR results with in-house TaqMan RT-PCR which was classified as a ‘need of Accurate and rapid dengue diagnosis is vital to triage and management. improvement’ method by EQA as part of a quality improvement effort. The World Health Organisation (WHO) proposed in 2009 an updated One hundred and eighty samples (60%) showed TaqMan positive results clinical definition of probable dengue replacing 1997 criteria for including 129 DENV-1 (43%), 39 DENV-2 (13%), 4 DENV-3 (1.3%) and 8 suspected dengue fever. Definitive laboratory diagnosis of dengue is not dual infections of DENV-1 and -2 (2.7%). Sequences of conventional PCR always possible, and newer methods such as testing NS1 antigen are primers binding sites of 9 TaqMan positive samples including 4 DENV-1, 3 undergoing evaluation. We prospectively enrolled 205 adult suspected DENV-2, and 2 DENV-3 revealed points of mismatch between primers and dengue cases referred to the Communicable Disease Centre, Singapore to templates that likely effected the sensitivity of the detection. These data comprehensively evaluate methods for rapid diagnosis of dengue. Clinical indicated that the genetic diversity of DENV impacts the sensitivity of RT- and laboratory criteria were evaluated, including daily PCR, NS1 antigen, PCR based methods, a critical concern during method development and and IgM/IgG serology for those positive by PCR or NS1 on presentation. quality control assessment. Confirmed dengue cases (n=142) were positive by PCR/NS1 or by IgM seroconversion by ELISA at 3-4 weeks. Non-dengue cases (n=20) were 1326 negative by PCR, NS1 and IgM ELISA in paired sera. Forty-three cases could not be assigned an acute dengue diagnosis because of a lack of CONSIDERATIONS FOR CHANGING PRNT DENGUE 4 paired sera or elevated IgM/IgG without seroconversion. The sensitivity REFERENCE VIRUSES: SUB-OPTIMAL IMMUNITY TO (Sn) of PCR at presentation (median fever duration 5 days, range 2-9 days) DOCUMENTED INFECTIONS was 70.4% and specificity (Sp) 100%. Median duration of viremia was 6 days (range 3-11 days). For NS1, Sn/Sp=89.4/100%, with median duration Butsaya K. Thaisomboonsuk1, Richard G. Jarman1, Robert V. of antigenemia of 7 days (range 2-10 days), significantly longer (p<0.001) Gibbons1, Ananda Nisalak1, Chonticha Klungthong1, In-Kyu Yoon1, than median viremic duration. Using <=5 days of fever as a cutoff for early Stephen Thomas1, Timothy P. Endy2, Alan L. Rothman3, Thomas W. illness, the Sn/SP of PCR (n=84) was 88.9/100% vs 51.4/100% late in Scott4 illness (n=78), compared to NS1 of 90.3/100% early and 88.6/100% late 1Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, in illness. Only 2 cases (1.4%) were detected only by IgM seroconversion 2SUNY Upstate Medical University, Syracuse, NY, United States, 3University with negative results by PCR and NS1. WHO 1997 criteria for dengue of Massachusetts Medical School, Worcester, MA, United States, fever had Sn/Sp=98.6/20.0% while the recent 2009 criteria for probable 4University of California, Davis, CA, United States dengue Sn/Sp=97.9/20.0%. Laboratory diagnosis using NS1 antigen had consistently high Sn/Sp, with markedly improved Sn compared with PCR The plaque reduction neutralization test (PRNT) is considered to be the after day 5 of fever (p<0.001), and was positive for a mean of 1.1 days “gold standard” to characterize and quantify circulating levels of anti- longer than PCR. Assessing seroconversion did not substantially increase dengue virus (DENV) neutralizing antibody. The PRNT is used to define the sensitivity of diagnosis in hyperendemic Singapore. Both clinical the immunogenicity of dengue vaccine candidates, support dengue guidelines had similar test characteristics: very sensitive but with poor seroepidemiologic and pathogenesis studies. Despite numerous efforts to specificity in a cohort of referrals for suspected dengue. standardize the assay and normalize data to better compare data between studies and natural and vaccine infections, there are several sets of reference viruses around the world. The Armed Forces Institute of Medical Sciences in Bangkok utilizes DENV-1 (16007), DENV-2 (16681), DENV- 3 (16562) and DENV-4 (1036) reference strains. In 2006 the dominate

www.astmh.org 399 serotype in circulation in Thailand was DENV-4. In cohort studies we 1328 observed poor or absent PRNT titers using the 1036 DENV 4, genotype 3 strain (originally isolated in 1976 in Indonesia) to documented DENV-4 DEVELOPMENT OF A RECOMBINANT TETRAVALENT DENGUE infections. New candidate DENV-4 reference viruses were selected from VACCINES (TDV) THAT LINKS INNATE AND ADAPTIVE isolates collected in the last 10 years. These viruses were tested using a IMMUNITY bank of sera from documented DENV-4 infections including homologous 1 2 3 1 sera from the individuals from which the strains were isolated. A candidate Ge Liu , David Beasley , Justin Julander , Jason Parent , John 1 1 1 1 reference strain was selected based on PRNT titers achieved, low cross Misczak , Hong Li , Langzhou Song , Lucia Reiserova , Shuliu 2 1 1 1 1 reactivity, and the ability of the virus to produce large well-formed plaques. Zhang , Scott Umlauf , Wenzhe Liu , Xiangyu Liu , Haijun Tian , 1 1 1 1 More than 300 samples were tested with the old and new reference virus. Nicole Chartrain , Sumin Cai , Youngsun Kim , Bruce Weaver , 1 Geometric mean titers were increased 4.2 fold. Using the new reference Lynda Tussey virus enabled identification of additional inapparent infections in cohort 1VaxInnate Corporation, Cranbury, NJ, United States, 2Sealy Center for studies and has enhanced our ability to characterize the DENV-4 immune Vaccine Development, University of Texas Medical Branch, Galveston, response. This study illustrates the need to continuously monitor the TX, United States, 3Institute for Antiviral Research, Utah State University, performance of viral strains in reference assays. Furthermore, this data Logan, UT, United States suggests that dengue viral evolution may have a profound effect on tests We have previously demonstrated that the domain III of West Nile virus that utilize reference strains. envelope antigen (EIII) fused to flagellin of Salmonella typhimurium (STF2, a TLR5 ligand) is immunogenic and efficacious against lethal 1327 WNV infections in mice (McDonald et al., 2007, J. Infect Dis. 195, 1607- SAFETY OF A RECOMBINANT LIVE ATTENUATED 1617). To develop a tetravalent dengue vaccine, we have designed, TETRAVALENT DENGUE VACCINE IN HEALTHY ADULT purified, and evaluated similar and alternative flagellin-EIII fusion vaccine formats, which differ in the site of antigen attachment to the flagellin. VOLUNTEERS These fusion proteins can be efficiently and economically manufactured Ivan D. Velez1, Cynthia Thomson2, Aurelia A. Haller3, Liliana in E. coli fermentation systems. Here we report immunogenicity results Lopez1, Esteban Echavarria4, Claire Y. Huang5, John Arguello3, of recombinant dengue vaccine candidates in monovalent, bivalent, Steven Erb3, Gilad Gordon3, Joseph Santangelo6, Mark E. Beatty7, and tetravalent formulations. BALB/c mice were immunized s.c. three Dan T. Stinchcomb3, Jorge E. Osorio8 times at 2 or 3 week intervals, and bled at various times post boost. 1Universidad de Antioquia, Medellin, Colombia, 2Inviragen, Singapore, In an efficacy study, AG129 mice lacking receptors of types I and II Singapore, 3Inviragen, Fort Collins, CO, United States, 4Clinica Las interferons were immunized with two or three doses of a monovalent DENV-2 vaccine candidate, and challenged with 2,100 LD of DENV-2 Americas, Medellin, Colombia, 5Centers for Disease Control and 50 Prevention, Fort Collins, CO, United States, 6Iniviragen, Singapore, (strain NGC). Serum neutralizing antibody titers were determined by 50% plaque reduction neutralization test (PRNT ). Survival rates, weight Singapore, 7Dengue Vaccine Initiative, Seoul, Republic of Korea, 8University 50 of Wisconsin, Madison, WI, United States changes, and viremia, as measured by qRT-PCR, of infected mice were determined. Our results indicated that immunizations of BALB/c mice Dengue (DEN) virus threatens over half the world’s population, causing with these vaccine candidates at doses of 2-15 μg elicit robust homotypic debilitating dengue fever, dengue hemorrhagic fever and dengue neutralizing antibody responses. Furthermore, a monovalent DENV-2 shock syndrome leading to over 20,000 deaths every year. DENVax is a candidate conferred partial protection against a lethal DENV-2 challenge tetravalent live attenuated dengue vaccine that is based on the DEN-2 and significantly reduced viremia and weight loss in infected AG129 PDK-53 genetic backbone. DEN-2 PDK-53 has been tested previously mice. The DENV-2 candidate was also found to elicit high PRNT50 titers in in humans and was found to be safe and immunogenic. Recombinant rabbits. Finally, BALB/c mice immunized with tetravalent dengue flagllin-EIII DENVax-1, DENVax-3 and DENVax-4 strains were generated in which the formulations developed strong neutralizing antibodies to all 4 serotypes of prM and E genes of PDK-53 were substituted with those of DEN- 1, -3 DENV (GMTs of PRNT50 = 200 - 3000). In conclusion, VaxInnate flagellin- or -4 viruses. These recombinant viruses retain the genetic attenuation EIII vaccine candidates are highly immunogenic in mice and rabbits and markers present in PDK-53 and direct the immune response to the other are effective in protecting AG129 mice against a lethal DENV-2 challenge, three serotypes. A single center, placebo-controlled, randomized study thereby justifying further development of a TDV. assessing the safety, tolerability of tetravalent DENVax formulations was performed in Rionegro, Colombia, a high altitude area with no Aedes 1329 aegypti and no dengue exposure. One of two dose levels (low or high) of DENVax was administered subcutaneously or intradermally to healthy IDENTIFICATION OF HOST FACTORS THAT INFLUENCE male and female subjects with no pre-immunity to flaviviruses. Two doses DENGUE VIRUS INFECTION IN HUMAN PRIMARY of DENVax or placebo were administered, separated by an interval of MONOCYTES AND MONOCYTE-DERIVED DENDRITIC CELLS 90 days. Safety was assessed as the frequency and severity of adverse events through physical examination, injection site examination, lab Timothy J. Savage, Dabeiba Bernal-Rubio, Avelino Teixeira, Ana examinations, and subject diary cards. Clinical laboratory assessments Fernandez-Sesma included serum chemistry, hematology and urinalysis. The safety data Mount Sinai School of Medicine, New York, NY, United States demonstrate that both tetravalent formulations were well-tolerated by Dengue virus (DENV) is a flavivirus in the family flaviviridae that infects either route of administration. To date, the most frequent adverse events up to 50-100 million people per year, with 2.5 billion people at risk. were local reactogenicity at the injection site for both dose levels and both The burden of disease is significant, with a clinical primary infection routes of administration. Systemic adverse events were mild to moderate manifesting as fever, rash, severe headaches, and intense myalgia and headache, muscle pain, nausea and fatigue. There were no meaningful arthralgia that persist for approximately one week. Elucidating the laboratory changes. This study highlights the safety of the tetravalent interactions between host cell proteins and the dengue virus is critical to DENVax formulations in healthy adults. Further clinical trials to assess the development of targeted and effective antiviral drugs. Learning the safety, tolerability, and immunogenicity in other age groups and in dengue details of these interactions will be essential to be able to rationally design exposed individuals are being planned. drugs targeting viral proteins, or to identify compounds that will interfere with host processes critical to DENV infection. The first step towards this end is to identify which host proteins interact with the dengue virus in a clinically relevant system. Proteomic evaluations of host cells following

www.astmh.org 400 dengue virus infection have been performed in liver cells and endothelial (GST). The anti-apoptotic effect plays a role as the second defense system cells but not the described primary target of dengue virus infection, on protection of mosquito cells from DENV infection. It was eventually primary human monocytes and dendritic cells. We used a proteomics- regulated by inhibitors of apoptosis (IAPs) that are the upstream regulators based approach to identify host factors relevant to dengue virus infection of caspase 9 and caspase 3. C6/36 cells with double knockdown of GST in primary human monocytes and monocyte-derived dendritic cells. After and IAP showed a synergistic effect on activation of caspases, causing a infecting these cells with DENV serotype 2 strain 16681 we compared higher rate of apoptosis rate (>20%) than those with knockdown of each their proteome to that of uninfected cells using the Beckman Coulter PF2D single gene (~10%), after infection by DENV. Compared with mammalian system, a fluid-based system analogous to a 2D-gel that separates proteins cells, residual H2O2 after anti-oxidation in DENV-infected C6/36 cells may by isoelectric point (pI) followed by hydrophobicity. After comparing serve as the signal up-regulating the expression of IAP. Taken together, two infected cells to uninfected cells we found approximately 75 proteins that defense systems including antioxidant defense and anti-apoptotic effects either increased or decreased in abundance by greater than 2.5 fold in exist in mosquito cells; which were linked by ROS, i.e., H2O2 signaling. the presence of DENV. These unidentified proteins were then subjected to Mass Spectrometry analysis. Proteins down-regulated in the presence 1332 of DENV in both monocytes and monocyte-derived dendritic cells were chosen for further analysis to elucidate their role in the pathogenesis of EPIDEMIOLOGY OF DENGUE IN MALAYSIA dengue virus. Balvinder S. Gill1, Xiao Jianguo2, Dagwin Luang-Suarkia1, Dewi 1 1 3 1 1330 Suryani , Geoffrey Shellam , David Smith , Allison Imrie 1The University of Western Australia, Crawley, Australia, 2Epidemiology SEROTYPE-SPECIFIC DENGUE VIRUS CIRCULATION AND Branch, Department of Health Western Australia, Perth, Australia, 3The DENGUE DISEASE IN BANGKOK, THAILAND FROM 1973 TO University of Western Australia, PathWest Laboratory Medicine, Western 2010 Australia, Crawley, Australia Ananda Nisalak1, In-Kyu Yoon1, Siripen Kalayanarooj2, The first major epidemic of dengue fever in Malaysia occurred in 1973, Chonticha Klungthong1, Butsaya Thaisomboonsuk1, Butsaya and since that time dengue epidemics have become more frequent, Thaisomboonsuk1, Piraya Bhoomiboonchoo1, Richard Jarman1, and more virulent. We analyzed data collected by the Ministry of Health Robert V. Gibbons1 Malaysia between 2001-2010 and describe increasing incidence from 68.2/100,000 in 2001 to 159.7/100,000 in 2010, with a spike of 1 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 176.5/100,000 in 2008. Analysis of surveillance and notification data 2 Queen Sirikit National Institute of Child Health, Bangkok, Thailand collected between 2005-2010 showed that the DHF/DSS:DF ratio was Since 1962, the Queen Sirikit National Institute of Child Health (QSNICH) 1:19 in 2005 and 1:10 in 2010, with higher rates of severe disease in and AFRIMS have cooperated in a mutual public health effort to secondary versus primary infections. The mean age of DHF/DSS cases was accurately diagnose dengue infections including serologic determinations 28 years. Age-specific incidence was highest in adults aged 20-29 years of antibody patterns and identification of dengue serotypes. The and incidence rates were higher in males, with a male to female rate ratio epidemiologic data included all patients admitted to the dengue ward of 1.397 (95% CI: 1.390 - 1.404; p<0.005). Between 2005-2010 there of QSNICH with suspected dengue fever and dengue hemorrhagic fever was a shift to increased transmission in urban settings, with an increase in who were subsequently proven to have dengue infection by serology or urban:rural rate ratios from 1.5 in 2005 to 2.0 in 2009, based on urban virus detection. Available data from 1973 t0 1999 have been analyzed incidence rates of 170.4/100,000 compared to 98.7/100,000 in rural areas and published previously (Nisalak et al, 2003). We report on data for with an urban and rural rate ratio of 1.727 (1.719 - 1.735; p<0.005). the expanded years from 1973 to 2010 including many more cases of Analysis of approximately 700 virus isolates collected between 2005-2010 DENV-4 infection than were observed previously. Findings that were showed that all 4 DENV serotypes circulated in Malaysia during this period, reconfirmed from the previous report: 1) primary cases are increasing with all four serotypes detected in each year. DENV-1 genotype I, a virus relative to secondary cases; 2) symptomatic primary cases were most which has circulated in Southeast Asia since 2003, reemerged as the likely due to DENV-1; 3) Primary non-infant hospitalized cases were less predominant serotype in 2010. We describe a marked increase in dengue severe than secondary non-infant hospitalized cases. The mean age of epidemic activity in Malaysia within the last decade, characterized by DHF cases are noted to be increasing. In 1973-1982, the mean age of hyperendemic transmission of all four dengue serotypes, increasing rates primary and secondary infection was 4.5 years and 8.0 years, respectively. of severe disease and increasingly urban transmission. In 2001-2010, it was 6.1 years and 8.0 years. These findings highlight the longitudinal epidemiology of dengue over a uniquely extended period of 1333 observation. Further spatial analysis is planned to elucidate transmission dynamics. AN ISLAND-WIDE DENGUE EPIDEMIC - PUERTO RICO, 2010 Tyler M. Sharp, Aidsa Rivera, Rosa Rodriguez-Acosta, Jorge L. 1331 Munoz-Jordan, Elizabeth Hunsperger, Luis M. Santiago, David F. Arguello, Harold S. Margolis, Kay M. Tomashek THE ROLE OF ROS SIGNALING IN MOSQUITO CELLS THAT SURVIVE DENGUE 2 VIRUS INFECTION Centers for Disease Control and Prevention, San Juan, PR, United States Dengue, a potentially fatal febrile illness caused by four mosquito- Tien-Huang Chen, Chao-Fu Yang, Shih-Hui Suen, Lin-Hsien Kao, transmitted dengue viruses (DENV-1-4), is endemic in Puerto Rico. In Wei-June Chen January, 2010, the number of suspected dengue cases reported to the Chang Gung University, Tao-Yuan, Taiwan Puerto Rico Department of Health/CDC passive dengue surveillance Dengue virus (DENV) is naturally transmitted by Aedes mosquitoes system exceeded the epidemic threshold. To characterize this epidemic, between humans and replicates efficiently in mosquito as well as in surveillance data were used to describe all reported cases. Suspected mammalian cells. However, the fate is distinct between the two types of cases were patients with a serum specimen submitted for dengue testing. cells in response to the infection. Cytopathic effects (CPE) in mosquito Laboratory-positive cases had (i) DENV identified via reverse transcriptase cells are generally trivial compared to that occur in mammalian cells that polymerase chain reaction (RT-PCR) in an acute specimen, and/or (ii) usually end up with apoptosis. In spite, production of ROS resulted from anti-DENV IgM detected in a convalescent specimen. Laboratory- mitochondria dysfunction occurs in both cell types. It was demonstrated negative cases had no anti-DENV IgM in a convalescent specimen and that the survival of mosquito cells is beneficial form up-regulation of an acute specimen that was either RT-PCR-negative or not submitted. genes related to antioxidant defense, such as glutathione S transferase Indeterminate cases were RT-PCR-negative in an acute specimen and had www.astmh.org 401 no convalescent specimen submitted. In 2010, 23,622 suspected dengue 1335 cases were reported, of which 10,956 (46.4%) were laboratory-positive, 2,588 (11.0%) were laboratory-negative, and 9,999 (42.3%) were ENHANCED SURVEILLANCE FOR FATAL DENGUE IN PUERTO indeterminate. Of 7,424 RT-PCR-positive specimens, DENV-1 (69.0%) and RICO DENV-4 (23.6%) were detected more frequently than DENV-2 (7.3%) and 1 1 1 DENV-3 (<0.1%). Of all laboratory-positive cases, nearly half (46%) were Aidsa Rivera , Kay Tomashek , Jorge Muñoz , Elizabeth 1 2 1 1 adults ≥ 20 years of age, 4,173 were hospitalized, and 254 had met the Hunsperger , Dianna Blau , Tyler Sharp , Jomil Torres , Infectious 2 3 3 1998 WHO definition of dengue hemorrhagic fever. Enhanced surveillance Diseases Pathology Branch , Lorenzo González , Carmen Deseda , 4 4 4 4 detected 38 laboratory-positive dengue deaths, yielding an incidence of Irma Rivera , Darío Sanabria , José Torres , Rosa Rodríguez , Javier 4 4 4 1 3.5 laboratory positive deaths per 1,000 laboratory positive cases; 89% Serrano , Francisco Dávila , Daniel López , Harold Margolis of these deaths were in adults. The 2010 epidemic was long in duration, 1Centers for Disease Control and Prevention Dengue Branch, San Juan, high in magnitude, and resulted in the most dengue-related deaths since Puerto Rico, 2Centers for Disease Control and Prevention Infectious surveillance for dengue began in Puerto Rico in the late 1960’s. For this Diseases Pathology Branch, Atlanta, GA, United States, 3Puerto Rico reason and using lessons learned from the 2007 epidemic in Puerto Department of Health, San Juan, Puerto Rico, 4Institute of Forensic Rico, CDC implemented an initiative to train Puerto Rico clinicians in the Sciences of Puerto Rico, San Juan, Puerto Rico management of dengue patients to minimize morbidity and mortality in Dengue has been endemic in Puerto Rico (PR) for four decades, and data future epidemics. suggests disease severity increased. Trends are monitored by the passive dengue surveillance system (PDSS) operated by the Centers for Disease 1334 Control and Prevention’s (CDC) Dengue Branch and the PR Department SEROLOGICAL SURVEY OF DENGUE INFECTIONS AMONG of Health (PRDH). Suspected cases, including fatals, are reported to the PRDH or the PDSS, and reporting to PDSS requires submission of a serum INDIVIDUALS IN RAYONG, THAILAND sample. Prior to 2010, fatal cases were also detected by review of death Rome Buathong1, Isabel Rodríguez-Barraquer2, Sopon certificates (DC) that had dengue as cause or contributing cause of death. Iamsirithaworn3, Justin T. Lessler2, Richard G. Jarman4, Robert V. However, a 2007 evaluation of PDSS found limited ability to detect deaths Gibbons4, Derek A. Cummings2 because it collects data early in the course of disease and few clinicians 1Bureau of Epidemiology, Ministry of Public Health Thailand, Nonthaburi, revise the PDSS report when a patient dies (<10%). In addition, <50% of Thailand, 2Johns Hopkins Bloomberg School of Public Health, Baltimore, laboratory-confirmed deaths identified by PDSS had “dengue” on their MD, United States, 3Bureau of Epidemiology, Ministry of Public Health, DC. Deaths are difficult to diagnose as many die on day 4 or 5 of illness Nonthaburi, Thailand, 4Armed Forces Research Institute of Medical when standard diagnostic tests are often unable to detect dengue virus Sciences, Bangkok, Thailand (DENV) or IgM anti-DENV. Although tissue diagnosis is more sensitive in fatal cases, <40% of cases in 2007 had tissue submitted and collection Dengue and DHF have been a major public health problem in Thailand was not systematic. To improve detection and diagnosis of fatal dengue over the past 50 years. Clinical dengue (DF and DHF) has traditionally cases, CDC developed an enhanced surveillance system in collaboration affected children with rare cases among adults. Even though the incident with Institute of Forensic Sciences of PR and CDC Infectious Diseases number of DHF cases does not seem to have decreased, a shift towards Pathology Branch. All patients who die with a dengue-like, acute febrile older age groups has been observed over the past years. The reasons illness are identified through weekly calls to hospitals and at death for this shift have not been elucidated. We report the results of an age- investigation, autopsy, or DC review, and tissue specimens and autopsy stratified serological study conducted among school aged children living findings are collected. During 2010 epidemic, 122 suspected fatal cases in the district of Mueang Rayong in Rayong, Thailand. Schools and were identified of which 38 were laboratory-confirmed, more than twice classrooms were sampled probabilistically from all schools serving the the number previously identified. The majority (65%) of suspected cases district. A total of 1812 children (approximately 140 per age group) from submitted tissue, a higher percent than in any other year which notably 25 schools were enrolled and provided a blood sample. Samples were reduced the proportion of laboratory-indeterminate cases. Only 16% of analyzed using single dilution neutralization testing (SDNT), an assay that the 38 laboratory-confirmed cases had dengue on their DC. In spite of differentiates between primary and secondary infection and is serotype conventional wisdom, dengue deaths appear to be under-reported even in specific for those subjects that have only been exposed to one dengue endemic areas. serotype. Preliminary results (n=720) show that 72% (95%CI 61-82%, n=71) of children have been exposed to dengue by age 10 years and that 1336 16% (95%CI 2-30%, n=25) of children have only undergone primary exposure by 18 years of age. These results are significantly different DENGUE FEVER DURING THE 2005 AND 2007 EPIDEMICS IN from a similar study conducted by Sangkawibha et al. in Rayong in PUERTO RICO: EXPERIENCE OF A TERTIARY LEVEL HOSPITAL 1980, where 97% (95%CI 93-100%, n=65) of sampled children were seropositive by age 10 years. This change is consistent with an overall Luisa I. Alvarado, Ivonne E. Galarza, Veronica Colón, Dimarys decrease in transmission intensity (force of infection) of dengue in Rayong Sanchez over the last years. Analysis of the full dataset will explore geographic Hospital Episcopal San Lucas, Ponce, Puerto Rico heterogeneity and factors associated with seropositivity. Dengue Fever is now the most important arthropod borne disease worldwide. It is caused by four serotypes of the Dengue virus. It can be a non-specific febrile illness without complications or it can progress to severe disease with plasma leaking, shock, bleeding and severe organ damage. In Puerto Rico there have been island wide epidemics since 1915 and these have increased in frequency and severity over the past 20 years. Until 2010, the 2007 epidemic had been one of the largest and most severe, presenting many diagnostic and management challenges. The goal of our study was to characterize the epidemiologic and clinical features of pediatric Dengue Fever admissions to the Hospital Episcopal San Lucas in Ponce during the 2007 epidemic and compare with the 2005 outbreak. The study was based at Hospital Episcopal San Lucas, a tertiary level hospital in southern Puerto Rico from January to December of 2007. Pediatric residents reviewed the records of 163 cases with a diagnosis www.astmh.org 402 of Viral Illness with Thrombocytopenia that met the World Health 1338 Organization definition of Dengue Fever. The study included a comparison with Dengue Fever admissions (n= 71) during the 2005 outbreak, when A RETROSPECTIVE GEOCODING STUDY ON THE IMPACT OF data was available. Female/male distribution was 49%/51% in 2007 and URBANIZATION ON INCIDENCE RATES OF DENGUE FEVER 44%/56% in 2005. About 90% of patients were between the ages of WITHIN BORNEO, KUCHING DIVISION, FROM 2009 TO 2010, 3 to 18 years in 2007 and in 2005. Platelet counts under 50,000 were USING ArcGIS more frequent in 2005 (74.3% versus 38.7%). During the 2007 epidemic 100% patients presented with a history of fever, 59% had vomiting Karen S Dindial and 39% had abdominal pain. Maintenance hydration therapy in 2007 University of South Florida, Tampa, FL, United States included fluids that are considered hypotonic in 65% and isotonic in 16% of cases. Our study reveals that the pediatric age group most frequently Review of available data indicates that the impact of urbanization has not affected in both epidemics was the school age and adolescent. Difficulties been extensively analyzed as it effects disease transmission of Dengue. in the diagnosis during the 2007 epidemic may be related to less Understanding urbanization, its meaning on the vector habitat, and how thrombocytopenia and the presence of gastrointestinal symptoms. Plasma it contributes to disease transmission is vital. The districts of Bau, Lundu leaking complications in 2007 may be due to disease severity complicated and Kuching, all have documented cases of Dengue and the repository of by hypotonic hydration fluids. epidemiological data at the Divisional Health Office in Kuching, Sarawak is extensive. By incorporating Geographic Information Systems (GIS) to retrospectively analyze urbanization within these districts of Sarawak, and 1337 their cases of DF and DHF from January 2009 to June 2010, projections of EVALUATION OF COMMERCIALLY AVAILABLE DENGUE the impact of urbanization on the vector distribution and density can be DIAGNOSTIC TESTS: NS1 AND IgM RAPID TESTS AND NS1 extrapolated and aid in the development of future prevention strategies. ELISAs Spatial and descriptive analyses methods were used as was two sample t-test. Specific to Kuching District there is a direct correlation between Elizabeth A. Hunsperger1, Sutee Yoksan2, Philippe Buchy3, urbanization and an increase in the vector responsible for dengue. Vinh Chau Nguyen4, Shamala Devi Sekaran5, Delia Enria6, Susana Additionally, urban areas experienced higher rates of disease transmission. Vazquez7, Elizabeth Cartozian1, Harvey Artsob8, Maria Guzman7, Three areas of the Kuching, Sarawak Division will be selected; Kuching, Susie Kliks9, Rosanna Peeling10, Martine Guillerm10, Julien Zwang10, Bau and Lundu Districts. The selection criterion is based on the presence Harold Margolis1 of Aedes mosquito breeding, and the frequent reporting of cases within 1Centers for Disease Control and Prevention, San Juan, PR, United States, these areas. All clinically diagnosed dengue cases, reported to the Health 2Mahidol University, Bangkok, Thailand, 3Institut Pasteur, Phnom Penh, Office from Jan 2009 - Jan 2010, from Kuching Division will be included Cambodia, 4Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam, for GIS geocoding. Only geocodable addresses using ArcGIS will be 5University of Malaya, Kuala Lumpur, Malaysia, 6Instituto Nacional de included. An interactive method using data abstraction from case records, Enfermedades Virales Humanas “Dr. Julio I Maiztegui”, Pergamino, use of additional maps and street reference will be used to determine Argentina, 7Instituto de Medicina Tropical “Pedro Kouri”, Havana, Cuba, geocode within ArcGIS. For Kuching Division, an overall description 8Public Health Agency of Canada, Winnipeg, MB, Canada, 9Pediatric of area (urban, semi-urban or rural), basic facilities, and geographical Dengue Vaccine Initiative, Seoul, Republic of Korea, 10World Health profiles, population, population density, and average annual population Organization, Geneva, Switzerland growth rate as provided by the Malaysian Census will be included. Incorporation into ArcGIS will be based on census tract files availability The World Health Organization (WHO) and the Pediatric Dengue Vaccine for 2009 - 2010. Clearly, supplemental GIS studies are warranted and Initiative (PDVI) established a network of 7 worldwide laboratories with should include habitat, climate and soil moisture modeling and variability dengue diagnostic expertise to provide an independent evaluation of in entomological parameters. In order to mitigate further disease currently available commercial kits for dengue diagnostics. Each laboratory transmission, an integrated approach is required for the endemic areas contributed serum samples to develop a well characterized panel for and developing areas of SE Asia. testing dengue non-structural protein 1 (NS-1) antigen and dengue virus IgM antibodies (IgM anti-DENV). Asia and America region panels of similar 1339 composition were developed to evaluate NS1 and IgM anti-DENV test kits. The NS1 combined panel consisted of 192 sera from 147 patients CHARACTERIZATION OF DENGUE FEVER IN SCHOOL defined as dengue positive and 142 negatives by culture or PCR. The IgM CHILDREN IN MEDELLIN, COLOMBIA anti-DENV panel had 228 positive and 155 negative sera as defined by 1 2 1 reference MAC-ELISAs at Mahidol University and CDC. Seven companies Diana Piedrahita , Jorge E. Osorio , Juliana Duque , Ivony 1 1 1 1 submitted 3 NS1 microplate ELISAs, 1 IgM microplate ELISA, 4 NS1 rapid Agudelo , Ruth Ramirez , Gabriel J. Parra , Berta N. Restrepo diagnostic tests (RDT) and 4 IgM anti-DENV RDTs for evaluation. All kits 1Instituto Colombiano de Medicina Tropical-Universidad CES, Sabaneta, were evaluated at the network sites using region-specific panels. The Colombia, 2University of Wisconsin, Madison, WI, United States panel was coded so that technicians performing the evaluation were Dengue fever is the arboviral disease with the most significant impact in blinded to the reference assay results. Evaluation results were analyzed to public health. In Colombia, the largest outbreak ever recorded occurred determine sensitivity, specificity, inter-laboratory agreement, inter-reader in 2010 with at least 151,774 cases. Understanding the factors and rates agreement, lot-to-lot variation and ease-of-use. Preliminary results showed of transmission in schoolchildren are needed towards characterizing the that the 3 NS1 ELISAs had sensitivities ranging 52-46% and specificities of burden of disease in the community and defining strategies for epidemic 71-80 %. In comparison, the NS1 RDT had sensitivities ranging 28-59% control. In this study, the incidence, seroprevalence and circulating and specificities of 71-76%. Of the 4 IgM RDTs, sensitivities ranged 52- serotypes of DENV were determined in schools from three different 95% with specificity from 83-90%. Sensitivity of IgM ELISA was 96% and neighborhoods of Medellin (San Javier, Poblado and Laureles). A cohort specificity 84%. The range of acceptable sensitivity or specificity from the containing 2,340 volunteer students from two public and one private combined Asia/America panels is being considered by a panel of experts school including primary and high schools (ages 5-19) was established. convened by WHO/PDVI. The threshold for acceptable performance In the cross sectional study, blood samples were obtained from all may vary by the purpose of testing and by local epidemiology and final admitted students and specific dengue IgM ELISA were performed. recommendations will be distributed as a report upon completion of the The longitudinal study involved surveillance of absenteeism of enrolled analysis. students due to febrile illness shorter than 7 days. Standardized physical exam were performed and venous blood samples were obtained from ill

www.astmh.org 403 students during both acute and convalescent stages. Dengue diagnosis 1341 was confirmed using RT-PCR and IgM ELISA. Among the 2340 students enrolled, 53% were women and students of all grades were represented. ASSOCIATION BETWEEN PRE-EXISTING DENV ANTIBODY In the cross sectional study, 69 (2.9%) students were positive for IgM AND THE OCCURRENCE OF SYMPTOMATIC ILLNESS DUE TO antibodies. Their mean age was 11.4 years (range = 5 to 19 years) and the DENV-4 INFECTION, IQUITOS, PERU distribution of cases by sex was the same. In the longitudinal study, among 1 2 1 the 146 students declared ill because of absenteeism due to febrile illness, Sandra M. Olkowski , Brett M. Forshey , Steven T. Stoddard , 3 2 3 12 (8.2%) had IgM antibodies against dengue and DENV-1 serotype was Amy C. Morrison , Stalin Vilcarromero , Kanya C. Long , Eric S. 4 5 6 detected by RT-PCR in three of them. The highest frequency of DENV Halsey , Moises Sihuincha Maldonado , Carlos Alvarez , Tadeusz J. 7 1 seropositive cases was detected in San Javier’s school (3.9%), followed Kochel , Thomas W. Scott by the Laureles’ school (3.4%). In this first phase of the study, a high 1University of California, Davis, Davis, CA, United States, 2Naval Medical incidence of dengue fever was found in school children mirroring the large Research Unit Six, Iquitos, Peru, 3University of California, Davis; Naval outbreak experienced in 2010. Medical Research Unit Six, Iquitos, Peru, 4Naval Medical Research Unit Six, Lima, Peru, 5Hospital de Apoyo Iquitos, Iquitos, Peru, 6Dirección Regional 1340 de Salud, Iquitos, Peru, 7Naval Medical Research Center, Washington, DC, United States ASSOCIATION OF POLYMORPHIC VARIANTS IN TNF-α, IL-6 Dengue fever is caused by infection with any of four distinct viral serotypes AND IFN-γ GENES IN PATIENTS AFRO-DESCENDANT AND (DENV-1 through DENV-4). Antibody induced by infection with one MESTIZOS WITH DENGUE INFECTION, COLOMBIA serotype can influence the clinical outcome of subsequent infections. In Berta N. Restrepo1, Efren Avendaño2, Juan C. Chacon2, Winston general, however, details of modifying effects are poorly understood, even Rojas2, Piedad Agudelo1, Ruth Ramirez1, Gabriel Bedoya2 though they are potentially critical determinants of transmission dynamics 1Instituto Colombiano de Medicina Tropical-Universidad CES, Sabaneta, and disease severity. We analyzed data from an on-going longitudinal Colombia, 2Universidad de Antioquia, Medellin, Colombia study of DENV transmission in Iquitos, Peru, to evaluate the relationship between infection history and disease outcome. Iquitos has been the site Dengue is an important problem of health public in tropical and sub- of intense DENV transmission since the early 1990s, with large outbreaks tropical countries. On the other hand, the response to dengue infection due to DENV-1, DENV-2 and DENV-3 since then. In 2008, DENV-4 was is influenced by the genetic background of the host. In this study was introduced into the city and became the dominant serotype over the evaluated the association of polymorphic in TNF-α, IL-6 and IFN-γ 2008/9 and 2009/10 transmission seasons (>99% of all cases). During genes between two ethnic groups with dengue. The study was carried that time, 1,397 participants in the longitudinal cohort seroconverted to in Antioquia and Chocó, two departments of Colombia. The study DENV-4 (35% of the study population). Of these, 5.7% experienced a population consisted of 122 Afro-descendents patients and 104 mestizos clinical case of dengue fever as detected by active door-to-door febrile patients with dengue infection. The ethnic group was based using 19 surveillance. No effect of pre-existing antibodies against heterologous ancestry informative markers (AIMs). The clinical form more frequently DENV serotypes was observed on DENV-4 seroconversion rates. However, was dengue fever (90.3%). Comparisons between ethnic groups showed we observed that primary and secondary infections resulted in a higher significant differences. In Mestizos was significantly more frequently the rate of symptomatic illness (8.7% and 9.0%, respectively) than third cases of dengue hemorrhagic fever (16.3% vs. 4.1%, p=0.003), more infections and fourth infections (3.1% and 3.3%, respectively). These patients had thrombocytopenia (66.7% vs. 47.3%, p=0.012) and more data suggest that although pre-existing antibodies did not confer sterile patients were hospitalized (63.5% vs. 23.8%, p< 0.000) compared immunity to heterologous serotypes, there may have been a cumulative with afro-descendents patients. The difference between the average of protective effect against symptomatic illness. Further analysis is being ancestral component was obtained with an ANOVA, showing that the conducted on the effects of specific serotypes, levels of neutralizing European component had effect above IL6 genotypes distribution, G/A antibodies and time interval between infections. (0.213 ± 0.135) G/G (0.3 ± 0.216) p=0.038. The African component was higher in dengue fever (0.526 ± 0.26) than in dengue hemorrhagic fever 1342 (0.376 ± 0.234), it confers protection until 50% (OR=0.49; IC=0.27- 0.9; p=0.023). Of the 5 candidate loci (IL64589, TNF -376, CD209-336, CIRCULATION OF DIFFERENT LINEAGES OF DENV-2 IN INF-Y4100, INF-Y 78), in INF_Y G4100T and TNFα G376A, minor allele GUATEMALA DURING RECENT DENGUE EPIDEMICS, THEIR frequency were lower to 5%; at this respect, in an association analysis, EVOLUTIONARY TIME-SCALE AND SELECTION PRESSURE was compared allelic and genotypic frequencies, finding significant ANALYSIS differences for IL6 4589 locus between predefined disease groups, been homozygous G/G frequency higher in DF than in DHF (83% vs. 68%, Germán Añez1, Maria E. Morales-Betoulle2, Maria Rios1 respectively). This was the first evidence that IL6 polymorphism can be 1U.S. Food and Drug Administration, Bethesda, MD, United States, implied in protection/susceptibility to the infection for dengue virus. These 2Universidad del Valle de Guatemala, Guatemala, Guatemala results provide evidence about the susceptibility genetic to the infection Dengue is the most common arboviral disease worldwide. Dengue is for dengue virus. However further studies are still necessary. caused by dengue virus (DENV), which exist in nature as a complex of four different viruses or serotypes (DENV-1 to 4), belonging to the genus Flavivirus. DENV serotypes have been classified into different genotypes based on phylogenetic analysis from sequences of different viral regions. Dengue is endemic in Central America and is present in Guatemala since at least 1978. Here we report the phylogenetic relationships of the first fully sequenced DENV-2 from Guatemala (GU/FDA-GUA09/2009) with strains representing all known DENV-2 genotypes. Phylogenetic analysis of the envelope (E) protein and whole coding region sequences by maximum-likelihood and Bayesian inference methods revealed that at least two lineages of the American/Asian genotype of DENV-2 have circulated in that country during the 2007 and 2009 epidemics, and have possibly co-circulated during these and other epidemic periods. We found that the time to most recent common ancestor for Central

www.astmh.org 404 American DENV-2 of American/Asian genotype existed about 18 years 1344 ago, and Bayesian Skyline analysis revealed that the genetic diversity of this DENV-2 genotype in the region has increased since 2005. Site-specific PHYLODYNAMICS AND CHARACTERIZATION OF NATURAL selection pressure analysis revealed positive selection in the NS2A, 4B ATTENUATION IN A SOUTH PACIFIC DENV-2 OUTBREAK and 5 proteins but none in E protein. The study of dengue evolution in 1 2 1 endemic regions is of importance, since nucleic acid technology (NAT) Argon Steel , Duane J. Gubler , Shannon N. Bennett assays has been developed and implemented in the detection of DENV 1University of Hawaii at Manoa, Honolulu, HI, United States, 2Program in these countries, as well as in places in which the disease is imported on Emerging Infectious Diseases, Duke-NUS Graduate Medical School, and can cause autochthonous transmission, as it happened recently in Singapore, Singapore the US. Even though primers and probes for these assays are designed Dengue is an arboviral disease that has seen a recent increase in activity to target the most conserved regions of the viral genome, there is always throughout the tropics in recent decades, marked by more frequent and a risk that the assay could fail to detect variants with mutations located severe epidemics. While the causes of this reemergence are probably at the target area. Therefore, the use of whole genomic sequences in multifold, including geographic expansion of both vectors and viruses, the molecular epidemiology studies appears to be more suitable for identifying importance of virus strains with greater fitness, epidemic potential and mutations that may occur throughout the viral genome, some of which possibly virulence, has been implicated. This prompted us to investigate could potentially impact the performance of detection assays. the role of virus molecular evolution in driving epidemics. Our study was a series of outbreaks of American genotype DENV-2 in the South Pacific 1343 beginning in 1971 in Tahiti and Fiji, which became increasingly severe EVALUATION OF LOW DOSE MONOVALENT DENGUE in New Caledonia and Niue Island in 1972. In Tonga in 1974, however, it became dramatically attenuated, with near-silent transmission. To VACCINES IN HUMAN VOLUNTEERS elucidate the relative contribution of viral genetic change in outbreak Kristen K. Pierce1, Beth D. Kirkpatrick1, Anna P. Durbin2, Janet dynamics, we conducted whole-genome phylogenetic analysis of DENV- C. Lindow1, Marya P. Carmolli1, D. Shaffer2, Elizabeth R. Colgate1, 2 strains collected during the South Pacific sweep paired with in vitro Catherine J. Larsson1, Kimberli A. Wanionek2, A. Andrada2, assays of comparative viral infection phenotype. Because all islands Stephen S. Whitehead3 were equally immunologically naive for dengue, this study offers an 1University of Vermont College of Medicine, Burlington, VT, United States, opportunity to isolate the effects of viral genetic variation from differential 2Johns Hopkins School of Public Health, Baltimore, MD, United States, herd immunity on epidemic behavior. We studied 17 low-passage DENV- 3National Institutes of Health, Bethesda, MD, United States 2 strains isolated during outbreaks on the islands of Fiji, Tahiti, New Caledonia, American Samoa and Tonga. Each isolate was subjected to Infections caused by the four serotypes of dengue virus represent a whole genome sequencing and phylogenetic analysis then compared for substantial burden of vector-borne disease. Globally, 3.6 billion persons infection efficiency, replication rate and productivity in cell culture. We are at risk for dengue infection; outcomes range from a self-limited febrile found variations in the coding portion of the dengue genome, particularly illness to a fatal shock syndrome. As part of the NIH dengue vaccine the pre-membrane gene (prM) and the nonstructural genes, NS2A and development program, we performed phase I clinical trials on two live NS4A that correlate with the attenuation of the Tongan strains of virus. attenuated monovalent dengue vaccines, DEN1Δ30 and DEN2/4Δ30, at Phenotypic characterization of viruses bearing these genetic substitutions, a dose of 101 PFU to further evaluate the safety profile and to determine in terms of their potential to account for different epidemic dynamics, will the human infectious dose 50% (HID50) of these candidates. Results were be discussed. Our analysis indicates a significant role for viral genotypic compared to prior studies at higher doses of each vaccine (103 PFU). change in dengue epidemic severity. Flavivirus-naïve healthy adult volunteers were dosed with 101 PFU of DEN1Δ30 or DEN2/4Δ30 (15 each) or placebo (3 each) and followed for 6 1345 weeks. Subjects were screened for viremia for 16 days and seroconversion was determined on days 28 and 42. For the two vaccines at lower dosages DENGUE VIRUS INFECTION MODULATES EXPRESSION OF relative to the higher dosages, no significant differences were observed REGULATORY COMPLEMENT FACTORS IN HepG2 CELLS for headaches, myalgias, arthralgias, rash, or neutropenia. Nearly identical 1 1 2 seroconversion levels were achieved for DEN1Δ30 at both doses: 93% Eduardo Nascimento , Bruno Douradinha , Renato Oliveira , 1 1 1 for 101 PFU and 95% for 103 PFU. However, peak geometric mean titers Sean P. Mcburney , Jared D. Evans , Kevin McCormick , Tianyi 1 1 were lower (91.5 vs. 160.6, p=0.029) at 101 PFU. Transient viremia was Wang , Ernesto T. Marques, Jr. similar for DEN1Δ30: 8 (53%) volunteers for 101 PFU and 9 (45%) for 103 1University of Pittsburgh, Pittsburgh, PA, United States, 2Centro de PFU. The duration of viremia was the same (2.8 days), but the mean day Pesquisas Aggeu Magalhaes/FIOCRUZ, Recide, Brazil of onset was delayed for the low dose cohort (D12 vs. D 10). In contrast, Several organisms, including flaviviruses, exploit the regulatory for DEN2/4Δ30, only 53 vs. 100% seroconverted at the 101 vs. 103 dose mechanisms of the complement system to evade innate immune (18.6 vs. 120, p=0.001 Peak GMT), though fewer vaccinees were viremic responses. Elevated activation of complement system is present in severe (33 vs. 55%). Viremia onset was delayed for the low dose (D13 vs. D9)s, dengue cases. Hepatocytes express several soluble complement factors but again the duration was similar (3.5 vs. 3.2 days). These data suggest and receptors; and liver damage is often found in severe dengue patients. that the human infectious dose (HID) of the DEN1Δ30 candidate vaccine 50 CD46 and CD55 are cellular regulatory proteins that inhibit complement is ≤10 1 PFU, however that of rDEN2/4Δ30 is approximately 10 PFU, activation on the host cell surface and protect healthy cells from indicating lower infectivity of this candidate. These data suggest a higher inflammation, whereas gC1qR is a C1q receptor that has been shown dose of DEN2/4Δ30, relative to other components, may be required in a to be involved in modulating cellular anti-viral responses by disrupting tetravalent formulation. MDA5 and RIG-I signaling. We hypothesized that dengue virus (DENV) exploits the complement regulatory mechanism to evade direct attack of the complement system compromising the homeostatic control of complement activation. Thus, we investigated the regulation of CD46, CD55 and gC1qR and complement deposition in a hepatoma cell line HepG2 infected with DENV serotypes 2 (Thailand 16681) or 3 (H87). The cells were analyzed both for virus infection and for the expression of above-mentioned receptors by flow cytometry. The frequency of infected cells (DENVpos) was approximately 50% for both viruses. The expression of complement receptors in DENVpos and bystander cells (DENVneg) www.astmh.org 405 revealed that, regardless of the serotype, the expression of CD46, CD55 in the country. Specifically, in Cucuta and Los Patios, transmission has and gC1qR on DENVpos cells was increased, whereas on DENVneg cells intensified. In cooperation with local hospitals and the health department the expression was decreased. Consistent with this finding, we observed of Norte de Santander, we tested serum collected from suspected dengue an increased deposition of complement factors, iC3b and C6, on cells cases. It was believed that dengue 3 was absent from the area, largely with lower expression of CD46, CD55 and gC1qR. These findings suggest based on the incidence of this serotype in the neighboring Venezuela. that DENVpos cells are protected from direct complement attack, whereas We show that not only is D3 circulating and infecting people in Norte de DENVneg cells are more susceptible. Overall, these results reveal a Santander, but it is at a comparable rate as dengue 1 and 2, speaking to mechanism to allow virus replication in DENVpos cells by protecting them the continuing expansion of the virus. Dengue 4 is largely overshadowed against complement attack and inhibiting the intracellular factors for virus by the other serotypes, though this report confirms the co-circulation of detection and suggest an indirect effect of virus infection on bystander all four serotypes in this area. Of particular interest was the presence of DENVneg cells making them more vulnerable to inflammation and liver four double positive individuals. Three individuals in Los Patios (D1/D2, D1/ damage. D3, D1/D2) and one individual in Cucuta (D2/D3) tested positive for two serotypes. Preliminary sequencing shows that D1 and D3 are genotypes 1346 that have been circulating in Colombia since 2008. D2, however, has 100% homology with a Venezuelan strain isolated in 2007. Further DIFFERENTIATING THE EFFECTS OF DENGUE VIRUS sequencing will inform phylogenetic relationships and possibilities of INFECTION AND AEDES AEGYPTI SALIVARY PROTEINS IN temporal and spatial patterns of viral movement. The presence of all DENDRITIC CELL IMMUNITY four serotypes indicates this is a hyperendemic area with high levels of transmission. These interactions require further investigation to further Michael K. McCracken, Rebecca C. Christofferson, Daniel M. inform public health officials on dengue transmission in this region. Chisenhall, Christopher N. Mores Louisiana State University, Baton Rouge, LA, United States 1348 Hematophagous arthropod saliva has been shown to possess a variety of SEASONAL PATTERNS OF DENGUE VIRUS TRANSMISSION IN effector functions that facilitate the acquisition of a blood meal. Mosquito saliva contains molecules with anti-inflammatory, anti-hemostatic, IQUITOS, PERU and immuno-modulatory capabilities. Arbovirus-infected mosquitoes Steven T. Stoddard1, Helen J. Wearing2, Brett M. Forshey3, Amy expectorate saliva and virus immediately prior to blood feeding and C. Morrison1, Helvio Astete4, Stalin Vilcarromero4, Carlos Alvarez5, this saliva may have the potential to aid the establishment of arbovirus Cesar Ramal-Asayag6, Claudio Rocha4, Alberto Laguna-Torres3, infection within the vertebrate host. Dengue virus (DENV), a mosquito- Moises Sihuincha7, Eric S. Halsey3, Thomas W. Scott1, Tadeusz J. borne flavivirus, is also known to modulate various components of the Kochel8 immune response to infection. As such, it is necessary to differentiate the 1University Of California, Davis, Davis, CA, United States, 2University of immuno-modulatory effects of the virus from those of the mosquito and New Mexico, Albuquerque, NM, United States, 3Naval Medical Research to examine their possible synergism. The effects of Aedes aegypti saliva Unit Six, Lima, Peru, 4Naval Medical Research Unit Six, Iquitos, Peru, on the immune response profile of monocyte-derived dendritic cells were 5Loreto Regional Health Department, Iquitos, Peru, 6Hospital Regional examined using multiplex cytokine immunoassays and ELISA. To reduce Iquitos, Iquitos, Peru, 7Hospital Apoyo Iquitos, Iquitos, Peru, 8Naval Medical the variance between samples often observed in immunological studies of Research Center, Washington, DC, United States multiple donors, the human monocytic leukemia cell line THP-1 was used in place of primary human peripheral blood mononuclear cells. Treatment Understanding periodicity in disease dynamics is fundamental to predicting with DENV type 2, strain 16803, results in down-regulation of numerous outbreaks and reveals factors involved in disease etiology. Dengue virus cytokines involved in the innate immune response as well as those that (DENV) has been shown to exhibit annual and multi-annual patterns of would shift the response toward Th2. Treatments with saliva and virus transmission, presumably driven by climate. We examine ten years of each raised the secretion levels of TNF-alpha and IL-8, and the combined laboratory-confirmed, acute DENV infections captured by passive, clinic- effects of treatment with saliva and virus further increased expression. based surveillance and routine entomological monitoring of adult Aedes Additionally, IL-1beta was significantly up-regulated in treatments aegypti population densities in Iquitos, Peru where there is minimal including saliva while virus alone had no such effect. The continued climatic variation (daily temperature = 25.9ºC ± 2.2). Wavelet analysis characterization of both the isolated and synergistic effects of saliva and of weekly DENV cases indicates a strong, annual signal of increased virus is vital to the understanding of the immunological environment transmission and weak evidence of periodicity on a three-year scale. The during infection establishment as well as developing possible therapeutic number of cases peaks annually in December, is lowest in July, and is applications. strongly correlated with mean daily temperatures, which consistently dip in early July. As has been observed in Thailand, however, mean temperatures 1347 do not significantly differ between high transmission and low transmission seasons (26.9ºC vs 25.6ºC), but neither does the daily temperature HYPERENDEMIC TRANSMISSION OF DENGUE IN NORTE DE range (10.3ºC vs 10.3ºC). Thus it appears that temperature variation is SANTANDER, COLOMBIA insufficient to explain annual periodicity in Iquitos, because although the short period of low temperatures in July could effectively slow viral 1 1 2 Christopher N. Mores , Berlin Londono , Carolina Cardenas , replication in mosquitoes, transmission rates decline well before that time 2 1 Lucio Cardenas , Ann-Marie F. Johnson , Rebecca C. (~March) and do not elevate until well after (~October). Furthermore, 1 Christofferson precipitation decreases during June and July (0.25 inches/day, annual 1Louisiana State University, Baton Rouge, LA, United States, 2Universidad mean 0.38 inches/day), but otherwise is constant throughout the year, de Pamplona, Pamplona, Colombia and variation in Ae. aegypti adult population densities does not show Dengue virus is a significant international public health threat with the any consistent pattern that fluctuates with trends in transmission. Viral potential to become a health security issue as it continues to emerge fade out in the human population appears to be strongly influenced by throughout the tropics and reaches across national borders. The incidence near-annual fumigation campaigns organized to control DENV outbreaks, of dengue infections in people is increasing within endemic regions across while annual amplification in the population remains unexplained. We the globe. Coincident with this emergence, dengue is also expanding into present and discuss a number of alternative models explaining annual less recently afflicted areas with increasing effect (including autochthonous amplification as well as inter-annual variation in the shape and magnitude transmission establishment). Currently the department of Norte de of epidemic curves. Santander, Colombia is experiencing the highest rate of DENV cases www.astmh.org 406 1349 1 DENV serotype). We modeled the mean rate of ascent, time to peak, and rate of decline for children in all three initial PRNT categories. We ENVIRONMENTAL AND AGING INFLUENCES ON ANTIBODY- observed a consistent response across all serotypes characterized by an ENHANCED DENGUE DISEASE OUTCOMES IN AN approximately linear rise in log titer followed by a peak 5-10 days after IMMUNOCOMPETENT MURINE MODEL return to normal body temperatures. Through analysis of the complete PRNT trajectory of those with a secondary infection, we find evidence that 1 2 1 Daniel G. Diniz , Cesar A. Foro , Maíra C. Turiel , Marcia C. the serotype with the highest titer at first sample dominates the secondary 3 1 3 Sosthenes , Sâmia Demachki , Giovanni F. Gomes , Carla M. neutralizing response thus supporting the theory of original antigenic 1 1 1 Damasceno Rego , Marina C. Magalhães , Brunno G. Pinho , sin. Furthermore, we find a rise in titer trajectories beginning two years 1 4 5 Juliana P. Ramos , Samir M. Casseb , Eliana V. da Silva , Márcio after secondary infection that may be explained by subclinical tertiary or 5 4 6 7 R. Nunes , José A. Diniz , Colm Cunningham , Victor H. Perry , quaternary infections. These results help characterize the longitudinal 5 1 Pedro F. Vasconcelos , Cristovam W. Diniz serologic immune response to infection with dengue virus and can assist in 1Universidade Federal do Pará, Belém, Brazil, 2Universidade Federal do the interpretation of PRNT results taken at different times from infection. pará, Belém, Brazil, 3Universidade Federla do Pará, Belém, Brazil, 4Instituto Evandro Chagas, Belém, Brazil, 5Instituto Evandro Chagas, Ananindeua, 1351 Brazil, 6School of Biochemistry and Immunology, Trinity College Institute of Neuroscience, Trinity College, Dublin, Ireland, 7Southampton Neuroscience WHAT DO PEOPLE KNOW AND DO ABOUT DENGUE AND Group, School of Biological Sciences, University of Southampton, PROTECTING THEMSELVES FROM IT IN IQUITOS, PERU? Southampton, United Kingdom Valerie Paz Soldan1, Jhonny J. Cordova2, Audrey Lenhart2, Amy T-lymphocytes are proposed to promote clearance during primary dengue C. Morrison3, John Elder4, Philip McCall2 virus (DENV) infection but contribute to immunopathology during 1Tulane University School of Public Health and Tropical Medicine, New heterologous infections. Since an enriched environment enhances T-cell Orleans, LA, United States, 2Liverpool School of Tropical Medicine, activity during viral infections and active older adults show less functional Liverpool, United Kingdom, 3University of California at Davis, Davis, CA, decline in T cell adaptive immunity, we hypothesized that enriched United States, 4San Diego State University, San Diego, CA, United States environment and aging would increase disease severity. To induce multiple infections of a single serotype or antibody-enhanced disease as it may As part of a community-randomized trial to evaluate the effectiveness of occur in human infection, serial i.p. injections with DENV3 (genotype III) insecticide-treated curtains (ITC) for dengue prevention in Iquitos, Peru, infected brain homogenate or anti-DENV2 hyperimmune serum followed we applied a survey between October-December 2009 to 1334 study 24h later by DENV3 (genotype III) infected brain homogenate were done. participants to examine their knowledge, attitudes and practices (KAP) Control mice received anti-DENV2 hyperimmune serum followed 24h associated with dengue and its prevention. Most of our respondents later by uninfected brain homogenate. Compared to antibody-enhanced were female (73.9%), had finished secondary school (78.4%), and had dengue disease, clinical signs after one serotype infection were less a median age of 39 (range: 16-88). Although most participants knew apparent and survival periods longer. After antibody-enhanced dengue that dengue was transmitted by a mosquito bite (85%), only 16.5% disease significant differences in the survival probability curves (p = 0.031) recognized that this mosquito bites during the daytime, 19.3% knew were found and both young and aged subjects from enriched environment that its legs have white stripes, and 14.7% knew dengue is transmitted showed higher mortality, intense clinical signs and hyperplasia of T cells by Aedes aegypti. The most commonly recognized symptoms of dengue in liver and lungs than subjects with impoverished environment. We were fever (86.5%), headache (76.4%), muscle or joint pain (57.2%), and propose that an enhanced immune response is occurring in the subjects nausea or vomiting (25.0%). The most commonly identified preventive of the enriched environment and in line with this concept glucocorticoids practices included getting rid of unusable items that might collect water reduced these adverse outcomes (37.3%), and use of products to kill or repel mosquitoes (13.5%). More than half the respondents knew someone who had had dengue at some 1350 point (65%), and amongst these individuals, the median number of people they knew was 2. When people were asked what one should do if LONGITUDINAL CHARACTERIZATION OF ANTIBODY one has dengue, only about half (54.1%) knew to take paracetamol. Most RESPONSE TO DENGUE VIRUS IN BANGKOK THAILAND common practices for mosquito control people mentioned were cleaning their homes (46.9%), picking up unusable items that might collect water 1 1 1 Andrew Azman , Henrik Salje , Isabel Rodriguez-Barraquer , (37.3%), covering water containers (26.2%), fumigating their homes 1 2 3 Benjamin Althouse , Timonthy P. Endy , Ananda Nisalak , Richard (17.8%) and using insecticides around their home (13.5%). Use of insect 3 3 1 Jarman , Robert Gibbons , Derek A. Cummings repellent was minimal (2%). Despite (1) dengue endemicity in Iquitos 1Johns Hopkins Bloomberg School of Public Health, Department of for the past decades, (2) that the Regional Health Authority routinely Epidemiology, Baltimore, MD, United States, 2State University of New York, fumigates and places larvicide in water containers, and (3) that there have Upstate Medical University, Department of Medicine, Syracuse, NY, United been different types of health education messages at the community level States, 3Armed Forces Research Institute of Medical Sciences, Department disseminated through various media (radio, signs on buses and other of Virology, Bangkok, Thailand public places), knowledge about dengue and its transmission, as well as household level practices to reduce dengue, could be improved in Iquitos. The plaque reduction neutralization test (PRNT) is the gold standard used to characterize the serologic immune response during and after infection with dengue virus (DENV). Few studies have described the trajectory of 1352 PRNT titers to all four serotypes after infection. We illustrate the antibody LEPTOSPIROSIS AND DENGUE FEVER CO-INFECTION: A response to infection in a cohort of Thai children through parallel analysis REPORT OF THREE REPRESENTATIVE CASES of longitudinal PRNT results for all four serotypes of DENV. One hundred and eighty children from 1 to 15 years old seen at 2 hospitals in Thailand David B. Bouslough, Robert Partridge, Tamara Thome with RT-PCR confirmed DENV infection were followed up for dengue virus Brown University, Providence, RI, United States antibody response. Blood samples were collected daily from enrollment to the day after defervescence. Subsequent samples were collected at one Leptospirosis and dengue fever cause overlapping symptom profiles week, six months, and yearly, with a maximum of three years of follow- leading to mis-diagnosis, higher morbidity and mortality. Concurrent up. Children were categorized based on their first PRNT measurement leptospirosis and dengue infections have not been widely studied. We report 3 representative cases of co-infection during the 2008 dengue as undetectable (PRNT50 < 10 to all DENV serotypes), monotypic (PRNT50 epidemic in American Samoa. Hospital infection control records from > 10 to only 1 DENV serotype) or, multitypic (PRNT50 > 10 to more than www.astmh.org 407 January to September were analyzed for dengue and leptospirosis co- 1354 infection cases. Three cases representing a spectrum of disease and treatment in outpatient, inpatient and critical care settings were identified. ECO-EPIDEMIOLOGICAL EVALUATION OF CHAGAS DISEASE Patient medical records were reviewed retrospectively. Data included PREVALENCE IN THE VILLAGE OF LAGARTERA GRANDE, demographic information, history and physical exam findings, laboratory REPUBLIC OF PANAMA and imaging results, treatment, length of hospital stay, severity of illness, complications and final outcomes. Of the 132 dengue IgM+ patients, Malika Atmakuri and 17 leptospirosis + patients identified during the study period, six Pennsylvania State University College of Medicine, Hershey, PA, United were identified to have a co-infection. Representative cases demonstrate States disease, treatment, and hospital setting variability. Case one is of a 41 Chagas disease has historically been endemic to the village of Lagartera year-old male presenting to the emergency department (ED) with fever, Grande, especially in children. Previous data collected in 2004 revealed chills, and headache. Laboratory analysis during two outpatient visits that 2.9% of the village children tested seropositive for Trypanosoma demonstrated leucopenia, hemo-concentration, thrombocytopenia, cruzi, the parasite responsible for Chagas. Once infection was detected elevated lactate dehydrogenase (LDH) and creatinine phosphokinase in this community, health officials began to implement preventive (CPK). Concomitant lower extremity cellulitis complicated decision measures. The Gorgas Memorial Institute conducted health education making. Symptoms resolved with supportive outpatient care. Case two projects in the community in cooperation with the Japan International is of a 33 year-old female presenting to the ED with chills, dizziness and Cooperation Agency (JICA) with particular focus on educating school nausea. Initial laboratory assessment demonstrated leucopenia, hemo- children on various aspects of the disease. The Department of Vector concentration, thrombocytopenia, and mild AST and ALT elevations. Control of the Ministry of Health began sporadic spraying of infested On day two she demonstrated a fine erythematous rash, critical houses and developed an efficient system of entomological surveillance thrombocytopenia, alkaline phosphatase (ALP), and LDH elevations. with community participation. This study serves to evaluate the progress Critical thrombocytopenia resolved with two days of supportive inpatient accomplished since 2004 by assessing the current prevalence of Chagas care. Case three is of 32 year-old female evaluated in the ED and admitted within Lagartera Grande. Seventy-seven members of the community to the Critical Care Unit for septicemia and multi-system organ failure. completed the Knowledge, Attitudes and Practices (KAP) survey for Critical thrombocytopenia developed with resultant hemorrhagic disease Chagas disease, the vector Rhodnius pallescens, and the vector’s arboreal and eventual death. Comparative case descriptions, laboratory analysis habitat the royal palm Attalea butyracea. Serum samples were also and treatment reviews are provided. In conclusion, challenges identifying collected from all the children in Lagartera Grande above the age of six dengue fever-leptospirosis co-infections result in treatment delays and months to assess the current prevalence of infection in the community. adverse outcomes. Dengue epidemics in American Samoa may require Samples were screened with 3 different serological tests: a commercial routine testing for both diseases. recombinant enzyme-linked ELISA (ELISA Chagatest, Wiener Laboratory, Argentina), a recombinant ImmunoComb commercial test (Orgenics, 1353 Israel), and an immunoblotting technique using a crude epimastigote SEA SURFACE TEMPERATURE MONITORING FOR DENGUE antigen preparation derived from a Panamanian T. cruzi strain. All samples EARLY WARNING IN ECUADOR collected tested negative, suggesting the successful implementation of preventive measures in the community. Continued surveillance and Anna Stewart1, Rachel Lowe2 monitoring was recommended since favorable conditions for transmission 1SUNY College of Environmental Science and Forestry, Syracuse, NY, are still present in Lagartera Grande. United States, 2International Centre for Theoretical Physics, Trieste, Italy 1355 Dengue fever, a mosquito-borne viral disease, is one of the most important emerging tropical diseases in Ecuador. We report a statistical model for IDENTIFICATION OF ANTHROPOLOGICAL AND assessing the importance of climate as a driver for inter-annual variability SOCIOCULTURAL RISK FACTORS FOR CUTANEOUS in dengue fever in southern coastal Ecuador. Climate variables from a local LEISHMANIASIS IN CAPIRA DISTRICT, PANAMA meteorology station (precipitation, relative humidity, min/max/mean air temperature) and Pacific sea surface temperature (SST) anomalies were Joanne Smucker1, V. Bayard2, J.A. Stoute1, A. Saldana2, J.E. used to predict annual dengue fever incidence (1993-2010). Non-climate Calzada2, M. Trefney3 confounding factors such as serotype introduction were also considered. 1Pennsylvania State University School of Medicine, Hershey, PA, United During El Niño events (positive Pacific SST anomalies), southern coastal States, 2The Gorgas Memorial Institute, Panama, Panama, 3The Kansas Ecuador experiences warmer and wetter conditions, while during La Niña State University School of Veterinary Medicine, Manhattan, KS, United events (negative Pacific SST anomalies), the climate is cooler and drier. States Preliminary results indicate that years with an above normal incidence Leishmaniasis is a disease with multiple clinical presentations that affects of dengue fever were associated with El Niño events and years with millions of individuals around the world. This parasitic disease is complex below normal incidence of dengue were associated with La Niña events. and difficult to control due to the intricacies of its transmission cycle, the Increased rainfall and warmer temperatures increase the availability of variety of animal host reservoirs, the many species of sand flies that act breeding sites and the development rate of the dengue mosquito (Aedes as transmission vectors, and diversity of ecosystems involved. In Panama, aegypti). Due to time lags involved in the climate-disease transmission there is an increasing incidence of the ulcerated cutaneous form of the system, monitoring El Niño / La Niña evolution in the Pacific Ocean could disease and it is suspected that there is an underestimate of the actual provide some predictive lead for forecasting dengue epidemics. This is number of people affected due to lack of access to healthcare by poor and the first study of dengue fever and climate in this region. This research disadvantaged populations. This project aimed at identifying risk factors provides the foundation to develop a climate-driven early warning system for cutaneous leishmaniasis that can be useful in establishing a prevention for dengue fever in Ecuador. plan tailored to the community where cutaneous lesihmaniasis is endemic. The population studied was from the village of Trinidad Las Minas in Capira District, Republic of Panama. One hundred and twenty-five individuals older than 12 years of age were surveyed between November and December 2009 in the village. Twenty-four households were randomly chosen and the characteristics of each household were recorded to identify risk factors. The variables studied included the floor material,

www.astmh.org 408 the roof material, the presence of cracks on the walls, the presence of 1357 surrounding royal palms, the presence of other animals around the home, and the presence of screens on the windows. The data was analyzed MULTIPLE ETIOLOGIC AGENTS; THE POSSIBLE CAUSE OF using descriptive statistics and univariate logistic regression. On univariate CUTANEOUS LEISHMANIASIS IN GHANA analysis the only variable associated with cutaneous leishmaniasis in the 1 2 3 home was the presence of dirt floors χ( 2 = 6.8, P = 0.01). Multiple logistic Godwin Kwakye-Nuako , Manal Jamjoom , Godwin Afebe , 4 2 regression will be carried out to determine whether this is an independent Patrick F. Ayeh-Kumi , Paul A. Bates factor. In conclusion, the floor material should be further studied as a 1Department of Human Biology, School of Biological Sciences, University potential risk factor for cutaneous leishmaniasis. The results may be useful of Cape Coast, Cape Coast, Ghana, 2Division of Biomedical and Life in developing a community prevention plan with the goal to decrease the Sciences, School of Health and Medicine, Lancaster University, Lancaster, incidence of the disease. United Kingdom, 3Disease Control Unit, Ministry of Health, Ho, Ghana, 4Department of Microbiology, University of Ghana Medical School, College 1356 of Health Sciences, Korle-Bu, Accra, Ghana SUDANESE LEISHMANIA DONOVANI POPULATION The emerging and/or re-emerging focus of cutaneous leishmaniasis (CL) STRUCTURE: UNCOVERED BY MULTILOCUS MICROSATELLITE in Ghana since 1999 has in recent times, seen more than one species of the parasite identified and implicated as the etiologic agent. Leishmania AND SEQUENCE TYPING major was first to be identified as the agent of the infection in 2006 in the Rania M. Baleela1, Sinead Fitzpatrick2, Katrin Kuhls3, Gabriele endemic focus in Ghana. An unknown and uncharacterised Leishmania Schönian3, Michael Miles2, Isabel Mauricio4 species was identified in 2007 but did not use species linked to African 1London School of Hygiene and Tropical Medicine and University leishmaniasis as positive control. The recent work done in 2008 using of Khartoum, London and Khartoum, Sudan, 2 London School of Leishmania species associated with infections in Africa as positive controls Hygiene and Tropical Medicine, London, United Kingdom, 3Institute of identified Leishmania contrary to the species previously recognized, as Microbiology and Hygiene (Parasitology), Charite´ Universita¨tsmedizin, one of the possible species influencing disease in Ghana. This study aimed Berlin, Germany, 4London School of Hygiene and Tropical Medicine and at the identification of species of Leishmania parasites responsible for Unidade de Parasitologia e Microbiologia Médicas, Instituto de Higiene e CL focus reported in Ghana. The endemic focus is located in the south- Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal eastern part of Ghana, which borders three countries in the West African sub-region. Twenty lesion aspirate and scraping samples were taken from The population structure of Leishmania donovani, the etiological agent active patents lesions for the study. Primers A1 and A2, were used to of visceral leishmaniasis (VL), was investigated. VL is highly endemic in amplify a fragment of ~1500 bp of the intergenic region between the countries such as India, Brazil and Sudan. Sudan has suffered serious ribosomal protein genes RPS7A and RPS7B on chromosome 1 and second epidemics in the past, particularly in the East and South. Here we had primers B1 and B2, were used to amplify an internal fragment of ~1350bp typed a large panel of Sudanese strains using multilocus microsatellite in a nested PCR. These nested PCR products obtained were digested using typing (MLMT) (n=14 markers, 103 isolates) and multilocus sequence restriction enzyme MspI and the products run on 2% agarose gel. The typing (MLST) (n=11 targets, 50 isolates). Both genetic typing methods bands produced from some samples showed a match to one of the control agreed on the presence of two main groups of L. donovani in Sudan: sample L. aethiopica, hitherto is found to be associated with leishmaniasis group R which contained 3 subpopulations as estimated by MLMT in the eastern part of Africa. Comparing this preliminary results to previous (SDA, SDB and SDD) or 2 subpopulations as estimated by MLST (SDA works by other investigators, one can somewhat say that there could be and SDB+SDD) and group G (~1:1 canine: human isolates) which more than one agent responsible for CL focus in Ghana. contained 2 subpopulation (SDC and SDC-outliers) as suggested by both typing methods. All subpopulations showed a significant deficiency of 1358 heterozygosity that cannot be explained by a Wahlund effect except SDA which showed higher than expected heterozygosity for MLMT TRYPANOSOMA SPP. OF RODENTS AND OPOSSUMS IN A markers only. In addition, subpopulation SDB of MLMT Group R was not CHAGAS DISEASE ENDEMIC REGION OF NORTHERN PERU significantly departed from Hardy-Weinberg equilibrium, was in linkage 1 2 3 equilibrium and had an inbreeding index (FIS= 0.20), selfing rate (s=0.33) Dawn M. Roellig , Michael Z. Levy , Robert Gilman , Vitaliano 1 and panmictic index (f= 0.02) values compatible with recombination. Cama In this study, it is tempting to suggest that this subpopulation is an 1Centers for Disease Control and Prevetion, Atlanta, GA, United States, intermediate between SDA and SDD, but this hypothesis needs to be 2University of Pennsylvania School of Medicine, Philadelphia, PA, United further investigated. Furthermore, all MLST subpopulations showed none States, 3Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, to minimal pairwise linkage disequilibrium after sequential Bonferroni United States correction ( = 0.05). The equal number of isolates from dogs and humans α An estimated 192,000 Peruvians are infected with Trypanosoma cruzi, in group G suggests a role in transmission for domestic dogs, at least the etiologic agent of Chagas disease. In an endemic region of northern in eastern Sudan. However, subpopulations SDC and SDC-outliers were Peru in the province of Cutervo, intervention methods, including spraying, moderately subdivided to effectively isolated from Group R subpopulations began in 2001 that seemingly decreased the incidence of Chagas disease (FST values ≥ 0.40). An association was suggested between the by the next year. Despite these efforts, new acute cases (2 months- 2 yrs) subpopulation and pathology, which if holds true after further were reported from 2004-2008 and prevalence from a 2004 study was investigations, may be very useful for disease diagnosis and pathology- still 27.7% in individuals <15 years of age. Many communities in Cutervo specific designed drugs. This study provided a plethora of new information are in close contact with the sylvatic environment having mud brick regarding the population genetics of Sudanese L. donovani, and raised homes in the middle of the Andean high jungle ecosystem. Considering several hypothesis regarding important aspects of the epidemiology of the presence of new cases and the sylvatic environment, the goal of the leishmaniasis in Sudan, and possibly other endemic areas. current study was to determine the role of wildlife reservoirs in T. cruzi transmission within the region. Rodents (Rattus norvegicus and R. rattus) and opossums (Didelphis albiventris) were live captured from within homes and surrounding areas in five communities (Casa Blanca, Pindoc, Esperanza, Rumiaco, and Nuevo Guayaquil). At the time of collection, trypomastigotes were observed by bright-field microscopy in 22 of 50 rodent whole blood samples, while no parasites were observed in opossums (n=7). A panel of PCR, including gene targets for the minicircle www.astmh.org 409 and 28S, 18S, and region from 18S to 5.8S rDNA, was run to identify false bitten by infective tsetse. Our results show that, contrary to expectation, negatives by microscopy and determine the Trypanosoma spp. present. people were more likely to be bitten by tsetse (G. morsitans morsitans, Of the rodents, 16/22 positive by microscopy were infected with T. lewisi, G. pallidipes) in the vicinity of their houses rather than in the woodlands while the Trypanosoma spp. of the remaining six rodents could not be (total catches = 375 vs. 264 tsetse caught over 221 days) where tsetse identified due to sample loss. T. cruzi was identified in 4/7 opossums; are apparently more abundant. For tsetse from houses, 44% were female these animals were hand-captured south of the Esperanza community. and of these, 30% were old enough to be able to carry mature infections. While the sample size of the current surveillance study is small, two main Moreover, natural repellents (human body odour, woodsmoke) produced observations were made: 1) Rats do not appear to play a role in the T. by humans which are highly effective outdoors, are ineffective against cruzi transmission cycle within Cutervo and 2) the opossum may have a indoor-biting tsetse. For humans in woodlands, the numbers of tsetse potential role as a wildlife reservoir for T. cruzi in the area. were greatest if the human was mobile and not in the vicinity of a natural non-human host: humans walking without an ox caught 20 tsetse/day 1359 compared to 0.2 tsetse/day for a stationary human accompanied by an ox, albeit the flies were younger and hence less likely to be infected (18% EPIDEMIOLOGICAL CHARACTERISTICS OF LEISHMANIASIS IN of females were old enough to be able to carry mature infections). We PERU 2004-11 suggest that for people in areas where Rhodesian sleeping sickness poses a risk, interventions designed to prevent or kill tsetse entering houses or Manuel J. Loayza, Gloria Cisneros, Micaela Lorenzo, Juan C. vehicles might reduce personal risk of HAT significantly. Velasco, Luis M. Loro Direccion de Salud V Lima ciudad/Ministerio de Salud, Lima, Peru 1361 Leishmaniasis is a parasitic disease of significant public health importance. Considered by WHO as one major Neglected disease, this disease is POPULATION STRUCTURE OF LEISHMANIA TROPICA IN Transmitted by sand fly vector. In Peru, leishmaniasis is an endemic disease NORTHERN PAKISTAN and NEIGHBORING COUNTRIES affecting several departments and is the second endemic tropical. Is Nazma Habib1, Colin Sutherland1, Syed Akram Shah2, Vanessa reported an annual average of 2500 cases. There is a need to provide Yardley1, Brighid O’Neill1, Rizwan Hashim3, Arfan Bari3, Akhtar information for the management of the disease. The study Focus on the Muneer4, Inamullah Khan5 determinate characteristics epidemiological and distribution of cases. 1London School of Hygiene and Tropical Medicine, London, United Notified leishmaniasis is near mandatory public health services. All Kingdom, 2University of Peshawar, Peshawar, Pakistan, 3Combined Military suspected cases were recorded in the Epidemiological sheet and reported Hospital, Peshawar, Pakistan, 4Kuwait Teaching Hospital, Peshawar, to the epidemiological surveillance system in the country. From 2004 Pakistan, 5Khyber Teaching Hospital, Peshawar, Pakistan until 2010, were reported 19113 cases of leishmania. Of all cases 92.6% were cutaneous leishmaniasis and the remaining 7.4% mucocutaneous In Pakistan, Anthroponotic Cutaneous Leishmaniasis (ACL) caused by leishmaniasis. Most have come from Cusco with 3318 cases (16.1%), Leishmania tropica, has a broad distribution occurring focally in the Ancash with 1836 cases (8.9%), Piura with 1816 cases (8.8%), Junín with Northern areas and Azad Kashmir. Reportedly, some isolates of L.tropica 1797 cases (8.7%) and San Martin with 1711 cases (8.3 %). Until week are heterozygous. This project aims to investigate the intra-specific epidemiology 13-2011, 1471 cases have been reported, which makes a diversity of L.tropica in Northern Pakistan using Multilocus microsatellite national incidence rate is 101.6 per 100 000 inhabitants. The endemic typing (MLMT). Further, the population structure and phylogenetics of area extends through the Andes and the valleys between 600 and 3 000 the parasite will be mapped by taking into account isolates from Pakistan meters above sea level, for cutaneous leishmaniasis, and areas of high and from other countries lying between the Mediterranean Sea and the and lowland forest below 2000 meters for mucocutaneous leishmaniasis. Bay of Bengal, namely Syria, Afghanistan, Iran and India. As a tool to The age group was most common entre 20 and 49 years old (67.56%). enhance our ability to identify L. tropica from L. major, which is sympatric The male / female ratio was 1.6. The majority of patients were farmers. in this region, we have developed a novel PCR that distinguishes between The unique lesion was The Most Frequent (69.8%) and the Majority these two species. Samples have been collected from 3 major hospitals of of injuries was in extremities (58.2%). In conclusion, leishmaniasis is Peshawar, Khyber Pukhtoon-Khwa (KPK),Northern Pakistan. These include endemic in tropical kind in Peru and is distributed in poor areas of various isolates in culture, biopsies and filter paper impressions. These samples departments. This produces a negative social and economic impact in have been typed for species and followed by MLMT for L. tropica isolates. the economically depressed. In addition, the destructive consequences it MLMT analysis of clinical isolates from Pakistan and other countries in the causes, particularly the mucocutaneous form the effect of isolating the region (confirmed as L. tropica), plus the WHO strains will be presented. individual, its irreversibility. Results providing an estimate of the ACL presentation rate at Peshawar hospitals will also be discussed briefly. 1360 1362 CAN WE REDUCE PERSONAL RISK OF RHODESIAN SLEEPING SICKNESS? ANALYSIS OF FACTORS CONTRIBUTING TO THE TRANSMISSION DYNAMICS OF TRYPANOSOMA CRUZI PROBABILITY OF BEING BITTEN BY TSETSE LINEAGE I IN TWO ENDEMIC PROVINCES OF ECUADOR Stephen J. Torr1, Clement Mangwiro2, Andrew Chamisa3, Glyn Alejandra P. Zurita-Lagos1, Juan J. Luthier2, Jaime A. Costales1, A. Vale1 Sofia Ocana-Mayorga1, Patricio Diosque2, Oscar Franzén3, Björn 3 4 1Natural Resources Institute, Chatham, United Kingdom, 2Bindura Andersson , Mario J. Grijalva University, Bindura, Zimbabwe, 3Tsetse Control Department, Harare, 1Infectious Disease Research Center, Pontifical Catholic University of Zimbabwe Ecuador, Quito, Ecuador, 2Instituto de Patología Experimental, Universidad Nacional de Salta-CONICET, Salta, Argentina, 3Karolinska Institutet, Rhodesian sleeping sickness, the zoonotic form of Human African Stockholm, Sweden, 4Tropical Disease Institute, Ohio University, Athens, Trypanosomiasis found in east and southern Africa, is often associated OH, United States with game parks and wilderness areas where tsetse flies (Glossina spp.) and wild reservoir hosts are abundant. People living and working within Trypanosoma cruzi infection affects an estimated 230,000 people in or near such areas have limited options for controlling HAT due to Ecuador. Recent reports indicate limited effectiveness of insecticide- concerns about the cost, feasibility and environmental impacts of tsetse based vector control interventions, due to re-infestation by sylvatic control. In the Mana Pools National Park of Zimbabwe, we carried out triatomines. Previous studies demonstrated that the lineage I (TcI) of T. studies to identify the circumstances in which people are most likely to be cruzi is the predominant lineage circulating in Loja (Southern Andes) and www.astmh.org 410 Manabi (Central Coastal) provinces. Furthermore, in southern Ecuador 1364 (Loja province) microsatellite analyses of TcI isolates showed two main parasite populations exist: one related with domestic and peridomestic ANTI-TRIATOMINE SALIVA IMMUNOASSAYS FOR THE environments and a second one related with sylvatic environments. The EVALUATION OF IMPREGNATED NETTING TRIALS AGAINST aim of this study was to evaluate TcI isolate divergence within and among CHAGAS DISEASE TRANSMISSION Loja and Manabí using Multi Locus Sequence Typing (MLST). We sampled 1 2 2 vectors and mammals and a wide geographic area within each province. Alexandra Schwarz , Jenny Ancca Juarez , Jean Richards , 2 3 3 Our results corroborate that the presence of two different parasite Bruno Rath , Victor Quispe Machaca , Yagahira E. Castro , Edith 3 4 5 6 populations in Ecuador, according to habitat: In Loja province, the previous Malaga , Katelyn H. Levy , Robert H. Gilman , Caryn Bern , 3 4 separation in two populations (domestic/peridomestic and sylvatic) was Manuela Verastegui , Michael Z. Levy confirmed. In Manabí province, this tendency was also seen, where the 1Institute of Parasitology, Biology Centre of the Academy of Sciences of sylvatic population was separated from the peridomesic population. Czech Republic, České Budĕjovice, Czech Republic, 2Universidad Peruana However, in both cases, limited genetic flow was evidenced. Interestingly Cayetano Heredia, Lima, Peru, 3Asociación Benéfica Prisma, Lima, Peru, sylvatic samples of both provinces cluster together, suggesting genetic 4University of Pennsylvania, Philadelphia, PA, United States, 5Johns Hopkins flow among sylvatic populations, while genetic separation was evident University, Balitmore, MD, United States, 6Centers for Disease Control and between domestic/peridomestic populations of both provinces. These Prevention, Atlanta, GA, United States results suggest that similar transmission dynamics are taking place in both Insecticide-impregnated nets can kill triatomine bugs, but it remains provinces where albeit at different rates, there is limited genetic flow unclear whether they can protect against Chagas disease transmission. In a between sylvatic and domestic/peridomestic T. cruzi populations within a field trial in Quequeña, Peru, sentinel guinea pigs placed into intervention small geographical area. Therefore, control strategies need to be adapted enclosures covered by deltamethrin-treated nets showed significantly to the intrinsic characteristics of a small geographic scale. lower antibody responses to saliva of Triatoma infestans compared to animals placed into pre-existing control enclosures. Our results strongly 1363 suggest that insecticide treated nets prevent triatomine bites and can DISTRIBUTION AND NATURAL INFECTION OF CHAGAS thereby protect against infection with Trypanosoma cruzi. Anti-salivary DISEASE VECTORS IN DOMESTIC, PERIDOMESTIC AND immunoassays are powerful new tools to evaluate interventions against Chagas disease. SYLVATIC HABITATS IN SOUTHERN ECUADOR Mario J. Grijalva1, Anita G. Villacis2, Maria V. Suarez-Davalos2, 1365 Sofia Ocana-Mayorga2, Cesar A. Yumiseva2, Esteban G. Baus2 PROXIMITY BETWEEN DOGS AND TRYPANOSOMA CRUZI 1Tropical Disease Institute, Ohio University, Athens, OH, United States, 2Center for Infectious Disease Research, Pontifical Catholic University of INFECTED TRIATOMINES AS A RISK FACTOR FOR THE Ecuador, Quito, Ecuador PERSISTENCE OF CHAGAS DISEASE 1 2 Chagas disease is endemic in 70% of the Ecuadorian territory. The main Ricardo Castillo Neyra , Victor R. Quispe Machaca , Jenny Ancca 2 2 3 vectors responsible for Trypanosoma cruzi transmission in the Southern Juarez , Lily Chou Chu , Fernando S. Malaga Chavez , Juan G. 3 2 1 Andean region of the country are Rhodnius ecuadoriensis, Triatoma Cornejo del Carpio , Cesar Naquira , Robert H. Gilman , Caryn 4 5 carrioni, Panstrongylus chinai and P. rufotuberculatus. This study aims to Bern , Michael Z. Levy describe the triatomine distribution and natural trypanosome infection 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United in domestic, peridomestic and sylvatic habitats in Loja Province. Active States, 2Universidad Peruana Cayetano Heredia, Lima, Peru, 3Dirección triatomine searches were conducted in domestic and peridomestic habitats Regional del Ministerio de Salud, Arequipa, Peru, 4Division of Parasitic in rural villages and in nearby sylvatic areas throughout the province. Diseases and Malaria, Center for Global Health, Centers for Disease 11,115 live triatomines were found infesting domestic units in 68% of Control and Prevention, Atlanta, GA, United States, 5Department of the 92 rural communities while 1,923 live triatomines were found in Biostatistics and Epidemiology, University of Pennsylvania, Philadelphia, PA, 52% of the 23 sylvatic localities examined. Nine percent of the domestic United States units (n = 3,191) were infested with one or more triatomine species and Chagas disease is a vector-borne disease transmitted by triatomine bugs 12% of the sylvatic nests (n=1,219) were infested with R. ecuadoriensis. and caused by the Trypanosoma cruzi parasite. It is one of the most Nymphs were observed in 80% of both infested domiciles and nests. neglected tropical diseases. Insecticide application campaigns to eliminate Triatomines were found in all ecological regions below 2,200 meters vectors are the most effective intervention to stop transmission of the above sea level. In the domicile R. ecuadoriensis and T. carrioni were parasite, and are routinely conducted in Arequipa, Peru. After these found mostly in bedrooms while in the peridomicile these species were campaigns, re-infestation can occur, and areas where vectors and T. abundant in chicken coops located near the house. Established colonies cruzi infected mammals overlap can be the starting point for re-initiation of P. chinai and P. rufotuberculatus were found restricted to the domicile. of disease transmission. Dogs have been described as reservoirs of T. Sylvatic triatomines were found mainly in squirrel and mouse/rat nests, cruzi in Argentina, Brazil, Mexico, and Venezuela. Our objective was to and to a lesser extent in bird nests. T. cruzi infection was found in 10% determine whether the presence of seropositive dogs can explain the of the domestic/peridomestic triatomines (n=775) and 64.7% (n=300) of clustered re-emergence of T. cruzi in vectors. The study was designed sylvatic triatomines analyzed. Mixed infections with T. cruzi and T. rangeli as a cross-sectional serological screening to detect antibodies against T. were found in 8% of sylvatic triatomines. To date, limited vector control cruzi in dogs, entomological collection of vectors from households to efforts have been implemented in this area. Although, the application of determine their infection status, and georeferencing of households to insecticide-based vector control could be effective in reducing domestic determine proximity between dogs and triatomines. The main outcome and peridomestic populations it must be complemented with constant was canine seropositivity. Its association with other factors was analyzed surveillance to detect and control post-intervention reinfestation by sylvatic with multivariate logistic regression for demographic and household triatomines. Our findings highlight the need for a systematic, sustained, risk factors and with spatial techniques for clustering and proximity and monitored vector control intervention in the region. correlation. Canine seroprevalence in the area was 12.3% (SE=2.6, N=154). The statistical results show that seropositivity in dogs was positively associated with proximity to T. cruzi infected triatomines, with proximity to high numbers of triatomines, regardless of their infection status, and with dog’s age. The presence of T. cruzi seropositive dogs

www.astmh.org 411 could explain the persistence of Chagas disease in endemic areas of Peru. in 38 households was unavailability of a net. In Kuhalong and Rajbila Massive insecticide campaigns allow for the collection of triatomine data respectively 89% and 87% of population slept under bed net at previous that are used to determine high-risk areas for humans. Interventions based night of the interview whereas use of insecticide treated bed net was on these entomological data should include the presence of dogs around about 80% in both. Lowest percentage of last night bed net users were houses where infected triatomines were collected. in C5 (63.5%) of Kuhalong and in C9 (80.2) of Rajbila. Insecticide treated net use was lowest in C11 (59.3%) and C4 (69%) of Kuhalong and Rajbila 1366 respectively. Person per bed net was as high as 3-4 in few areas of the unions. Nontribals, household heads, females, married persons, children GENETIC DIVERSITY OF RHODNIUS ECUADORIENSIS <5 years of age, individuals from family size <5 were more likely to use (HEMIPTERA: REDUVIIDAE) POPULATIONS IN THE CENTRAL bed net in previous night of the interview (P <0.001). Use of insecticide AND SOUTHERN ANDEAN REGIONS OF ECUADOR treated bednet was significantly higher among tribal counterpart and individuals from family size <5 (P<0.001). Last night bed net users were 1 2 2 Anita G. Villacis , Paula L. Marcet , Ellen M. Dotson , Mario J. higher among persons from households that own radio/tape recorder 3 Grijalva (<0.001) or dwelling unit ≤2 (P <0.001). However, insecticide treated bed 1Center for Infectious Disease Research, Pontifical Catholic University of net users were also significantly higher among members of households Ecuador, Quito, Ecuador, 2Centers for Disease Control and Prevention, having bamboo floor/wall (P <0.001) or using oil/kerosene lamp ((P Division of Parasitic Diseases and Malaria, Entomology Branch, Atlanta, <0.001). In both unions, there was no significant variation between GA, United States, 3Tropical Disease Institute, Ohio University, Athens, OH, malaria positives and last night bed net user. Gap between insecticide United States treated bed net user and nonuser should be reduced and more emphasis Rhodnius ecuadoriensis is the most widespread vector of Chagas Disease should be given to seek malaria associated risk factors in these areas. in Ecuador. Effective control of this disease requires a good understanding of the epidemiological cycles, including a reliable analysis of the genetic 1368 structure of populations of this important vector. R. ecuadoriensis COMMUNITIES IN NETWORKS MAPPING MALARIA occupies domestic, peridomestic and sylvatic habitats and is a widely distributed species in the central Coastal (Manabí province) and southern MOVEMENT IN KENYA Highlands (Loja province) regions of Ecuador. These two regions are Deepa Pindolia1, Amy Wesolowski2, Andres Garcia1, Nathan phytogeographically and climatically different and correspondingly, Eagle3, Caroline O. Buckee3, David L. Smith1, Andrew J. Tatem1 bugs collected from these areas demonstrate differences in several 1University of Florida, Gainesville, FL, United States, 2Carnegie Mellon phenotypic characters (i.e., body size, antennal sensilla and wing geometry University, Pittsburgh, PA, United States, 3Harvard School of Public Health, morphometrics), as well as in behavioral traits (feeding and defecation Boston, MA, United States patterns, and life cycle). To evaluate the genetic relationships among R. ecuadoriensis populations between these regions, we sequenced the With malaria eradication back on the global agenda and subsequent mitochondrial cytochrome b (Cytb) gene in 168 insects collected from elimination targets for various low endemic countries, control strategies both regions (n=95 in Loja) and (n=73 in Manabí). We found 34 Cytb require a strong quantitative evidence base. The failure of previous haplotypes determined by 53 variable sites. Only three haplotypes were elimination efforts has shown that human population movements are shared between the two provinces (15 were exclusive for Loja and 16 important for infection exchange between different transmission areas. For for Manabí). A moderate genetic differentiation was observed between countries, that have overall low transmission but a few high transmission hotspots, population movements from high to low transmission zones the two geographical regions (GST=0.05622) and remarkably, a third genetically different group within the Loja province was found. Our results may threaten imported infections, therefore local control agendas and support the hypothesis of disruptive selection acting upon R. ecuadoriensis challenge larger scale elimination efforts. Here, a unique and extensive populations, probably due to geographical isolation. The genetic patterns mobile phone records dataset was analyzed with network analysis tools, observed in this work contribute to the knowledge of genetic variability a countrywide Plasmodium falciparum transmission map and previously of R. ecuadoriensis across different geographical regions and provides developed transmission models to assess communities within Kenya linked background for interpretation of routes of dispersion and isolation. by infection flows. The likely principle sources of imported infections within national boundaries, which may threaten onward transmission or 1367 have clinical significance, were mapped at a settlement level. Clusters of settlements were identified and compared to approximate “natural” DEMOGRAPHIC AND SOCIOECONOMIC DETERMINANTS OF malaria-relevant migration boundaries, splitting the country into regions MOSQUITO NET USAGE IN MALARIA ENDEMIC REGION OF that share malaria-relevant movement characteristics. With elimination BANGLADESH as the ultimate goal for Kenya, we provide a quantitative platform for strategic control planning, by targeting control resources at defined spatial 1 1 1 1 Sabeena Ahmed , Wasif A. Khan , Chai S. Prue , Jacob Khyang , and temporal scales. Malathi Ram2, Md. Sharif Hossain1, Myaing M. Nyunt2, Rashidul Haque1, David A. Sack2, David Sullivan2 1369 1International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, Bangladesh, 2Johns Hopkins Malaria Research institute, Johns Hopkins SOCIO-DEMOGRAPHIC RISK FACTORS FOR MALARIA IN Bloomberg School of Public Health, Baltimore, MD, United States A NEWLY ESTABLISHED SURVEILLANCE REGION IN THE Use of mosquito net is a proven strategy for malaria control. One of the CHITTAGONG HILLS TRACTS OF BANGLADESH vector control components of the National Malaria Control Program Heather Scobie1, David A. Sack1, Wasif A. Khan2, Sabeena in Bangladesh was to promote insecticide treated bed net to ensure Ahmed2, Chai S. Prue2, Jacob Khyang2, Malathi Ram1, Myiang prevention or reduction of malaria mortality and morbidity. However, M. Nyunt1, Gregory Glass1, Timothy Shields1, Md Z. Huq2, David use of mosquito net as well as insecticide treated bed net varies among Sullivan1 households. We investigate associated factors in bed net usage at 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United Kuhalong and Rajbila unions under Bandarban Upazila, south-east region States, 2International Centre for Diarrhoeal Disease Research, Bangladesh, of Bangladesh. We utilized data of an ongoing demographic surveillance Dhaka, Bangladesh from 4567 households with population size 20755. Both the unions were divided into 12 clusters (C). Overall 99.2 % (4529) households possessed Until recently, the Chittagong Hills have been hyperendemic for malaria mosquito nets. Most cited reason for not using bed net while sleeping but in recent years has been hypoendemic. A new study initiated in two www.astmh.org 412 unions (population=20,563) near Bandarban, Bangladesh in 2009 was and the mean total IgG levels to GLURP RO at birth, 3 mo and 6 mo were designed to improve knowledge of malaria transmission, to monitor 257 95% CI 187.38, 352.46, 38.5 95% CI 28.38, 52.21, 36.9 95% CI malaria interventions, and serve as an area for developing new control 26.56, 51.20 respectively. In conclusion, the incidence of malaria among strategies. The project included: (a) demographic surveillance system, infants in the first year of life is low while the incidence of anemia is high. (b) periodic surveys of knowledge, attitude, and practice, (c) geographic In addition IgG levels at birth was higher to CSP than GLURP and decayed information system, (d) weekly active and continuous passive surveillance to lower levels at month 3 and month 6th in the infant cohort. for malaria infections, (e) monthly mosquito surveillance, (f) daily weather measures. The program included both standard and molecular methods 1371 for case detection. Between October 2009 and January 2011, 151 cases of malaria were detected. 83% were symptomatic infections (97% ASSESSING THE CLINICAL EFFICACY OF MALARIA chloroquine resistant Plasmodium falciparum, 2.5% P. vivax, and 0.8% VACCINES IN TRIALS AND THEIR IMPACT IN THE FIELD: A mixed infection). Malaria infections were highly clustered geographically MULTIFACETED APPROACH TO USING SIMULATION MODELS and seasonally. Risk factors associated with higher malaria rates included FOR PREDICTION age, pregnancy, education and occupation. In univariate analysis, there was increased odds of high-season malaria disease in children aged Melissa A. Penny, Alan Brooks, Olivier Briët, Nakul Chitnis, 1-4 (95%CI 1.1, 3.8) and 5-14 (95%CI 1.7, 4.0) compared with those Amanda Ross, Nicolas Maire, Diggory Hardy, Marcel Tanner, aged >14. Tribal people had a 4-fold higher odds compared to Bengalis Thomas A. Smith (p=0.019) living in the same unions. These effects were independent Swiss Tropical and Public Health Institute, University of Basel, Basel, and of similar magnitude and significance in a multivariate model. After Switzerland controlling for the effects of union of residence, age and sex, there was Assessing vaccines against Plasmodium falciparum in clinical trials and an increased odds of malaria amongst people doing day labor (2.5-fold, predicting their impact after licensure comes with many complications p=0.046) and a hillside agriculture practice by tribal groups called “jhum” and open questions. RTS,S, the vaccine furthest in clinical development (4.3-fold, p=0.002), but not other agricultural occupations (p=0.304). with initial Phase III trial results expected before the end of the year, may These results reveal socio-demographic risk factors for symptomatic be licensed in the next few years. During trials of this type of vaccine malaria during the high-season in this Chittagong Hills surveillance region and others (such as vaccines directed towards the blood-stage cycle of that will: (1) serve as a basis for future hypothesis-driven epidemiological malaria) important questions are raised concerning how to adequately studies, or (2) target future intervention strategies to high risk groups. define endpoints to assess efficacy, and if the impact of the vaccine will vary with transmission setting, age-group immunized, or delivery 1370 strategy. In this work, we examine these questions using an ensemble of INCIDENCE OF MALARIA IN THE FIRST YEAR OF LIFE IN A simulation models for malaria epidemiology and control. We discuss the advantage of model ensembles to quantify uncertainty about predictions HIGH MALARIA TRANSMISSION AREA IN GHANA and show how uncertainty in predictions varies with transmission setting, Kwaku Poku Asante1, Daniel Dodoo2, Ellen Boamah1, Ben with simulations suggesting greater confidence in predictions of health Gyan2, Micheal Ofori2, Emmanuel Mahama1, Grace Manu1, effect for lower transmission settings than for higher ones. We discuss Kingsley Osei-Kwakye1, Jones Opoku-Manu1, George Adjei1, simulation results that show the choice of clinical endpoints used to Mohammed Adams1, Miecks Twumasi1, Kwadwo Koram2, Seth assess the efficacy of vaccines in trials, especially for blood-stage vaccines, Owusu-Agyei1 impacts the perception of the success of a vaccine. In addition, we show 1Kintampo Health Research Centre, Ghana Health Service, Brong Ahafo how ensemble models might be used to study new approaches for Region, Ghana, 2Noguchi Memorial Institute for Medical Research, Greater delivery of pre-erythrocytic vaccines like RTS,S, with results indicating Accra Region, Ghana mass vaccination strategies, even at modest coverage, substantially reduce transmission compared to immunization of infants alone and contribute Malaria in pregnancy is a risk for abortions and low birth weight. to much greater health effects per dose. Our multifaceted approach to Placental malaria may also have an impact on the infants susceptibility to modelling and simulation of malaria vaccines, with multiple models for parasitemia, clinical malaria and anemia. A prospective birth cohort study predictions at the population level and within-host level, offers not only was carried out among pregnant women and their infants to determine a pragmatic way to predict their impact and cost-effectiveness, but also the incidence of malaria and its effects on the infant’s health in a high allows decision makers to appraise alternatives for delivery and for efficacy malaria transmission area in Ghana. At birth, placental biopsy was taken assessment to those considered in trials. to determine placental malaria. The infants were followed up for a period of one year. Each infant was followed up monthly for malaria parasitemia, 1372 anemia and passively for clinical malaria. The mean total IgG levels to the Plasmodium falciparum CSP antigen (NANP6) were also determined by MICRO-GEOGRAPHIC ENVIRONMENTAL RISK FACTORS ELISA in a subset of the birth cohort at birth, 3 and 6 months of age. A FOR CHILDHOOD MALARIA DURING THE DRY SEASON IN total of 2810 pregnant women were identified and followed up till birth. LIWONDE, MALAWI, 2010 The average age of mothers was 27 years with 25% and 75% being 1 2 1 primigravidae and multigravidae, respectively. Twenty-two (22) percent Lindsay R. Townes , Don P. Mathanga , Mark L. Wilson of the pregnant women were in the very poor quintile of socioeconomic 1University of Michigan, Ann Arbor, MI, United States, 2University of status. The coverage of intermittent preventive treatment with at least Malawi, Blantyre, Malawi one dose of SP was 95% and insecticide treated net (ITN) use was 38%. Despite the importance of environmental factors to Plasmodium The prevalence of placental malaria was 37.3%, 95% CI 35.22-39.44. A transmission and the re-emergence of spatial epidemiologic methods for total of 1605 infants contributed to 1079.0 PYs of followed up. The mean studying malaria, the role of fine-scale environmental heterogeneity in coverage of ITN use among the infant cohort at any point of contact was rural locations has been relatively unexplored, particularly during periods 36%. The incidence of malaria parasitemia was 0.50PYRS (95% CI 0.46 of seasonally low transmission. During the dry season of 2010 in Liwonde, -0.55 unadjusted), 0.68PYRS.( 95% CI 0.47-0.97 adjusted); incidence of Malawi, children attending Machinga District Hospital’s (MDH) under- anemia was 3.34 95% CI 3.17-3.52 unadjusted, 4.13 95% CI 2.22 -7.68 5s clinic were studied for household environmental characteristics that adjusted and incidence of clinical malaria was 0.22 95% CI 0.20 - 0.25 predicted malaria. P. falciparum infection in children were determined unajusted, 0.45 95% CI 0.16 -1.23 adjusted. The mean total IgG levels using Parachek® Pf Rapid Diagnostic Test (RDT), with demographic and to CSP at birth, 3 mo and 6 mo were 1091.20 95% CI 952.21 -1249.51; environmental data collection occurring at the residence. House location 203.9 95% CI 174.11 -238.89, 179.4 95% CI 147.53 -218.18 respectively

www.astmh.org 413 and elevation were recorded with a global positioning system (Garmin We conducted a population census during the low transmission season eTrex Venture® HC), as were locations of water sources. Four distinct land prior to distribution and tested all children under five for malaria parasites cover categories, along with materials used in house construction, were and severe anemia. A repeat of the enumeration and testing is planned assessed by direct observation. In multivariate logistic regression, children after the campaign. The census was carried out using a shortened version who lived within a 25 m radius of actively cultivated agricultural land were of the MIS questionnaire programmed on personal digital assistants. more likely to have malaria when compared to children who did not, after We geo-referenced all households to analyze spatial patterns. We controlling for age and elevation (odds ratio = 2.39, 95% confidence enumerated 2,189 households, which included a total population of interval: 1.12, 5.10). No spatial clustering or autocorrelation of malaria 11,079 including 1330 under five years children. Insecticide-treated-net cases was found, perhaps indicating that increased risk from proximity (ITN) coverage was higher on Likoma Island (64.9%) than the national to agriculture is independent of geographic location across the study estimate from the 2010 MIS (58.2%). ITN use among under five years old region (Moran’s I = -0.20, P value = 0.66). This study provides preliminary children, was lower (45.9%) than the 2010 MIS estimate (55.4%). ITN evidence of continued Plasmodium transmission during the dry season, use among pregnant women was 35% compared to 49.4% in the 2010 and of various environmental factors that influence malaria risk in rural MIS. Bio-marker samples were collected from 904 out of 1330 children Malawi. The implications for attempting to eliminate malaria are explored. (68.0%) aged 6 to 59 months, of which 835 (62.8%) were included in parasitological reading. Samples were not collected from over 30% of 1373 children due to frequent travel to the mainland. Malaria prevalence was 9.2% and severe anaemia (Hb < 8g/dl) prevalence was 7.5%. Spatial STANDARDIZING A NATIONAL MALARIA BULLETIN analyses are currently being conducted to assess disease clustering. As FOR TAPPING THE POTENTIAL OF ROUTINE HEALTH National Malaria Control Programmes is planning universal coverage of MANAGEMENT INFORMATION SYSTEM DATA IN AFRICA: LLINs and other interventions, it is important to explore new and creative PROCESS AND RESULTS FROM ZAMBIA ways to best measure the impact of interventions. In locations with small confined populations, a population census that includes malaria parasite 1 1 2 Mercy Mwanza Ingwe , Mulakwa Kamuliwo , Jason Pickering , testing of all children under five provides important data on the spatial 3 4 5 John M. Miller , Freddie Masaninga , Benjamin Winters , Mac distribution of disease. We conducted this census in the dry season, when 2 Otten malaria parasite infections are more likely to be spatially clustered. When 1Ministry of Health, Lusaka, Zambia, 2Consultant, Lusaka, Zambia, 3PATH combined with health facility incidence data, census data such as ours MACEPA, Lusaka, Zambia, 4World Health Organization, Lusaka, Zambia, can help elucidate disease dynamics and support operational research into 5Akros Research, Lusaka, Zambia how best to improve control. For high-burden African countries, routine information systems offer many often untapped opportunities to present local-level information on 1375 malaria impact, logistics and service delivery. The information needs and MALARIA CONTROL INTERVENTIONS: A COST- opportunities of the Zambia National Malaria Control Programme are EFFECTIVENESS ANALYSIS reviewed with respect to the routinely-reported health information system. A standard indicator set, methods of data collection, data systems for Sarah R. Kaslow, Darren Filson storage and most importantly, analysis and presentation of results were Claremont McKenna College, Claremont, CA, United States developed from national HMIS system data to support the information needs of an ever-complex malaria epidemiological situation over the With a human burden of more than 500 million people every year, the period 2001-2010. The resulting national malaria bulletin serves, based on need for cost-effective malaria control is outstanding. The objective of a consolidated District Health Information System 2.0 information system malaria control is to significantly reduce the rate and number of cases platform, as a model for malaria endemic-African countries to improve of parasitic infections and clinical malaria. This study analyzes a number analytic capacity of the Ministry of Health and malaria control partners to of control interventions and presents a template for decision makers promote best-practice malaria monitoring, evaluation, and surveillance for looking to reduce the burden of malaria in a cost-effective manner.Twenty improved decision making for malaria control at national and local levels. two malaria control interventions are analyzed. Cost data from WHO- CHOICE were used to calculate the current DALY value was calculated 1374 using twelve scenarios, differing in β (age weighting parameter), K (age weighting modulation factor) and r (rate of discounting). Standard life USING CENSUS METHODS TO PREPARE FOR EVALUATING expectancy was calculated using WHO tables. Total current DALYs were THE IMPACT OF UNIVERSAL SCALE-UP OF MALARIA calculated using the current DALY value and age group population data INTERVENTIONS: CASE OF LIKOMA DISTRICT, MALAWI for each country from the United Census Bureau International Data Base. DALYs averted were then calculated by multiplying the coverage rate 1 2 Bertha M. Nhlema Simwaka , Misheck Luhanga , Jessica of an intervention and its efficacy with the total current DALY. Efficacy 3 4 2 5 1 Oyugi , Wilfred Dodoli , John Zoya , Rick Steketee , John Miller , rates were determined using WHO cited literature but were varied to 6 2 Adam Bennett , Doreen Ali model three scenarios for each intervention. The cost-effectiveness of 1Malaria Control and Evaluation Partnership for Africa, Lusaka, Zambia, interventions was analyzed by considering the average cost per DALY 2National Malaria Control Programme, Community Health Sciences Unit, averted. The most cost-effective intervention under every scenario is Ministry of Health, Lilongwe, Malawi, 3Centers for Disease Control, case management with artemisinin-based combination therapy; the least Lilongwe, Malawi, 4World Health Organisation, Lilongwe, Malawi, 5Malaria cost-effective intervention is insecticide-treated bed nets. Results from Control and Evaluation Partnership for Africa, PATH, Atlanta, GA, United the cost-effectiveness analysis suggest that marked increases in funding States, 6Tulane University School of Public Health and Tropical Medicine, for and supplies of insecticide treated bed nets may be misguided from a New Orleans, LA, United States cost-effectiveness standpoint. Recent pushes to scale up artemisinin-based combination therapy policies fall in line with the findings of this study, Malaria Indicator Surveys (MIS) and Demographic Health Surveys (DHS) especially in sub-Saharan Africa. are traditionally used to document coverage and impact of interventions on disease burden. Malawi carried out its first MIS in 2010, but sampling excluded Likoma Island, which is situated in Lake Malawi. The Malawi National Malaria Control Programme planned to pilot a campaign to achieve universal coverage of long-lasting insecticide treated nets (LLINs) in 2010 in Likoma district; comprising Likoma and Chizumula Islands.

www.astmh.org 414 1376 treated mosquito nets (ITNs) and indoor residual spraying (IRS) has increased since 2006. However, while malaria infection and anemia UNEXPECTED PATTERNS OF PRIMARY SYMPTOMATIC prevalence in children younger than 5 years dropped in 2008, results MALARIA AND NON-MALARIA ILLNESS EMERGE IN A BIRTH from 2010 indicate that levels have rebounded in several parts of the COHORT OF GHANAIAN CHILDREN DESPITE WIDESPREAD country. We sought to ascertain the relative effects of ITN coverage, IRS, USE OF INSECTICIDE-TREATED BEDNETS (ITNS) AND and climate variability on the spatial distribution of malaria infection and STANDARD ARTESUNATE COMBINATION THERAPY (ACT) anemia in 2006, 2008, and 2010. We fit Bayesian geostatistical models to assess the effect of intervention coverage on malaria infection and David J. Fryauff1, Frank Atuguba2, Kwadwo A. Koram3, Timothy severe anemia prevalence, while adjusting for climatic and socioeconomic Awine2, Thomas Anyorigiya2, Tracy Nolen4, Abraham R. Oduro2, factors. We assessed the spatial dependence of disease distribution Victor Asoala2, Abraham Hodgson2, Thomas L. Richie1, Francis through time with spatial random effects for each survey. Model fit was Nkrumah3 conducted with Markov chain Monte Carlo simulation. Malaria infection 1Naval Medical Research Center, Silver Spring, MD, United States, and severe anemia prevalence rose from 2008 in six of nine provinces, 2Navrongo Health Research Centre, Navrongo, Ghana, 3Noguchi Memorial and from 12% to 20% across rural areas nationally. Parasite prevalence Institute of Medical Research, Accra, Ghana, 4Research Triangle Institute, increased by the largest percentage in Luapula (132%), Northern, (97%), Raleigh, NC, United States and Eastern (137%) provinces. Household ITN possession fell 31% in Luapula province and 29% in Northern province, but remained constant To gauge the extent and impact of a national malaria prevention/control in Eastern province. Parasite prevalence also increased in Central (by strategy based on insecticide-treated bednets (ITNs) and Artesunate 19%), Western (96%), and Copperbelt (22%) provinces, even though Combination Therapy (ACT), all-cause illness, symptomatic malaria, and ITN coverage also increased, by 45%, 121%, and 9%, respectively. 20- death was monitored in a cohort of 2,279 live births from March, 2006 day cumulative rainfall estimates two months before each survey were to October, 2008, in the Kassena-Nankana District of northern Ghana. positively associated with odds of malaria parasite infection; rainfall was Ownership of ITNs in this cohort rose from 76% at 4 mos. to 95% at 16 highest preceding the 2010 survey, and lowest in 2008. In 2010, greater mos. and malaria prevalence in young infants was 15% in households ITN age was associated with greater odds of malaria parasite infection. with an ITN compared to 31% in households without (OR: 2.7; 95% Spatial dependence increased with each survey year. These results suggest CI: 2.0, 3.6). Despite high ITN coverage and free, standard Artesunate that a combination of climatic factors, lower ITN coverage, and ITN age + Amodiaquinetreatment for uncomplicated malaria, Plasmodium contributed to the rebound in parasite infection prevalence in some areas. falciparum was detected in 44% (608/1,375) of all inpatient (IP) and Unusual rainfall patterns in the early part of 2010, perhaps related to 35% (6,327/18,223) of all outpatient (OP) visits. Several unexpected moderate El Niño conditions, may have contributed to this increase. We findings emerged: 1) Firstborns had a significantly lower incidence of emphasize the importance of accounting for climate variability and spatial primary symptomatic malaria and a longer time to this event than infants heterogeneity when using cross-sectional data for malaria evaluation of multigravid mothers, but suffered a higher incidence of non-malaria efforts. illness; 2) Between the two dominant ethno-linguistic groups (Kassem and Nankam), onset of primary malaria illness in infants of Nankam ethnicity 1378 came two months earlier and infections amounted to an estimated 500 more OP malaria cases/thousand, but paradoxically, these children accrued FACTORS AFFECTING ACCESS TO ACT TREATMENT FOR grossly lower annualized rates of severe malaria and non-malaria illness UNCOMPLICATED MALARIA IN WESTERN KENYA requiring hospitalization; 3) More high parasitemias were seen in females, but males accounted for a significantly greater proportion of severe Guofa Zhou1, Guiyun Yan1, Yaw A. Afrane2, Andrew K. Githeko2 malaria anemia cases (2.6% vs. 1.9%; P = 0.04). Malaria was associated 1University of California at Irvine, Irvine, CA, United States, 2Centre for with 36% of cohort deaths that occurred in the hospital but case fatality Global Health Research, Kenya Medical Research Institute, Kenya rate for children with malaria was 2.5% compared to 5.7% (P = 0.02) for admissions with no detectable parasitemia. All symptomatic malaria Effective case management is central to reducing malaria mortality and was seen in the post-neonatal period, associated with 20% of IP deaths morbidity worldwide, but only a minority of those affected by malaria, in infants, 53% of IP deaths in children >12 mos., and collectively in 36% have access to prompt effective treatment. In Kenya, the treatment of all IP deaths. Our cohort all-cause infant mortality rate of 37/thousand, policy for malaria has changed from chloroquine (CQ) to sulphadoxine- which may owe ~20% of its deaths to malaria, is well below the infant pyrimethamine (SP) as the first-line antimalarial drug for uncomplicated mortality rate of 68/thousand for the Upper East Region, and the national malaria in 1998, and then from SP to artemisinin-based combination rate of 71/thousand calculated by the Ghana Multiple Indicator Survey therapies (ACTs) in 2004. Despite this, these three classes of antimalarial in 2006. These results, derived from real-life practices and outcomes in a drugs are being used by residents in endemic areas in Kenya; however the rural community may be indicative of new trends in malaria prevention extent of ACT usage is unknown. We have surveyed 1,100 households of and child survival that are occurring widely in sub-Saharan Africa. 5,775 individuals and 1,930 antimalarial prescriptions/treatments in three district in western Kenya in 2003 and 2010. We found that the SP and amodiaquine (AQ) based antimalarials accounted for 88% of prescriptions/ 1377 treatments in 2003, 4% of the cases were treated with quinine (QN) ASSOCIATION BETWEEN VECTOR CONTROL COVERAGE, and the rest were with CQ. Malaria treatment-seeking occurs mostly in CLIMATE VARIABILITY AND THE SPATIAL DISTRIBUTION OF the formal sector, i.e., government-run health centers and hospitals. In MALARIA AT THREE TIME POINTS IN ZAMBIA 2010, 58% of the cases were treated in government-run health centers/ hospitals. Overall, only 60% of the antimalarials used in 2010 were Adam F. Bennett1, John Miller2, Moonga Hawela3, Busiku first-line government recommended drugs. Shortage of ACTs in stock at Hamainza3, Penelope Vounatsou4, Mulakwa Kamuliwo3, Rick government hospitals and clinics and less cost for CQ and SP drugs in Steketee5, Thomas P. Eisele1 private sectors are the major reasons for the patients to obtain them from 1Tulane University School of Public Health and Tropical Medicine, New the private sectors. Orleans, LA, United States, 2PATH/MACEPA, Lusaka, Zambia, 3National Malaria Control Centre, Lusaka, Zambia, 4Swiss Tropical and Public Health Institute, Basel, Switzerland, 5PATH/MACEPA, Ferney-Voltaire, France Three malaria indicator surveys (MIS) have been conducted in Zambia since 2006 to evaluate intervention scale-up. Coverage of insecticide-

www.astmh.org 415 1379 among single or concurrent P. falciparum, P. malariae, P. vivax and P. ovale infection. By LDR, 38% of Milalani samples (267/704) and 19% HOUSEHOLD SURVEYS OF MALARIA INFECTION, FEVER of Nganja samples (92/481) were positive for at least one Plasmodium AND ANEMIA IN A BIRTH COHORT OF GHANAIAN INFANTS species. Of these positives, 44% and 48% were positive for P. falciparum REVEAL TRENDS IN ITN OWNERSHIP, BENEFIT AND alone in Milalani and Nganja respectively, 20% and 17% positive for P. PARENTAL ACTION IN SEEKING HEALTH CARE FOR THE BABY malariae alone, 7% and 2% positive for P. vivax, and 7% and 4% for P. ovale alone. Pediatric cases were more common in Milalani (32%) than David J. Fryauff1, Frank Atuguba2, Kwadwo A. Koram3, Timothy Nganja (23%, p=0.014), however Milalani sampling occurred in July 2009 Awine2, Thomas Anyorigiya2, Abraham R. Oduro2, Victor Asoala2, (early dry season) and Nganja collection in April 2009 (mid wet season). Abraham Hodgson2, Francis Nkrumah3 The difference between villages in adult cases was not significant. As a 1Naval Medical Research Center, Silver Spring, MD, United States, screening tool, ICT cards (designed for detection of P. falciparum) had 2Navrongo Health Research Centre, Navrongo, Ghana, 3Noguchi Memorial sensitivity of 43% for falciparum, 10% for vivax and 0% for ovale and Institute for Medical Research, Accra, Ghana malariae. Specificity was 99% for all four species. Specificity remained the same for falciparum in single vs. multi-species infections (99%), but Prospective study of illness and death in a birth cohort of 2279 Ghanaian sensitivity lowered to 29% when non-falciparum species were present. infants was complemented by cross-sectional surveys timed to correspond ICT positive predictive value (PPV) for P. falciparum was 86% and negative with the end of low (April-May) and high (Nov.-Dec.) malaria transmission predictive value (NPV) was 90%. For single-species P. malariae infection, seasons. Home visits during Oct.-Nov. 2006, when infant’s age averaged the PPV was 0% and NPV was 92%. PPV for isolated P. vivax was 15% 4 mos., found ITN ownership in 75% of households (range: 62-83%). and NPV was 98%, and PPV for P. ovale was 0% and NPV was 97%. We Malaria prevalence associated with ITN ownership was 15%, ranging conclude that accurate attribution of infection-associated morbidities will from 3% (town) to 21% (non-irrigated, rural). Malaria prevalence among require the more sensitive and comprehensive PCR approach to molecular infants lacking ITNs averaged 31% and ranged from 12.5% (town) to detection of infection. 42% (irrigated rural). Although ITN ownership was similar in the four ethnic groups (74-89%), malaria prevalence was significantly higher among Nankam infants (22%; P = 0.011). Fever was reported by mothers 1381 in 29% of Plasmodium falciparum infections but actually measured in INTERVENTION IS NOT ENOUGH: FUNCTIONAL SOCIAL 13.5% of these cases. Prevalence of shaking, diarrhea, vomiting, cough, NETWORKS ARE CRITICAL TO THE SUCCESS OF COMMUNITY or breathing difficulty was similar in infants with and without parasitemia LEVEL MALARIA CONTROL but fever and anemia was more prevalent in children with P. falciparum. Records showed that only 27% of infants with parasitemia were brought Lucy Smith Paintain1, Lamine Gueye2, Mbayang Gueye Sall2, to clinic for treatment within 7 days of the home visit. Severe anemia, Alexandra Hyde1, Manuela Claite1, Babacar Gueye2, Jayne fever and older age distinguished those brought to clinic. Analysis also Webster1, Caroline Jones1 revealed that malaria in these infants was associated with multiple 1London School of Hygiene & Tropical Medicine, London, United Kingdom, maternal factors: older age, less education, no education, fewer ANC 2IntraHealth International, Dakar, Senegal visits, and no use of ITN during pregnancy. By Oct.-Nov. 2007, when children averaged 15 months old, ITN ownership had increased to 95% Despite recent trends showing a decrease in the malaria burden in but prevalence of P. falciparum infection was nearly twice greater than in Senegal, there is still substantial regional variation. In the Tambacounda the previous year, gametocyte carriage rate was more than doubled (9.6% region malaria accounts for 13.5% of all outpatient consultations vs. 3.7%; P < 0.001), high parasitemias >20,000/µL were four times more and 22.4% of mortality; parasite prevalence in children under five prevalent (13% vs. 3%; P < 0.0001), and anemia (Hb <8.0) had increased is 23.4% compared to the national figure of 5.7%. Coverage with in both parasitemic (45.5% vs 10.3%; P < 0.0001) and parasite-free (15% key interventions, including access to prompt and effective treatment vs. 5.2%; P < 0.0001) children. Records showed that 38% of children with artemisinin combination therapy (ACT), is low. The population of with parasitemia were brought to clinic for treatment within seven days Tambacounda is widely dispersed and access to health care is limited. The of the home visit. Among 259 children with malaria who did not report health system in Senegal provides community outreach services at the there were 64 parasitemias >10,000/µL, 27 with fever, and 6 with severe village level through health huts staffed by community health workers anemia. It appears that the benefit of ITNs was offset by heightened (CHW) and managed by village health committees (VHC). However, in immunological susceptibility of these young children. Tambacounda most community level services are no longer functional. The Mobilise Against Malaria programme rehabilitated 24 non-functional 1380 health huts and implemented an intervention to improve access to prompt and effective malaria treatment in these health huts. Midterm household PERFORMANCE OF A RAPID DIAGNOSTIC CARD TEST FOR survey data found limited overall improvement in prompt and effective DETECTION OF SINGLE- OR MULTI-SPECIES PLASMODIUM treatment of febrile children under five. Contextual data suggest several INFECTIONS AMONG RESIDENTS OF SOUTHERN COAST reasons for these results but monitoring data indicate that some health PROVINCE, KENYA huts perform better than others despite sharing many contextual factors. We conducted case studies of 3 well performing and 3 poorly performing Laura J. Sutherland1, Amaya L. Bustinduy1, Peter L. Mungai1, Eric health huts to investigate the facilitating factors and barriers to their M. Muchiri2, Uriel Kitron3, Peter A. Zimmerman1, Charles H. King1 effective operation. Health huts were identified using a combination 1Case Western Reserve University, Cleveland, OH, United States, 2Ministry of objective and subjective functionality criteria (including treatment of Public Health and Sanitation, Nairobi, Kenya, 3Emory University, Atlanta, indicators, utilisation rates, ACT stock, completeness of reporting). GA, United States In-depth interviews were conducted with the CHW and VHC of the 6 health huts as well as their district supervisors. The health hut system is As part of a study of polyparasitism, we examined the utility of rapid centred on the CHW; however the CHWs are themselves at the centre of diagnostic testing for classification of malaria infection status. Diagnostic a complex network of social relationships that function to influence the cards are readily deployable but may fail to accurately identify all prevalent success of the health hut as a community-level service provider. These cases and may miss non-falciparum or multi-species Plasmodium critical relationships and the broader implications of the findings are infections. In our study, adult and pediatric samples were collected in discussed. two villages in Msambweni District, Kenya, and tested for malaria both by ICT card and multiplex PCR-Ligase Detection Reaction (LDR). LDR is an accurate and sensitive method to detect malaria that can discriminate

www.astmh.org 416 1382 (35.1%). The majority of nets observed were LLIN: 90.2% (N=147) in Abia and 93.2% (N=811) in Plateau. The percentage of persons using nets the MALARIA INFECTION IN KENYAN PREGNANT WOMEN IS previous night were: Abia: 2.9% of all ages, 5.5% of children under 5 ASSOCIATED WITH LOW FETAL AND NEONATAL BIRTH years and 4.7% of pregnant women; Plateau: 15.5% of all ages, 20.7% WEIGHTS of children under 5 years, and 24.7% of pregnant women. Despite lower net use, the overall crude malaria prevalence (by CareStart PAN/Pf RDT) 1 2 Elizabeth M. McClure , Christopher L. King , Steven R. was lower in Abia (36.2%, 95% CI 34.3-38.0%, N=2614) than in Plateau 1 2 Meshnick , Arlene Dent (45.2%, 95% CI 43.8-46.8%, N=4212). Age-adjusted prevalence was 1University of North Carolina Chapel Hill, Chapel Hill, NC, United States, 29.7% in Abia and 36.9% in Plateau. Age specific prevalence peaked 2Case Western Reserve University, Cleveland, OH, United States in the 5 to 9 year age group at 52% (95%CI 48.0-55.9%) in Abia and 61% (95% CI 58.3-63.8%) in Plateau, with second highest prevalence Low birth weight is associated with malaria exposure in pregnancy; in the 10-14 year age group (Abia 48.9%,95% CI 42.2-55.6% ; Plateau however, little is known about fetal growth in utero among malaria- 54.9%, 95% CI 50.0-59.8).The percentage of children <5 with anemia exposed fetuses, which has implications for outcomes. We sought to (hemoglobin < 8 g/dl) was higher in Abia (20.5%, 95% CI 17.7-23.5%, evaluate fetal growth, birth weight and anthropometrics associated with N=785) than Plateau (9.9%, 95% CI 8.3-11.6%, N=1367). The results malaria exposure among infants born to Kenyan pregnant women. From indicate that malaria is highly prevalent in these two states, and that LLIN 2005 to 2007, pregnant women in Kenya were recruited at antenatal care ownership is low. The national campaign now underway to provide 2 LLIN (ANC) and those with term live births delivering at the study hospital who to every household in Nigeria is a welcome development. consented enrolled. Women received IPTp and were tested for malaria by microscopy at presentation and delivery. At least 1 fetal ultrasound (US) exam which measured fetal head circumference (HC), biparietal diameter, 1384 abdominal circumference, and femur length to generate fetal weight was NOVEL STRATEGIES LEAD TO NEAR ELIMINATION OF performed. Fetal growth measures stratified by gestational age at first MALARIA IN PREVIOUSLY HIGH-RISK AREAS IN SURINAME, US were compared using t-tests between fetus/infants with or without SOUTH AMERICA evidence of maternal malaria infection at first ANC. At birth, neonatal weight, length and HC were obtained and tested between those with and Helene Hiwat without malaria infection at first ANC. 485 women were enrolled and 29 Ministry of Health Malaria Program, Paramaribo, Suriname were malaria-positive at first ANC. Fetal weights were lower in fetuses of mothers with malaria infection in early pregnancy (<22 wks) compared Suriname was a high malaria risk country before the introduction of a to <22 wks fetuses of mothers without malaria infections (324 vs 395 new 5-year malaria control program in 2005, the Medical Mission Malaria g, respectively, p=0.04,). At > 23wks, while the estimated fetal weights Program (MM-MP). Malaria was endemic in the forested interior, where were lower for malaria-exposed, no statistically significant differences the stabile village communities were most affected. The interventions of were found. Birth weight of neonates born to mothers with vs. without the MM-MP included new strategies for prevention, case management, evidence of malaria infection at first ANC was significantly lower, 2836 behavioral change communication (BCC)/ information, education and g (SD 397) vs 3019 g (SD 432), respectively, p=0.03. Neonatal HC and communication (IEC), and strengthening of the health system (surveillance, length were not significantly different between malaria exposed and monitoring and evaluation and epidemic detection system). The malaria not exposed neonates. Our results suggest that infants exposed interventions of the MM-MP are reviewed and related to the Performance to malaria in utero had lower fetal and birth weights compared to infants Indicators established by the donor. The changes in the national malaria born to mothers with no evidence of malaria infection. Given the small situation during the years of the MM-MP, based on analysis of the national sample size, further research is needed. databases, are discussed. After a slow first year with non-satisfying scores for the performance indicators, the MM-MP truly engaged in its 1383 intervention activities in 2006 and kept its performance up until the end of 2009. A total of 69.994 long-lasting insecticide treated nets were MALARIA AND ANEMIA PREVALENCE AND LONG LASTING distributed and more than 15.000 nets re-/impregnated. Residual spraying INSECTICIDAL NET (LLIN) OWNERSHIP AND USE MEASURED was performed in high risk areas and over 10.000 people were screened IN REPRESENTATIVE HOUSEHOLD SURVEYS IN PLATEAU with Active Case Detection in outbreak or high risk areas. Additional AND ABIA STATES, NIGERIA IN 2010 notification points were established and the national health system was strengthened. Malaria vector populations, monitored in sentinel Amy E. Patterson1, Adamu Sallau2, Frank O. Richards1, sites, collapsed after 2006 and the number of national malaria cases Emmanuel Emukah3, Emmanuel Miri2, Abel Eigege2, Iheanyichi decreased from 8618 in 2005 to 1509 in 2009. Malaria transmission Okoroafor4, Mary Umar5, Olusola B. Oresanya6, Masayo Ozaki1, risk has shifted from the stabile village communities to the mobile gold Elizabeth Cromwell1, Patricia M. Graves1 mining communities, especially those along the French Guiana border. 1The Carter Center, Atlanta, GA, United States, 2The Carter Center Nigeria, The novel strategies for malaria control introduced in Suriname have led Jos, Nigeria, 3The Carter Center Nigeria, Owerri, Nigeria, 4Abia State to a significant decrease in the national malaria burden. The challenge is Ministry of Health, Umuahia, Nigeria, 5Plateau State Ministry of Health, to further reduce malaria using the available strategies as appropriate in Jos, Nigeria, 6Federal Ministry of Health, Nigeria, Abuja, Nigeria the affected areas and populations. Elimination of malaria in the country will require a thorough understanding of transmission dynamics and a There have been few recent surveys of malaria prevalence and net dedicated investment in key effective interventions. coverage in Nigeria. In September 2010, The Carter Center worked with the ministries of health of Abia (south east Nigeria) and Plateau (north central Nigeria) states to conduct a modified Malaria Indicator Survey. The survey was completed prior to mass LLIN distribution campaigns in these states. In 60 systematically selected clusters (census enumeration areas or segments thereof) of 25 households each per state, the average household size was 4.4 persons in Abia (1429 households, 5764 persons) and 6.2 in Plateau (1379 households, 8331 persons). Children <10 years of age in all households were tested for malaria and anemia, and all persons in every third household were tested for malaria. The percentage of households owning at least one net was much lower in Abia (7.2%) than Plateau

www.astmh.org 417 1385 and expanding coverage of community funds in rural areas could offer protection to rural households from paying more than they earn and when MALARIA FORECASTING: PAST WORK AND FUTURE they are not able to work. DIRECTIONS 1387 Kate Zinszer1, Aman Verma1, John Brownstein2, Timothy Brewer1, David Buckeridge1 FORECASTING AND MONITORING MALARIA RISK IN THE 1McGill University, Montreal, QC, Canada, 2Harvard University, Boston, MA, AMHARA REGION, ETHIOPIA United States Alemayehu Midekisa1, Gabriel Senay1, Geoffrey M. Henebry1, Since 1911, when Christophers quantified the strong correlation between Paulos Semuneguse2, Abere Mihretie2, Michael C. Wimberly1 malaria incidence and rainfall, researchers have sought to discover other 1South Dakota State University, Brookings, SD, United States, 2Anti-Malaria sources of spatial and temporal variability of malaria. The field of malaria Association (AMA), Addis Ababa, Ethiopia incidence forecasting has incorporated predictors responsible for this variability although the approaches are quite diverse. We conducted Malaria is a major health problem in most sub-Saharan Africa countries, a scoping review to summarize the heterogeneous field of malaria including Ethiopia. Mosquito populations and malaria risk are affected forecasting and describe the modeling approaches and methods of model by environmental triggers, including rainfall, temperature, and humidity. evaluation. Two reviewers identified articles by using medical subject The main objectives of this study are to compare alternative statistical headings and key terms to search electronic databases and grey literature, forecasting models and quantify the lead time of satellite derived including articles that presented models predicting human Plasmodium environmental variables with malaria outbreaks. Daily rainfall data were falciparum malaria incidence or prevalence. The initial search captured acquired from the Tropical Rainfall Measuring Mission (TRMM) with 0.25 213 different citations, 46 (22%) of which were reviewed. Most models degree spatial resolution. Land surface temperature (LST), normalized predicted malaria incidence in Africa (72%), with 10 studies (23%) difference vegetation index (NDVI), and enhanced vegetation index (EVI) conducted in Kenya. Models mostly included two predictors (52%), were derived from MODIS 8 day and 16 day composites with a 1 km which were the previous month’s cases of malaria and rainfall. Typically, spatial resolution. Actual evapotranspiration (ETa) was estimated by using researchers used ARIMA-based models, which performed optimally the simplified surface energy balance method. Monthly malaria case data within the first 3 months and evaluated using magnitude of correlation for the Amhara region were acquired from the Anti Malaria Association between predicted and observed incidence or prevalence. Nearly all (AMA), Ethiopia. Satellite derived indices were aggregated at a monthly malaria prediction models have narrowly focused on a small number of resolution to match malaria cases. Using the historical satellite and case environmental predictors despite the importance of other malaria risk data, we explored a variety of time series modeling approaches with factors (land use, bednets, indoor residual spraying, drug resistance). To combinations of the different variables. We used environmental variables advance malaria prediction forecasting, we need more comprehensive with lags ranging from 1 to 6 months and examined the temporal models as coarse models will not provide the precision necessary to guide cross correlations between outpatient malaria cases and environmental targeted intervention efforts. variables. The results showed that there were significant correlations based on a two standard error limit with rainfall at a lag of 3 months; nighttime 1386 LST and ETa at 1 month; and NDVI and EVI at 2 months. We found that rainfall and nighttime LST were the strongest predictors for malaria risk MONITORING MALARIA IMPLEMENTATION COST - THE CASE in the Amhara region of Ethiopia. Forecasts were validated using data OF ARTEMETHER-LUMEFANTRINE withheld from the model fitting. The results showed that the models provide indications of future outbreaks with a lead time of 1 to 3 months. 1 1 1 Fatuma Manzi , Gumi Abdallah , Rashid Khatibu , Patricia The findings can be used to enhance future operational Malaria Early 2 Akweongo Warning Systems in the Amhara region, Ethiopia. 1Ifakara Health Institute, Dar es Salaam, United Republic of Tanzania, 2University of Ghana School of Public Health, Accra, Ghana 1388 Tanzania has adopted Artemether-Lumefantrine (Alu) as the first line THE DYNAMICS OF NATURAL PLASMODIUM FALCIPARUM treatment for malaria. INESS has introduced a series of studies in Rufiji INFECTIONS and Kilombero/Ulanga Health and Demographic Surveillance System (HDSS) sites to evaluate safety and effectiveness of this drug and their Thomas A. Smith1, Michael Bretscher1, Seth Owusu-Agyei2, related factors. The collection of treatment related costs has provided Hans-Peter Beck1, Ingrid Felger1 an opportunity to analyse the household level costs for getting malaria 1Swiss Tropical and Public Health Institute, Basel, Switzerland, 2Kintampo treatment. Here we report findings of assessment of treatment costs in Health Research Centre, Kintampo, Ghana Rufiji HDSS. Data were collected using household survey conducted in Rufiji DSS site. Members with recent fever from the sampled households Natural immunity to Plasmodium falciparum malaria has been widely were asked questions about professions, economic activities, treatment studied, but its effects on parasite dynamics are poorly understood. seeking, costs involved and day lost due to illness. 29.6% of patients Acquisition and clearance rates of untreated infections are key elements interviewed reported to be actively engaged in income generating of the dynamics of malaria, but estimates of these quantities for endemic activities. Farming is the main economic activity in the area accounting for areas is challenging because of frequent super-infection and imperfect 76.3%. A daily median income in the study area was USD.1.4. However, detectibility of parasites. Consequently, information on the effects of 22.7% of patients and 59% of accompanying persons reported to host immune status or age on these parameters is fragmentary. An age- have lost their income due to illness or escorting patients respectively. stratified cohort of 349 individuals from Northern Ghana were sampled The number of days lost due to a single malaria episode ranged 1-14 six times at 2 month intervals. High-throughput capillary electrophoresis days for patients. The direct medical costs (drugs, laboratory tests and (CE) was used to genotype the msp-2 locus of all detectible P. falciparum consultation) were paid by 81% of patients with median payment of infections. Force of infection (FOI) and duration were estimated for each USD0.8. Equally, 11.5% of patients paid non medical cost (including age group using an immigration-death model that allows for imperfect food, lodging, telephone and gifts) with median payment of USD0.4. detection. Effects of naturally acquired immunity on the FOI and duration Only 10.4% of patients reported paying transport cost to get to health should be reflected in age dependence in these indices, but FOI tends to providers with median payment of USD1.0. A single malaria episode was increase with age, and persistence and chronicity of individual parasite observed to cost households more than their income. Alternative payment clones is characteristic of all age-groups. Duration peaked in 5-9 year old mechanism in the form of insurance should be considered. Strengthening children, (average duration 319 days, 95% confidence interval 318;320). www.astmh.org 418 FOI tended to increase with age. The estimated multiplicity of infection 656 children across three villages were tested for malaria antigenemia (MOI) was considerably higher than the observed number of clones, by rapid diagnostic test and parents simultaneously completed survey especially in older ages. The main age-dependence is on parasite densities, questionnaires related to demographics, socio-economic status, maternal and acquired immunity therefore appears to control transmission mainly education, as well as bednet use and recent febrile illness. 19% of by limiting parasite densities in the human circulation. children were parasitemic (11%, 22% and 23% in Goma, Butembo and Beni, respectively; p=0.009). Increasing levels of maternal education 1389 were associated with a lower risk of malaria antigenemia in their children (p=0.001). Children from households with higher numbers of children MALARIA PARASITE GAMETOCYTEMIA IN A under 5 years of age were also more likely to be parasitemic (p=0.035). MIXED MALARIA SPECIES HYPOENDEMIC IN PERU: On the other hand, socio-economic index was not statistically associated ENVIRONMENTAL FACTORS AND GENETIC MARKERS with malaria antigenemia (p=0.27). In a multivariable model adjusting THAT PREDICT EFFICIENT PLASMODIUM FALCIPARUM for age, recent febrile illness, and bednet use, maternal education and TRANSMISSION number of children under five in the household remained significant predictors of malaria antigenemia. In summary, children of mothers with 1 1 2 Julia Johnke , Patrick L. Sutton , Jean N. Hernandez , OraLee H. low education level and living in households with large numbers of young 1 Branch children appear to be at higher risk of asymptomatic malaria carriage. 1Department of Medical Parasitology, New York University, New York, NY, Social determinants of malaria parasitemia may influence transmission United States, 2Laboratorio de Investigaciones de Productos Naturales y patterns and may be useful tools for targeted control efforts. Antiparasitarios, Universidad Nacional de la Amazonia Peruana, Iquitos, Peru 1391 Malaria parasites must convert to its sexual stage for transmission from LARGE DECLINE IN PLASMODIUM FALCIPARUM AND P. human to mosquito. Factors regulating if asexual stage trophozoites MALARIAE INFECTION RATES AMONG WOMEN AND THEIR switch to the sexual stage gametocytes are not known, but in Plasmodium OFFSPRING IN A COMMUNITY ON THE SOUTH KENYAN falciparum (PF) it is clearly not a linear function of within host trophozoite density. The rate of conversion might be attributable to environmental COAST FROM 1997-2010 factors and/or a heritable genetic trait. We have a study in Peru where Benjamin C. Kalayjian1, Indu Malhotra1, Peter Mungai2, Eric a high proportion of infections have gametocytes despite overall low Muchiri2, Christopher L. King1 transmission, enabling us to study the density and the proportion of 1Case Western Reserve University, Cleveland, OH, United States, 2Division gametocytes to trophozoites. In active and passive case detection of of Vector Borne Diseases, Nairobi, Kenya c.1,900 individuals from 2003-2007, we detected 456 PF and 956 P. vivax (PV) infections. This study focus was PF. We surveyed for PV by Expanded malaria control initiatives in Kenya include intermittent microscopy to determine mixed species infections. We also considered preventive treatment during pregnancy, insecticide treated bed-net usage, febrile illness, age (>14.5 considered adult), and hematocrit levels as and availability of artemisinin combination therapies for case management they relate to gametocytaemia. We genetically characterized PF parasites of malaria illness has resulted in dramatic decreases in reported hospital using 14 microsatellite (MS) markers. The proportion of infections with admission for severe malaria. The overall malaria infection rates have gametocytaemia ranged from 18% (year-2004) to 38% (2007). In the presumably decreased as well, however no studies have reported adults not in children, there was a stepwise increase in proportion with longitudinal malaria infection rates in the same population of women gametocytes each year. Gametocyte densities were higher in mixed species and young children, using sensitive and specific molecular diagnostic infections. Using the program Structure, we found six families of related methods over the past decade of malaria control interventions in Kenya. parasites based on the MS markers. One of the six clusters included 56% Randomly selected archived blood samples obtained from four birth of infections with gametocytemia. Moreover, with principle component cohorts of pregnant women and their children (starting in 1997 [n=54], analysis we found that PF infections with a ratio of trophozoite to 2000 [n=174], 2006 [n=444], 2009 [n=76]) enrolled from the Msambweni gametocyte density was >25% were caused by a genetically distinct District hospital located in Coast Province, Kenya were examined by the group of PF parasites. Therefore, in addition to environmental factors same PCR- based molecular diagnostic assays for infection with the four for conversion there appears a genetic signature for parasites with high human malaria parasites. Maternal Plasmodium falciparum (Pf) infections gametocyte conversion in our study population. We are determining if at delivery (1997 and 2000 cohorts where malaria chemoprophylaxis was there is a genetic association with PF gametocytemia independent of infrequently used) and at first antenatal clinic visit (2006 and 2009) were environment and antibody responses. Our work provides insight into the 37%, 40%, 15% and 17% respectively. P. malariae (Pm) infections in transmission of PF in Peru and suggests that eradication campaigns could mothers also declined from a peak of 12% in 2000 to 8% in 2009. Peak create a reservoir of more transmissible PF parasites. Pf and Pm malaria infection rates in children occurred at 30-36 months of age in all cohorts. At this age infections rates with Pf declined from 58% 1390 in 2000 to 10% in 2006. Similarly Pm declined from 9% to 3% over the same period. Peak P. ovale infections remained at ~2% and P. vivax was SOCIAL DETERMINANTS OF ASYMPTOMATIC MALARIA not observed in the cohorts. These results show a profound reduction in ANTIGENEMIA IN TROPICAL AFRICA malaria transmission consistent with an overall reduction in the burden of malaria in the coastal regions of Kenya. Michael Hawkes1, Claude Kasereka MC2, Tsongo Kibendelwa2 1University of Toronto, Toronto, ON, Canada, 2HEAL Africa, Goma, The Democratic Republic of the Congo In the context of intensifying efforts toward global malaria eradication, understanding social patterns of malaria transmission from asymptomatic populations in endemic zones will become increasingly important. Social determinants of health have previously been linked with child and maternal mortality, but not to asymptomatic malaria carriage. We conducted a cross-sectional study of afebrile, healthy children aged 2 months to 14 years attending well-child and/or immunization visits in the North Kivu province in eastern Democratic Republic of Congo. A total of

www.astmh.org 419 1392 was 85% in the first and 60% in the second infection (p=0.012). The probability of not detecting an infection in the second year (despite having PLASMODIUM SPECIES UNDETECTED BY PLASMODIUM at least 6 active detection visits to find even asymptomatic infection) FALCIPARUM SPECIFIC RAPID DIAGNOSTIC TESTS AMONG ranged from 21 to 62%, increasing with number of infections prior to FEVER PATIENTS IN ZANZIBAR these two successive infections. Febrile illness was associated with having a different MSP1-B2 genotype in the second infection (p=0.041). To further 1 2 1 Kimberly A. Baltzell , Deler Shakely , Michelle Hsiang , consider genetic diversity and immunity, we genetically characterized 303 1 3 2 Jordan Kemere , Abdullah S. Ali , Anders Björkman , Andreas PF infections using the MS markers. The overall genetic diversity increased 2 2 3 Mårtensson , Kristina Elfving , Bakari J. Mohammed , Makame H. over time, with parasites in the later years having markedly different MS 3 1 1 Makame , Bryan Greenhouse , Philip J. Rosenthal haplotypes. We calculated the number of genetic differences between 1University of California San Francisco, San Francisco, CA, United States, the first and second infection in each individual. MS markers are not 2Karolinska Institutet, Stockholm, Sweden, 3Zanzibar Malaria Control considered under immune selection, but in low transmission they would Programme, Zanzibar, United Republic of Tanzania indicate potential allelic differences in antigens in linkage disequilibrium with (nearby) the MS markers. Infections spaced by <18 months (mos) had Malaria incidence has decreased markedly in Zanzibar, and the island an average of 5.3 (se: 0.3) differences. Infections spaced by >=18 mos had is considering elimination. However, non-falciparum malaria may be an average of 4.0 (se: 0.3) differences. The significant association with particularly difficult to eliminate. Importantly, the routine diagnostic time separating infections suggested that parasites causing reinfection standard in Zanzibar has been a rapid diagnostic test (RDT) based on within 18 mos had to be more genetically different than those occurring histidine-rich protein-2, which is produced by Plasmodium falciparum but after the immune response might no longer exert a selective pressure not other malaria species. The use of P. falciparum specific RDTs therefore against a similar parasite reinfecting. Other tests of genetic differences in may prevent comprehensive malaria case finding in Zanzibar. We report reinfections versus differences expected by chance indicated selection for species data on RDT negative, PCR positive malaria cases in Zanzibar parasite diversity. Our results suggest immunity develops and at least some during the 2010 rainy season. We collected blood on filter paper from of this immunity is directed to antigens that have different allelic forms in a random sample of febrile patients who tested negative for malaria by this population. RDT in 6 primary health care facilities in North A and Micheweni District, respectively (N = 595). DNA was extracted with Chelex from dried blood spots in pools of 10, and nested PCR targeting the cytochrome 1394 b gene was performed. Samples from positive pools were reextracted INCUBATION PERIOD OF PLASMODIUM FALCIPARUM individually. An AluI restriction digest and gel electrophoresis were then MALARIA IN ADULT TRAVELERS IN THE UNITED KINGDOM performed on positive individual samples for species identification. Of the 595 RDT negative samples, 13 (2%) were positive by PCR. Nine out T. Deirdre Hollingsworth, Christl A. Donnelly, Amy Pinsent, of the 13 positives (69%) were identified as P. falciparum, 3/13 (23%) Geoffrey Pasvol as P. malariae, and 1/13 (8%) as P. vivax. Three of 4 subjects with non- Imperial College London, London, United Kingdom falciparum infections were from Micheweni and the remaining subject was from North A. This study is the first, to our knowledge, to report The incubation period (from infection to onset of symptoms) for P. vivax in Zanzibar. With improved control of P. falciparum infection on Plasmodium falciparum malaria has been shown to vary widely from the island and with challenges to control of other species, infection with days to months in adults. Estimation of the incubation period distribution non-falciparum species may make up an increasing proportion of malaria informs both diagnosis criteria in non-endemic areas and the design of cases on Zanzibar. In this context RDTs including both P. falciparum specific passive surveillance in areas where local elimination is been achieved. Data and pan-species specific antigens should be considered for improved on the duration of holiday, date of arrival in the UK and date of onset of overall malaria case detection and to ensure that elimination efforts are symptoms for 413 adults (over 16 years old) with P. falciparum malaria comprehensive. reporting to the Infectious Diseases Unit at Northwick Park Hospital, London, UK between April 1991 to May 2006 were analysed. The mean 1393 incubation period was estimated using interval censored survival analysis assuming a Gamma distributional form. The role of self-reported previous RAPID DEVELOPMENT OF PLASMODIUM FALCIPARUM malaria, antimalarial use, severity of disease, age and ethnic origin (a STERILE, INFECTION BLOCKING IMMUNITY OVER key determinant of severity in this population) was investigated. 17% of SUCCESSIVE INFECTIONS IN PERU SELECTING FOR PARASITE cases had onset of symptoms prior to arriving in the UK, 40% became GENETIC DIVERSITY ill within the first week of arrival and 5% reported first symptoms more than a month after arrival. The mean incubation period was 20 days (95% OraLee Branch1, Patrick L. Sutton1, Jean Hernandez2, Julia confidence interval 11-41days). Important determinants of the incubation Johnke1, Gisely Hijar3 period (p<0.05) were self reporting of previous malaria (mean 17 days for 1Department of Microbiology and Medical Parasitology, New York no previous malaria, 23 days with previous malaria) and severity of disease University School of Medicine, New York, NY, United States, 2Laboratorio (mean 16 days in patients with severe disease, 21 days in other patients). de Investigaciones Productos Naturales Anti-parasitarios, Universidad This analysis has been performed in a group of adults resident in the UK Nacional Amazonia Peruana, Iquitos, Peru, 3Instituto Nacional de Salud, and abroad who had severe enough symptoms to report to hospital, and Ministerio de Salud, Lima, Peru these estimates may therefore be biased. Severity is associated with a short incubation period in these patients, and therefore incubation periods may The malaria paradigm is that resisting symptomatic and high-density be much longer for mild cases and in immune populations. The degree Plasmodium falciparum (PF) infections requires years of frequent exposure of variability in incubation periods has only rarely been reported and to develop and continued exposure to maintain. In contrast, in a setting therefore this study gives useful insights for both the UK and international of recent and low transmission in Peru, we see a rapid onset of clinical context. and anti-parasite immunity. Immunity might be related to the PF genetic diversity. We followed 456 individuals who had a PF infection in one year between 2003 and 2007 and were in our active and passive case surveillance in the following year. We genotyped parasites using the Merozoite Surface Protein-1 (MSP-1 B2) and using 14 MS markers scattered across the 14 PF chromosomes. Considering the 175 individuals who had two infections in these two years, the probability of febrile illness

www.astmh.org 420 1395 annual increment ranging from 15-44%, in the this non endemic region. Although the total number is modest, when compared with those HIGHER MALARIA PREVALENCE IN CHILDREN FIVE YEARS recorded in endemic areas in Brazil, two challenges are set for this scenario AND ABOVE IN LAGOS, NIGERIA to avoid increase in morbidity and mortality: the difficulty of rapid and accurate diagnosis; and the training on the management of potentially Wellington A. Oyibo, Oladipo O. Oladosu severe malaria including multidrug resistant P. falciparum, which is College of Medicine of the University of Lagos, Nigeria, Lagos, Nigeria extremely different from the observed in the Amazon region. Malaria is almost invariably ranked as the leading cause of morbidity and mortality in Africa. There is growing evidence of a decline in malaria 1397 transmission, morbidity and mortality over the last decades especially in CHARACTERISTICS OF THE INFECTIONS BY PLASMODIUM Africa. Reports on malaria prevalence in children in Nigeria are divergent in SPP. DETECTED BY ACTIVE SEARCH OF CASES IN FEBRILE the literatures. This study therefore reports the trend in malaria prevalence in febrile children in Lagos, Nigeria. This study was conducted at the St. AND NON-FEBRILE INDIVIDUALS OF THREE ENDEMIC Kizito Primary Health Centre, Lekki, and Massey Street Children’s Hospital COMMUNITIES IN OLANCHO, HONDURAS, SEPTEMBER 2010 Lagos State, Southwestern Nigeria. This was a cross-sectional study with Jackeline Alger1, Jorge Garcia2, Ofelia Martinez3, Martin a total of 1,211 children, aged 0-12years who presented with fever or Ramirez3, Ricardo Aviles4, Miguel Quintana5, Eric Garges6 history of fever in the last 24 hours at the Outpatient’s Department of 1Hospital Escuela; Faculty of Medical Sciences, Universidad Nacional the Clinics. Among the children tested, 658(54.4%) were males and Autonoma de Honduras, Tegucigalpa, Honduras, 2Hospital Escuela, 553(45.6%) were females. Of the total children tested microscopically, Tegucigalpa, Honduras, 3Region Sanitaria Departamental, Olancho, 251 (20.7%) were positive for malaria parasites. Children >1-12years Juticalpa, Honduras, 4Elemento Médico, Fuerza de Tarea Conjunta Bravo, in Age Group III had a malaria prevalence of 25.8%, 11.0% in the Comayagua, Honduras, 5U.S. Army Public Health Command Region - Age Group I (0-≤1year) (p=0.001), 16.9% in 0-≤5years and 42.1% in South, San Antonio, TX, United States, 6Preventive Medicine Residency >5-12years in Age Group II (p=0.001). The highest mean parasite density Program, Army POC - Military Tropical Medicine, Silver Spring, MD, United (43,097.6 p/μl) was reported in Age Group III (>1-12years). Most of the States malaria positive children (33.9%) had parasite density between 1-500p/μl. There was no significant association in monthly malaria prevalence in the Public health agencies in Honduras implement malaria control activities studied children. This study reported a decline in malaria prevalence, which by responding to reports of active cases. Previous investigations suggest may be attributed to large-scale implementation of malaria interventions. that subclinical cases of malaria exist in communities throughout the There was a shift in malaria prevalence from the well reported prevalence country, but these studies were limited in scope. Failure of health in 0-<5years to >5-12years. The shift in malaria prevalence to >5-12 year authorities to include subclinical cases in planning malaria interdiction olds may reflect successful implementation of malaria control interventions efforts minimizes the effectiveness of control programs. In this study, in under-fives, and underscored the need to extend such interventions to investigators performed malariometeric surveys to estimate the frequency older children and indeed implement universal target in malaria control. of subclinical cases in populations of three semi-rural communities located in the Department of Olancho, Honduras. Medical teams visited 30 homes 1396 in the communities of Sosa Lobo, Chacon and Villa Antonia, Olancho, to perform the survey. The visits took place from August 29 - September OCCURRENCE AND PATTERN OF IMPORTED MALARIA 2, 2010. Participants were interviewed and a cursory physical exam was CASES IN RIO DE JANEIRO, A NON-ENDEMIC STATE IN THE carried out. Blood samples were taken using finger sticks and a rapid BRAZILIAN EXTRA-AMAZONIAN REGION diagnostic tests (RDT) was performed during home visits. Additionally,

1 1 1 thick film slides were prepared for microscope evacuation and dried Otilia Lupi , Ana Claudia Vidigal , Cecilia Longo , Anielle de Pina blood filter paper collections were used for evaluating malaria parasites Costa1, Margarete Tavares1, Carolina Romero1, Patricia Moza2, 3 1 for polymorphic molecular markers. Seventy-one individuals participate Claudio Tadeu Daniel-Ribeiro , Patricia Brasil in the survey with 19 participants experiencing an episode of febrile 1IPEC/FIOCRUZ, Rio de Janeiro, Brazil, 2SESDEC/RJ, Rio de Janeiro, Brazil, illness (26.7%) within 30 days period prior to the home visit. Technicians 3Center for Malaria Diagnosis and Training, FIOCRUZ, Rio de Janeiro, Brazil detected one Plasmodium vivax (1.4%) positive when evaluating In Brazil, almost all (99.4%) of malaria reported cases are seen in the specimens with RDT. Subsequent microscopic evaluation of the thick films Amazon region where P.vivax accounts for 82.2% of them. The low resulted in the detection of four additional cases of P. vivax (7.0%). Two specificity of initial clinical presentation, overlapped with other acute of the participants that were thick film positive (2.8%) did not have a febrile diseases and the potential risk of severe malaria represents an extra clinical history suggestive of malarial infection and could contribute to the challenge for unaffected regions. The high lethality rates of malaria in persistence of transmission in their communities. The molecular analysis the extra-Amazon (70.8 times higher than in the Amazon region) drew of parasite genomic material from the five positive specimens detected the attention of authorities. A retrospective study was carried out from one genotype based on DNA fragment size: markers MSP1 5/6 (~380bp), Jan/07 to Apr/11 in a Reference Center that handles 23% of the malaria CSP (~600bp) and GAM1 (~500bp). The analyzed samples demonstrated cases seen in Rio de Janeiro. From the total of 291 admitted malaria genetic homogeneity and an absence of polyclonal infections. suspect patients, 83(28.5%) had the confirmed diagnosis of malaria. The distribution according to Plasmodium species was 58 (69.9%) P. vivax, 1398 18(21.7%) P. falciparum, 3(3.6%) P. malariae or P. ovale and 4 (4.8%) ECONOMICAL RABIES POST-EXPOSURE PROPHYLAXIS: A P. vivax/P. falciparum co-infection. Most of suspected cases were primo- infected men over 25 years old and the total lethality rate was 2.4%. SIMPLIFIED 4-SITE INTRADERMAL 3-VISIT REGIMEN ON DAYS Africa or Asia countries were the likely source of infection for 33(37.5%) 0, 7 AND 28 between 2007-08 and 77(38.1%) between 2009-11 (OR:1.04; IC:0.59- Mary J. Warrell, David A. Warrell 1.82%). This pattern of imported malaria in Rio de Janeiro is different University of Oxford, Oxford, United Kingdom from the one recorded in the Amazon region and may result from the recent Brazilian economic growth that has increased the presence of Human encephalitis caused by a dog rabies virus remains 100% fatal, construction, mining and oil companies in endemic areas as well as the although on rare occasions, patients have recovered from infection with presence of Brazilian enterprises in major international project, especially less pathogenic American bat rabies viruses. Post-exposure prophylaxis is in Africa. The authors note the recent increase in the absolute number often unaffordable or unavailable in Asia and Africa. The intradermal (ID) of suspect cases and in the percentage of P. falciparum infections, with www.astmh.org 421 route of vaccination has immunological and economical advantages over controls, seven were identified as AstV-canonical, one as AstV-MLB and IM. Two multiple-site ID regimens (8-site and 2-site) requiring <40% of the one as AstV-VA, respectively. Our results indicated that a real-time RT-PCR standard vaccine have been recommended by WHO for 14 years. They are assays can be used to detect and genotype human astroviruses. used successfully in a few places in Asia, but rarely in rural areas where 80% of rabies occurs. ID vaccination is practically unknown in Africa. 1400 Pharmaceutical and practical difficulties inhibit ID use together with failure to explain and promote the regimens. A new simplified 4-site ID vaccine KINETICS OF CHIKUNGUNYA INFECTION DURING AN regimen could overcome many of these problems (Warrell MJ et al. PLoS OUTBREAK IN SOUTHERN THAILAND, 2009 NTD 2008; 2(4):e224). The regimen employs the same doses of vaccine Sarunyou Chusri1, Pisud Siripaitoon1, Kachornsak Silpapojakul1, and the same schedule as the 8-site method, but can be used with rabies Tanaporn Hortiwakul1, Boonsri Charernmak1, Ananda Nisalak2, vaccines reconstituted to volumes of 1 ml or 0.5 ml per ampoule. The Butsaya Thaisomboonsuk2, Chonticha Klungthong2, Robert V. 4-site regimen consists of a whole ampoule of vaccine divided between Gibbons2, Richard G. Jarman2 4 ID sites, one on each limb on day 0. Two ID injections are given on day 7 and one on day 28. The ID dose is 0.2 ml or 0.1 ml for vaccines of 1 ml 1Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand, or 0.5 ml per ampoule respectively. The 4-site regimen is as immunogenic 2Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand as the ‘gold standard’ 5 dose IM course of vaccine. It requires a total of The Indian Ocean Chikunguna epidemic reemerged in Southern Thailand less than 2 ampoules, only 3 clinic visits and does not rely on expert ID in September of 2009. We enrolled forty-five adults with laboratory injection technique and so is safer in inexperienced hands. It meets all confirmed chikungunya. Serial blood collections and clinical assessments the WHO requirements and is very economical for both the clinic and were performed every two-three days through the acute and convalescent patients compared with all other regimens (Hampson K et al. PLoS NTD phase of the disease until day 30. Patient symptoms were recorded and 2011; 5(3):e982 Note extra data in ‘Comments’). Rabies vaccines do antibody responses with viral kinetics were evaluated using PCR and not contain a preservative and are not licensed as multi-dose ampoules serological assays. The patients mean age was 49 years with a male to unless authorised by a national regulatory body. If ampoules of vaccine female ratio of 1:1.4. Thirty-five (77.8 %) patients were rubber planters. are shared, ID rabies vaccination is given ‘on the doctor’s responsibility’ All patients experienced joint pain and 42 (93%) of them involved more in most countries. However without any sharing of ampoules, the 4-site than one joint. Interphalangeal joints were the most common affected regimen is economical even if only one person is treated: using a total of in 41 (91%) patients. The mean duration of severe joint pain was 5.8 3 doses with some wastage. The method is applicable globally wherever days with 11 (25%) experiencing discomfort through the duration of the financial resources or vaccine supplies are limited or if the number of clinic study. Rash was observed in 37 (82%) patients, a mean 3.5 days after visits is critical. the onset of symptoms. Patients were positive by PCR for a mean of 5.9 days and the peak viremia was at day 5 with 6.24 log PFU/ml. IgM 1399 antibodies appeared on day 4 and peaked at day 7. IgG antibodies first appeared at day 5 and rose steadily through day 24. The understanding of DETECTION AND GENOTYPING OF HUMAN ASTROVIRUS chikungunya disease clinical manifestation, antibody responses and viral IN NEPAL USING REAL-TIME REVERSE TRANSCRIPTASE kinetics are important for diagnosis and treatment of the disease. POLYMERASE CHAIN REACTION Pimmnapar Neesanant1, Ladaporn Bodhidatta1, Sanjaya K. 1401 Shrestha2, Jyoti Ratna Dhakhwa3, Bhola Ram Shrestha4, Carl J. PRE-CLINICAL AND CLINICAL DEVELOPMENT OF A VACCINE Mason1 FOR THE PREVENTION OF HAND FOOT AND MOUTH 1 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, DISEASE CAUSED BY ENTEROVIRUS 71 2Walter Reed/Armed Forces Research Institute of Medical Sciences Research Unit Nepal (WARUN), Kathmandu, Nepal, 3Kanti Children Cynthia Thomson1, Ping Ping Tong1, Yock Ann Lee1, Shi Hsia Hospital, Kathmandu, Nepal, 4Bharatpur Hospital, Bharatpur, Nepal Hwa1, Andrew Thomson1, Joseph Brewoo2, Charalambos D. Partidos2, Gilad Gordon3, Dan T. Stinchcomb3, Jorge E. Osorio2, Human astroviruses (HuAstVs) cause gastrointestinal disease; eight (1- Joseph D. Santangelo1 8) classic human astrovirus (AstV-canonical) serotypes are responsible for acute, nonbacterial diarrhea in children. Recently, two new human 1Inviragen (Singapore) Pte Ltd, Singapore, Singapore, 2Inviragen Inc., astrovirus variants (MLB and VA) have been described in humans. We Madison, WI, United States, 3Inviragen Inc., Fort Collins, CO, United States detected HuAstVs by a real-time reverse transcription-polymerase chain Hand Foot and Mouth Disease (HFMD) is caused by viral pathogens of reaction (RT-PCR). One step TaqMan multiplex real-time RT-PCR assays the enterovirus genus such as enterovirus 71 (EV71) or Coxsackie A16 were developed to broadly detect HuAstVs. The most conserved sequence (CA16). HFMD is generally a self-limiting disease characterised by fever, between ORF1 coding for the non-structural protein and ORF2 encoding small blisters in the mouth, and a rash with blisters. However in a small for capsid protein junction was chosen as the assay target. Three sets number of cases, HFMD caused by EV71 can lead to viral meningitis, of primer and probe were designed with specificity for AstV 1-8, AstV- encephalitis, interstitial pneumonitis or poliomyelitis-like paralysis, and MLB1- 2, and AstV-VA1-3. Primers were optimized with known positive may be fatal. The disease can infect any age group, but is rare in children specimens identified by RT-PCR from a previous study using SYBR Green over the age of 10. EV71 and HFMD are endemic in the Asia Pacific based real-time RT-PCR. TaqMan multiplex RT-PCR assay was applied to region causing millions of cases in recent years. We are developing a well screen stool samples with no previously identified enteric pathogens by characterized, multiple dose, highly purified, inactivated EV71 vaccine standard microbiology for enteric bacteria; EIA for rotavirus, adenovirus, formulated with alum adjuvant. Pre-clinical studies have shown that this astrovirus, giardia and cryptosporidium; or RT-PCR for norovirus and vaccine is highly immunogenic and generates strong neutralizing antibody rotavirus. These stool samples were collected from children aged 3 months responses in mice, rats and rabbits. We also have demonstrated that to 5 years with diarrhea and non-diarrhea controls in a hospital based these antibodies are capable of cross neutralizing EV71 sub-genogroups study in Nepal. All positive samples were tested for HuAstV typing by currently circulating in Asia. A double blind, placebo controlled Phase 1 individual sets of primers and probe. A total of 634 stool samples, 284 clinical study is being conducted in Singapore to assess the safety and cases and 350 controls, were screened. HuAstVs were detected in 9/284 immunogenicity of two different dose levels of the inactivated EV71 (3.2%) of cases and 9/350 (2.6%) of controls. All 9 samples from cases vaccine in healthy adults. An update of the clinical trial data will be were identified as AstV-canonical. On further testing of the samples from presented.

www.astmh.org 422 1402 1404 SENTINEL SURVEILLANCE FOR INFLUENZA IN SMALL MOLECULE INHIBITORS OF VENEZUELAN EQUINE PHRAMONGKUTKLAO HOSPITAL IN BANGKOK THAILAND ENCEPHALITIS VIRUS INFECTION IDENTIFIED USING HIGH THROUGHPUT PHENOTYPIC SCREENING OF THE VACCINE Richard G. Jarman1, Sriluck Simasathien2, Veerachai Watanaveeradej2, Phirangkul Kerdpanich2, Danabhand STRAIN TC83 Phiboonbanakit2, Tundorn Chirabandhu2, Piraya Gene G. Olinger, Jr.1, Corinne Scully1, Jennifer Wichterman2, Bhoomiboonchoo1, In-Kyu Yoon1, Robert V. Gibbons1 Ronald L. Johnson2, Pam Glass1, Ruili Huang2 1Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 1U.S. Army Medical Research Institute for Infectious Diseases, Frederick, 2Phramongkutklao (PMK) Hospital, Bangkok, Thailand MD, United States, 2National Institutes of Health Chemical Genomics Phramongkutklao (PMK) Hospital with the Armed Forces Research Institute Center, Bethesda, MD, United States of Medical Sciences conducted surveillance to identify, characterize and Currently, there is no approved vaccine or therapeutic for prevention or determine the prevalence of influenza and other respiratory pathogens treatment of infection by Venezuelan equine encephalitis virus (VEEV) in Bangkok. Patients ≥6 months who meet the criteria of influenza-like in humans. Antiviral therapeutics are highly desired for treatment of illness (fever ≤38oC and cough or sore throat within 3 days of onset), VEEV infection. We have developed a screening assay in 96- and 1536- without tuberculosis and who were not immunocompromised were well formats to identify antiviral compounds against TC-83, a BSL-2 live eligible. History, physical examination, and nasal swabs for rapid kits attenuated vaccine strain (IND stage) of VEEV. Briefly, primate kidney were performed. Throat swabs were sent for PCR and viral isolation/ Vero cells are infected with TC-83 and after 48 hours, cell viability is characterization. 919 subjects were enrolled, 688 (75%) were ≤ 18 years assessed by measuring cellular ATP levels. The assay was miniaturized old; 319 (35%) subjects tested positive for influenza by PCR (72% for to 1536-well plate format and screened against 7,243 unique bioactive influenza A). The pandemic strain was the most prevalent (197 cases). small molecules to identify 33 initial actives. The activities of a subset of The first peak of influenza occurred in Jan/Feb 2010 with >40% of the compounds have been tested in orthogonal in vitro assays against VEEV cases testing positive, the majority were the pandemic strain. The second TC-83, VEEV Trinidad (TrD) strains and additionally EEEV and WEEV. We peak occurred in Aug 2010 with 56% testing positive; the pandemic have demonstrated that the approach of using authentic virus, even a strain remained dominant, but influenza B accounted for nearly 40% of surrogate vaccine strain can identify inhibitory compounds against VEEV. the cases. There were 106 admissions; 19% were influenza and none had Furthermore, the approach can identify molecular targets critical for VEEV received influenza vaccine. Of 863 subjects reporting vaccination status infection that could be further developed for human-use antiviral drugs. only 113 had received it within the previous 12 months, and 27 were influenza positive (17 with the pandemic strain). HA gene sequencing on 1405 selective samples revealed that the strains in circulation were similar to the 2009 southern hemisphere vaccine strains. In conclusion, influenza is a BURDEN AND EPIDEMIOLOGY OF ROTAVIRUS DIARRHEA: significant cause of morbidity at PMK Hospital. The population was largely RESULTS OF A PREVALENCE STUDY IN NIGER unvaccinated and the majority of influenza cases were caused by the 1 2 2 pandemic strain. Vaccination would likely significantly reduce morbidity. Anne-Laure Page , Viviane Jusot , Abdul-Aziz Mamaty , Lagare Adamou3, Jérôme Kaplon4, Pierre Pothier4, Ali Djibo5, Mahamane 1403 Laouali Manzo5, Brahima Toure2, Céline Langendorf2, Jean-Marc Collard3, Rebecca F. Grais1 A CHIKUNGUNYA VIRUS HIGH FIDELITY VARIANT LOSES 1Epicentre, Paris, France, 2Epicentre, Niamey, Niger, 3CERMES, Niamey, FITNESS IN MOSQUITOES AND MICE Niger, 4Reference Centre for Enteric Viruses, Dijon, France, 5Ministry of Health, Niamey, Niger Lark L. Coffey, Yasnee Beeharry, Marco Vignuzzi Institut Pasteur, Paris, France Diarrhea is still the second leading cause of death in children under 5 years of age in developing countries, representing nearly one in five child The error rate of RNA dependent RNA polymerases (RdRp) strongly affects deaths. Rotavirus is the most common etiologic agent of severe diarrhea the mutation frequency in a population of viral RNAs. Previously, we and vaccines are readily available. However, knowledge about the disease used a high fidelity variant of an RNA virus (poliovirus) to illustrate the burden as well the circulating strains is still lacking in many countries to importance that mutation frequency plays in virus fitness and adaptability make informed decisions about vaccine introduction. The regional hospital in vivo. Since arboviruses replicate within very different hosts, the need and ten health centers of Maradi region, and the three main hospitals to generate such genetic diversity may be even more significant than for in Niamey, Niger, were selected for a one-year prevalence study. Stool single ost RNA viruses. Using chikungunya virus (CHIKV), we describe samples were collected from all children under 5 having diarrhea with an arbovirus fidelity variant isolated in mutagen treatment with a moderate or severe dehydration and tested for the presence of rotavirus single C483Y amino acid change in the NSP4 RdRp gene that increases using a rapid diagnostic test. Genotyping was performed on a subset of replication fidelity. The increase in fidelity does not have significant costs rotavirus positive stools. In addition, a sample of the collected stool was in terms of replication and RNA synthesis, but shows significant fitness used for bacteriology analyses in Maradi. From December 2009 until costs in vivo. Compared to wild type CHIKV, the higher fidelity population December 2010, 5247 children were included in the study. The median presents reduced infection and dissemination in mosquitoes. Furthermore, age was 9 months [IQR: 7-11 months]. Overall, the proportion of rotavirus viremias in newborn mice are truncated and organ titers are significantly positive diarrhea was 26.8% (95% CI: 25.6-28.0) and varied monthly lower. These results indicate that increased replication fidelity and reduced from 10.6% (95% CI: 7.6-13.6) in May to 48.5% (95% CI: 44.7-52.4) genetic diversity negatively impact arbovirus fitness in invertebrate and in November. Around 65% of the rotavirus cases were children 6 to 12 vertebrate hosts, a factor that may explain why RNA viruses maintain months old. The most frequent genotypes were G2P[6], G2P[4] and error-prone RdRp genes. G1P[8]. Coprocultures performed on stools from 1988 children showed Salmonella spp. in 10.5% of the cases, Campylobacter jejuni in 8.1%, Shigella spp. in 3.0%. More than 10% of the Salmonella spp. identified carried an extended-spectrum beta-lactamase. This study points out that rotavirus is a major cause of diarrhea with dehydration in Niger, particularly in young children under 1 year of age, and during the cool and dry season. The variety of circulating genotypes in Africa should

www.astmh.org 423 be taken into consideration for the development of better adapted 200x mean coverage depth of the entire genome. Assisted assembly vaccines. Bacterial pathogens are also responsible for an important part methods using reference genomes, as well as de novo assemblies have of diarrhea, and point to the emergence of resistance to third generation been created with high success. This method can be used and adapted cephalosporin. to sequence many known pathogens for high- and low-throughput sequencing initiatives. 1406 1408 SYNERGISTIC ACTION OF ROTAVIRUS AND COINFECTING PATHOGENS: EVIDENCE FROM A COMMUNITY-BASED CASE SURVEILLANCE OF MEASLES AND RUBELLA INFECTIONS IN CONTROL STUDY IN NORTHWESTERN ECUADOR RWANDA: 2003-2011 Darlene Bhavnani1, Jason E. Goldstick1, William Cevallos2, Jean-Frederic Flandin, Zena Uwimana, John Baptist Gatabazi, Gabriel Trueba2, Joseph N. Eisenberg1 Odette Mukabayire, Odette Mukabayire 1University of Michigan, Ann Arbor, MI, United States, 2Universidad San National Reference Laboratory, Kigali, Rwanda Francisco de Quito, Quito, Ecuador Measles and Rubella viruses are still important viral infections in tropical Diarrheal disease is a leading cause of morbidity in children under five. In countries including Rwanda. Since 2003, the National Reference developing countries, where diarrheal disease burden is greatest, enteric Laboratory of Rwanda is accredited by WHO/AFRO as a national Measles coinfection is common. There is little understanding, however, of the laboratory, has been involved in the surveillance of Measles and Rubella biological interaction between coinfecting pathogens. We investigated infection throughout the country. Cumulative data show that of the the potential for synergistic action by coinfecting pathogens on diarrhea approximately 1,894 samples suspected of Measles, 163 were positively pathogenesis using an epidemiological framework. We conducted a identified by ELISA (8.61%). In Rwanda, geographical data indicates that community-based case control study in 22 villages in northwestern the Rubavu district in the West province, which is at the border with Ecuador. Risk ratios of diarrhea associated with single and coinfections Democratic Republic of Congo is particularly prone to Measles infection. were estimated. Biological interaction of coinfecting pathogens was Since 2003, Rwanda has been hit by 3 major epidemics of Measles. The assessed through the interaction contrast ratio (ICR) and departure of the first 2 epidemics, in 2005 and 2006, were limited to the Rubavu district risk ratios from multiplicativity (MD). After adjusting for age, we found while in 2010, infections were found in various locations, including both departure from risk difference additivity and departure from risk ratio Kigali city. In collaboration with Uganda Virus Research Institutes, multiplicativity in the effects of rotavirus coinfections on acute diarrhea. circulating strains were genotyped. It has been observed that in 2005 The ICR was 11.9 (95% CI = 4.7-29.9) for rotavirus-Giardia coinfections, and 2006, epidemics strains were mostly of the type B2 which is a strain and 22.9 (95% CI = 10.5-46.8) for rotavirus-E. coli/Shigella coinfections. characterized as Congolese. However, during 2010 epidemics, most of the The MD for these coinfections was 11.8 (95% CI = 3.9-29.4) and 20.1 cases were of the type B3, an indigenous strain predominantly found in (95% CI = 2.2-44.2), respectively. This research provides epidemiological Burundi. In addition to Measles testing, negative samples were tested for evidence for biological synergism between rotavirus and other enteric Rubella infection. Of these 1,731 samples tested, 282 were found to be pathogens. During coinfection, the pathogenic potential of each organism positive (16.29%) and were distributed equally throughout the country. appears to be enhanced. 1409 1407 IMMUNOLOGICAL FEATURES AND PROVIRAL LOAD IN CREATING A PIPELINE FOR NEXT GEN SEQUENCING OF PATIENTS WITH OVERACTIVE BLADDER ASSOCIATED WITH EBOLAVIRUS ZAIRE HTLV-1 ARE INDICATORS OF AN EARLY STAGE OF HTLV-1 ASSOCIATED MYELOPATHY Stephen K. Gire1, Zach Bjornson2, Elizabeth Ryan3, Kristian Andersen1, Lisa Hensley2, Pardis Sabeti1 Edgar M. Carvalho1, Silvane B. Santos1, Paulo Oliveira1, Tania 1Harvard University, Cambridge, MA, United States, 2United States Army Luna1, Anselmo Souza1, Marcia Nascimento1, Isadora Siqueira1, Medical Research Institute of Infectious Diseases, Fort Detrick, MD, United Davi Tanajura1, André Luiz Muniz1, Marshall Glesby2 States, 3The Broad Institute, Cambridge, MA, United States 1Federal University of Bahia, Salvador, Brazil, 2Weill Cornell Medical College, New York, NY, United States With Next Generation sequencing becoming a more cost-effective method of sequencing full-length microbial genomes, established pipelines for The majority of HTLV-1 infected subjects are considered as carriers but sequencing pathogens of high impact are greatly needed. These validated a high frequency of urinary manifestations of overactive bladder (OB) pipelines will exponentially increase sequence data available for studies in has been documented in these individuals. The aim of this study was population dynamics, viral evolution and genetics, and the identification to determine if viral and immunological factors that are associated with of novel targets for therapeutics, vaccines and rapid diagnostics. Here we development of HTLV-1 associated myelopathy/tropical spastic paraparesis developed two methods for full-length sequencing of ebolavirus Zaire (HAM/TSP) are also observed in patients with overactive bladder associated using Roche 454 pyrosequencing and Illumina RNA-seq technologies. For with HTLV-1. Participants (n=135) were classified as HTLV-1 carriers, 454 sequencing, a “demi-hemi” approach, originally developed at the HTLV-1 associated overactive bladder (HTLV-1 OB) defined by the criteria Broad Institute, was used to design 5-prime amine-modified primers to of International Continence Society (ICS) and HAM/TSP patients. We create long-range PCR amplicons along the entire ebolavirus genome. demonstrated that peripheral blood mononuclear cells from HTLV-1 OB Amplicons were designed with approximately 500bp overlapping regions patients produce higher spontaneous levels of proinflammatory cytokines to aid in downstream assembly. Illumina sequencing technology was also (IFN-γ, TNF-α and IL-17) than HTLV-1 carriers and similar levels of TNF-α used to confirm sequence data and further refine primer sequences to and IL-17 to patients with HAM/TSP. Proviral load was higher in HTLV- capture potential divergent populations within the Zaire strain. While 1 OB and HAM/TSP than HTLV-1 carriers and correlated positively with Illumina sequencing provides unbiased sequencing data invaluable to production of proinflammatory cytokines. In contrast to HAM/TSP, patients identifying divergent populations, 454 technology can cheaply sequence with overactive bladder had serum levels of Th1 chemokines (CXCL-9 and isolates at a high-throughput capacity with longer sequencing reads that CXCL-10) similar to HTLV-1 carriers and exogenous addition of regulatory aid de novo assembly. Together, these methods have produced a validated cytokines (IL-10 and TGF-β) decreased IFN-γ production in cell cultures 454 sequencing pipeline that has been used to successfully sequence full- from patients with HTLV-1 OB. We conclude that HTLV-1 infected patients length ebola genomes from prepared viral seed stocks and time-course with overactive bladder have some immunological features and proviral infections of nonhuman primates, with as little as 0.1ng cDNA and over load profiles in common with HAM/TSP patients. However as they are www.astmh.org 424 still able to down modulate the inflammatory immune response and the in the E2 glycoprotein while the other two, 7004 and 7006 had amino recruitment of activated T cells to the central nervous system (CNS) is not acid substitutions in the NSP1 and E1 proteins, respectively. In alpha/beta enhanced, they present overactive bladder, an oligosymptomatic form of interferon receptor deficient mice (A129) the two E2 mutation viruses HAM/TSP. showed a delay in mortality by three days compared to the other two mutants, which showed similar mortality kinetics as wild-type CHIKV, 1410 indicating a lack of attenuation. Reduced viremia, pro-inflammatory cytokines, and footpad swelling were also seen in the E2 mutants. In NOVEL VACCINE CANDIDATE PROTECTS MACAQUES C57bl/6 mice, an immunocompetent arthritis model, the E2 mutants again AGAINST CHIKUNGUNYA FEVER proved to be more attenuated, behaving similarly to CHIKV 181/25 with less footpad swelling and lower viremias than the E1 and NSP1 mutants. Scott C. Weaver1, Eryu Wang1, Alison P. Adams1, Robert While the single mutations in the E2 gene were not sufficient to create the Seymour1, Stephanie Z. Killeen2, Chad J. Roy2 attenuation seen in CHIKV 181/25, it is clear they resulted in significant 1University of Texas Medical Branch, Galveston, TX, United States, 2Tulane attenuation to the parental CHIK virus compared to the mutations in the National Primate Research Center, Covington, LA, United States E1 and NSP1 genes, which resulted in a wild-type-like phenotype. Further Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes studies with viruses containing multiple mutations should be performed to an acute febrile illness typically accompanied by rash and severe, often probe the genetic cause of the attenuating phenotype observed in CHIKV persistent arthralgia. The virus has emerged since 2004 to cause major 181/25 vaccine virus. epidemics in the Indian Ocean, India and Southeast Asia, involving millions of persons. Autochthonous transmission in Italy and France 1412 after CHIKV introductions via travelers also underscored the risk that CHIKV poses to the U.S. Because no licensed vaccine exists to protect CHALLENGES AND BREAKTHROUGHS IN THE DEVELOPMENT against chikungunya fever, we used an alphavirus attenuation approach OF SEQUENCING TECHNOLOGY FOR CLINICAL ISOLATES OF involving a picornavirus internal ribosome entry site (IRES), which replaces LASSA FEVER VIRUS the subgenomic promoter, to produce a live vaccine strain capable of Eleina M. Zaitsev1, Kristian G. Andersen2, Stephen Gire2, Lina inducing protective immunity after a single administration. This vaccine Moses3, Robert Garry3, Christian Happi4, Pardis Sabeti2 provides robust immunity and protection in murine models, and is 1The Broad Institute of MIT and Harvard, Cambridge, MA, United States, incapable of infecting mosquito vectors, an important safety feature 2Harvard University, Cambridge, MA, United States, 3Tulane University, for use in nonendemic locations. For further preclinical testing, we New Orleans, LA, United States, 4University of Ibadan, Ibadan, Nigeria subcutaneously or intradermally vaccinated 3 cohorts of 4 cynomolgus macaques, monitored their responses telemetrically to evaluate safety, Lassa Virus (LV) is the causal agent of Lassa fever, a severe hemorrhagic then challenged them with wild-type CHIKV to assess efficacy. None of the fever endemic to West Africa. It is responsible for thousands of deaths 12 animals developed fever or detectable changes in respiratory or cardiac each year, and evidence suggests that LV acted as a selective agent in function after vaccination, and all developed robust neutralizing antibody recent human evolution. This, combined with observations that it can responses. After challenge, most sham-vaccinated animals developed fever infect and replicate in its natural host, the rodent Mastomys natelensis, followed by hypothermia, acute viremia, and many also exhibited changes without causing severe disease, make it a desirable topic for studies of in respiratory and cardiac function. In contrast, all vaccinated animals host-pathogen evolution. In an effort to compile a large dataset of full- remained completely normal in all physiological and clinical parameters, length LV genomes, we have applied 454 and Illumina next-generation and showed no development of viremia from challenge. These results sequencing technology on clinical samples collected from patients and indicate that these new IRES-based vaccine candidates show great promise rodents in West Africa. While we have successfully used both to generate for use in humans to control chikungunya fever in endemic locations, as full-length sequences, each platform presents unique challenges and well as to reduce the risk of further spread, including into the Americas. benefits for viral sequencing. 454 technology can generate data from small input volumes, ideal for clinical samples of low viral titers. 454 1411 also produces longer sequence reads, allowing for a wider range of downstream analyses than Illumina-generated sequences. A downside MUTATIONS IN THE E2 GLYCOPROTEIN GENE OF of 454 is that it requires high quality, undegraded starting material - a CHIKUNGUNYA VIRUS ASSOCIATED WITH VIRAL-INDUCED problem frequently encountered on samples stored under sub-optimal ARTHRITIS IN MOUSE MODELS conditions in the field. Illumina sequencing does not require as high quality material but can only be used when other input requirements James D. Weger1, Charalambos D. Partidos2, David M. Reinitz1, are met (i.e., low host-contamination, high viral titer). Here we discuss Rodion Gorchakov3, Kenneth Plante3, Robert Seymour3, Scott C. the pros and cons associated with each platform as well as the technical Weaver3, Dan T. Stinchcomb4, Jorge E. Osorio1 developments that have contributed to their successful application. Our 1University of Wisconsin-Madison, Madison, WI, United States, 2Inviragen, initial results from sequencing have allowed us to make conclusions Inc., Madison, WI, United States, 3University of Texas Medical Branch, about new outbreaks of novel LV strains in Northern Sierra Leone, as well Galveston, TX, United States, 4Inviragen, Inc, Fort Collins, CO, United as better catalogue circulating strains. Such knowledge will aid in our States understanding of viral evolution, allow us to better predict patterns of Chikungunya virus (CHIKV) is an emerging arbovirus associated with disease spread, and lay the foundation for better diagnostic development. explosive outbreaks of febrile illness often accompanied by rash and arthralgia. The United States Army Medical Research Institute of Infectious Diseases (USAMRIID) developed a live attenuated vaccine virus, 181/ clone25 (hereafter 181/25), which despite producing transient arthralgia in a small subset of patients, was shown to be highly efficacious and well tolerated in phase 1 and 2 clinical trials. CHIKV 181/25 was produced by 18 plaque-to-plaque passages of CHIKV strain 15561 in human lung cells (MRC-5). The resulting virus contains 9 nucleotide substitutions, 5 of which result in amino acid changes. In order to probe the specific genetic effects of the mutations, single nucleotide mutation-containing viruses were made that correspond to the amino acid substitutions present in CHIKV 181/25. Two viruses, 7005 and 7014, had amino acid substitutions www.astmh.org 425 1413 and norovirus as well as determine the genotype of rotavirus and norovirus. It may therefore be of great utility in epidemiologic surveillance MOLECULAR CHARACTERIZATION BY DEEP SEQUENCING post rotavirus vaccine implementation OF DIVERSE MEMBERS OF THE GENUS ORTHOBUNYAVIRUS FROM MOSQUITOES COLLECTED IN THE AMAZON BASIN OF 1415 PERU HUMAN CELLULAR RESPONSES TO RIFT VALLEY FEVER Loreen L. Lofts, Michael J. Turell, Jeffrey R. Kugelman, Chris A. VIRUS Whitehouse Amy M. Edwards1, Zachary P. Traylor2, Amy G. Hise2, Charles United States Army Medical Research Institute for Infectious Diseases, King2, James W. Kazura2, A. Desiree LaBeaud3 Frederick, MD, United States 1University Hospitals Case Medical Center, Cleveland, OH, United States, Members of the genus Orthobunyavirus (family Bunyaviridae) are 2Case Western Reserve University, Cleveland, OH, United States, 3Children’s segmented, negative-sense, single-stranded RNA viruses that are Hospital Oakland Research Institute, Oakland, CA, United States responsible for mild to severe disease in humans. As part of a long-term study of arbovirus ecology in the Amazon basin of Peru, more than Rift Valley Fever (RVF) virus is an arbovirus in the Bunyaviridae family that 160 viral isolates were made from mosquitoes captured near Iquitos, was first isolated in the Rift Valley region of Kenya in the 1930’s. The virus Peru. Preliminary analysis using immunofluorescent antibody assays (IFA) is a significant cause of morbidity and mortality in humans associated with identified many of these viruses as members of the Group C, Guama, devastating epizootic outbreaks in livestock. While endemic to the Rift and Bunyamwera serogroups within the genus Orthobunyavirus. Follow- Valley region, the virus has shown an ability to spread to virgin territory up IFAs using complex or virus-specific antisera in complement fixation such as Egypt and the Arabian Peninsula. Furthermore, RVFV has emerged and hemagglutination-inhibition assays identified some of the viruses as a target for bioterrorism in recent years. RVFV can cause diverse as Caraparu, Guama, Itaqui, Mirim, Murutucu, Oriboca, and Wyeomyia pathology in humans from non-specific viral illness to severe hemorrhagic viruses; others remain uncharacterized. Additionally, basic knowledge disease, encephalitis and death. While it is known that RVFV can cause a is lacking in regard to the potential reassortment of segments among range of diseases, the pathogenesis is still not well understood. A number these viruses in nature. Therefore, we determined the whole genome of factors such as route of transmission and host immune response nucleic acid sequences of these viruses in relation to the viral quasispecies are thought to play a role. Animal studies have shown that a type 1 to investigate the extent of intra- and inter-reassortment among RNA interferon response in the infected individual affects viral clearance. In segments (S, M, and L) of these viruses. Implications of the reassorted addition, a delayed IFN response is associated with worse morbidity and genomes will be discussed. mortality. In this study we infect human peripheral blood mononuclear cells (PBMC) with attenuated MP-12 and MP-12 Nss knockout strains 1414 of RVFV to determine time to viral uptake and to monitor inflammatory production. Time to viral uptake was determined using plaque assays and DEVELOPMENT OF A MULTIPLEX PCR/LDR ASSAY TO DETECT qPCR for the L segment of the viral genome and showed that viral uptake AND GENOTYPE ROTAVIRUS happens occurs as early as two hours post infection. The production of various inflammatory mediators, such as TNF alpha and IFN alpha was Sanchita Das1, Aashiq Hussain1, Maneesh Pingle1, Mark S. monitored using ELISA. We show using naïve North American donors that Rundell1, George E. Armah2, Ben Gyan2, Jon Gentsch3, Vinje inflammatory responses begin as early as 6 hours after introduction of the Jan3, Davise Larone1, Eric D. Spitzer4, Francis Barany1, Linnie M. virus. Additional studies will elucidate which cells are primarily responsible Golightly1 for the inflammatory response and define innate receptor utilization. 1Weill Medical Center, New York, NY, United States, 2Noguchi Memorial Research Institute, Accra, Ghana, 3Centers for Disease Control and Prevention, Atlanta, GA, United States, 4Stony Brook University Medical 1416 Center, Stony Brook, NY, United States ECOLOGY OF VENEZUELAN EQUINE ENCEPHALITIS IN THE Rotavirus and enteric caliciviruses are the most common etiologic agents GULF COAST REGION OF MEXICO of diarrhea in children and adults worldwide. The implementation Jose G. Estrada-Franco1, A. Paige Adams1, Francisco J. Ramirez- of rotavirus vaccines holds the promise of significantly reducing the Aguilar2, Grace Leal1, Irene Lopez-Gonzalez3, Roberto Navarro- associated morbidity and mortality in children. However the vaccines Lopez3, Amelia Travassos Da Rosa1, Ann Powers4, Robert Tesh1, has shown variable efficacy in Africa and Asia attributed to different Richard Bowen5, Scott C. Weaver1 circulating genotypes than those targeted and/or viral co-infection, 1 which is common in these areas. The WHO has therefore recommended University of Texas Medical Branch, Galveston, TX, United States, 2 3 continued surveillance post vaccine implementation. We report the Universidad Autonoma de Chiapas, Tapachula, Mexico, CPA-SAGARPA 4 development of a comprehensive molecular assay that can simultaneously Mexican Agriculture Ministry, Tuxtla Gutierrez, Chiapas, Mexico, Centers detect and genotype the enteric viruses. One-step reverse transcriptase for Disease Control and Prevention, Fort Collins, CO, United States, 5 PCR amplifies virus specific targets. Ligase detection reaction and Colorado State University, Fort Collins, CO, United States subsequent hybridization of the fluorescent products to beads then To characterize the ecology of Venezuelan equine encephalitis virus (VEEV) detects sapovirus and identifies genotypes of rotavirus, and norovirus. The in endemic regions of the Mexican Gulf Coast, serosurveys in humans and assay was optimized using previously characterized viral culture supernates equids, vector incrimination studies, studies of natural infection in equine and stool specimens and then used to analyze 296 clinical specimens hosts, and phylogenetic characterization of isolates were conducted in obtained in the US and Ghana. The assay was found to be 97% sensitive 2008-2010. Human serosurveys from suspected dengue patients (N=237) and 100% specific with a 100% concordance for genotype determination were VEEV positive in 32 individuals (13.5% seroprevalence), including of norovirus. The rotavirus G- and P-type were determined in 98.6% 5 IgM positives. Equine serosurveys of unvaccinated animals revealed and 92.3% of the samples, respectively. Mixed genotypes were found widespread endemic VEEV from the southernmost region in Tabasco State in 11.8% of the samples. A significant finding was the identification of to a northern municipality in Tamaulipas State, located adjacent to the two isolates of rare genotypes G6P[6] and G6P[8], both genotypes being Texas border. Using rodent serosurveys, we identified putative reservoir reported for the first time in Ghana. The PCR/LDR assay is a sensitive, species of the genera Sigmodon and Oryzomys in Minatitlan, Veracruz. specific and high-throughput method that can detect rotavirus, sapovirus Using hamster-baited traps in Minatitlan, high-titered mosquito pools

(4.9-6.4 log10 PFU/pool) of Culex (Melanoconion) taeniopus were identified

www.astmh.org 426 in at least 3 transmission events as the likely principal vector of enzootic 1418 subtype IE VEEV. As previously observed on the Pacific Coast of Mexico, naturally infected horses developed neurologic disease while producing INTERNAL RIBOSOME ENTRY SITE (IRES)-DRIVEN high-titered viremia (2.4-3.6 log10 PFU). Phylogenetic analysis was EXPRESSION OF THE CAPSID PROTEIN IN VENEZUELAN performed on isolates from horses, sentinel hamsters, and mosquitoes in EQUINE ENCEPHALITIS VIRUS INCREASES THE ATTENUATION Veracruz State. Based on the glycoprotein precursor sequence, all isolates AND SAFETY OF THE TC-83 VACCINE STRAIN grouped within the Gulf/Caribbean IE genotype and were temporally distinct from the 1963-1969 isolates from the same region. These Mathilde Guerbois1, Eugenia Volkova1, Naomi A. Forrester1, findings suggest that endemic subtype IE VEEV is currently circulating in Joseph A. Rodriguez1, Eryu Wang1, Kenneth S. Plante1, Ilya Frolov2, widespread regions of the Mexican Gulf Coast, including areas located Scott S. Weaver1 near the Texas border. Although we implicated Cx. taeniopus as the main 1University of Texas Medical Branch, Galveston, TX, United States, subtype IE VEEV vector in the Gulf Coast region, Aedes (Ochlerotatus) 2University of Alabama at Birmingham, Birmingham, AL, United States taeniorhynchus mosquitoes were also abundant in this region and have The live-attenuated TC-83 strain is the only licensed veterinary vaccine been associated with equine outbreaks on the Pacific Coast of Mexico. available to protect equids against Venezuelan Equine Encephalitis Thus, the continuous circulation of endemic VEEV in Mexico has the Virus (VEEV) infection and to protect humans indirectly by preventing potential for developing into an outbreak that could rapidly spread into equine amplification. VEEV is a mosquito-borne virus endemic to several the US via equine amplification. areas of Central and South America, where both endemic disease and periodic epidemic outbreaks affect hundreds-of-thousands of humans. 1417 However, TC-83 vaccine has previously been isolated from mosquitoes RSV, SEASONAL INFLUENZA AND H1N1: CLINICAL collected in the wild. Because it relies on only two point mutations for its MANIFESTATIONS AND CO-INFECTIONS IN 2009 attenuation, transmission of revertants represents a major risk to initiate an epidemic or to circulate enzootically. To improve its attenuation and Rosa M. Delgado-Ayala, Zoe Gonzalez, Annette Pietri stability, and to prevent infection of mosquitoes, recombinant TC-83 was Hospital Episcopal San Lucas-Ponce, Ponce, Puerto Rico previously engineered by placing the expression of the viral structural proteins under the control of the Internal Ribosome Entry Site (IRES) of Respiratory Syncytial Virus (RSV) and Influenza primordially affect encephalomyocarditis virus (EMCV), which drives translation inefficiently in children. RSV is a major cause of Bronchiolitis and Pneumonia in children mosquito cells. However, this vaccine candidate was poorly immunogenic. <1 yo, Influenza is characterized by abrupt onset of constitutional and Here we describe the second generation TC-83 recombinant in which respiratory signs and symptoms. Both manifest in colder months of the only the capsid protein gene is translated from the IRES, while the viral year in temperate climates. In April 2009 influenza virus, H1N1 (swine flu) surface envelope glycoproteins are expressed from a subgenomic message created an outbreak reaching pandemic status in 6 weeks. United States in a cap-dependent manner. This TC-83/IRES/C vaccine does not infect experienced first wave on May 2009 and a second wave, that Puerto mosquitoes, is stable in its attenuation phenotype after serial passages in Rico also experienced, peaking by end of October. Due to similar clinical vivo, and is more attenuated in newborn mice but still protective as well presentations of RSV, Seasonal Influenza and Influenza H1N1, physicians against VEEV challenge. Thus, by using the IRES to modulate TC-83 capsid differentiated them by Influenza rapid test and RSV nasopharyngeal test. protein expression, we generated a vaccine candidate that combines Aim of study: describe clinical and epidemiologic characteristics of RSV, efficient immunogenicity and efficacy with lower virulence and a reduced Seasonal Influenza and H1N1. After IRB approval, records of bacteriology potential for spreading in nature. and epidemiology from Hospital Episcopal San Lucas were reviewed for positive results of RSV, Influenza, and H1N1 in patients 0y/o - 4y/o, 1419 admitted to Pediatrics from October-December 2009 (peak of H1N1 epidemic). Patients with respiratory problems at admission were included, EPIDEMIOLOGICAL MODELING AND RISK ANALYSIS OF bacterial sepsis was excluded. Records were reviewed for: age, gender, VENEZUELAN EQUINE ENCEPHALITIS IN THE HUMAN past medical history, symptoms prior to hospitalization, admission or POPULATION OF COASTAL CHIAPAS, MEXICO IN 2007-2009 transfer to PICU, supplemental oxygen and days of hospitalization. Forty seven patients tested positive for RSV and/or Influenza. RSV was the Francisco J. Ramirez-Aguilar1, Carlos Martinez-Alfaro2, Neftaly most frequently reported (29) with majority of admissions to PICU (5). Vazquez-Pimentel2, Roberto Navarro-Lopez3, Pamela L. Phillips4, Seventeen patients had Influenza, 9 H1N1. Most patients with RSV had Scott C. Weaver5, Jose G. Estrada-Franco5 term birth history while those with Influenza were mostly pre-term. Of 9 1Universidad Autonoma de Chiapas, Tapachula, Mexico, 2ISECH, Instituto patients positive for H1N1 past medical history revealed pre-term birth in de Salud de Chiapas, Tuxtla Gutierrez, Chiapas, Mexico, 3CPA-SAGARPA, 6, and conditions where asthma predominated. One case was co-infected Mexican Agriculture Ministry, Tuxtla Gutierrez, Chiapas, Mexico, 4U.S. (RSV/Influenza A), admitted with no serious morbidity. Epidemic H1N1 Department of Agriculture, ARS, Kerrville, TX, United States, 5University of (2009) caused difficulties in differentiation of respiratory illnesses requiring Texas Medical Branch, Galveston, TX, United States admission. In our experience most children under 4 years had RSV. RSV was associated with most morbidity. Analysis of 101 febrile illness patients seropositive for Venezuelan equine encephalitis (VEEV) was carried out in a retrospective study along 18 municipalities and endemic VEEV pacific coastal regions of the State of Chiapas in southern Mexico. Geographic information systems (GIS), satellite imagery and a detailed questionnaire were used in the analysis. Using ESRI ArcGIS 10.1 software and spatial statistics tools we measured the geographic distribution of VEEV cases along coastal Chiapas. The distribution of VEE cases were principally located along the Pacific coastal plain with the mean center of positive cases to be in the municipality of Huixtla. The directional distribution of cases around the mean were dispersed in a pattern between the Pacific coastline and coastal mountain range. Temporal and spatial dynamics showed clear separation between cases in the southern and northern regions during the dry season with a peak of positive cases for both regions during the wet season. The analysis was based on Euclidian distances identifying spatially significant clusters

www.astmh.org 427 as hot spots. All the spatial analysis was complemented with relative risk 1421 (OR) bivariate and multivariate statistical analytical models showing neck stiffness (OR=6.03; 1.2318-29.5801, p=0.027) multivariate; (OR=13.87, SEASONALITY, TIMING, AND CLIMATE DRIVERS OF IC 3.1861-60.4217, p=0.000), bivariate, muscle weakness (OR=10.12; IC INFLUENZA ACTIVITY WORLDWIDE 2.1633-47.4002, p=0.003) multivariate; (OR=23.05, IC5.3134-100.0692, 1 1 2 p=0.000) bivariate, taste dysfunction (OR=3.54; IC 1.1380-11.0234, Eduardo Azziz-Baumgartner , Christine Dao , Sharifa Nasreen , 2 2 2 p=0.029) multivariate; (OR=6.40; IC 2.3449-17.5045, p=0.000) bivariate, Mejbah Uddin Bhuiyan , Mah-e Munir , Abdullah Al Mamun , 2 2 1 and photophobia (OR=16.98, IC 2.2321-129.2713, p=0.006) bivariate M. A. Sharker , Rashid Uz Zaman , Po-Yung Cheng , Alexander 1 1 1 and conjunctivitis (OR=3.25, IC 1.3273-7.9948, p=0.010) bivariate, as the I. Klimov , Marc-Alain Widdowson , Timothy Uyeki , Stephen P. 1 1 1 most important clinical manifestations associated with VEEV for individuals Luby , Anthony Mounts , Joseph Bresee of coastal Chiapas. Overall, our results indicate a coastal band of endemic 1Centers for Disease Control and Prevention, Atlanta, GA, United States, VEE extending from the Guatemalan border through the State of Chiapas 2International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, to the adjacent State of Oaxaca. Bangladesh Influenza is a vaccine preventable disease which annually causes 1420 substantial morbidity and mortality, but data on influenza virus activity THE ROLE OF THE INTERFERONS VERSUS THE ADAPTIVE in tropical countries are limited. We analyzed publicly available influenza data to better understand the global circulation of influenza viruses. IMMUNE RESPONSE IN A MOUSE MODEL OF O’NYONG- We searched for laboratory-confirmed influenza surveillance data in NYONG VIRUS INFECTION FluNet, Google™, and PubMed using the key words: “influenza,” Robert L. Seymour, Shannan L. Rossi, Krista Versteeg, Hannah E. “epidemiology,” “season,” and “surveillance” to abstract data on Romo, Nicholas Bergren, Scott C. Weaver the percent of samples testing positive for influenza during each epidemiologic week. The start of influenza season was defined as University of Texas Medical Branch, Galveston, TX, United States the first week when the proportion of samples that tested positive O’nyong-nyong virus (ONNV) is an alphavirus transmitted by mosquitoes remained above the annual median for at least 6 weeks. We assessed that shares 90 percent nucleotide sequence identity with Chikungunya changes in the relationship between percent of samples testing positive virus (CHIKV) and has been the cause of two major epidemics in Africa and average monthly temperature using linear regression models.We during the past 20 years. These viruses produce very similar acute febrile identified data on laboratory-confirmed influenza virus infection from illnesses characterized by rash and debilitating arthralgia. While there 84 countries comprising 5.4 billion (83%) of the world’s population. are many studies into the pathogenesis of CHIKV, little is known about While (44 [94%] of 47) temperate and all four subtropical countries had the pathogenesis of ONNV. To determine which portions of the immune one annual epidemic, 24 (72%) of 33 tropical countries had one annual system are important in protection during initial infection with ONNV we influenza epidemic, seven (21%) had biannual epidemics, and 2 (6%) inoculated several different strains of mice with two different strains of had insufficient data to analyze. Influenza was identified every week in ONNV (SG650 and MP30). These mice included wild type C57BL/6J and 4 (9%) of 47 temperate, 0 subtropical countries versus, and 10 (30%) of mice knocked out for the following genes: STAT1 (STAT1 KO; defective 33 tropical countries (p=0.04). Peak influenza activity occurred within two in both type I and type II interferon signaling), Type I interferon receptor months after the lowest temperatures in 36 (82%) of 44 temperate, 1 (A129), interferon g receptor (IFNgR KO), recombinase activation gene 1 (25%) of 4 subtropical, and 6 (27%) of 22 tropical countries with available (RAG1 KO). Mice were inoculated subcutaneously with 100-10000 PFU of data (p<0.001). Influenza activity peaked in Southeast Asia and Oceania ONNV or sham inoculated with phosphate buffered saline. Mice were bled during June-July; Australia and China during August; Middle East, North to assess viremia, weighed for at least 14 days unless infection was lethal, Africa and Mexico during December, Europe and North American during and observed for illness daily. Tissue samples were collected to test for viral February-March and South America and South Africa during May-June. load and for histopathologic evaluation. The C57BL/6J, RAG KO and IFNgR In conclusion, annual influenza epidemics occur in consistent temporal KO mice showed no statistical difference in weight compared to control patterns depending upon climate. Local influenza surveillance and climate mice, and never generated detectable viremia. A129 mice demonstrated data may best inform influenza prevention activities and focus efforts morbidity but survived. STAT1 KO mice demonstrated an age dependent during periods of highest local activity. mortality with 6-week-old mice succumbing to illness by day 12, while 8-14 week old mice developed morbidity as evidenced by weight loss 1422 and clinical signs of illness but survived. Tissues of STAT1 KO mice demonstrated a monocytic infiltrate in all major organs and infectious CHARACTERIZATION OF THE NATURAL HISTORY OF LASSA virus was detected in brain and skeletal muscle. We conclude that the FEVER VIRUS DISEASE IN CYNOMOLGUS MACAQUES adaptive immune system is not necessary for protection against initial FOLLOWING AEROSOL EXPOSURE ONNV infection in the mouse model while the type I interferon response demonstrates an age-dependent phenotype. These results will be valuable A.N. Honko, W.D. Pratt, J.C. Johnson, C.I. Shaia, C.R. Reid, E.M. for designing animal models for testing candidate vaccines or therapeutics Mucker, J. Shamblin, H. Esham, G. Olinger, L.E. Hensley against ONNV infection. U.S. Army Medical Research Institute for Infectious Diseases, Ft. Detrick, MD, United States Viral hemorrhagic fever (VHF) in humans is caused by members of four families of enveloped, negative-sense or ambisense RNA viruses; Arenaviridae, Bunyaviridae, Filoviridae and Flaviviridae. These viruses are considered possible biothreats due to their potential for airborne and person-to-person transmission, making characterization of the aerosol route of infection paramount. First described in Nigeria in 1969, LASV is endemic to the West African countries of Sierra Leone, Liberia and Guinea. In Africa, members of the Mastomys genus of mulimammate rats are persistently infected with LASV from birth. Humans become infected via the respiratory route through exposure to virus in rat excreta, as well as by preparing and eating infected animals. It is estimated LASV may cause 5,000-10,000 fatalities annually; however, there are currently no FDA-approved therapeutics or vaccines. Examining the natural history of www.astmh.org 428 a disease, from exposure through resolution, permits the description of an important role in genomic stability. We hypothesize that SOD1 is stages of disease course, the immune responses, as well as mechanisms diminished in cells from HTLV-1-infected subjects with high DNA damage, of pathogenesis or correlates of survival. The objectives of this study were and that hbz expression is associated with the expansion of these cells. to 1) identify early clinical signs that can be used to indicate or predict To test this hypothesis we measured Proviral Load (PVL), hbz mRNA, infection, 2) identify markers that indicate disease progression as well as and SOD1 protein levels in cells from HTLV-1-infected subjects with development of severe disease and 3) determine the impact of anesthesia high (HD) and low (LD) DNA damage. Peripheral blood mononuclear on clinical disease progression. The natural history of LASV infection cells (PBMCs) were isolated to estimate DNA damage by alkaline comet following aerosol exposure was examined in the cynomolgus macaque assay. Two groups of HTLV-1-infected subjects were defined: HD (≥ 51 model. Animals were surgically implanted with telemetry providing comets/100 nucleoids; n=18); and LD (≤ 50 comets/100 nucleoids; n=9). simultaneous real-time monitoring of pressures, ECG, and temperature We measured PVL and hbz mRNA levels by real time PCR, and plasma as well as daily blood sampling occurred via central venous catheters SOD1 levels by a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). beginning from Dy -3. Based on these results, a sequential sampling PVL was expressed as HTLV-1 tax copy number /104 PBMCs. Samples were study was designed to collect presymptomatic, early, intermediate and classified as hbz+ (detectable) or hbz (undetectable). Statistical analyses late stages of disease. Summary results will be presented, including were based on non-parametric tests. We did not find differences in PVL hematologic changes (complete blood counts), blood chemistry analysis (p=0.959) or SOD1 (p=0.064) between HD and LD. However, all hbz+ (Piccolo and iSTAT), coagulation assays, and plasma cytokine responses, as samples came from a cluster of HD subjects with high PVL (3168 ±980), well as pathology findings. who also showed lower levels of SOD1 than the remaining HD group (p=0.039) and the LD group (p=0.01758). In conclusion, DNA damage is 1423 not always associated with low SOD1 levels in HTLV-1-positive subjects. Our results suggest that hbz expression supports survival and expansion SUSCEPTIBILITY OF MARMOSETS (CALLITRIX JACCHUS) TO of HTLV-1-infected cells bearing DNA damage associated with low SOD1 MONKEYPOX VIRUS levels. Eric Mucker, John Huggins, Joshua Shamblin, Sarah McCarthy, Jennifer Chapman, Lisa Hensley 1425 U.S. Army Medical Research Institute for Infectious Diseases, Fort Detrick, OLIGONUCLEOTIDE MICROARRAYS FOR THE DETECTION MD, United States AND CONFIRMATION OF ARBOVIRAL PATHOGENS IN THE Although current nonhuman primate models of monkeypox and smallpox FIELD diseases provide some insight into disease pathogenesis, they require John S. Lee1, Nathan Grubaugh1, Lawrence Petz1, Lewis Long1, a high titer innoculm, use an unnatural route of infection, and/or do Vanessa Melanson2, Sarah Pisarcik1, Ampornpan Kengluecha3, not accurately represent the entire disease course. In our studies, we Boonsong Jaichapor3, Prasan Kankaew3, Alongkot Ponlawat3, altered half of the test system by using a New World primate species, Brian Evans3, Monica O’Guinn4 the common marmoset. Adult male marmosets were intravenously 1U.S. Army Medical Research Institute for Infectious Diseases, Frederick, infected with 2.4x107, 9.5x105, and 7.8x104, 5.0x103, 510, and 48 MD, United States, 2Walter Reed Army Institute of Research, Silver Spring, PFU. Clinical, hematological, and viral load data were assessed. Animals MD, United States, 3Armed Forces Research Institute of Medical Sciences, were euthanized or succumbed to disease between 6 and 15 days post- Bangkok, Thailand, 4MIDRP, Frederick, MD, United States infection, in a dose dependent manner. The animals exhibited signs of hemorrhage, had high genome viremia, and altered hematological With the emergence and re-emergence of arthropod-borne diseases parameters. At the lower doses, rash was more demarcated and some throughout the world, it is critical to be able to detect arthropod- short-lived macules were observed. As is, our model is 6 logs lower than transmitted pathogens in a timely manner. Standardized field-diagnostic the current intravenous cynomalgus model and 4-6 logs lower than protocols for identifying arthropod-borne pathogens within any given respiratory models. The aggressive nature of the disease manifested in region of the world are indispensable tools for obtaining real-time these animals implicates an even lower dose and warrants exploration of information for health-care providers and preventive medicine specialists other infection routes. Also, these data should invoke consideration for in field settings. Arboviruses are responsible for major outbreaks of variola experimentation in marmosets. acute, febrile disease throughout most areas of the world. Dengue (DEN), Japanese encephalitis (JE), yellow fever, Chikungunya, and tick-borne 1424 encephalitis complex viruses are but a few of the viruses that account for a majority of the arboviral infections that cause morbidity and SURVIVAL AND EXPANSION OF HTLV-1-INFECTED CELLS mortality in humans. Here we report the development and field-testing WITH HIGH DNA DAMAGE: RELATIONSHIP BETWEEN SOD1 of an oligonucleotide microarray for the detection and confirmation AND GENOMIC STABILITY of arboviruses in pools of field collected mosquitoes. During the field evaluation, mosquito pools were screened by using generic PCR assays Karina Flores1, Giovanni López1, Michael Talledo2, Carolina and then by using virus specific real-time PCR assays. The phylogenetic Álvarez2, Elsa González2, Kristien Verdonck2, Johan Van relationships among these viruses, their mosquito hosts, and their possible Weyenbergh3, Eduardo Gotuzzo2, Daniel Clark1 role in causing human disease is also presented. 1Universidad Peruana Cayetano Heredia, Lima, Peru, 2Instituto de Medicina Tropical “Alexander von Humboldt-UPCH, Lima, Peru, 3Rega Institute for 1426 Medical Research, KULeuven, Leuven, Belgium HTLV1 may exert an initial pro-apoptotic stimulus via the oncoprotein MOLECULAR DIAGNOSIS AND ANALYSIS OF IMPORTED Tax-induced DNA damage. On the other hand, the viral protein Hbz CHIKUNGUNYA VIRUS STRAINS, JAPAN, 2006-2010 antagonizes some Tax effects and recent studies show that hbz is Chang-Kweng Lim, Tomohiko Takasaki, Meng Ling Moi, Akira always expressed in leukemic cells, suggesting its involvement in the Kotaki, Ichiro Kurane, Masayuki Saijo maintenance of malignity. Tax has been implicated in the initiation of Department of Virology 1, National Institute of Infectious Diseases, Tokyo, cellular transformation, chromosomal instability, and induction of cellular Japan DNA damage by reactive oxygen species (ROS). Superoxide dismutase 1 (SOD1) is an antioxidant enzyme present in the cytoplasm, nucleus, and Chikungunya (CHIK) virus has re-emerged as an important mosquito- intermembrane space of mitochondria; SOD1 catalyzes the dismutation borne pathogen causing epidemics in several parts of the world. The of superoxide to hydrogen peroxide and molecular oxygen, thus playing CHIK virus belongs to the Alphavirus genus in the family Togaviridae. www.astmh.org 429 A large-scale epidemic of CHIK fever started in Kenya in 2004 and 1428 spread to Reunion Island, other Indian Ocean islands, India, Sri Lanka, Singapore, Thailand and Malaysia. One of the main vectors responsible THE POPULATION STRUCTURE OF AMAZONIAN for transmission between humans is Aedes albopictus, which is widely PLASMODIUM FALCIPARUM distributed in urban areas of Europe, the USA and East Asia. This fact 1 2 raises concern that the virus could be introduced and become established Sean M. Griffing , Giselle Rachid Viana , Tonya Mixson- 1 1 1 in these areas. During 2007-2010, 19 imported CHIK cases were detected Hayden , Sankar Sridaran , Md. Tauqeer Alam , Andrea M. 1 1 1 in Japan from South and Southeast Asia. The samples were tested for McCollum , Alexandre Macedo de Oliveira , John W. Barnwell , 3 2 dengue virus as well as CHIK virus by IgM-capture ELISA, real time RT- Ananias A. Escalante , Marinete Marins Povoa , Venkatachalam 1 PCR, virus isolation with Vero and C6/36 cell, and plaque reduction Udhayakumar neutralization tests. In this study, we report two cases of imported 1Centers for Disease Control and Prevention, Atlanta, GA, United States, infection in patients who had returned to Japan from Malaysia and 2Instituto Evandro Chagas/Funesa, Belem, Brazil, 3Arizona State University, Indonesia. Both viruses were successfully isolated from the cases by using Phoenix, AZ, United States a plaque purification technique. Phylogenetic analysis showed that the We previously showed that population structure of Peruvian Plasmodium strain from Indonesia was grouped into the Asian genotype. However, falciparum during the peak expansion of malaria in 1990s, following the isolate from Malaysia was identical to the Central/East/South African almost two decades of very low malaria transmission, was confined genotype and was clustered with currently reported Indian isolates. The to five major clonal lineages that we referred to as clonets. We have strain from Malaysia also had the E1-A226V mutation. These data suggest also shown that Venezuelan P. falciparum parasites appeared to have that the CHIK virus circulating in Malaysia is related to that currently somewhat more diverse clonets. Given the shared borders between epidemic in South and Southeast Asia. Further characterization of these Brazil and these two countries, we compared the population structure isolates is in progress. of P. falciparum parasites in the Amazon basin, and coastal Peru, using neutral microsatellite markers. The microsatellite data was also combined 1427 with drug resistance genotypes and microsatellite markers flanking genes THE APPLICATION OF PYROSEQUENCING TO IDENTIFY associated with drug resistance (pfcrt, dhfr, dhps and pfmdr1) in an aggregate analysis. A total of 190 samples from Brazil were examined MULTISPECIES PLASMODIUM INFECTIONS IN HUMANS AND from three states (Amapá, Pará, and Rondônia), representing multiple sites APES IN WEST CENTRAL AFRICA and time periods since the 1980s in this study. We analyzed our data using Sesh A. Sundararaman1, Weimin Liu1, Brandon F. Keele2, Scott network diagrams, estimates of heterozygosity, and tests for bottlenecks A. Sherrill-Mix1, Yingying Li1, Julian C. Rayner3, Paul M. Sharp4, and recent population expansions. Like Peru and Venezuela, Brazil Martine Peeters5, Frederic D. Bushman1, Beatrice H. Hahn1 exhibited low parasite diversity and the distance between collection sites was not significantly correlated with increasing genetic differentiation, 1University of Pennsylvania School of Medicine, Philadelphia, PA, but unlike these countries there were no obvious clonets. We interpreted United States, 2AIDS and Cancer Virus Program, SAIC-Frederick, Inc., the apparent Brazilian P. falciparum population structure to be the logical National Cancer Institute, Frederick, MD, United States, 3Sanger Institute outgrowth of multiple waves of internal migration, admixture, and Malaria Programme, The Wellcome Trust Sanger Institute, Cambridge, sexual recombination over past decades. We described the evidence for United Kingdom, 4Institute of Evolutionary Biology, University of population bottlenecks at various sites within Brazil. Furthermore, we Edinburgh, Edinburgh, United Kingdom, 5Institut de Recherche pour le related the parasite populations from Peru and Venezuela to the Brazilian Développement (IRD), University of Montpellier 1, Montpellier, France Amazon basin. At least two of the clonets in the Pervian Amazon were Recent studies have identified several new species of Plasmodium in linked with Brazilian Amazon isolates, while the Peruvian coastal lineages wild-living chimpanzee and gorilla populations. Using single genome were only related by way of samples collected in the Peruvian Amazon. amplification (SGA), we have shown that most apes are co-infected with While Venezuelan isolates generally seemed to have more in common multiple divergent strains and traced the origin of human P. falciparum with each other, they were also directly linked to Brazilian isolates. This to western gorillas. This has raised the question whether wild-living apes underscored the remarkable clonality of Peru and the intermediate serve as a recurring source of human infection. While SGA methods clonality of Venezuela. Our findings suggest that, if Brazilian patterns of are useful in precluding in vitro recombination, this approach is not internal migration continue in the future, future drug resistance might designed to identify low abundance strains in multi-species infections. rapidly spread throughout the country. Here, we describe a novel method of addressing this question by using pyrosequencing technologies. We developed a set of 454 FLX Titanium 1429 pan-Plasmodium primers that amplify a 510 bp fragment of mitochondrial DNA containing sufficient diversity to differentiate all previously identified POPULATION GENETICS OF COPY NUMBER VARIATION IN ape and human Plasmodium species. These primers, in conjunction with PLASMODIUM FALCIPARUM the GS FLX System, generate over 1 million reads in a single run, providing Becky Miller1, Ian Cheeseman2, John C. Tan1, Asako Tan1, Shalini both depth of sequencing and high sample throughput. Primers are 2 2 1 2 tagged with sample-specific 12-mer barcodes allowing us to identify the Nair , Standwell Nkhoma , Michael T. Ferdig , Tim Anderson sample origin of each read. To process our data, we have developed a 1University of Notre Dame, South Bend, IN, United States, 2Texas Institute method of rapidly classifying reads by alignment to multiple reference for Biomedical Research, San Antonio, TX, United States sequences from all known primate Plasmodium lineages. Amplicon Genome rearrangements, such as copy number variation (CNV), are -3 sequencing by this method has an error rate of 1.5x10 mismatches/ ubiquitous in eukaryotic genomes. Gene dosage changes resulting nucleotide, low enough to accurately distinguish even the most closely from gene duplications or deletions may play an important role in related lineages. In a pilot run of 90 human buffy coat samples from adaptive evolution. However, the role of DNA rearrangments has been Cameroon, 59 were positive for P. falciparum alone, while the remainder largely ignored in malaria biology despite the fact that the Plasmodium represented multiple species infections with P. falciparum and P. malariae karyotype is highly variable, many rearrangements have been reported in (n=23), P. falciparum and P. ovale (n=5), or all three species (n=3). Read laboratory lines and CNVs are known to influence drug resistance. This ratios give an indication of the relative abundance of each Plasmodium project examines extent and functionality of genome rearrangements in species in a sample. These data demonstrate that deep sequencing the malaria parasite Plasmodium falciparum, as well as their origins and technology can be used to identify mixed-species infections, even when evolutionary dynamics. We detected genome-wide CNV in more than one or more species is present at ratios below 1:2000. 100 parasites from SE Asia, (Cambodia, Lao PDR and Thailand) and Africa (Malawi and Gambia) using comparative genomic hybridization (CGH) on www.astmh.org 430 a custom Nimblegen microarray (the CNV-SNP array) that assays both CNV 1431 and SNP variation at high resolution genome-wide. The parasites examined were prescreened to exclude multiple clone infections, and patient derived GENOME SEQUENCING ASSOCIATION STUDIES: A NEW material was used to avoid artifacts caused by laboratory adaptation. We APPROACH FOR UNDERSTANDING ANTIMALARIAL determined the size, gene content, and population frequency of genome RESISTANCE IN PLASMODIUM FALCIPARUM rearrangments within and between parasite populations and evaluate 1 2 1 the roles of drift and selection (positive and purifying) in shaping the Daniel J. Park , Daria Van Tyne , Hsiao-Han Chang , Meghan 2 2 3 3 CNV distributions. Geographical variation in SNPs and CNVs and linkage Galligan , Amanda K. Lukens , Kevin Gallinsky , Lauren Young , 4 4 4 disequilibrium in regions flanking CNVs were used to better understand Daouda Ndiaye , Papa Diogoye Sene , Souleymane Mboup , 3 3 1 the evolution of genome rearrangements in P. falciparum. Roger C. Wiegand , Daniel E. Neafsey , Daniel L. Hartl , Sarah K. Volkman2, Pardis C. Sabeti1, Dyann F. Wirth2 1430 1Harvard University, Cambridge, MA, United States, 2Harvard School of Public Health, Boston, MA, United States, 3Broad Institute, Cambridge, HIGH COVERAGE GENOME SEQUENCING OF FIELD ISOLATES MA, United States, 4Cheikh Anta Diop University, Dakar, Senegal PROVIDES UNIQUE INSIGHTS ON PLASMODIUM VIVAX Plasmodium falciparum malaria’s rapid adaptation to new drugs allows BIOLOGY it to remain one of the most devastating infectious diseases of humans. David Serre1, Ernest R. Chan1, Didier Menard2, Pheaktra Chim2, Understanding the genetic basis of these adaptations is critical to Melinda Blood3, Arsene Ratsimbasoa4, Peter David5, Odile successful intervention. Using next-generation sequencing and drug- Mercereau-Puijalon5, Peter A. Zimmerman3 sensitivity testing, we performed genome sequencing association studies 1Cleveland Clinic, Cleveland, OH, United States, 2Pasteur Institute of (GSAS) on 25 recently isolated parasites from Senegal against and 13 Cambodia, Phnom Penh, Cambodia, 3Case Western Reserve University, antimalarial drugs_including amodiaquine, artemisinin, atovaquone, Cleveland, OH, United States, 4Madagascar Ministry of Health, chloroquine, dihydroartemisnin, halofuganone, halofantrine, lumefantrine, Antananarivo, Madagascar, 5Pasteur Institute, Paris, France mefloquine, piperaquine, primaquine, pyrimethamine, and quinine_and 20 synthetic compounds. This novel use of whole genome sequence data The biological diversity of Plasmodium vivax is poorly understood, greatly expands our power to detect associations in low LD populations, partly because the parasite cannot be easily propagated in vitro. As an as it assays nearly a thousand-fold more positions in the genome than our alternative it is becoming increasingly feasible to characterize the genetic previous 17,582 SNP array (average coverage of 17.2 Mbp per parasite). diversity of P. vivax isolates across their genomes and to associate genetic However, it comes with some new analysis challenges, including the variation with biological traits. We present here whole genome sequences reliable characterization of marker genotypes from read data (127,362 generated directly from the blood of three patients and show how robust SNPs and 7,919 microsatellites are polymorphic in this population), characterization of the genomic diversity can provide unique insights handling missing data and more LD in the denser marker set. We about P. vivax biology. We analyzed blood samples from one Cambodian adapted recent mixed-model GWAS tools, such as EMMA and GCTA, and two Malagasy patients harboring parasitemias between 0.1 to and selection tools, such as HLR and XP-EHH, to study the heritability of 0.35% and confirmed P. vivax mono-species infection by Plasmodium drug response phenotypes and identify known and novel loci associated species PCR-based diagnosis. After leukocyte depletion of whole blood with drug resistance at genome-wide significance. This demonstrates (5 ml; using CF11-packed columns), we extracted P. vivax DNA from improvements of the GSAS approach over traditional, array-based GWAS parasitized red cells and prepared libraries from each individual sample for understanding the genetic basis for antimalarial drug resistance in after fragmentation of the DNA into 250-300 bp. We sequenced each the wild, potentially identifying important biomarkers for surveillance as library on individual lanes of an Illumina HiSeq 2000. We were able to map elimination and eradication efforts are pursued. 20-60 % of the 80 million 100 bp paired-end sequences generated from each sample to the SalI P. vivax genome sequence while the remaining 1432 reads (40-80%) mapped to the human genome. The very high coverage (100-300X) generated by our sequencing effort provides both a robust GENETIC ANALYSIS OF PLASMODIUM FALCIPARUM description of the genetic diversity across the entire P. vivax genome, and GAMETOCYTOGENESIS allows us to identify and differentiate multiple P. vivax strains within each 1 2 3 infected patient. In addition, we show that we can analyze variations in Hiromi Ikadai , Kathryn Shaw Saliba , Stefan Kanzok , Kyle 2 3 2 sequence coverage along each genome to identify gene duplications and McLean , Kim C. Williamson , Marcelo Jacobs-Lorena deletions. Finally, we describe multiple DNA sequences shared among 1Kitasato University Japan, Towada, Japan, 2Johns Hopkins Bloomberg the newly sequenced genomes that are missing (for either technical or School of Public Health, Baltimore, MD, United States, 3Loyola University biological reasons) from the SalI reference genome. Overall, our analyses Chicago, Chicago, IL, United States confirm that sequencing of P. vivax genomes from field isolates is very Within the mammalian host, the Plasmodium parasite has two feasible following leukocyte depletion and illustrates that substantial developmental fates: cyclic asexual replication or terminal sexual information can be generated regarding genome structure, sequence differentiation (gametocytogenesis). The sexual forms of the parasite polymorphism, and complexity of infection. (gametocytes) are the only form that is able to survive and propagate in the mosquito vector. Therefore, gametocytes are absolutely essential for parasite transmission. Very little is known about the mechanisms involved in the commitment of Plasmodium to sexual differentiation. To gain insight into these mechanisms, we conducted a piggyBac transposon-mediated insertional mutagenesis and screened for parasites that no longer formed mature gametocytes. Of 736 parasites (clones) screened in 3 independent transfection experiments, 29 clones did not form gametocytes. For each clone, insertion of piggyBac was verified by Southern blot analysis and the disrupted genes were identified by inverse PCR. This led to the identification of 16 putative gametocytogenesis-disrupting genes. Genetic complementation for 4 of the 16 genes was successfully carried out showing that these genes are essential for gametocytogenesis. To epistatically order the 16 genes, we measured their expression pattern along with the expression pattern of other known gametocyte-specific www.astmh.org 431 genes in each of the gametocyte-minus mutants using RT-PCR. We found and 2009. Results: We observed improvements in the availability (from a subset of the genes that are likely to act very early in commitment to 0.24 shops per 1,000 people in 2004 to 0.39 in 2008) and accessibility gametocyte differentiation, another subset likely to act just after the (from 71% of households within 5 km of a shop in 2004 to 87% in committed merozoite invades the red blood cell, and a third set likely to 2008) of drug shops. After the introduction of ALu stock levels of the act early (stage I) gametocytes. Thus, we have carried out a comprehensive drug were relatively high in public health facilities (over 80% months screen for genes essential to commitment and early differentiation of in stock), but the drug could only be found in 30% of drug shops. the Plasmodium gametocyte. This line of investigation may lead to novel The proportion of children treated with an antimalarial within 24hrs of strategies to reduce parasite transmission and disease burden. onset of fever increased from 66% to 89% between 2004 and 2008. However, only 51% were treated with the newly introduced ALu in 2008. 1433 Under-five mortality decreased from 28.4 cases per 1000 person years (c/1000py) in the years before 2004 to 18.5 c/1000py in 2008 and 2009. VARIANT ANTIGEN EXPRESSION IN PEDIATRIC MALARIA The ACCESS interventions were independently associated with decreases in mortality, controlling for other malaria interventions and contextual Jacqui Montgomery1, Dumizulu Tembo1, Danny Milner2, Fingani factors (incidence rate ratio comparing before 2004 vs. after 2008=0.84, Mphande1, Matthew Berriman3, Stephen Rogerson4, Terrie Taylor5, 95%CI=0.72 to 0.99). In conclusion, an integrated approach which tackles Malcolm Molyneux1, Alister Craig6 both users and providers, recognising the important role of the private 1 Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, retail sector, can lead to improvements in terms of access to malaria 2 Malawi, Harvard School of Public Health, Boston, MA, United States, treatment and can contribute to decreases in mortality in rural African 3 4 Wellcome Trust Sanger Institute, Hinxton, United Kingdom, The settings. University of Melbourne, Melbourne, Australia, 5Michigan State University, East Lansing, MI, United States, 6Liverpool School of Tropical Medicine, 1435 Liverpool, United Kingdom The ability of Plasmodium falciparum-infected erythrocytes to sequester STRENGTHENING MALARIA INFORMATION SYSTEMS IN from the circulation into organ microvasculature is associated with much SOUTH AFRICA: MOVING TOWARDS ELIMINATION of the lethality of this species. We have investigated genetic variation Mbavhalelo Shandukani1, Tej Nuthulaganti2, Jennifer and antigen expression of parasites in organ biopsies from 25 Malawian Drummond3, Daniel Williams3, Gerdalize Kok4, Samson Katikiti5, paediatric malaria patients. Patients were autopsy-confirmed cerebral Rajendra Maharaj6, Eunice Misiani1, Devanand Moonasar1 malaria cases or parasitaemic controls with an incidental or mild P. 1South Africa National Department of Health, Pretoria, South Africa, falciparum infection and another identified cause of death. Cerebral 2Clinton Health Access Initiative and Global Health Group – University of malaria cases had low multiplicity of infection with often a single genetic California San Francisco, Pretoria, South Africa, 3Clinton Health Access variant dominating the infection throughout the organs. Expression of the Initiative, Pretoria, South Africa, 4Mpumalanga Department of Health variant surface antigen, P. falciparum erythrocyte membrane protein-1, and Social Services, Nelspruit, South Africa, 5World Health Organization, was investigated by quantitative PCR and analysis of expressed sequence Harare, Zimbabwe, 6Medical Research Council of South Africa, Durban, tags. Cerebral malaria infections and parasitaemic controls showed South Africa similar patterns of antigen expression in host tissues, with particular antigens often being expressed at dominant levels (>33%) by parasites South Africa has made significant progress in controlling malaria during in the brain, heart or gut. These dominant antigens can vary between the past decade. Between the years 2000 to 2010 malaria incidence organ populations within a single host. There was high overlap in the has been reduced from 8.79 to 0.63 local cases per 1000 population at antigens observed in different patients, with 22% of the 644 antigens risk, respectively. Total locally transmitted malaria cases have declined by being detected in multiple patients, and 30% of antigens in the brain 92% and malaria deaths have declined by 82% in 2010 as compared also observed in brain biopsies from other patients. This finding was to 2000. As a result, South Africa is progressing towards achieving unexpected given the negligible overlap in antigen diversity seen globally malaria elimination and will need to develop and manage a robust and in endemic sites. Our findings suggest that a restricted number of malaria information system (MIS) that will form the basis for effective antigens are implicated in sequestration in the paediatric host. management by enabling evidence-based decision-making on appropriate use of human, technical and financial resources. To monitor and evaluate 1434 progress towards elimination, a comprehensive MIS will need to capture epidemiological information on cases and deaths, GIS mapping of malaria AN EVALUATION OF THE IMPACT OF INTEGRATED foci and breeding sites and entomological and parasitological data. The INTERVENTIONS TO IMPROVE ACCESS TO MALARIA purpose of this review is to determine the 2010 baseline for intervention TREATMENT IN TANZANIA - THE ACCESS PROGRAM coverage rates stratified among targeted municipalities as well as to determine reporting breakdowns within the current MIS. This paper Sandra Alba1, Rose Nathan2, Mathew Alexander3, Angel Dillip3, describes South Africa’s National MIS and reviews challenges and best Alexander Schulze4, Flora Kessy3, Christian Lengeler1 practices in developing an integrated rapid notification malaria database 1Swiss Tropical and Public Health Institute, Basel, Switzerland, 2Ifakara that is unified and standardised across provinces. This standardisation is Health Institute, Dar es Salaam, United Republic of Tanzania, 3Ifakara essential in order to increase information flow and inform programmes Health Institute, Ifakara, United Republic of Tanzania, 4Novartis Foundation of potential outbreaks, allowing for prompt response and investigation. for Sustainable Development, Basel, Switzerland As countries in sub-Saharan Africa move towards elimination, a vigorous The ACCESS Programme was implemented between 2004 and 2008 surveillance program is necessary to rapidly identify new and reignited in two Tanzanian districts to improve access to malaria treatment with foci of transmission, as well as track the movement of vectors, parasites a set of integrated interventions at three levels: 1) community level; 2) and parasite carriers in this fluid environment. Within the context of public heath facilities; and 3) commercial drug sector. The study period South Africa the lack of a functional, standardised information system, saw the switch from Sulphadoxine-Pyrimethamine (SP) to Arthemeter and the absence of timely notification of changes in the epidemiological Lumefantrine (ALu) as first line treatment for malaria in 2006. This study landscape, has led to an inability to inform targeting of interventions due aims at evaluating the ACCESS Programme’s interventions. We conducted to undetermined foci of transmission. The obstacles encountered and yearly censuses in all health facilities and drug shops between 2004 and overcome in the restructuring of South Africa’s MIS provides necessary 2008 and treatment seeking surveys on approximately 150 individuals in guidance to other countries who seek to strengthen control programmes 2004, 2006 and 2008 in the Ifakara Demographic Surveillance Site (DSS). and lay a strong foundation for the eventual development of elimination The DSS provided yearly estimates of under-five mortality between 1997 strategies. www.astmh.org 432 1436 (RDT) or microscopy were enrolled and provided travel history, location of residence, and fingerprick blood samples. We performed genotyping MALARIA BURDEN AND COVERAGE ESTIMATES FROM of 9 P. falciparum microsatellites to see if genetic information would THE 2010-2011 MONTHLY ‘ROLLING’ MALARIA INDICATOR provide insight into transmission patterns. We enrolled 906 patients, 712 SURVEY (rMIS) IN CHIKHWAWA DISTRICT, MALAWI: A (79%) in the wet season (May-July). 35% of patients presenting in the dry POTENTIAL DISTRICT-LEVEL MALARIA MONITORING AND season reported travel outside of Zanzibar in the past month versus 3% EVALUATION (M&E) TOOL FOR PROGRAM MANAGERS of those in the wet season (RR 11, 95%CI 7-17, p <.001), suggesting that a higher fraction of dry season cases may be imported. Thus far, we have Arantxa Roca-Feltrer1, Kamija Phiri2, David Lalloo3, Dianne 511 complete multilocus genotypes from 384 patients. Closely related Terlouw3 parasites (100%, 80-99%, or 50-79% identity) were more likely to come 1MLW, Blantyre, Malawi, 2CoM, Blantyre, Malawi, 3Liverpool School of from patients living in the same shehia (smallest administrative unit) than Tropical Medicine and Malawi Liverpool Wellcome Trust Programme, less related parasites, indicating fine-scale spatial clustering of related Liverpool, United Kingdom parasites (RR: 18, 15, and 3 respectively, vs. <50% identity, p<.001 for all). 91 clusters of parasites ≥80% identical were identified, with cluster size Novel malaria M&E tools are urgently needed to complement the current ranging from 2 to 19 parasites. In the wet season, 57% of patients were ‘gold standard’ Malaria Indicator Surveys (MIS). Rapid up-scaling of infected with a clustered parasite versus 13% in the dry season (p<.001), malaria control efforts is resulting in substantial reductions in malaria demonstrating that local transmission occurs and likely dominates in the burden across sub-Saharan Africa. As transmission goes down, timely, wet season. Exploratory visualization of clusters of parasites on a map accurate, sub-national and district level burden estimates are needed suggested specific patterns of local transmission. Local transmission to guide increasingly targeted, sub-national control efforts in remaining appears to account for a high proportion of malaria cases during the wet hotspot areas. To test a novel district level M&E tool, we started a monthly season in Zanzibar, but imported malaria may play a significant role during ‘rolling’ MIS (rMIS) in May 2010 covering 51 villages in Chikhwawa the dry season. Analysis of parasite genetics may provide insight into district. This is one of the National Malaria Control Programme (NMCP) malaria control strategies in low-endemic settings. focus districts with high insecticide treated net coverage and annual indoor residual spraying with round 1 in January-February 2011. During 1438 the first year, approximately 1,200 households were randomly selected using a probability proportional to (village) size. Approximately 100 MALARIA ACTIVE INFECTION DETECTION IN AN AREA households were visited each month, and approximately 60 under-fives OF LOW PARASITEMIA PREVALENCE: LUSAKA DISTRICT, were tested for anaemia and parasitaemia. Parasitaemic children (by RDT ZAMBIA in the field) were treated as per national guidelines. Each month, data was collected in a one-week period by two teams of two people. Data Anna M. Winters1, Zunda Chisha2, Mercie Mwanza3, Mulakwa was collected using Personal Digital Assistants, and uploaded daily for Kamuliwo3, Clara Mbwili4, Matimba Chiko-Like4, Moonga quality checking. Results were usually available within two weeks from Hawela3, Peter Mumba5, Jacob Chirwa3, Benjamin Winters1, completion of data collection. Data quality was maintained throughout Matthew Burns1, Daniel Bridges2, Kathrine R. Tan6, Oliver the whole period. Standard malaria impact indicators (moderate anaemia Lulembo7, John Miller8, Allen S. Craig6 (Hb<8g/dL) and malaria prevalence in under-fives) as well as intervention 1Akros, Laramie, WY, United States, 2Akros, Lusaka, Zambia, 3National coverage indicators will be presented for the first 12 months of the study Malaria Control Centre, Lusaka, Zambia, 4Lusaka District Health Office, and by season. The strengths and weaknesses of burden estimates from Lusaka, Zambia, 5Zambia Integrated Systems Strengthening Program, monthly data collections will be discussed. There will be a particular focus Lusaka, Zambia, 6Centers for Disease Control and Prevention, Atlanta, GA, on whether short-term changes in malaria indicators can be detected United States, 7President’s Malaria Initiative, Lusaka, Zambia, 8MACEPA, following district-wide indoor residual spraying after accounting for Lusaka, Zambia seasonality. Small-scale, rolling sub-national MIS surveys could be a viable complementary malaria M&E approach for district level program managers Malaria surveillance in Zambia has been via passive case detection. The and control efforts. 2010 Malaria Indicator Survey reported a very low prevalence of malaria in Lusaka and it is suspected that cases of malaria in the urban parts of this 1437 district are primarily imported. As transmission rates continue to decline as this area progresses towards malaria elimination, it is necessary to find EVIDENCE FOR LOCAL MALARIA TRANSMISSION IN THE WET and treat asymptomatic malaria cases implying the need for cost-effective, SEASON AND IMPORTED MALARIA IN THE DRY SEASON IN focal intervention and surveillance strategies. To address this need, the ZANZIBAR National Malaria Control Centre, Lusaka District Health Office and partners began malaria active infection detection (AID) where laboratory-confirmed, Jordan Kemere1, Michelle S. Hsiang2, Abdullah S. Ali3, Mwinyi passively-detected malaria cases are followed-up in the community I. Msellem3, Makame H. Makame3, Kimberly A. Baltzell1, Stacy through testing and treating of households (n=9) immediately surrounding Salerno1, Tanya Libby4, Alanna Schwartz1, Edmund Seto4, Andreas the house of each index case. Since March 2011, community-based teams Mårtensson5, Peter D. McElroy6, Bryan Greenhouse1 have led weekly AID responses in 5 health facility catchment areas within 1University of California, San Francisco, San Francisco, CA, United Lusaka District with expansion to additional clinics forthcoming. Thus far, States, 2University of California, San Francisco, Global Health Group, a total of 21 index cases have received AID response, with a total of 678 San Francisco, CA, United States, 3Zanzibar Malaria Control Programme, individuals tested for malaria during these responses. Of the 678 tested, Zanzibar, United Republic of Tanzania, 4University of California, Berkeley, 20 (2.9%) were found to be positive. Of the 20, 14 reported a recent Berkeley, CA, United States, 5Karolinska Institutet, Stockholm, Sweden, (within 1 month) history of malaria indicating their rapid diagnostic test 6President’s Malaria Initiative, Center for Disease Control, Dar es Salaam, (RDT) results were potential false positives. Remaining positives were United Republic of Tanzania likely to be imported as they reported recent travel to malarious areas outside Lusaka. Of the index cases reporting no travel history that were Zanzibar is considering malaria elimination. A critical component of this followed-up, there was no indication of ongoing transmission within strategy is better understanding transmission. To investigate malaria their household or among sampled surrounding households. Initial transmission in Zanzibar, we combined molecular tools with passive results indicate that as low-prevalence areas in Zambia progress towards surveillance at all 47 public health facilities in three contiguous districts malaria elimination, it is important to collect patient travel history and of Zanzibar from May 2010 to April 2011. Subjects presenting to these to provide public health messaging encouraging individuals to practice facilities with fever who tested positive for malaria by rapid diagnostic test

www.astmh.org 433 malaria prevention especially when traveling to malaria endemic areas. indoor residual spraying, mass drug administration, IPTi, IPTc and a pre- Furthermore, results indicate AID may be a reasonable, focal intervention erthyrocytic vaccine. This model has been parameterised by an extensive for application in low prevalence areas such as urban Lusaka District. literature review and Bayesian fitting to multiple data sources. The tool is a downloadable user-friendly interface to the model which can be run 1439 on any standard Windows PC (each model scenario running in under a minute). The model includes pre-computed estimates of current parasite VALIDATING LAMP FOR IDENTIFYING TRANSMISSION ‘HOT prevalence (as a marker of transmission intensity), estimates of LLIN SPOTS’ FOR MALARIA ELIMINATION AND ERADICATION use in the past 10 years, a seasonal pattern of transmission determined by rainfall data and expert-maps of malaria vectors (An.gambiae s.s., Nahla B. Gadalla1, Jacklin Mosha2, Sharan Atwal1, Chris An.funestus, An.arabiensis) at the first administrative level. The user Drakeley1, Teun Bousema1, Colin J. Sutherland1, Daniel can modify these inputs based on local knowledge and data: if they do, Chandramohan1, Roly Gosling3 the program will recalculate the natural transmission potential taking 1London School for Hygiene and Tropical Medicine, London, United account of the fact that conditions are changing both from one year to Kingdom, 2National Institute of Medical Research, Mwanza Centre, another and seasonally The user can run scenarios with any combination Mwanza, United Republic of Tanzania, 3Global Health Group, University of of interventions, different coverage levels and staggered timings for their California, San Francisco, CA, United States introduction. Outputs include projected EIR, parasite prevalence and Recent trends in reducing malaria transmission in sub-Saharan Africa and incidence of clinical malaria over time, which are plotted visually and can an interest in the steps to reach pre-elimination levels of malaria control be output for further analysis. Thus users can quickly and easily visualise mean a paradigm shift in surveillance for malaria is needed. Surveillance and compare the possible impacts of various control strategies. Future and monitoring of all infections including asymptomatic infections updates will include the ability to enter data on individual program costs is required to find foci of ongoing transmission and to detect when so that the cost-effectiveness of different scenarios can be compared. reintroduction of malaria has occurred in an area of local elimination. For this purpose it is likely that a more sensitive test is needed than microscopy 1441 or currently available Rapid Diagnostic Tests (RDTs) can deliver. In this study we compare the use of 3 different molecular methods for the purpose of EXPULSION OF THE DENGUE VIRUS GENOME BY A PEPTIDE screening blood samples from a large cross sectional survey in a moderate INHIBITOR to low transmission setting in north western Tanzania. This study was Joshua M. Costin1, Shee-Mei Lok2, Yancey M. Hrobowski3, carried out in Misungwi District, Mwanza, Tanzania, to identify potential Dawne K. Rowe1, Petra Kukkaro2, Heather Holdaway4, Paul P. falciparum transmission hotspots by mapping parasite prevalence within Chipman4, Krystal A. Fontaine5, Michael R. Holbrook6, Robert house holds. Participants were recruited from 4 villages in Kanyelele F. Garry7, Victor Kostyuchenko2, Sharon Isern1, Michael G. namely Mwakalima, Kanyelele, Gambajiga and Budutu. The villages have Rossmann4, Scott F. Michael1 a total of 33 sub-villages with 1,600 households and a population of 1Florida Gulf Coast University, Fort Myers, FL, United States, 2Duke-NUS, about 11,000 people. These villages were characterized for transmission Singapore, Singapore, 3Center for Naval Analyses, Alexandria, VA, United intensity as low transmission with less than 5% parasitaemia and high States, 4Purdue University, West Lafayette, IN, United States, 5University of transmission with more than 10% parasitaemia, using RDTs. A pilot Washington, Seattle, WA, United States, 6National Institute of Allergy and study was conducted of 180 finger prick blood samples collected from Infectious Diseases, Fort Detrick, MD, United States, 7Tulane University, participants on Whatman® filter paper, representing 90 randomly selected New Orleans, LA, United States from each of two villages, one low and one high transmission. Three molecular methods were performed on chelex-extracted DNA samples: A 33 amino acid peptide (DN59) of identical sequence to the highly LAMP using a mitochondrial target sequence, qPCR with SybrGreen conserved amphipathic stem region of the E protein of the dengue 2 virus detection, and nested PCR. Taking the nested PCR as the established gold is able to inhibit DENV entry into multiple cells types with independent standard, the sensitivity and specificity of LAMP were 85.7% and 91.5%, entry pathways and at similar low micromolar concentrations. DN59 respectively while the qPCR gave a sensitivity and specificity of 70% and inhibits entry of all representative flaviviruses tested and is non-toxic 90%, respectively. In this context of asymptomatic individuals LAMP to cells. Cryoelectron microscopic images of cell-free, peptide-treated has performed better than qPCR in terms of sensitivity. The difference dengue virions suggest that the virions are empty. Three-dimensional in detecting parasite prevalence between LAMP and qPCR was highly reconstructions of these images reveal hollow particles with holes at the significant p=0.0000 (OR 29.7; 95% CI [9.07 - 123.11].These results five-fold vertices. Peptide treatment of dengue virions in the absence coupled with the rapidity of obtaining results (<1hour) and the potential of cellular receptors renders their genomes sensitive to RNase digestion ability to perform LAMP in field conditions by direct boiling of blood spots over a wide range of input virus. Western blot analysis suggests that the renders this technique a useful tool for identifying hot spots of parasite capsid is still associated with treated virions. Thus, a peptide mimicking transmission for malaria elimination. a conserved sequence in the membrane-associated stem region of the flavivirus E protein induces expulsion of the viral RNA genome resulting in 1440 the inhibition of infection. MALARIA TOOLS: A USER-FRIENDLY SOFTWARE PACKAGE FOR EXPLORING THE IMPACT OF COMBINATIONS OF INTERVENTIONS IN AFRICAN COUNTRIES Jamie T. Griffin, Danail Stoyanov, Neil M. Ferguson, Azra C. Ghani Imperial College, London, United Kingdom In the current era of intensified efforts to control or eliminate malaria, individual countries need to prioritise which combinations of interventions they will introduce to reduce transmission and to set realistic expectations about their likely impact. We describe a tool developed for African countries to help guide such decision-making. Underlying the tool is an individual-based dynamic malaria transmission model incorporating current interventions, namely long-lasting insecticide treated nets (LLINs), www.astmh.org 434 1442 effective (net savings per DALY averted) for very conservative scenarios, even if vaccination does not reduce the need for vector control. Paediatric GLUCOSIDASE INHIBITOR AS BROAD-SPECTRUM ANTIVIRAL vaccination is preferred to mass vaccination for prices per dose greater AGAINST MULTIPLE HEMORRHAGIC FEVER VIRUSES than $187 and below $365. Mass vaccination, however, presents greater potential of avoided costs and will be preferred for vaccine prices below 1 1 1 2 Jinhong Chang , Lijuan Wang , Tina Gill , Wouter Schul , Travis $187 per dose. If the price of the vaccine is above $45 per dose and 3 3 4 4 4 K. Warren , Sina Bavari , Wenquan Yu , Hong Ye , Yanming Du , below $187, a serology test to administer the vaccine only to non-exposed 4 4 1 Xiaodong Xu , Andy Cuconati , Ju-Tao Guo and Timothy M. individuals should complement the mass vaccination program. Block1, 4 1Drexel University College of Medicine, Doylestown, PA, United States; 1444 2Novartis Institute for Tropical Diseases, Singapore, Singapore; 3U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD, United DIFFERENTIAL EFFECT IN MONKEYS AND MAN OF States; 4Institute for Hepatitis and Virus Research, Hepatitis B Foundation, PARTIAL IMMUNITY TO THE CYD DENGUE VACCINE, AND Doylestown, PA, United States IMPLICATIONS FOR THE SAFETY OF TETRAVALENT CYD Viral Hemorrhagic fever (VHF) designates a group of diseases, caused by VACCINATION enveloped, single-stranded RNA viruses from four different virus families: Bruno Guy1, Gustavo Dayan2, Anke Harenberg1, Nathalie Arenaviridae, Bunyaviridae, Filoviridae and Flaviviridae. Because many VHF Mantel1, Veronique Barban1, Jean Lang1 initially do not present with distinguishing symptoms and are difficult to 1sanofi pasteur, Marcy l’Etoile, France,2 sanofi pasteur, Swiftwater, PA, clinically diagnose at early stages, it is important to develop a drug that United States is universally active against all or most of these agents. Although viruses causing VHF differ in certain features, from the virology point of view, The sanofi pasteur tetravalent dengue vaccine (TDV) candidate is they all have enveloped virions, with viral glycoprotein(s) as envelope. The composed of four recombinant, live, attenuated viruses (CYD-1-4). This host ER α-glucosidases are considered to be essential for the maturation, vaccine, given in a 3-dose, 0-6-12-month regimen, is currently being secretion, and function of viral envelope glycoproteins. In this study, we investigated in clinical phase 3 trials. In a monkey model we previously provided genetic evidence that both α-glucosidase I and II are essential observed that pre-existing immunity conferred by bivalent CYD vaccination host factors for dengue virus and tacaribe virus. Consistent with this enhances immune responses to subsequent bivalent CYD vaccination notion, we have demonstrated that, imino sugar, the known inhibitor performed 2 months later against the remaining two serotypes, without of glucosidases, inhibited multiple hemorrhagic fever viruses from four increasing viremia, an indirect indicator of safety. We explored such a families (Junin, Dengue, Rift valley fever and Ebola), in tissue culture. complementary, bivalent vaccination regimen as part of a phase 2 clinical Furthermore, treatment with one of our lead imino sugar significantly trial (Clinicaltrials.gov: NCT00740155): flavivirus-naïve adult volunteers protected animal death in two mouse models with lethal dengue virus were vaccinated with a bivalent mixture of CYD-1 and -3, and 3.5 months infection. Currently, our extensive SAR study has led to the discovery later with a complementary bivalent mixture of CYD-2 and CYD-4. In of several more potent imino sugar derivatives in tissue culture. The in contrast to findings in monkeys, immunity conferred by CYD-1 and vivo efficacy test of these compounds in multiple VHF animal models is -3 vaccination neutralized the subsequent CYD-2 and CYD-4 vaccine underway. viruses (absent/lower viremia compared to that observed after tetravalent CYD vaccination) and dampened the serotype 2 and 4 specific immune 1443 responses. This inhibition was less marked at the serotype-specific cellular immunity level. From these findings we draw two important conclusions. ECONOMIC IMPACT OF DENGUE ILLNESS AND THE COST- Firstly, as the first bivalent vaccination provided ‘protection’ against the EFFECTIVENESS OF FUTURE VACCINATION PROGRAMS IN second (rather than enhancing viremia and immunogenicity) we propose SINGAPORE that in the case of natural infection before the completion of the 3-dose

1 2 1 2 TDV vaccination regimen, the partial immunity elicited by the first one L. Roman Carrasco , Linda K. Lee , Vernon J. Lee , Linn T. Tun , or two doses will not enhance disease, even if antibody responses to Eng Eong Ooi3, Alex R. Cook1, David Lye2, Lee Ching Ng4, Leo Yee 2 the infecting serotype are low; furthermore, these findings suggest that Sin heterologous immunity induced by CYD vaccination may be broadly cross- 1National University of Singapore, Singapore, Singapore, 2Tan Tock Seng protective between immunizations. Secondly, the interval of time during Hospital, Communicable Disease Centre, Singapore, Singapore, 3Duke-NUS which cross-neutralization occurs is longer in humans than in monkeys, Graduate Medical School, Singapore, Singapore, 4National Environment further highlighting the importance of interspecies differences in terms of Agency, Singapore, Singapore immune responses and immunization regimens. Dengue fever and dengue hemorrhagic fever cause 50-100 million cases worldwide and threaten 2.5 billion people in the tropical and subtropical 1445 regions. Little is known about the disease burden and economic impact SAFETY EVALUATION AND IMMUNOGENICITY OF FIVE of dengue in resourced countries and the cost-effectiveness of potential dengue vaccines. We estimate the direct and indirect costs of dengue TETRAVALENT ADMIXTURES OF THE NIH LIVE ATTENUATED from hospitalized and ambulatory cases in Singapore. We consider inter DENGUE VACCINE CANDIDATES alia the impacts of dengue on the economy using the human-capital and Anna P. Durbin1, Beth D. Kirkpatrick2, Kristen K. Pierce2, Janet the friction cost methods. Disease burden was estimated using disability- Lindow2, Dan Elwood1, Kimberli Wanionek1, Andrew Andrada1, adjusted life years (DALYs) and the cost-effectiveness of a potential vaccine Cathy Larsson2, Marya Carmoli2, Stephen S. Whitehead3 program was evaluated. The average economic impact of dengue illness in 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United Singapore from 2000 to 2009 ranged between US 2010 $0.95 billion and States, 2University of Vermont College of Medicine, Burlington, VT, United 1.25 billion, with 59-63% corresponding to control costs. We estimated States, 3National Institute of Allergy and Infectious Diseases, National an annual average disease burden of 16-27 DALYs per 100000 habitants, Institutes of Health, Bethesda, MD, United States making it comparable to diseases such as meningitis or tuberculosis (22 and 36 DALYs per 100000 habitants, respectively). The rate of Dengue virus (DENV) has become the most important arbovirus worldwide symptomatic dengue cases detected by the national surveillance system with approximately 36 million cases of dengue fever and more than 2 was estimated to be low, decreasing with age (e.g. ratio of infected million cases of severe dengue occurring annually. Because a secondary symptomatic individuals per detected individual of 1.7-3.8 for 0-24 years, DENV infection with a serotype different from that which caused the 12.2-50 for >55 years). Potential vaccines are estimated to be highly cost- www.astmh.org 435 primary infection is a significant risk factor for DHF/DSS, a DENV vaccine 1447 must induce a long-lived immune response to all four DENV serotypes. The goal of the National Institutes of Health (NIH) DENV vaccine program PRECLINICAL AND CLINICAL TESTING OF A RECOMBINANT is to produce a minimally reactogenic, highly immunogenic, genetically SUBUNIT VACCINE FOR DENGUE stable, live attenuated DEN vaccine that is cost-effective and safe for the 1 2 1 community. Over the past 10 years, the NIH has tested 8 monovalent Beth-Ann Coller , Sarah George , Andrew J. Bett , David E. 1 1 1 1 vaccines in 15 Phase I clinical trials to identify DENV-1, DENV-2, DENV- Clements , Michele L. Yelmene , Michele Coia , Susan Manoff , 1 1 3, and DENV-4 candidate vaccine viruses that are safe and maintain Sangeetha L. Sagar , Jan H. ter Meulen the optimal infectivity and immunogenicity profiles for inclusion in a 1Merck and Co., West Point, PA, United States, 2St. Louis University, St tetravalent formulation. Each monovalent candidate was well tolerated Louis, MO, United States by volunteers with no volunteer experiencing a dengue-like illness. Six Dengue viruses are a major cause of morbidity and mortality throughout monovalent DENV candidate vaccines (a DENV-1 candidate, a DENV-2 the tropics and subtropics with an estimated 50-100 million infections candidate, two DENV-3 candidates, and two DENV-4 candidates) were annually. To date no specific vaccine or therapy has been licensed to evaluated in five different tetravalent admixtures in healthy adult flavivirus- combat this important disease. Live attenuated vaccines for dengue naïve subjects to identify those admixtures with the most favorable safety have faced issues with interference between the four viral components. and immunogenicity profiles. Safety, infectivity, and immunogenicity data To overcome this issue Merck and Co. is evaluating a tetravalent of the five admixtures following administration of a single subcutaneous recombinant subunit vaccine to protect individuals against dengue virus- dose will be presented. Up to 90% of subjects had at least a trivalent induced disease. Preclinical studies conducted in mice and non-human antibody response following a single vaccination, with an excellent safety primates have demonstrated the immunogenicity and efficacy of both profile similar to that observed in the monovalent trials. Preliminary data monovalent and tetravalent formulations adjuvanted with alum or from a second dose administered 6 months after the first dose will also ISCOMATRIX™ adjuvant. These studies have shown the capacity of the be presented. Factors contributing to the immunogenicity profiles of the recombinant proteins to induce balanced tetravalent responses without different admixtures will be discussed. evidence of interference. Formal preclinical safety assessment studies have demonstrated the acceptable safety of the antigens in various 1446 formulations in rats and rabbits. A Phase 1 clinical study of monovalent A RECOMBINANT LIVE ATTENUATED TETRAVALENT DENGUE recombinant protein (DEN1-80E) adjuvanted with alum has been conducted in healthy volunteers and the data from this study will be VACCINE INDUCES NEUTRALIZING ANTIBODIES TO ALL FOUR presented. DENGUE VIRUSES IN HEALTHY ADULT VOLUNTEERS Jorge E. Osorio1, Ivan D. Velez2, Liliana Lopez2, Cynthia 1448 Thomson3, Aurelia Haller4, Claire Y. Huang5, Shawn J. Silengo4, Jaclyn C. Scott4, John Arguello4, Steven M. Erb4, Joseph D. MICRODAM CONTRIBUTION TO THE PRODUCTION OF Santangelo3, Dan T. Stinchcomb4 ANOPHELES SPP. IN WESTERN, LOWLAND KENYA 1University of Wisconsin, Madison, WI, United States, 2Universidad de Robert S. McCann1, John E. Gimnig2, John M. Vulule3, Joseph P. Antioquia, Medellin, Colombia, 3Inviragen Singapore Pte Ltd., Singapore, Messina1, Edward D. Walker1 4 5 Singapore, Inviragen, Inc., Fort Collins, CO, United States, Division of 1Michigan State University, East Lansing, MI, United States, 2Centers Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort for Disease Control and Prevention, Atlanta, GA, United States, 3Kenya Collins, CO, United States Medical Research Institute, Kisumu, Kenya The tetravalent live attenuated dengue vaccine (DENVax) is based on the Anopheles funestus is an important vector of human malaria in Africa DEN-2 PDK-53 virus. Recombinant DENVax-1, DENVax-3 and DENVax-4 yet the production of adults of this species from larval habitats is poorly strains were generated in which the prM and E genes of PDK-53 were understood. In western Kenya, this species remains an important malaria substituted with those of DEN- 1, -3 or -4 viruses. This approach retains vector and has been increasing recently in the Asembo Bay region, where the genetic attenuation markers present in PDK-53. A randomized, insecticide-treated bed nets are in widespread use. One hypothesis for placebo-controlled phase 1 clinical trial was performed to evaluate the this resurgence is the increased larval habitat provided for A. funestus safety and immunogenicity of tetravalent DENVax formulations in healthy, by small dams (“microdams”) built in the area. Constructed simply with flavivirus negative adults. The study was completed in Rionegro, Colombia, earthen dikes, the microdams create small reservoirs (mean surface area a high altitude area with no Aedes aegypti and no dengue exposure. ~ 0.2 ha) by impounding stream water and rain run-off from upslope Low or high dose formulations of DENVax were administered at 0 and 3 in the water sheds of this gently rolling landscape. They are important months by either intradermal or subcutaneous injection. The vaccine was sources of water for both humans and livestock in the local communities well-tolerated with mostly mild and transient local or systemic reactions. In and conserve soil. Naturally growing vegetation, water-logged hoof prints addition, DENVax induced significant neutralizing antibody responses to all from livestock, and pools of standing water near the microdams create four dengue viruses after one or two administrations. This study highlights ideal habitats for anopheline larvae. We georeferenced the microdams and the safety and immunogenicity of the tetravalent DENVax formulations; used a combination of larval and adult mosquito collections to quantify the vaccine warrants further evaluation in clinical trials in dengue endemic the contribution of these microdams to the population of malaria vectors areas. in this region. Microdam architecture revealed an up-slope mud plane providing hoof print habitat for A. gambiae and A. arabiensis larvae, as did the muddy edges of the dams; while vegetated zones provided habitat for A. funestus and A. coustani larvae. Collections of adult Anopheles indoors using the pyrethrum knockdown method showed that houses closer to the microdams had more A. funestus than did houses farther away. While the microdams are important sources of water for the communities, they also contribute to the production of malaria vectors in the area, creating a conflict in this rural landscape.

www.astmh.org 436 1449 high nightly use. Such a shift in the feeding of An. arabiensis is of interest since the vector is reported to be highly zoophagic and endophilic. A total ARE TRENDS IN HOUSING AFFECTING VECTORIAL CAPACITY of 2000 samples of the three vector species from the remaining study AND MALARIA TRANSMISSION IN AFRICA? areas are under investigation and results of the study will be presented and discussed. Jo Lines1, John Gimnig2 1London School of Hygiene and Tropical Medicine, London, United 1451 Kingdom, 2Entomology Branch Division of Parasitic Diseases and Malaria Centers for Disease Control and Prevention, Atlanta, GA, United States EVIDENCE FOR NEW MALARIA VECTOR SPECIES IN THE WESTERN KENYAN HIGHLANDS Changes in housing design and structure are believed to have made a significant contribution to the elimination of malaria from the USA and Jennifer Stevenson1, Brandy St. Laurent2, Neil Lobo2, Mary Northern Europe. Could something similar happen in Africa? Housing Cooke1, Samuel Kahindi1, Robin Oriango3, Jonathan Cox1, Chris changes are occurring, and not only in urban areas: even in villages that Drakeley1 a few years ago contained only mud-and-thatch houses, brick walls and 1London School of Hygiene and Tropical Medicine, London, United metal or tile roofs are now becoming more and more common. Numerous Kingdom, 2University of Notre Dame, South Bend, IN, United States, studies have reported an association between such structural features of 3KEMRI/Centers for Disease Control and Prevention, Kisumu, Kenya house design (screening, ceilings, metal roof) and a reduction in mosquito entry and/or the risk of malaria for the human occupants. Here we review In Africa current malaria control interventions such as indoor residual these studies, and discuss the hypothesis that housing improvements may spraying and long-lasting insecticidal bednets rely heavily on targeting cause a reduction not only in house entry by malaria vectors, but also in the endophilic and late biting nature of a few key mosquito vectors. vector survival and longevity, and hence vectorial capacity. Such effects The presence of vectors that do not conform to these behaviours may could have considerable public health value, because they represent limit the effectiveness of such interventions. In a recent entomological a means by which national authorities could plan to reduce malaria study carried out in Kisii, an area prone to malaria epidemics in Western receptivity, as part of a very-long-term elimination strategy. Further studies Kenya, the majority of Anopheles caught were morphologically distinct of such effects are needed in order to give evidence-based advice to from previously described vectors. For 81% of 475 samples caught, householders, local authorities and governments about how best to “build no amplification product was obtained following an An. gambiae out” malaria. complex species diagnostic PCR. The ITS2 region of ribosomal DNA was successfully sequenced for 395 samples, of which 73% (n= 289) 1450 could not be matched to known sequences. The samples were grouped according to similarity of sequences and phylogenetic trees were made. IMPACTS OF INSECTICIDE-TREATED BEDNETS (ITNs) AND The most abundant group of samples could not be matched to known INDOOR RESIDUAL SPRAYING (IRS) ON HOST SELECTION sequences (173 of 395 samples sequenced) and could not be identified PATTERN BY ANOPHELES GAMBIAE S.S., AN. ARABIENSIS to species level using conventional morphological keys. Sequencing of AND AN. FUNESTUS IN WESTERN KENYA the mitochondrial DNA CO1 gene also indicated that the majority had no published sequence, and the resulting phylogenetic tree groupings Bernard O. Abong’o1, Nabie M. Bayoh2, Collins Ouma1, Edward were similar to the rDNA ITS2 trees for the same samples. Of all samples D. Walker3, Maurice Ombok2, John Vulule2, John Gimnig4 sequenced, 5 were found to be sporozoite positive for Plasmodium 1Maseno University, Maseno, Kenya, 2Centre for Global Health Research, falciparum by ELISA, all of which were caught outdoors. These 5 had no KEMRI/Centers for Disease Control and Prevention Collaboration, Kisumu, previously published ITS2 or CO1 region sequence and 3 fell into the most Kenya, 3Michigan State University, East Lansing, MI, United States, 4Centre abundant sequence group. 86% of this group (149/173) were caught for Disease Control and Prevention, Atlanta, GA, United States outdoors and the majority (77%) were caught before 22:30, prior to when people enter their houses. Preliminary morphological identification of Malaria continues to be a global public health priority and control house spray catches and larval collections from 2009 and 2011 in 6 other interventions are undergoing scale-up of historical proportions. Emphasis villages across highland Nyanza Province, also revealed the presence of this has been placed on vector control with insecticides due to its effectiveness, species. These results indicate the presence of a novel malaria vector in the especially when used in nets as insecticide-treated nets (ITNs) and indoor highlands of Nyanza with early, outdoor biting behaviour. The implications residual spraying (IRS). The WHO has approved vector control using of of these findings in relation to malaria control in the area will be discussed. ITNs and IRS as one of the key malaria control strategies. The use of ITNs and implementation of IRS in selected districts in western Kenya is on the rise, however, mosquitoes are still able to feed and continue to transmit 1452 malaria. These interventions may result in marked changes in the vector COLLAPSE OF ANOPHELES DARLINGI POPULATIONS IN population structure and behavior, manifested in alteration of biting time SURINAME AFTER INTRODUCTION OF INSECTICIDE-TREATED and host preference. Knowledge of the biology and behavioral changes of NETS (ITNs); MALARIA DOWN TO NEAR ELIMINATION LEVEL Anopheles mosquitoes is important in enhancing understanding of ways of malarial transmission and can further aid in evaluation and designing of Helene Hiwat appropriate control interventions. A study aimed at determining the actual Ministry of Health Malaria Program, Paramaribo, Suriname blood feeding and host preference by An. gambiae s.s., An. arabiensis and An. funestus in the presence of ITNs, IRS or both interventions in western A longitudinal study of malaria vectors, aiming to study Anopheles darlingi Kenya was conducted in Nyando, Rarieda, Busia and Bungoma districts in population dynamics, man-biting and sporozoite rates, was carried out western Kenya. Molecular techniques involving Polymerase Chain Reaction in three villages in the Interior of Suriname between January 2006 and (PCR), sequencing and a BLAST search in the GeneBank database were April 2010. During 13,392 man hours of human landing collections, a used to test for host blood. In an initial analysis of host blood type in total of 3,180 female mosquitoes were collected of which 33.7 % were 196 mosquitoes from Asembo in Rarieda district, all samples being An. anophelines. An. darlingi and An. nuneztovari accounted for 99.2 % arabiensis, 13.3% had cow blood, 25% human blood, 5% goat blood of the total anophelines collected. The highest mean human biting rate and 0.5 % rat blood. A number of the analyzed mosquitoes failed either (HBR) observed per survey for An. darlingi was 1.43 bites/man/hour PCR or sequencing, however, the percentages of host blood show a shift outdoors and 1.09 bites/man/hour indoors. Individual ELISA assays of 683 in the host selection by An. arabiensis with more selections for human anophelines yielded two An. darlingi females infected with Plasmodium blood than cattle in a region with high ITN coverage (above 75%) and falciparum. The anopheline HBR decreased to zero in all sites after the onset of malaria intervention activities in 2006, which included the mass www.astmh.org 437 distribution of ITNs. Malaria transmission decreased significantly and samples of wild-caught infected mosquitoes and to assess their age. In Suriname reached the Millennium Development Goal for malaria in 2007. our study, we exposed A. gambiae females of a colony from Burkina Faso It is concluded that the combination of ITN introduction and climatic to P. falciparum isolates from naturally infected patients of the same area. events have led to the disappearance of malaria vectors in the study sites An adequate negative control was determined to obtain a corresponding in the interior of the country. pool of non infected mosquitoes fed on the blood of the same donor. Females exposed and non-exposed to the parasites were maintained until 1453 death and dissected immediately after to measure the level of infection. In optimal environment for the mosquitoes, we observed an effect of DIFFERENCES IN ANOPHELES GAMBIAE GENE infection on mosquito survival in interaction with the origin of the blood. REGULATION IN RESPONSE TO INGESTION OF LOCAL AND This interaction could be the result of variation in parasites (more or less GEOGRAPHICALLY DISTANT ISOLATES OF PLASMODIUM virulent genotypes; single vs. multiple infections), human patients, or less FALCIPARUM probably in mosquitoes. In conditions of limited nutritional resources after the blood meal, the infected mosquitoes had significantly shorter lifespan 1 2 Caroline Harris , Isabelle Morlais , Karolina-Anthoula than non-infected mosquitoes. This is the first clear evidence of a negative 3 2 4 Akinosoglou , Louis Clement Gouagna , Parfait Awono-Ambene , effect of P. falciparum sporogony on the survival of its natural vector A. 5 2 3 Roch K. Dabire , Didier Fontenille , George Christophides , Anna gambiae. In malaria transmission, longevity is the most important factor of 5 3 Cohuet , Dina Vlachou vectorial capacity. The results have important implications in understanding 1Liverpool School of Tropical Medicine, Liverpool, United Kingdom, 2Institut evolutionary forces that maintain susceptible and resistance alleles de Recherche pour le Développement, Montpellier, France, 3Imperial to parasite infections in natural mosquito populations. Moreover, the College London, London, United Kingdom, 4IRD-OCEAC, Yaoundé, effect of the parasite on mosquito survival might impact malaria control Cameroon, 5Institut de Recherche en Sciences de la Sante, Bobo Dioulasso, strategies that would aim at increasing mosquito vector refractoriness or Burkina Faso specifically target infected mosquitoes. On ingestion of malaria parasites, Anopheles gambiae is known to regulate a suite of genes and launch an immune response. Most research 1455 on this host-parasite interaction has been done using either model CHARACTERIZATION OF THE VIRAL FITNESS OF NORTH systems or mosquito and parasite strains colonised many years ago. AMERICAN WEST NILE VIRUS ISOLATES BY IN VIVO With little research using natural malaria systems the details of host- parasite interactions in the wild remain poorly understood. This study COMPETITION IN BIRDS AND MOSQUITOES aimed to determine An. gambiae gene regulation in recently colonised Gabriella Worwa1, Sarah S. Wheeler1, Christy C. Andrade1, Payal strains after ingestion of wild Plasmodium falciparum isolates. Mosquito D. Maharaj2, Aaron C. Brault2, William K. Reisen1 strains from Cameroon and Burkina Faso in Central and West Africa were 1Center for Vectorborne Diseases (CVEC), Davis, CA, United States, experimentally infected in parallel with their local and distant parasite 2Division of Vectorborne Diseases, Centers for Disease Control and isolates for comparison. Whole genome microarrays were completed Prevention, Fort Collins, CO, United States on the mosquitoes 24 hours after infected blood meal ingestion. Eight biological replicates were completed with both the local and distant West Nile virus (WNV) is a mosquito-transmitted flavivirus of global public, parasite. Gene regulation was found to be highly diverse across all veterinary and wildlife disease importance. Genetic changes in the genome infections, suggesting that the natural genetic polymorphism present in of the invading WNV strain (NY99) have given rise to new seemingly the samples used here but absent from most previous studies has a great better mosquito-adapted genotypes (WN02) that may represent a key effect on host-parasite interactions, leading to specific gene regulation. evolutionary mechanism enabling the persistence of WNV. Although some Previous work comparing these local and distant infections showed that newly emerged WNV genotypes have been genetically characterized, no local mosquito-parasite combinations produce lower infection intensities. in-depth study has compared the phenotypic performance and in vivo The current study highlights a small number of mosquito genes potentially competition fitness of these genotypes in biologically relevant avian and involved in these interactions that are consistently regulated in local mosquito hosts. Our study addresses the viral fitness of several North infections but not in distant ones. It also shows that the most important American WNV strains utilizing an in vivo competition fitness assay factor in determining gene regulation patterns is the parasite population in House finches and Culex tarsalis mosquitoes. The founding 2003 rather than the mosquito strain or local/distant pair. These findings California WNV isolate belonging to the current dominant WNV genotype have great impacts on future malaria control methods via the mosquito, (WN02) was genetically marked by site-directed mutagenesis and used as especially those which aim to interfere with host-parasite interactions. The the reference strain for competition against the original invading strain fact that these interactions may be differentially evolving over space and (NY99) and post invasion California isolates. Starting with a 1:1 virus or time means that any such control method must be vigorously tested ratio for inoculation, the outcome of competition between the two virus before implementation. populations was analyzed based on individual replication data resulting from a novel RT-PCR approach, specifically designed for genetic distinction 1454 and quantification of mixed virus competition samples. Fitness in birds was determined by examining sera obtained throughout the viremia period and PLASMODIUM FALCIPARUM DOES AFFECT SURVIVAL OF ITS from tissues collected post mortem. In mosquitoes, fitness was evaluated NATURAL VECTOR ANOPHELES GAMBIAE based on the ability of each virus to infect, disseminate and be transmitted after extrinsic incubation. Here, we report a novel diagnostic approach for 1 2 3 Ibrahim Sangare , Yannis Michalakis , Roch Dabiré , Anna the quantitative detection of nucleotide polymorphisms, facilitating viral 1 Cohuet competition studies. Furthermore, we present data on the fitness of North 1IRD-IRSS, Bobo Dioulasso, Burkina Faso, 2CNRS, Montpellier, France, 3IRSS- American WNV isolates in their natural hosts and discuss our findings in Centre Muraz, Bobo Dioulasso, Burkina Faso the context of WNV evolution and persistence. The question of the effect of malaria parasites on vector survival has been raised for decades but is still not properly resolved. Several studies addressed the subject but most of them used unnatural Plasmodium- Anopheles combinations and therefore disregarded the coevolution between the parasite and the vector. In the most relevant couple for human malaria, A. gambiae- P. falciparum, the number of studies is very limited and results are contrasted due to difficulties to obtain large www.astmh.org 438 1456 isolate, and evaluated morbidity, mortality and virus persistence. These studies allow a clearer understanding of how genomes that contain large TEMPORAL AND SPATIAL FLUCTUATIONS IN ARBOVIRUS in-frame deletions can influence virus replication and pathogenesis, and MUTANT SWARMS may shed light on mechanisms for WNV persistence in vertebrates, an emerging health concern in particular regions where the virus is enzootic. Alexander T. Ciota, Dylan J. Ehrbar, Graham G. Willsey, Greta V. Jerzak, Jean DeMarco, Laura D. Kramer 1458 Wadsworth Center, New York State Department of Health, Slingerlands, NY, United States GENETIC DIVERSIFICATION AND DYNAMICS OF WEST NILE VIRUS IN A NORTHERN TEMPERATE REGION: CONNECTICUT Arboviruses often exist within hosts as a swarm of closely related minority genotypes. Although the size and composition of these mutant 1999-2008 swarms can have direct phenotypic consequences, the specifics of how Philip M. Armstrong1, Charles R. Vossbrinck1, Theodore G. stochastic and selective pressures shape arboviral swarms is not fully Andreadis1, John F. Anderson1, Kendra N. Pesko2, Ruchi M. defined. Although host-specific differences in mutant swarm breadth have Newman3, Niall J. Lennon3, Bruce W. Birren3, Gregory D. Ebel2, been identified with arboviruses, the influence of genetic bottlenecks Mathew R. Henn3 in mosquitoes during initial virus infection of midgut cells, egress from 1The Connecticut Agricultural Experiment Station, New Haven, CT, United midgut tissue, salivary gland infection and, ultimately, host transmission States, 2University of New Mexico School of Medicine, Albuquerque, remain uncharacterized. In addition to these spatial influences, temporal NM, United States, 3Broad Institute of MIT and Harvard, Cambridge, MA, changes are also important in shaping swarm dynamics both within United States hosts and throughout seasons. In order to better define swarm dynamics, we’ve combined experimental studies with genetic analyses of natural West Nile virus (WNV) has become firmly established in northeastern isolates and evaluated changes to West Nile virus (WNV) and St. Louis U.S., reemerging every summer since its introduction into North America encephalitis virus (SLEV) through time and space. Specifically, studies with in 1999. To determine whether WNV overwinters locally or is reseeded WNV in Cx. pipiens following feeding on artificial swarms have begun to annually, we examined the patterns of viral lineage persistence and define within-host bottlenecks. Results indicate that when infecting with replacement in Connecticut over 10 consecutive transmission seasons by phylogenetic analysis. In addition, we compared the full protein coding equal proportions and high input titers (>8.0 log10 pfu/ml) mutant swarm breadth is maintained spatially throughout the mosquito, yet a significant sequence among WNV isolates to search for evidence of convergent and decline in diversity occurs over time. Results from feeding on different adaptive evolution. Viruses sampled from Connecticut segregated into a variant ratios indicate also that proportions of variants are maintained number of well-supported subclades by year of isolation with few clades in early midgut infection and replication, yet stochastic pressures may persisting ≥2 years. Similar viral strains were dispersed in different locations result in the dominance of rare minority variants (<2.5%). In addition, across the state and divergent strains appeared within a single location analyses of primary mosquito isolates of SLEV from TX and CA indicate during a single transmission season, implying widespread movement and a decline in intrahost diversity over time, possibly reflecting the role of rapid colonization of virus. Numerous amino acid substitutions arose in the seasonal bottlenecks in limiting swarm breath. Studies with WNV isolates population but only one change, V to A at position 159 of the envelope from NY are also providing insight into the genetic consequences of local protein, became permanently fixed. Several instances of parallel evolution maintenance. These data significantly advance our understanding of how were identified in independent lineages, including one amino acid change intra- and interhost dynamics alter viral swarms over time and space and in the NS4A protein that appears to be positively selected. Our results how such fluctuations could impact virus evolution and adaptation. suggest that annual reemergence of WNV is driven by both reintroduction and local-overwintering of virus. Despite ongoing diversification of WNV, 1457 most amino acid variants occurred at low frequencies and were transient in the virus population. NATURALLY OCCURRING WEST NILE VIRUS DELETION MUTANTS: ROLE AS DEFECTIVE INTERFERING PARTICLES 1459 AND INFLUENCE ON PATHOGENESIS IN VIVO CHARACTERIZATION OF WEST NILE VIRUSES ISOLATED 1 1 2 Kendra Pesko , Kelly Fitzpatrick , Ruchi Newman , Niall J. FROM CAPTIVE AMERICAN FLAMINGOES (PHOENICOPTERUS 2 2 1 Lennon , Matthew Henn , Gregory Ebel RUBER) IN MEDELLIN, COLOMBIA 1University of New Mexico, Albuquerque, NM, United States, 2Broad 1 1 1 Institute, Boston, MA, United States Jorge E. Osorio , Karl A. . Ciuoderis , Juan G. Lopera , Darby Murphy1, James LeVasseur1, Diana Piedrahita1, Lina Carrillo2, Due to error-prone replication, RNA viruses such as West Nile virus Martha C. Ocampo3, Erik Hofmeister4 (WNV; Flaviviridae, Flavivirus) exist in individual hosts as heterogeneous 1University of Wisconsin, Madison, WI, United States, 2Universidad populations of related genomes. Genomes that contain deletions are de Antioquia, Medellin, Colombia, 3Zoologico Santa Fe de Antioquia, frequently overlooked components of these populations because they are Medellin, Colombia, 4USGS-National Wildlife Health Center, Madison, WI, inefficiently detected by conventional approaches to sequencing the virus United States genome and they are widely considered to be replication-incompetent, dead-end byproducts of infection. We have identified three WNV isolates Serum samples were collected from captive otherwise healthy wild birds from birds that have as a portion of their population mutants with in the summer of 2008 at the zoological collection in Medellin (Colombia) large (~2.5 kb), in-frame, internal deletions to their structural coding and tested for the presence of flaviviruses. Total RNA was extracted from sequences. To determine whether these mutants function as defective- sera and tested by reverse transcription-polymerase chain reaction (RT- interfering (DI) genomes, we infected Vero cells at a range of multiplicities PCR) using both universal flavivirus and West Nile virus (WNV) specific of infection (MOI) and evaluated virus output and whether the deletions primers. Eight serum and swab pools from groups of 12-14 wild birds were maintained through subsequent passage. At high MOIs, deletion and containing 20 species were evaluated. Interestingly, eighteen samples mutants persist through several passages and reduced production of full of twenty five tested from American Flamingoes (Phoenicopterus ruber) length virus, relative to an infectious clone derived control. Therefore, were positive for WNV. Selected samples were then inoculated onto the mutants appear to act as DI genomes in vitro. To determine whether subconfluent monolayers of Aedes albopictus C3/36 cells and three serially deletion mutants can influence pathogenesis and/or WNV persistence in blind passages were conducted before confirmation of virus presence vivo, we infected C3H and C57 Bl/6 mice and day old chickens with virus by immunofluorescence. Four isolates (524, 739, 928, and 9835) were containing deletion mutant or a full length virus purified from the same further selected for full sequence analysis as well as in vitro and in vivo www.astmh.org 439 phenotypic characterization. All RT-PCR products revealed West Nile 1461 viruses. In addition, sequence analysis showed a total of 15 nucleotide changes resulting in 6 amino acid substitutions in comparison to the WNV WEST NILE VIRUS (WNV) EPIDEMIOLOGY IN NEW YORK New York 1999 strain. Further analysis has confirmed that these viruses STATE: DO HYDROGEOGRAPHY AND CLIMATE VARIABLES are more closely related to Louisiana isolates from 2002. All viruses were INTERACT IN THE OCCURRENCE OF WNV IN HUMAN CASES highly cytopathic on BHK21 cells while no cytopathic effect was observed AND MOSQUITO POOLS? on Vero cells. Viruses were highly pathogenic in embryonated chicken eggs and newborn mice. Surprisingly, these isolates diverged in their Michael G. Walsh pathogenicity in 4 week-old Balb/c mice. The epidemiological implications SUNY Downstate, Brooklyn, NY, United States of this new West Nile virus in Colombia and the potential effect on wild The complex ecology that determines West Nile virus (WNV) epidemiology and domestic animals as well as human populations are currently being is not completely understood. The interaction between climate and investigated. hydrogeography (HG) in WNV occurrence has not been identified. This study examines the distribution of mosquito pools positive for WNV 1460 (WNVm) and human WNV cases (WNVh) across the 62 counties of New URBAN ECO-EPIDEMIOLOGY OF WEST NILE VIRUS IN York State (NYS) during the 2006 WNV season (May through October). ATLANTA, GEORGIA Climate data were obtained from the National Climate Data Center, HG data came from the USGS National Hydrology Dataset, and WNVh Rebecca Levine1, Daniel Mead2, Gabriel Hamer3, Paula Marcet4, and WNVm surveillance data were obtained from the NYS Department David Hedeen5, Meghan Hedeen5, Christopher Showalter6, James of Health. The distributions of total precipitation (PR) during the WNV Ballance7, Juanette Willis8, Uriel Kitron1 season, mean July temperature (TM) adjusted for mean temperature 1Emory University, Atlanta, GA, United States, 2University of Georgia, standard deviation for May through October, HG, and WNVh and WNVm Athens, GA, United States, 3Michigan State University, East Lansing, MI, were mapped in ArcGIS, and specific county clustering was identified United States, 4Centers for Disease Control and Prevention, Atlanta, GA, using the local Moran’s Index (LMI). Poisson regression was used to United States, 5Georgia Department of Transportation, Atlanta, GA, United model the associations between each of the 2 outcomes, WNVh per States, 6Fernbank Science Center, Atlanta, GA, United States, 7Zoo Atlanta, county population and WNVm per square mile, and the climate and HG Atlanta, GA, United States, 8DeKalb County Board of Health, Decatur, GA, variables. WNVh and WNVm were concentrated in the coastal counties United States corresponding to the Atlantic Ocean/Long Island Sound and Lake Ontario Tributaries watersheds in the southeastern and western counties, Since its introduction in 1999, West Nile Virus (WNV) has become the respectively. The LMI indicated a high degree of local WNV clustering. The most important mosquito-borne disease in the USA. WNV activity in the HG parameter, total surface water area (SWA), was an effect-modifier of mosquito vectors and reservoir hosts (birds) is clustered in space and the association between the presence of WNV and the amount of PR per time, with transmission focused in certain urban centers (in the East county. A significant inverse relationship was observed between total PR and Midwest) during the summer. However, not all urban areas with and WNVh (IRR=0.99; p<0.0001) among counties with low SWA (< the intensive enzootic activity see corresponding human cases of disease. In median SWA), whereas high SWA (≥ the median SWA) counties showed Georgia, substantial WNV presence in the vector and host species has no association between PR and WNVh (IRR=1.0; p=0.5). The same effect- not translated into a large number of human cases, reflecting a similar modified associations between PR and WNVm were observed for low pattern seen throughout the Southeast, one that is in sharp contrast to (IRR=0.99; p<0.0001) and high (IRR=1.0; p=0.1) SWA. The associations some urban areas in the Northeast and Midwest. In a study conducted in between TM and WNVh and WNVm were modified in the same way by Atlanta, Georgia’s major urban center, we are addressing the question: in SWA. These findings suggest the possibility that associations between the face of abundant reservoir hosts, disease vectors, and viral presence, precipitation and WNV may depend on the surface water present. why is spillover transmission of WNV (beyond the enzootic) suppressed? We perform comprehensive avian and mosquito sampling in a variety of 1462 urban microhabitats, over multiple seasons, to determine the distribution, density, and prevalence of WNV infection in the host and vector species NITRIC OXIDE MEDIATES EXPERIMENTAL HOOKWORM of Atlanta. We focus on sampling in four habitat types within the urban INFECTION center: mixed-use parks, old-growth forest patches, residential areas, and outdoor animal-holding facilities. Fine-resolution aerial imagery is Amanda L. Berndt1, Stephanie M. Moesh1, Sarah E. McNutt1, used to characterize habitat types, percent tree cover, and height of the Lisa M. Harrison2, Michael Cappello2, Blaise Dondji1 tree canopy. Avian point counts are conducted at each site to estimate 1Laboratory of Parasitology and Immunology, Department of Biological bird species richness and abundance. Using these data, we evaluate the Sciences, Central Washington University, Ellensburg, WA, United States, role of Atlanta’s diverse urban habitats in disease transmission, focusing 2Department of Pediatrics, Yale School of Medicine, New Haven, CT, on differences in percent tree cover and height of the tree canopy in United States constraining WNV transmission in time and space. We also explore the Hookworm infection is a major cause of anemia, malnutrition, and extent to which the diversity of avian host species in Atlanta contributes growth delay in resource poor countries, where more than 500 million to a WNV “dilution effect.” This study targets some of the complex people are infected. Human and animal studies confirm that infection ecological factors governing vector-borne disease transmission in urban with these intestinal nematodes is associated with suppression of the settings, combining ecological, epidemiological, and general public health host immune response. We have previously reported higher levels of nitric approaches. oxide (NO) from supernatants of spleen cells harvested from Ancylostoma ceylanicum infected hamsters compared to uninfected animals. In order to further characterize the role of NO in hookworm pathogenesis and pathology, in vivo experiments involving inhibition of NO secretion using N-Monomethyl-L-Arginine (L-NMMA) were conducted. Ten male golden Syrian hamsters were infected with 100 third stage larvae (L3) of the hookworm A. ceylanicum. Five infected hamsters received a daily intraperitoneal (IP) injection of L-NMMA starting on day 0 post-infection (PI), while five others had a daily IP injection of PBS. Uninfected control animals received a daily IP injection of either L-NMMA or PBS. At day 36

www.astmh.org 440 PI, infected L-NMMA-treated hamsters showed reduced intestinal worm 1464 burdens (4 ± 2) compared to PBS-control treated animals (21 ± 4, p < 0.005). Flow cytometry analysis using splenocytes at day 36 PI revealed a MATERNAL GEOHELMINTH INFECTIONS INCREASE higher proportion of CD4+ T cells in infected L-NMMA-treated hamsters SUSCEPTIBILITY TO INFECTION IN CHILDREN than in control animals (12.5 ± 1.2 % vs. 6.8 ± 0.7 %, p= 0.001); a similar 1 2 2 difference was also observed for surface IgG+ B cells (32.0 ± 1.4% vs. 23.1 Raaj S. Mehta , Alejandro Rodriguez , Martha Chico , Irene 2 2 3 4 ± 4.3 %, p= 0.03). There was no difference between uninfected PBS- Guadalupe , Carlos Sandoval , Edward Mitre , Philip J. Cooper treated controls and L-NMMA-treated hamsters in the proportion of CD4+ 1Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, T cells (27.8 ± 1.8 % vs. 27.7 ± 1.8 %) or surface IgG+ B cells (44.83 ± Quito, Ecuador, 2Laboratorio de Investigaciones FEPIS, Quininde, Ecuador, 1.0 % vs. 46.7 ± 3.1 %). Infected L-NMMA-treated hamsters also had 3Uniformed Services University of the Health Sciences, Bethesda, MD, higher blood hemoglobin levels than infected PBS-treated control animals. United States, 4Liverpool School of Tropical Medicine, Liverpool, United Together, these data demonstrate that NO modulates both hookworm Kingdom infection intensity and anemia in vivo. Experiments are underway to In utero exposure to helminth infections may affect newborn immunity identify the mechanism(s) by which NO mediates hookworm disease and influence susceptibility to infection during childhood. To test the pathogenesis and blunts host cellular immune responses. hypothesis that maternal geohelminth infections increase susceptibility to infection in children, we conducted a nested case-control study in 1463 an area of Ecuador where geohelminths are endemic.1004 children HOOKWORM INFECTION AMONG SCHOOL AGE CHILDREN with infection data from between 7 months and 3 years of age were selected from an ongoing cohort study. Cases were children with Ascaris IN THE KINTAMPO NORTH MUNICIPALITY, GHANA: lumbricoides and/or Trichuris trichiura, controls without. Exposure was NUTRITIONAL RISK FACTORS AND RESPONSE TO SINGLE defined as the presence of maternal infection with A. lumbricoides or DOSE ALBENDAZOLE TREATMENT T. trichiura detected in a stool sample collected in the third trimester of Benjamin Simms1, Debbie Humphries1, Dylan Davey2, Joseph pregnancy. To control for risk of infection, the study was restricted to Otchere3, Josephine Quagraine3, Samuel Newton4, Elyssa Berg2, households with at least one family member infected with A. lumbricoides Lisa M. Harrison2, Daniel Boakye3, Michael Wilson3, Michael and/or with T. trichiura, as determined by stool samples collected after Cappello2 the child’s birth. Children of mothers with geohelminth infections had a significantly greater risk of infection relative to children of uninfected 1Yale School of Public Health, New Haven, CT, United States, 2Yale School mothers (46.3% exposed vs. 30.7% unexposed, adjusted OR 2.62, 95% of Medicine, New Haven, CT, United States, 3Noguchi Memorial Institute CI: 1.90-3.62, p<0.001). This effect was particularly strong in children for Medical Research, Accra, Ghana, 4Kintampo Health Research Center, of co-infected mothers (62.6% exposed vs. 30.7% unexposed, adjusted Kintampo, Ghana OR: 5.57, 95% CI: 3.46-8.98, p<0.001). No significant differences were A cross-sectional study of hookworm infection and household nutrition observed between intensities of infection or between distributions of egg was conducted in the Kintampo North Municipality, Brong Ahafo Region, counts. Our data suggest that maternal geohelminth infections increase Ghana. Children (N=844) between the ages of 6 and 11 years attending susceptibility to infection during early childhood. This may be due to in 16 schools were screened using anthropometry. Study participants were utero immune modulation by maternal geohelminth infections that induce selected from the tails of a normal distribution of stunting (Height for greater neonatal and childhood tolerance. Age Z-score (HAZ) <= -1.80 or HAZ >= -0.10). Hookworm prevalence at baseline was 39% (109/279), while the overall prevalence of anemia 1465 was 62% in the study population. Of children who provided fecal and blood samples (N=248), 35% were co-infected with both hookworm and IMMUNE REGULATION BY Treg SUBSETS DURING HUMAN malaria, 50% were infected with malaria alone, and 3% were infected GEOHELMINTH INFECTION: EFFECT OF ANTI-HELMINTH with hookworm alone. Nearly all (96%) of the hookworm infections TREATMENT were light (<2000 eggs/gram of feces). When controlling for age, gender, 1 2 3 household absolute wealth index, and history of recent deworming, Linda Wammes , Aprilianto E. Wiria , Firdaus Hamid , Kit Yeng Liu1, Erliyani Sartono1, Adrian J. Luty1, Taniawati Supali2, Hermelijn statistically significant risk factors for baseline hookworm infection 1 1 included malaria co-infection (p<0.05), access to health care (p<0.01), H. Smits , Maria Yazdanbakhsh lower weekly consumption of protein-rich food groups (p<0.05), and 1Leiden University Medical center, Leiden, The Netherlands, 2University household geographic location (p<0.05). The degree of stunting based of Indonesia, Jakarta, Indonesia, 3University of Hasanuddin, Makassar, on HAZ did not correlate with hookworm infection status at baseline. Indonesia Hookworm-infected children (N=109) received a single oral dose of Chronic helminth infections induce profound regulatory immune albendazole (400mg) and follow-up fecal analysis revealed an overall egg responses, leading to strong T cell hyporesponsiveness. The reduced reduction rate of 88% in the study population, with a cure rate of 44%. immune reactivity does not only affect parasite antigens, but may also be No single variable collected in this analysis was significantly associated extended to vaccines or to other coinciding pathogens, such as malaria with remaining hookworm positive after receiving a single dose of parasites. Helminths induce regulatory T (Treg) cells, characterized by albendazole. These results confirm prior observations of treatment efficacy high expression of CD25 and FOXP3, which are able to downregulate for single dose albendazole against hookworm in Kintampo, and offer effector T cell responses. The question arises whether Tregs are responsible insight into the potential for current school based deworming programs for the immunosuppressive state during human helminth infection and to impact hookworm disease in areas of moderate prevalence and low might have consequences on bystander responses. To investigate Tregs infection intensity. Future studies are needed to more clearly define in helminth infections, we set up a study in a rural area on Flores island, the role of nutritional parameters, including dietary protein intake, in Indonesia where schoolchildren with or without helminth infections mediating susceptibility to human hookworm infection. were selected. CD4CD25hi T cells were magnetically depleted from PBMC and subsequently, mock- and Treg-depleted PBMC were cultured with BCG, Plasmodium falciparum parasitized RBC (pRBC) or control RBC. Proliferation and cytokine responses were analyzed as function of helminth infection status. Proliferation to BCG and pRBC was reduced in geohelminth-infected compared to uninfected children. This difference was not reflected in changes in Treg frequencies in peripheral blood. However, following CD4CD25hi T cell depletion proliferation in the www.astmh.org 441 helminth-infected group was restored to levels similar to those seen in 1467 helminth negatives. Removal of CD4CD25hi T cells also increased the IFN-γ production in response to BCG and pRBC only in helminth positive SEROPREVALENCE OF ANTIBODIES TO STRONGYLOIDES children. Although numbers of Treg were not increased, Tregs displayed STERCORALIS NIE AS A TOOL TO IDENTIFY COMMUNITIES a higher capacity to suppress proliferation and IFN- γ production to FOR ANTHELMINTIC INTERVENTIONS IN NORTHERN bystander antigens in geohelminth-infected compared to uninfected ARGENTINA children. In an ongoing anti-helminth treatment trial, we are investigating Treg phenotype and function in a larger cohort at Flores island. Treg Ruben O. Cimino1, Silvana Cajal1, Marisa Juarez1, Adriana Di depletion assays were carried out and repeated 1 and 2 years after anti- Paolo1, Norma Acosta1, Carlos Villalpando1, Eugenia Socias2, helminth treatment, enabling us to see differences in helminth-infected, Noelia Floridia1, Julio Nasser1, Thomas Nutman3, Monica Carlos1, -uninfected and -treated individuals. Cytokine and proliferation data from Patrick Lammie4, Alejandro Krolewiecki1 the longitudinal Treg study are still under analysis, but this is expected 1Universidad Nacional de Salta, Salta, Argentina, 2Fundación Mundo to be finalized by September 2011. Treg depletion in human helminth Sano, Buenos Aires, Argentina, 3Laboratory of Parasitic Disease, National infection is a new concept and this helminth-induced effect on the Institutes of Allergy and Infectious Diseases, Bethesda, MD, United States, immune system may have major consequences in vaccine implementation 4Division of Parasitic Diseases and Malaria, Centers for Disease Control and and malaria elimination strategies. Prevention, Atlanta, GA, United States Infections with Strongyloides stercoralis (Ss) are cosmopolitan and 1466 mostly subclinical; new diagnostic approaches are needed to define the INCREASED LOCAL REGULATORY T CELLS AND DIMINISHED prevalence and distribution of infections and to monitor intervention IgE EXPRESSION IN DUODENAL MUCOSA OF SS/HTLV-1 activities. The identification of Ss-specific recombinant antigens increases COINFECTED PATIENTS the availability of serologic assays for these efforts. We evaluated the performance of Ss-NIE-1 by ELISA previously shown to be sensitive and Luis Malpica1, Cristina Leguia1, Natalia Freundt1, Nicolas Barros1, specific in the setting of a mass deworming program in a rural community E. Antonio Antunez de Mayolo2, A. Clinton White3, Martin of northwestern Argentina (S 22°53´60´´; W64°20´06´´) with a total Montes1 population of 618 people. Prior to drug administration for community- 1Instituto de Medicina Tropical ‘Alexander von Humboldt’ Universidad wide treatment, a subset of individuals was randomly selected for soil Peruana Cayetano Heredia, Lima, Peru, 2Departamento de Patologia transmitted helminth (STH) assessment at baseline evaluation. Of the Universidad Peruana Cayetano Heredia, Lima, Peru, 3University of Texas 80 persons (mean±SD age: 28.5± 20) who participated in the Ss study, Medical Branch, Galveston, TX, United States 48 (mean±SD age: 30.5± 21) were also evaluated by stool analysis that included 4 techniques (concentration-sedimentation, agar plate, Harada- Strongyloides stercoralis (SS) is an intestinal nematode unique in its Mori and Baermann with bone charcoal culture). Serum samples were ability to replicate in the human host, permitting ongoing cycles of analyzed with the Ss-NIE-1 ELISA. The results demonstrated Ss larvae autoinfection, persisting for decades within host. Although usually in 16% (8/48) stool samples; in contrast, 31% (25/80) were positive by asymptomatic, overwhelming infections can occur in SS and HTLV-1 co- serology (mean±SD age: 32.8.5± 20.8). Thus, there was a significant infected individuals (SS/HTLV-1). Regulatory T cells (Treg) are able to blunt difference between the stool and more sensitive serum analysis (p<0.05 specific Th2 response necessary to control the parasite. We previously by Fisher’s exact test). Although three stool positive individuals (mean±SD reported that peripheral blood Treg are increased in SS/HTLV-1 and age: 31.3± 22.4) were Ss-NIE seronegative, the known improved sensitivity correlate with low Th2 responses. We hypothesized that local Tregs are of the SS-NIE ELISA for Ss infection with its higher throughput and ease of increased in duodenal mucosa of SS/HTLV-1 patients. Paraffin embedded performance provides an epidemiologic tool to identify Ss-endemic regions duodenal biopsies were obtained from 10 HTLV-1/SS subjects and 3 of the world in need of anthelmintic therapy. Moreover, the Ss-NIE ELISA control samples from non-parasitic chronic duodenitis (NPCD) subjects. confirmed the high prevalence of this STH in the study area, which aids in Immunohistochemistry was performed with CD3, IgE and FoxP3 human region-wide strategies for community intervention for this important STH. mAbs. The numbers of cells were counted using a conventional light microscope. 400x magnification images were taken and the area was 1468 measured using the ImagePro Plus software. The numbers of CD3+, FOXP3+ and IgE positive cells per 0.35 mm2 were assessed. Patients STAGE-SPECIFIC GENE EXPRESSION BY STRONGYLOIDES with HTLV-1/SS had higher T lymphocyte counts in areas non-adjacent STERCORALIS FOLLOWING HOST INVASION: A to the parasite (NAP) compared to areas adjacent to the parasite (AP) MICROARRAY-BASED ANALYSIS (NAP [P50]: 24.5 IQR: 20.3-36.3, AP [P50]: 6.5 IQR: 2.8-12.3, p=0.0003 Mann-Whitney). The number of IgE expressing cells was higher in areas Roshan Ramanathan1, David Abraham2, Thomas J. Nolan3, non-adjacent to the parasite (NAP [P50]: 35 IQR: 17.5-57.5, AP [P50]: 3 James B. Lok3, Sudhir Varma1, Thomas B. Nutman1 IQR: 0.5-6.0, p=0.001 Mann-Whitney). Patients with HTLV-1/SS show an 1National Institutes of Health, Bethesda, MD, United States, 2Thomas increased counts of Tregs cells (FoxP3+ expressing cells) when compared Jefferson University, Philadelphia, PA, United States, 3University of with patients with non-parasitic chronic duodenitis (SS/HTLV-1 [P50]: 3.3 Pennsylvania, Philadelphia, PA, United States IQR: 0.6-8.4, NPCD [P50]: 0.0 range 0-1.2, p=0.099 Mann-Whitney). In The key molecular factors that enable Strongyloides stercoralis (Ss) larvae conclusion, our data shows an increased Treg cell count in the duodenum to initiate infection have not been elucidated to date, and are critical for of SS/HTLV-1 patients. In addition, T lymphocytes and IgE expressing cells developing new insights into the biology of this nematode and identifying were diminished in parasite adjacent areas. Altogether, this study suggests novel drug and vaccine targets. We describe here a microarray analysis of an important role for Tregs in down-regulating local parasite effector gene expression by infective third stage larvae before (L3i) and 72 hours responses during SS/HTLV-1 co-infection. after (L3+) host invasion. Differentially labeled cDNA obtained from RNA extracted from these larvae was hybridized to a Ss DNA microarray. Genes that were more highly expressed in either stage were identified based on a cutoff of 2 fold increased gene expression and adjusted p-value < 0.01 [false discovery rate of 1%]. Using this method, 96 differentially expressed genes were identified. Expression of genes putatively encoding extracellular matrix proteins, such as collagen, was notably increased in L3i compared to L3+ larvae (p = 0.0003). By contrast, an enzyme homologous

www.astmh.org 442 to Brugia malayi chitinase (BLAST E value 4E - 009) was significantly 1470 expressed in the L3+ stage (p = 0.001). Following invasion, genes putatively encoding components of the ubiquitin proteasome pathway DO LIMES ACCELERATE SOLAR DISINFECTION OF WATER were highly expressed. L3+ larvae additionally demonstrated a significant (SODIS)? A LAB/FIELD STUDY COMPARING EFFICACY WITH increase in the number of differentially expressed genes encoding MOUSE NOROVIRUS, E. COLI AND MS2 BACTERIOPHAGE enzymes with putative catalytic activity (such as hydrolases, ligases and 1 2 transferases ; p = 0.02). Stage-specific differences in metabolism were Alexander S. Harding , Kellogg J. Schwab found. A significantly higher number of differentially expressed L3i genes 1Johns Hopkins University School of Medicine, Baltimore, MD, United were putatively involved in energy metabolism (p = 0.042), while twice States, 2Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, as many differentially expressed L3+ genes were putatively involved in United States amino acid, carbohydrate and lipid metabolism. These data indicate that Two million people in the world die annually of diarrheal disease, mostly Ss larvae downregulate expression of extracellular matrix and energy among the 850 million people who do not have access to sources of metabolism genes, and increase expression of genes encoding catalytic improved drinking water. Household water treatment (HWT) can prevent enzymes following host invasion, and provide important clues regarding waterborne illness. Solar disinfection of water (SODIS) is a HWT method differentiation to the L3+ stage and establishment of infection following that involves exposing water bottles to sunlight for 6 hours. A potential host invasion. mechanism to increase use of SODIS would be to reduce the treatment time. Psoralens and acids both interact synergistically with ultraviolet 1469 radiation to accelerate inactivation of microbes. This study replicated THE IMPACT OF A COMMUNICATION CAMPAIGN ON USE OF field-based SODIS conditions by using 2L bottles and dechlorinated tap ORAL REHYDRATION SOLUTION BY MOTHERS OF CHILDREN water in solar experiments (n=8). SODIS + lime juice bottles contained approximately one-half lime in 2L water. 5-Methoxypsoralen was used as UNDER FIVE IN MALAWI a control for naturally occurring psoralens. Treatment efficacy (n=3) was Abel Irena1, Cecilia Kwak1, Justin Buszin1, Megan Littrell2 evaluated for E. coli, MS2 bacteriophage, and mouse norovirus. E. coli 1Population Services International, Washington, DC, United States, was ablated >6.08log by SODIS + lime slurry, >5.98log by SODIS + lemon 2Population Services International, Nairobi, Kenya juice, and 5.59log by SODIS + lime juice in 30-minute solar exposures, compared with 1.50log for SODIS alone. MS2 was inactivated >5.77log The use of oral rehydration solution (ORS) has stagnated at 40% in most by SODIS + lime slurry, 3.10log by SODIS + lime juice, and 2.73log by countries since 1995. Population Services International (PSI) began a social SODIS alone in the sunniest of three 2.5-hour solar exposures. In contrast, marketing program in 2005 in Malawi to improve access to and use of mouse norovirus, a surrogate for human norovirus, was highly resistant Thanzi ORS for children under five. PSI uses targeted communications to to all forms of SODIS. Using simulated sunlight in laboratory ultraviolet bring about behavior change in early recognition and treatment seeking experiments, SODIS + lime slurry and SODIS + lime juice inactivated mouse during diarrheal illness. Repeated household surveys were used to evaluate norovirus 2.03log and 1.84logs, respectively, after 6 hours lamp exposure, the impact of the communications campaign. Nationally representative while conventional SODIS achieved only a 0.30log reduction. In field-based household surveys were conducted in 2005 and 2008, and are conditions, all treatment bottles (n=3) showed <2log reductions in mouse complemented by 2010 preliminary DHS results. Guided by a conceptual norovirus after a 6-hour solar exposure. A pH of <4 facilitated inactivation framework for understanding caregiver behavior, multi-item scales of E. coli, while psoralens were more effective than pH in inactivating were developed to measure opportunity, motivation and ability factors. viruses. SODIS + citrus dramatically reduced E. coli levels in just thirty Factor and reliability analysis guided scale development. At baseline, minutes, a treatment time on par with boiling. Furthermore, familiarity multivariate logistic regression was used to test for adjusted associations with citrus juice may make this method appealing to potential users. between hypothesized behavioral determinants and diarrhea treatment. Mouse norovirus, a human norovirus surrogate, was highly resistant to all At follow-up, significant changes in behavior and behavioral determinants SODIS treatments. The efficacy of SODIS against human norovirus should were examined. At baseline (2005), perceived availability of ORS and be investigated further. knowledge of causes of diarrhea were found to be key determinants that differentiated users of ORS from non-users. Baseline results guided 1471 development of a communication campaign involving mass media and interpersonal communication. In 2008, perceived availability, knowledge, AN ACCEPTABILITY AND FEASIBILITY PILOT OF HOUSEHOLD and self efficacy improved significantly. Similarly, attitudes toward the LEVEL WATER TREATMENT IN URBAN BANGLADESH Thanzi brand improved. In 2005, 58.1% of children with diarrhea in 1 2 3 the past two weeks were reported to have taken ORS. This increased Shaila Arman , Elli Leontsini , Pavani Kalluri Ram , Leanne 1 1 1 1 to 64.3% in 2008 and to 69% in 2010. Thanzi ORS made up 75% and Unicomb , Fazlul Kader Chowdhury , Md. Al Mamun , Smriti Roy , 1 4 1 83.7% of the ORS utilized in 2005 and 2008 respectively. Although a Subas Chandra Biswas , Peter Winch , Stephen P. Luby number of factors could have resulted in the increased use of ORS to treat 1International Center for Diarrheal Diseases Research, Bangladesh, Dhaka, diarrhea in under-five children in Malawi, Thanzi’s proportionally large Bangladesh, 2Johns Hopkins Bloomberg School of Public Health, Baltimore, ORS market share, marketed by PSI using a social and behavior change MD, United States, 3University at Buffalo, State University of New York, approach, is indicative of the role that evidence-based social marketing Buffalo, NY, United States, 4Johns Hopkins Bloomberg School of Public interventions can play in improving diarrhea treatment. Health, Baltimore, MD, United States Household water treatment prevents diarrhea. However, most household water treatment interventions fail to be used consistently. We conducted a small trial in urban Bangladesh to assess the acceptability of various approaches for point of use water treatment. In May and June 2010, we used the trial of improved practices methodology in 15 low income compounds with a total of 88 individual households. Of 3 available water treatment options, we introduced and provided 1 option in each compound for a free 30 day trial. These options included liquid sodium hypochlorite from a dispenser for communal use (n=58), liquid sodium hypochlorite from a dropper bottle for household use (n=20), and a double chamber ceramic filter for household use (n=10). For the 78 households that received chlorine-based options, we provided www.astmh.org 443 standardized water containers to 51households. In each of 5 follow- across all wealth index quintiles. Our observations indicate that shared up visits we conducted either semi structured interviews, household sanitation facilities can increase the risk of diarrhea, regardless of the type observations, and/or group discussions. The double chamber ceramic filter of facility, and supports their classification as “unimproved”. Increasing had the most (10/10), and the chlorine dispenser had the least (10/58) access to private sanitation facilities may reduce diarrhea incidence among sustained self reported use. Most of the participants were able to follow young children. the operating instructions as observed during follow up visits. However, 25/27 participants without the standardized water container compared 1473 with only 5/51 participants with the standardized container had difficulty achieving the correct concentration of chlorine. Frequently mentioned ACCESS TO WATERLESS HAND SANITIZER IMPROVES HAND benefits of using the various technologies included ease of use, the clarity CLEANING BEHAVIOR AFTER TOILET USE AT PRIMARY of treated water and being less time consuming than boiling water. SCHOOLS IN KIBERA, KENYA Barriers in using the technologies included the strong smell of chlorine, 1 1 1 lack of a standardized storage container with the chlorination options, Amy J. Pickering , Jennifer Davis , Annalise Blum , Jenna 1 2 2 3 and the leaking chlorine dispenser’s valve. The double chamber water Scalmanini , Beryl Oyier , George Okoth , Pavani K. Ram filter was most acceptable because the filtered water did not have any 1Stanford University, Stanford, CA, United States, 2KEMRI, Nairobi, Kenya, odor, whereas, water chlorination was less acceptable due to the strong 3University at Buffalo, SUNY, Buffalo, NY, United States smell of chlorine. Odor minimizing techniques, including better models of Hundreds of millions of school days are estimated to be lost each year dispenser that do not leak and standardized containers should be tested. due to diarrheal illness. Handwashing is an established effective strategy Prospective users should be reassured that the odor is inconsequential to to reduce diarrheal and respiratory illnesses. However, promoting their health and well-being. In addition, the reported benefits, such as handwashing is challenging in settings with limited water access. This clarity of treated water compared to supply water and the short treatment study investigated the impact of providing waterless hand sanitizer on time required can be used to promote sustainable water treatment student hand cleaning behavior and health in six primary schools within behavior change interventions. Kibera, Kenya. Two schools received a waterless hand sanitizer (HS) intervention, two received a handwashing with soap (HW) intervention, 1472 and two received no intervention (controls). Hand cleaning behavior was monitored for 8 weeks through structured observation and by surveillance THE RISK OF MODERATE AND SEVERE DIARRHEA IN cameras placed at school toilets. Students were interviewed weekly CHILDREN LESS THAN FIVE YEARS OLD IS INCREASED to monitor diarrheal and respiratory illness. The average rate of hand AMONG FAMILIES WHO SHARE A SANITATION FACILITY cleaning after toileting was 82% of 2589 events observed in HS schools Kelly K. Baker1, Ciara E. O’Rielly2, Eric D. Mintz2, Tamer Farag1, (OR=8.1, 95% CI=2.8-23.0), 38% of 3607 events observed in HW schools Dilruba Nasrin1, Sandra Panchalingham1, William Blackwelder1, (OR=1.1, 95% CI=0.2-7.5), and 37% of 3031 events observed in control Yukun Wu1, James P. Nataro1, Karen L. Kotloff1, Pedro Alonso3, schools. At HW schools, soap was used for 97% of handwashing events, Robert F. Breiman4, Dipika Sur5, A.S.G. Faruque6, Anita Zaidi7, while at control schools, soap was used for only 6% of events. Water was Debasish Saha8, Samba O. Sow9, Myron M. Levine1 unavailable in 39% and 30% of observations at HW and control schools, respectively. When water was available, handwashing rates were 62% 1Center for Vaccine Development, Baltimore, MD, United States, 2Centers at HW schools and 53% control schools. Students at HS schools were for Disease Control, Atlanta, GA, United States, 3Manhiça Health 26% less likely to report diarrhea (p=0.005), 20% less likely to report Research Centre, Maputo, Mozambique, 4Centers for Disease Control cough (p=0.013), and 30% less likely to have a runny nose observed by and Prevention-Kenya, Niarobi, Kenya, 5National Institute of Cholera enumerators (p=0.009) compared to control schools. Students at HW and Enteric Diseases, Kolkata, India, 6International Centre for Diarrhoeal schools were 19% less likely to report diarrhea (p=0.04) and 32% less Disease Research, Dhaka, Bangladesh, 7Aga Khan University, Karachi, likely to have an observed runny nose (p<0.001) compared to control Pakistan, 8Medical Research Council (UK) The Gambia, Banjul, Gambia, schools. Among Kenyan primary schools, provision of waterless hand 9Center for Vaccine Development - Mali, Bamako, Mali sanitizer markedly increased rates of hand cleaning after toilet use, while The WHO classifies sanitation facilities shared by two or more families as provision of soap and water tanks did not. These findings suggest that use “unimproved” for tracking progress towards the Millennium Development of waterless hand sanitizer is a promising option for reducing infectious Goals. However, little is known about the health impact of shared disease in schools with limited water access. sanitation. We examined associations between household sanitation facilities and moderate to severe diarrhea (MSD) among children <5 1474 years old participating in the Global Enteric Multicenter Study (GEMS) in 7 developing country sites in sub-Saharan Africa and South Asia. Cases ASSOCIATIONS WITH HANDWASHING IN THE HOME AND seeking care for MSD (defined as ≥3 loose stools in 24 hrs with one or RESPIRATORY AND DIARRHEAL ILLNESS IN CHILDREN more of the following: sunken eyes, skin tenting, dysentery, IV rehydration, UNDER FIVE YEARS OLD IN RURAL WESTERN KENYA or hospitalization) were enrolled at health facilities. Age-, gender- and 1 2 2 2 community-matched controls were enrolled at home. Sanitation facilities Kelly B. Kamm , Daniel R. Feikin , Godfrey Bigogo , George Aol , 2 3 4 1 were observed during follow-up household visits (between 60-90 days) to Allan Audi , Adam L. Cohen , Melisa Shah , Jihnhee Yu , Robert F. 2 1 all GEMS participants. Among 8,977 cases and 12,278 matched controls, Breiman , Pavani K. Ram 5.7% practiced open defecation; the remainder used pit latrines (55.5%), 1University at Buffalo, State University of New York, Buffalo, NY, United VIP latrines (3.6%), pour flush toilets (32.4%) and flush toilets (1.6%). States, 2Centers for Disease Control and Prevention, Kenya and the Kenya No type of sanitation facility was statistically associated with MSD overall Medical Research Institute/Centers for Disease Control and Prevention or at any site. However, families of case children more commonly used Public Health and Research Collaboration, Kisumu, Kenya, 3Centers for shared sanitation facilities than control families (47.5% vs. 41.2%, mOR Disease Control and Prevention - South Africa, Pretoria, South Africa, = 1.2; 95% CI: 1.1-1.3), overall and in Pakistan (mOR=1.7; 1.4-2.0), Mali 4Emory University School of Medicine, Atlanta, GA, United States (mOR=1.2; 1.1-1.4), India (mOR=1.3; 1.0-1.6), and Kenya (mOR=1.2; Diarrhea and pneumonia are leading causes of death in children 1.0-1.5). The odds of MSD for shared sanitation were increased two-fold worldwide. Handwashing with soap is effective in reducing childhood if feces was present (mOR=2.2; 1.6-3.2) than if was absent (mOR=1.2; diarrhea and pneumonia in resource poor areas in south Asia, but data 1.1-1.3). While access to unshared sanitation facilities was more common are sparse for Sub-Saharan Africa. We observed presence of a designated among higher-income households, shared sanitation facilities were handwashing (HW) location, soap at a designated HW location, or soap in consistently more common among case than among control households the home, and examined the association with longitudinal prevalence of www.astmh.org 444 diarrhea and acute respiratory illness (ARI) in children <5 years old in rural 1476 Asembo, Kenya. Syndromic surveillance for diarrhea and ARI in children <5 years old is conducted biweekly in Asembo. In April 2009, we assessed FILARIAL-EXPANDED HUMAN NATURAL REGULATORY T households in the surveillance area for handwashing indicators. Syndrome CELLS DO NOT DIRECTLY MODULATE ANTIGEN PRESENTING data collected in the four months preceding the survey was used. We used CELL RESPONSES TO MALARIA ANTIGEN IN A FILARIA/ generalized linear regression models to estimate differences in longitudinal MALARIA CO-ENDEMIC REGION prevalence of illness associated with living in households with each HW indicator, adjusted for household wealth, age, sex, and within household Simon Metenou1, Yaya I. Coulibaly2, Housseini Dolo2, Siaka intraclass correlations. The sample included 2547 children in 1745 Konate2, Abdallah A. Diallo2, Lamine Soumaoro2, Michel E. households. Overall longitudinal prevalence of diarrhea and ARI were Coulibaly2, Amy Klion1, Thomas B. Nutman1 2.5 and 12.7 days of illness per 100 child-days, respectively. Longitudinal 1National Institutes of Health, Bethesda, MD, United States, 2Filariasis prevalence was 30.7% lower (95% CI 24.4%, 34.4%) for diarrhea and Unit, Faculty of Medicine, Pharmacy and Dentistry, University of Bamako, 20.5% lower for ARI (95% CI 12.2%, 25.8%) in children in households Bamako, Mali with observed soap compared to children in households with no observed We have previously shown that patent filarial infection is associated with soap. A designated HW location was identified in 3.3% of households, an expansion of regulatory T cells (both adaptive [aTreg] and natural and 1.2% had a designated HW location with observed soap and/or water [nTreg]) and the IL-10 mediated suppression of malaria-induced production present, and neither was associated with a difference in diarrhea or ARI of IL-12, CXCL-9 (MIG), CXCL-10 (IP-10) and IFN- . Because nTregs prevalence after adjustment. The presence of observed soap in the home, γ have been shown to inhibit antigen presenting cell (APC) function by a proxy measure of handwashing behavior, is associated with reduced altering their maturation and modulating the expression of co-stimulatory illness in children in rural western Kenya. A minority of households molecules, we sought to determine whether the filarial-induced expansion had a designated HW location, which, based on previous studies of of nTregs contribute to the diminished APC cytokine production following handwashing behavior, offers substantial opportunity to increase malaria antigen (MalAg) stimulation in a filarial/malarial co-endemic handwashing behavior by providing a visual cue to stimulate handwashing population of Mali. From filarial-infected (Fil+; n=18) or -uninfected (Fil-; at critical times for pathogen transmission. n=19) individuals, purified nTregs (CD4+CD25+CD127low) and effector T (Teff) (CD4+CD25-CD127low) cells were cultured with purified APCs in 1475 the presence or absence of MalAg, and APC-derived cytokines measured. THE ASSOCIATION OF SOAP IN HOUSEHOLD ON CHILD APCs from Fil+ individuals produced significantly lower levels of CXCL-9 MORTALITY FROM RESPIRATORY INFECTIONS (p = 0.0006) and IL-12 (p = 0.02) compared to APC from Fil- subjects. Although MalAg induced CXCL-9 (p = 0.0009), and CXCL-10 (p = 0.0009) Nadira K. Sultana1, Emily S. Gurley1, Mizanur Rahman1, P. K. as well as IFN-γ (p = 0.0011) in the APC:Teff co-culture of all subjects, the Ram2, Stephen P. Luby1 addition of nTreg to APC cultures did not directly alter the APC cytokine 1International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, response to MalAg (p > 0.05) in either group. Surface marker blockade, Bangladesh, 2University at Buffalo, Buffalo, NY, United States previously implicated in the delivery of negative signals by nTregs (e.g GITR, PD1, LAG-3, IL-10R and CTLA-4) to APC failed to alter cytokine Respiratory disease is the leading cause of childhood death globally. Hand production when nTregs were added to APCs. In contrast, blocking PD1 washing with soap prevents respiratory disease morbidity but there is or IL-10R in APC:Teff cocultures markedly augmented the production of no direct evidence of its impact in preventing deaths from respiratory IFN-γ, IL-12p70, CXCL-9 and CXCL-10 (P <0.05 for all comparisons) in Fil+ infection. We explored the contribution of household handwashing subjects. This augmented cytokine production was significantly higher in behavior to child mortality from respiratory diseases in rural Bangladesh. Fil+ subjects compared to Fil- subjects. Our data clearly show that nTregs We obtained data on deaths among children <5 years of age from a large do not directly inhibit APC cytokine production in response to MalAg scale, sanitation, hygiene education and water supply intervention in rural stimulation and suggest that expansion of nTregs is not responsible for the Bangladesh which spanned 16 districts. To assess cause of death, we diminished cytokine production by APCs in filarial infections. conducted verbal autopsies approximately 1 year after child death. Each death among a child <5 years (ICD-9 code 869.3-869.6) was age matched 1477 to three children from the same community who did not have respiratory symptoms at the time of interview. We interviewed family members, HISTAMINE RELEASE DURING LITOMOSIDES SIGMONDONTIS conducted 5 hours of household structured observation including INFECTION ENHANCES ADULT WORM BURDEN observation for the presence of soap in the household. We constructed a socioeconomic index using principal component analysis of parental Ellen C. Mueller1, Marc Huebner2, Paul Morris1, Edward Mitre1 education, household assets, number of rooms, and housing material. 1Uniformed Services University of the Health Sciences, Bethesda, MD, We used logistic regression to adjust for immunization, socio-economic United States, 2Institute for Medical Microbiology, Immunology and status, family size, and number of people sleeping in the same bed as Parasitology, University Hospital, Bonn, Germany child. Compared to 315 age matched healthy children, the 105 cases were Numerous studies have demonstrated that helminth antigens induce more likely to have smaller families (p<0.001) and less educated mothers release of histamine from basophils and mast cells of infected hosts. To (p=0.012). Cases (29%) were more frequently assigned to the poorest date, however, the role histamine plays in the immune response against wealth quintile than controls (18%). In multivariate analysis, having had helminths has not been well characterized. In this study, we evaluated any childhood vaccination [AOR: 0.26, CI: 0.11-0.58] and having any the role of histamine in mice infected with Litomosoides sigmondontis, bar soap in the home [AOR: 0.53, CI: 0.29-0.98] was independently a tissue-invasive filarial infection of rodents that lives for months in protective. Household participants washed hands in only 10 (2%) of 523 immunocompetent Balb/c mice. Extended time-course studies revealed observed respiratory events such as coughing, sneezing, or nose cleaning that histamine in plasma peaked at 8 weeks of infection (mean 30nM or blowing. All 10 washed with only water. Household ownership of at 4 wks, 325nM at 8 wks, and 100nM at 12 wks compared to 20nM soap, a proxy hand washing measure was associated with deaths from in uninfected age-matched controls) whereas expression of histidine respiratory disease. However, handwashing with soap was not practiced decarboxylase mRNA in circulating blood cells increased throughout the after contact with respiratory secretions. Understanding optimal strategies course of infection (fold increase over uninfected age-matched controls = within households to interrupt respiratory pathogen transmission by hand 20 at 8 wks and 35 at 12 wks). Mice vaccinated with irradiated L3 larvae washing would help decrease respiratory disease mortality as well as demonstrated substantial increases in circulating histamine levels 30 morbidity. minutes after challenge infection (mean 400nM vs 20nM in unchallenged,

www.astmh.org 445 p<0.001), but administration of HR1 and HR2 receptor blockers did 1478 not attenuate the protective efficacy of vaccination (82% protection in untreated groups, 82% protection in HR1 treated, 80% protection in HR2 MICROFILARIAE OF BRUGIA MALAYI INDUCE BOTH AN treated). Interestingly, short time course measurements demonstrated ALTERNATIVELY ACTIVATED AND PROINFLAMMATORY that primary infection of unvaccinated mice with L3s also causes PHENOTYPE IN HUMAN MONOCYTES histamine release into the bloodstream 30 minutes following infection (mean 200nm vs 20nM in uninfected, p<0.05), indicating a non-specific Vanessa M. Moore, Lily Mahapatra, Vivornpum Sanprasert, mechanism of histamine release. To evaluate the role histamine may play Thomas B. Nutman, Roshanak Semnani during infection, mice were chronically administered HR1, HR2, and a National Institutes of Health, National Institute of Allergy and Infectious combination of HR1 and HR2 blockers in their drinking water and assessed Diseases, Bethesda, MD, United States for adult worm survival after inoculation with 40 L3 larvae. Surprisingly, Monocyte dysfunction has been proposed as a cause of the reduced at 8 weeks post-infection all groups of mice treated with antihistamine parasite-antigen specific T-cell response seen in patients with chronic antagonists had significantly reduced numbers of adult worms compared filarial infections. Monocytes from these infected individuals internalize to untreated controls (mean number of adult worms = 5 in HR1 group, 12 filarial antigens and express markers associated with alternatively activated in HR2 group, 6 in HR1/HR2 group, and 20 in untreated infected control macrophages (Mφ). To understand the role of filarial antigens in monocyte group, p values of <0.05 when comparing each treatment group against differentiation, human monocytes from healthy volunteers were exposed infected controls). Taken together, these data indicate that histamine, to either live microfilariae (mf) of Brugia malayi or cytokines known to rather than being involved in vaccine-mediated protection, may be produce classically activated (combination of LPS and IFN-γ or MCSF) or induced by filarial parasites for their growth and/or survival in vivo. alternatively activated (IL-4) phenotypes. The cells were then assessed for their expression of markers associated with alternative activation 1477 and their ability to phagocytose. Our data indicates that, similar to IL- HISTAMINE RELEASE DURING LITOMOSIDES SIGMONDONTIS 4, mf significantly (p<0.05) upregulated mRNA expression of CCL15, INFECTION ENHANCES ADULT WORM BURDEN CCL17, CCL18, CCL22, PDL1, and PDL2 but not Arg-1 in monocytes. Secreted products from mf or IL-4 significantly downregulated the Ellen C. Mueller1, Marc Huebner2, Paul Morris1, Edward Mitre1 monocyte mRNA expression of TLR3 and TLR7 resulting in decreased 1Uniformed Services Univeristy, Bethesda, MD, United States, 22Institute for production of IL-6 following TLR ligand stimulation. Unlike IL-4, but similar Medical Microbiology, Immunology and Parasitology, University Hospital, to LPS/IFN-γ, mf significantly (p<0.05) upregulated the production of Bonn, Germany proinflammatory cytokines IL-1β, IL-6, neopterin, soluble ICAM-1, and TNF-α. Furthermore, both live mf and soluble factors from mf enhanced Numerous studies have demonstrated that helminth antigens induce the cell surface expression of ICAM-1 on monocytes. Interestingly, mf release of histamine from basophils and mast cells of infected hosts. To significantly upregulated (4 fold) the mRNA expression of Indoleamine2, date, however, the role histamine plays in the immune response against 3-dioxygenase (IDO) as well as its activity. Functionally both IL-4- and mf- helminths has not been well characterized. In this study, we evaluated exposed monocytes showed a significant decrease in their phagocytic the role of histamine in mice infected with Litomosoides sigmondontis, ability compared to MCSF-cultured cells. Our data suggest that exposure a tissue-invasive filarial infection of rodents that lives for months in of human monocytes to live mf of Brugia malayi induce monocytes to immunocompetent Balb/c mice. Extended time-course studies revealed have characteristics of both alternative and classical (‘proflammatory’) that histamine in plasma peaked at 8 weeks of infection (mean 30nM activation. Further studies directed toward defining, at the single cell at 4 wks, 325nM at 8 wks, and 100nM at 12 wks compared to 20nM level, the multifunctionality of a given monocyte subset induced by filarial in uninfected age-matched controls) whereas expression of histidine worms are ongoing. decarboxylase mRNA in circulating blood cells increased throughout the course of infection (fold increase over uninfected age-matched controls = 1479 20 at 8 wks and 35 at 12 wks). Mice vaccinated with irradiated L3 larvae demonstrated substantial increases in circulating histamine levels 30 ONCHOCERCA VOLVULUS RECOMBINANT ANTIGENS Ov- minutes after challenge infection (mean 400nM vs 20nM in unchallenged, 103 AND Ov-RAL-2 ARE ASSOCIATED WITH PROTECTIVE p<0.001), but administration of HR1 and HR2 receptor blockers did IMMUNITY IN HUMANS AND INDUCE RESISTANCE TO not attenuate the protective efficacy of vaccination (82% protection in INFECTION IN MICE untreated groups, 82% protection in HR1 treated, 80% protection in HR2 treated). Interestingly, short time course measurements demonstrated Sara Lustigman1, Bin Zhan2, Maria Elena Bottazzi2, Thomas R. that primary infection of unvaccinated mice with L3s also causes Klei3, Jessica A. Hess4, David Abraham4 histamine release into the bloodstream 30 minutes following infection 1Lindsley F. Kimball Research Institute, New York Blood Center, New York, (mean 200nm vs 20nM in uninfected, p<0.05), indicating a non-specific NY, United States, 2Department of Microbiology and Tropical Medicine, mechanism of histamine release. To evaluate the role histamine may play The George Washington University, Washington, DC, United States, during infection, mice were chronically administered HR1, HR2, and a 3School of Veterinary Medicine, Louisiana State University, Baton Rouge, combination of HR1 and HR2 blockers in their drinking water and assessed LA, United States, 4Department of Microbiology and Immunology, Thomas for adult worm survival after inoculation with 40 L3 larvae. Surprisingly, Jefferson University, Philadelphia, PA, United States at 8 weeks post-infection all groups of mice treated with antihistamine antagonists had significantly reduced numbers of adult worms compared Onchocerca volvulus (Ov) remains an important cause of blindness and to untreated controls (mean number of adult worms = 5 in HR1 group, 12 chronic disability. While mass drug administration with ivermectin is in HR2 group, 6 in HR1/HR2 group, and 20 in untreated infected control ongoing, evidence is building for the existence of resistance to the drug. group, p values of <0.05 when comparing each treatment group against The development of an anti-larval vaccine would reduce adult worm infected controls). Taken together, these data indicate that histamine, burdens and thereby diminish microfilariae numbers in the skin resulting in rather than being involved in vaccine-mediated protection, may be reduced pathology and the interruption of transmission. Two O. volvulus induced by filarial parasites for their growth and/or survival in vivo. vaccine candidates, Ov-103 and Ov-RAL-2 were chosen for study based in part on their homologues being highly protective in other nematode animal models. The antigens are expressed by the larvae in the basal layer of the cuticle and hypodermis. Ov-103 is also present in the basal lamina, channels connecting the esophagus to the cuticle and multivesicular bodies within the hypodermis. Ov-103 and Ov-RAL-2 are highly antigenic

www.astmh.org 446 in Ov exposed and infected populations with significant correlations 1481 between the IgG1 and/or IgG3 cytophilic antibodies responses and the development of protective immunity. The anti-Ov-103 IgG3 responses in FATTY ACID AND RETINOL-BINDING (FAR) PROTEINS OF the putatively immune individuals (PI) and the infected (INF) were elevated FILARIAL NEMATODES: POTENTIAL VACCINE CANDIDATES and similar while the anti-IgG1 responses were significantly higher in the FOR ONCHOCERCIASIS AND LYMPHATIC FILARIASIS INF. The anti-Ov-RAL-2 IgG3 responses in the INF are significantly increased 1 2 3 with age, while the IgG1 is highly elevated regardless of age. To test the Sridhar Arumugam , Bin Zhan , Malcolm Kennedy , Jessica A. 4 1 1 4 antigens’ potential as vaccine candidates, mice were immunized with the Hess , Danielle T. Ward , Thomas R. Klei , David Abraham , Peter 2 5 antigens in alum. Yeast derived Ov-103 induced significant protection Hotez , Sara Lustigman (31%) against larval Ov, while antigen from E. coli did not. Immunization 1Louisiana State University, Baton Rouge, LA, United States, 2The George with E. coli derived Ov-RAL-2 induced significant protection (44%), Washington University, Washington, DC, United States, 3University of whereas the yeast produced antigen did not. These results suggest that Glasgow, Glasgow, United Kingdom, 4Thomas Jefferson University, the expression system has an effect of the potency on the recombinant Philadelphia, PA, United States, 5Lindsley F. Kimball Research Institute, New vaccine antigens. We conclude that Ov-103 and Ov-RAL-2 are associated York Blood Center, New York, NY, United States with protective immunity in humans and that both antigens induced The FAR proteins of filarial nematodes are helix-rich, fatty acid and significant levels of protective immunity in mice, thus making them retinol-binding (FAR) proteins that appear to be specific to nematodes potential candidates for a prophylactic vaccine against onchocerciasis. and secreted into the surrounding tissues of the host. These proteins may play an important role in scavenging fatty acids and retinoids from 1480 the host and are probably essential for the survival of filarial nematodes PROTECTIVE RESPONSE OF HOST NON-HOMOLOGOUS and hence are considered as potential vaccine/drug targets. In previous CONFORMATIONAL EPITOPES OF WUCHERERIA BANCROFTI studies, immunization with a fragment of the Onchocerca volvulus retinol- binding protein (Ov-RBD-1/Ov-FAR-1) induced significant resistance to THIOREDOXIN (Wb-TRX) challenge infection with third-stage larvae. Ov-FAR-2 was cloned by using Kaliraj Perumal1, Prince Rajaiah Prabhu1, Madhumathi serum from individuals who have developed concomitant immunity and Jeyaprakasam1, Nasser Yousef2, Christian Betzel2, Rao D.N3, Reddy Bm-FAR-2 was identified by bioinformatics. To further the development M.V.R.4 of recombinant vaccines against O. volvulus and lymphatic filariae, we 1Anna University, Chennai, India, 2University of Hamburg, Hamburg, focused on two members of the filarial FAR protein family. Recombinant Germany, 3All India Institute of Medical Sciences, New Delhi, India, FAR-1 and FAR-2 proteins from O. volvulus and Brugia malayi were 4Mahatma Gandhi Institute of Medical Science, Sevagram, India expressed in Escherichia coli and Pichia pastoris and their ligand binding properties were compared using a fluorescence-based assay. The O. Vaccine studies in parasitic diseases have been a long term struggle due volvulus and B. malayi FAR-1 proteins bind to both retinol and DAUDA to the complex life cycles and immune evasion mechanisms. The present and can be displaced from DAUDA by structurally-related oleic acid. study explores the avenues of structural analysis on parasitic antigens to Surprisingly, both filarial FAR-2 proteins bind only to retinol and not to qualitatively improve protective immune response in mammalian hosts DAUDA, which suggests that FAR-2 and FAR-1 are different in structure against nematode parasitic infections. In this regard, Wuchereria bancrofti and potentially also in function. The recombinant FAR proteins were thioredoxin (Wb-TRX) which protects filarial worms from radical-mediated tested for their efficacy as vaccine in the O. volvulus - mouse chamber damage of the host was selected. Accordingly, recombinant Wb-TRX was model and in the B. malayi - jird animal model. Mice immunized with Ov- purified without fusion tag for structural studies. The enzyme activity FAR-1 in alum, expressed in either E. coli or P. pastoris, killed 54% of the was demonstrated using insulin reduction assay. The structure of Wb-TRX O. volvulus challenge infection. Immunization of jirds with recombinant was solved by X-ray crystallography and was utilized for conformational Bm-FAR-1 expressed in E. coli and formulated with alum resulted in 41% epitope analysis. Since Wb-TRX shares sequence homology (~40%) reduction in worm count and immunization with Bm-FAR-1 formulated with mammalian proteins, certain putative host-non-homologous with the Montanide-720 adjuvant resulted in 74% reduction in worm regions from the protein sequence was selected and analysed by in silico count. Further vaccination experiments investigating the FAR-2 molecules immunoinformatic tools. These selected regions were located and analysed in these two animal models are underway. Correlation between the in the protein structure of Wb-TRX. Additionally, these regions were also immune responses in humans to the FAR proteins vs. those in the validated against online conformational epitope databases. Based on protected mice and jirds will be presented. In conclusion, the FAR proteins these analyses, putative Discontinuous Epitope Peptide regions (DEP) were of filarial nematodes are excellent candidates for use in prophylactic selected and synthesized. The activity assay performed for Wb-TRX in the vaccines against infection with O. volvulus and B. malayi. presence of anti-sera raised in mice against these Discontinuous Epitope Peptides (DEP) characteristically reduced activity proposing a mechanism of 1482 enzyme inhibition depriving antioxidant ambience and challenging parasite survival. Further, a linear Peptide Conjugate (PC1) derivative of the DEP FILARIAL LYMPHATIC PATHOLOGY REFLECTS ELEVATED was evaluated for vaccine efficacy in permissive Mastomys coucha model. LEVELS OF CIRCULATING INFLAMMATORY BIOMARKERS OF Interestingly, PC1 showed 75% greater protection compared to 63% LYMPHATIC AND IMMUNE DYSFUNCTION protection of rWb-TRX. Hence, the current study reports the utilization 1 1 1 2 of conformational epitope enrichment through structural validation as R. Anuradha , Jovvian George , Pavan Kumar , V. Kumaraswami , 3 1 a novel strategy for enhancing the efficacy of parasite antigens in host Thomas B. Nutman , Subash Babu protection. 1NIH-ICER, Chennai, India, 2TRC, Chennai, India, 3National Institutes of Health, Bethesda, MD, United States Infection with Wuchereria bancrofti can be associated with development of serious pathology in the form of lymphedema, hydrocele, and elephantiasis in a subset of infected patients. Dysregulated host inflammatory responses, lymphatic dysfunction, endothelial activation and extracellular matrix remodeling play central roles in filarial disease pathogenesis. To identify factors contributing to pathogenesis of disease in lymphatic filariasis, we examined the role of microbial translocation markers (LPS, LBP, EndoCAb and sCD14); acute phase proteins [α-2

www.astmh.org 447

Macroglobulin (α-2 m), Haptoglobulin, C-reactive proteins (CRP) and development. This is a significant advancement in our understanding of Serum Amyloid protein-A (SAA)]; angiogenic factors (VEGF - A, C, D, R1, the temporal and pathophysiological events that may contribute to FGR R2 and R3 and Angiopoietin -1 and 2); pro- and/or anti-fibrotic factors due to Pf malaria in pregnancy. (MMP - 1, 7, 8 and 9 and TIMP - 1, 2, 3 and 4) and pro-inflammatory cytokines (IFNγ, TNFα, IL-12, IL-1β, IL-6, IL-17 and GM-CSF) in chronic 1484 filarial pathology with (CP Ag+ (n=24)) or without (CP Ag- (n=65)) active infection as well as in asymptomatic, infected (INF; n=84); and ANTENATAL CLINIC ATTENDANCE, INTERMITTENT uninfected, endemic normal (EN; n=64) individuals. Markers that were PREVENTIVE TREATMENT USE AND PREGNANCY OUTCOME significantly elevated in CP Ag+ compared to INF but not in CP Ag- AMONG PARTURIENT WOMEN ATTENDING ADEOYO compared to EN individuals were considered to truly reflect biomarkers of TEACHING HOSPITAL, IBADAN, OYO STATE, NIGERIA pathogenesis. CPAg+ individuals had significantly elevated plasma levels of LPS (p=0.0001), α-2m (p=0.0003), haptoglobulin (p<0.0001) and SAA Damilola C. Olorunda (p=0.0385) among the microbial translocation and acute phase panels. University Of Ibadan, Nigeria, Ibadan, Nigeria Among the angiogenic growth and fibrotic factors, we found significantly Intermittent Preventive Treatment in pregnancy with Sulfadoxine elevated levels of VEGF-A (p=0.0031) and C (p<0.0001), VEGF-R1 Pyrimethamine (IPTp-SP) is currently the recommended method for control (p=0.0033), R2 (p<0.0001), R3 (p=0005) and Angiopoietin-1 (p=0.0481) of malaria in pregnancy. However, in Nigeria the proportion of women but not the MMP/TIMP family. In addition, a variety of pro-inflammatory who take the recommended two doses during Antenatal Clinic (ANC) cytokines including IFNγ, IL-12, GM-CSF (p<0.0001 for all) and IL-1β is low. In this study, we investigated the relationship between number (p=0.0073) were significantly elevated in CP Ag+ individuals. The elevated of ANC visits and IPTp coverage, relationship between IPTp use and levels of these factors suggest quite strongly that the alteration of prevalence of malaria parasite at parturition in both mother and baby lymphatic integrity and peri-lymphatic inflammation should be implicated as well as pregnancy outcome.339 mother-baby pairs were enrolled in the pathogenesis of lymphatic filarial pathology. at delivery in a secondary health care facility in Ibadan, south western Nigeria. An interviewer administered questionnaire was used to collect 1483 information on demographic details, history of index pregnancy. Thick DOES MALARIA IN PREGNANCY IMPAIR PLACENTAL blood films from maternal finger prick and neonatal heel prick were prepared and stained with fresh Geimsa stain. Data was summarized DEVELOPMENT? EVIDENCE FROM AN IN VITRO MODEL using frequency tables and means while differences in proportion were Alexandra J. Umbers1, Danielle I. Stanisic2, Francesca Baiwog3, compared using Chi-square test. There were six twin delivery giving 339 Ivo Mueller3, Peter Siba3, Christopher L. King4, James G. Beeson5, mothers and 345 neonates. Mean age of mother was 28.1 ± 0.281 years, Stephen J. Rogerson1 126/339 (37.2%) mothers were primigravida, 80/339 (23.6%) were 1University of Melbourne, Parkville, Victoria, Australia, 2WEHI, Melbourne, secundigravidae, 133/339 (39.2%) had had 3 pregnancies and above. Use Australia, 3Papua New Guinea Institute of Medical Research, Goroka, of IPTp-SP was reported by 88/339 (26%) parturient women and 17/88 Papua New Guinea, 4Case Western Reserve University, Cleveland, OH, (19.3 %) of them received two doses before delivery. Coverage of IPTp-SP United States, 5Macfarlane Burnet Institute of Medical Research and Public was significantly higher among women who had ≥4 ANC visits (32.2%) Health, Melbourne, Victoria, Australia as compared with women who <4 visits (15.6%). There was no significant relationship between number of ANC visits and transportation cost as In malaria endemic areas, Plasmodium falciparum (Pf) malaria during well as distance of residence to clinic. The prevalence of malaria parasite pregnancy is the leading preventable cause of low birth weight and in the parturient women and neonates were 13.3% and 3.5% (12/345). neonatal mortality, often due to fetal growth restriction (FGR). The Prevalence of malaria parasitemia was lower (0%) in those who used 2 or underlying pathogenic mechanisms are poorly characterized, but more doses of SP than in those who used less than 2 doses 4/71 (5.6%). may include impaired placental development. The peak prevalence of There was no significant relationship between the number of doses maternal Pf infections between 13-18 weeks gestation coincides with the of SP taken and maternal and neonatal haematocrit, birth weight and establishment of the placental circulation, when extravillous trophoblasts gestational age at parturition. Compliance with IPTp-SP use is ensured by (EVT) invade the maternal uterus and transform maternal spiral arteries frequent ANC visits and reduces level of parasiteamia in both mother and increasing placental blood supply. Adequate trophoblast invasion is child. Effort at malaria control should be targeted more at encouraging essential for the establishment of appropriate placental function and women to attend antenatal clinic regularly to achieve completion of successful fetal growth and impairment of this process is associated with recommended dose of intermittent preventive treatment. other causes of human FGR. To address the impact of malaria infection early in pregnancy on placental development, we tested serum from 1485 Papua New Guinean women with Pf in peripheral blood at their first antenatal presentation (between 16 and 22 weeks gestation) for the PARASITOLOGIC ASSESSMENT OF TWO-DOSE AND ability to inhibit first trimester EVT-cell line invasion and viability in vitro. MONTHLY INTERMITTENT PREVENTIVE TREATMENT OF Compared to uninfected controls, serum from malaria-infected women MALARIA DURING PREGNANCY WITH INTERMITTENT significantly reduced trophoblast invasion (P <0.001). This phenomenon PREVENTIVE TREATMENT WITH SULPHADOXINE- could not be explained by changes in trophoblast viability (P =.2). Because PYRIMETHAMINE (IPTp-SP) IN LAGOS, NIGERIA trophoblast invasion is enhanced by a number of hormones, cytokines and chemokines, and is inhibited by pro-inflammatory cytokines, many Chimere O. Agomo1, Wellington A. Oyibo2, Funke Odukoya- of which are dysregulated in malaria in pregnancy, we further compared Maije3 concentrations of known modulators of trophoblast invasion in maternal 1Nigerian Institute of Medical Research, Lagos, Nigeria, 2Department of blood between the groups. Serum collected from malaria-infected women Medical Microbiology and Parasitology, College of Medicine, University had significantly lower levels of trophoblast invasion promoting factors of Lagos, Lagos, Nigeria, 3Department of Obstetrics and Gynaecology, (Insulin like growth factors -1 and -2, P =.0001, P =.01 respectively, and Ajeromi General Hospital, Ajegunle, Lagos, Nigeria IL-8 P = 0.02) and higher levels of invasion inhibitory modulators (human chorionic gonadotrophin P =.002, IL-10 P =.01). Although malaria- Intermittent preventive treatment of malaria with sulphadoxine- induced elevated pro-inflammatory cytokines and reduced fetal growth pyrimethamine (IPTp-SP) is a key strategy in the control of malaria in hormones have been reported at delivery, this study is the first to describe pregnancy from the second trimester. However, there is no data on altered levels of such factors early in pregnancy. These inflammatory the protective efficacy of IPTp-SP in Lagos, Nigeria. High sulphadoxine- and hormonal disturbances in early pregnancy may impair placental pyrimethamine (SP) resistance reported among Plasmodium falciparum

www.astmh.org 448 isolates has been reported from clinical trials and molecular studies in the biggest magnitude of effect seen with VAR2CSA-DBL5 compared to children. This has necessitated the continuous monitoring of the efficacy merozoite responses. At the population level, antibody titres increased of SP in pregnant women. Reports of malaria prevalence in Nigeria with gestation time in those with concurrent P. falciparum parasitaemia suggest that malaria is hyperendemic in most areas, thus raising concerns most likely reflecting boosting of responses with each successive on the adequacy of the standard two-dose IPTp-SP strategy adopted infection. This study provides the most comprehensive analysis, to date, for HIV-negative pregnant women. This study was done to determine of antibody dynamics towards two Plasmodium spp. and contributes to the protective efficacy of IPTp-SP; and to assess the equivalence of our understanding of malaria during pregnancy and immune responses to monthly-dose to the standard two-dose IPTp-SP in Lagos. The study was infectious diseases during pregnancy. a longitudinal study. The women were randomly allotted to two arms: two-dose IPTp-SP (Arm A) and monthly dose IPTp-SP (Arm B). A total of 1487 259 pregnant women [Arm A=122; Arm B=137] attending antenatal clinics in two hospitals in Lagos, Nigeria were recruited. Eligibility criteria MALARIA IN EARLY PREGNANCY ADVERSELY AFFECTS IN were the absence of symptomatic malaria, HIV and multiple pregnancy. UTERO FETAL GROWTH Outcome measures were: absence of malaria parasites in peripheral blood; Jennifer B. Griffin1, Victor Lokomba2, Sarah H. Landis1, Amy proportion of live births and low birth weight. Baseline parasitaemia (M0) H. Herring3, John M. Thorp Jr.4, Antoinette K. Tshefu5, Steven R. in the two group was 5(4.1%) and 3(2.2%) in Arms A and B respectively. Meshnick1 The overall protective efficacy of IPTp-SP was 98.4% (Arm A, 98.3% and Arm B, 98.5%) at M1(P=0.636). Similar result was obtained at the second 1Department of Epidemiology, Gillings School of Global Public Health, month (M2) (P = 0.466). However, none of the women in the monthly University of North Carolina, Chapel Hill, NC, United States, 2University IPTp-SP (Arm B), developed parasitaemia after M1; while a woman of North Carolina-DRC Programme, Kinshasa, The Democratic Republic became parasitaemic at M2 in the 2-Dose IPTp-SP group (Arm A). The of the Congo, 3Department of Biostatistics, Gillings School of Global monthly dosing was not superior to the two-dose regime. The proportion Public Health, University of North Carolina, Chapel Hill, NC, United States, of live births and low birthweight were similar in the two study arms 4Department of Obstetrics and Gynecology, University of North Carolina, (P>0.05). Intermittent preventive treatment of malaria during pregnancy Chapel Hill, NC, United States, 5Ecole de Sante Publique, Faculte de with SP is effective in protecting pregnant women from malaria infection Medecine, University de Kinshasa, Kinshasa, The Democratic Republic of in Lagos. Monthly-dose IPTp-SP is equivalent to the standard two-dose the Congo IPTp-SP in Lagos, Nigeria. Maternal malaria produces adverse maternal and fetal outcomes, including maternal anemia and intrauterine growth restriction (IUGR). There is a 1486 scarcity of research regarding the effects of early malaria on pregnancy outcomes despite, typically, a lack of prevention and control measures ACQUISITION AND MAINTENANCE OF IgG RESPONSES during this critical time of placental development. We evaluated the effect TO PLASMODIUM FALCIPARUM AND P. VIVAX DURING of malaria parasitemia prior to 21 weeks’ gestation on estimated fetal PREGNANCY weight (EFW) and IUGR among a sample of 128 pregnant women enrolled Freya J. Fowkes1, Rose McGready2, Mirja Hommel1, Nadia Cross1, prior to 21 weeks’ gestation in a longitudinal ultrasound study from Julie Simpson3, Kurt Lackovic4, Jack Richards1, SJ Viladpai-nguen2, May 2005 to May 2006 in Kinshasa, DR Congo. Malaria exposure and Marion Avril5, Joseph Smith5, David Narum6, Robin Anders7, ultrasound estimated fetal weight were measured monthly until delivery. Takafuni Tsuboi8, Francois Nosten2, James G. Beeson1 Intermittent preventive treatment in pregnancy (IPTp) was provided twice (from 16-27 and 28-32 weeks’ gestation), insecticide treated nets were 1Burnet Institute, Melbourne, Australia, 2Shoklo Malaria Research Unit, provided, and slide-positive malaria cases were treated. Linear mixed Mae Sot, Thailand, 3University of Melbourne, Melbourne, Australia, models were fitted to estimate beta coefficients and 95% confidence 4Walter and Eliza Hall Institute, Melbourne, Australia, 5Seattle Biomedical intervals (CIs) to describe the effect of early malaria parasitemia on the Research Institute, Seattle, WA, United States, 6National Institute of Allergy mean difference in subsequent EFW; log-binomial general estimating and Infectious Diseases/National Institutes of Health, Rockville, MD, United equation (GEE) regression models were fitted to estimate risk ratios States, 7LaTrobe University, Melbourne, Australia, 8Ehime University, Ehime, (RRs) and 95%CIs for effect of early malaria on risk of subsequent IUGR. Japan Twenty-one percent of pregnant women had malaria parasitemia prior Pregnant women are more susceptible to, and more severely affected to 21 weeks’ gestation and 43% ever experienced an IUGR episode after by, malaria and other infectious diseases. In malaria endemic regions 21 weeks’ gestation. Primigravidae with early malaria parasitemia had an pregnant women typically develop high parasite densities, placental approximately 80 gram decrease in EFW compared to primigravidae with infection and associated complications, despite substantial immunity no early malaria (95%CI: -116, -34). Primigravidae with early malaria had to malaria that may have been acquired prior to pregnancy. This has 3.6 times the risk of subsequent IUGR compared to the referent group of largely been attributed to both the modulation of maternal immune multigravidae with no early malaria parasitemia (95%CI: 2.1, 6.2). Early responses and the sequestration of Plasmodium falciparum parasites in the malaria was also associated with a small, non-significant increased risk placenta. However, data on the acquisition and maintenance of antibody of IUGR among multigravidae (RR: 1.4; 95%CI: 0.8, 2.5). Our findings responses throughout pregnancy, and their relation to malaria is unclear. indicate that appropriate malaria prevention and control efforts should Furthermore, there are limited data on malarial immunity among pregnant begin earlier in pregnancy in order to prevent the adverse effects of women in low transmission settings, in Asia, and in a setting where P. malaria on fetal growth. falciparum and P. vivax are prevalent. In a nested case-control study of pregnant women on the Thai-Burmese border, we measured IgG levels to P. falciparum merozoite antigens (AMA1, EBA-175, MSP2, MSP3, schizont extract) and P. vivax merozoite antigens (Pv-AMA1) and the P. falciparum pregnancy-specific antigen VAR2CSA-DBL5 at 2-weekly intervals during pregnancy until delivery in 136 malaria cases and 124 controls (over 2000 samples in total). ELISAs were performed using novel high-throughput technology to facilitate determination of antibody levels in a large number of samples. Longitudinal analysis revealed that at the individual level, antibody responses could be grouped as dynamic or relatively stable during gestation. The most dynamic species-specific responses were seen in women who experienced active infection during pregnancy and

www.astmh.org 449 1488 be neither a sensitive nor a specific indicator of placental infection. To improve estimation of the prevalence of infection directly from placental EVALUATION OF SULFADOXINE-PYRIMETHAMINE FOR prevalence, a mathematical model linking the progression of placental INTERMITTENT PREVENTIVE TREATMENT OF MALARIA IN infection throughout pregnancy to an existing model of the prevalence PREGNANCY--- MANSA, ZAMBIA, 2010 of malaria within the general population was developed which includes both age and parity as factors which affect the dynamics of infection. The 1 2 3 Kimberly E. Mace , Victor Chalwe , Allen S. Craig , Bonnie model was fitted to placental histology data (prevalence of acute, chronic 4 2 2 L. Katalenich , Michael Nambozi , Luamba Mubikayi , Chikuli and past infection by age and parity) from two transmission settings (Kilifi, 5 6 1 1 Mulele , Mulakwa Kamuliwo , Scott J. Filler , Kathrine R. Tan Kenya, 1995/96, EIR≈5 and Ifakara, Tanzania, 1994/95, EIR≈365) using 1Centers for Disease Control and Prevention, Atlanta, GA, United States, Bayesian MCMC methods to provide estimates of the rate of progression 2Tropical Disease Research Center, Ndola, Zambia, 3Centers for Disease of infection within the placenta. Our results suggest that parity-specific Control and Prevention, Lusaka, Zambia, 4United States Peace Corps, immunity which results in faster clearance of placental infection is more Lusaka, Zambia, 5Wusakile Mine Hospital, Kitwe, Zambia, 6National consistent with patterns of placental infection in different parities than Malaria Control Centre, Lusaka, Zambia an immunity mechanism that decreases the probability that an acquired peripheral infection sequesters the placenta. Furthermore, we estimate Intermittent preventive treatment of malaria in pregnancy (IPTp) with that the duration of chronic infection decreases rapidly with exposure to sulfadoxine-pyrimethamine (SP) decreases placental parasitemia and infection in previous pregnancies with a single prior infection reducing maternal anemia, thus improving birth outcomes, especially among this time by over two-thirds. From these parameters we derived the primigravidae. Zambian policy recommends three SP doses, given relationship between transmission intensity and age- and parity-specific presumptively to pregnant women, spaced one month apart after prevalence of placental infection which can be used to aid the assessment 16 weeks of gestation. Given increased parasite resistance to SP, we of appropriate intervention strategies in areas of differing transmission evaluated the effectiveness of SP for IPTp. HIV-negative pregnant women intensity. were enrolled during delivery at two facilities in an area of high malaria transmission. Women were interviewed, SP exposure was determined from antenatal records and self-report, and blood and placental samples 1490 were taken. We evaluated outcomes of maternal parasitemia, maternal LONGITUDINAL MOLECULAR EPIDEMIOLOGY OF anemia (Hb<11 g/dl), and placental parasitemia in women who took PREGNANCY-ASSOCIATED MALARIA IN BLANTYRE, MALAWI ≥2 (n=286) versus <2 (n=149) SP doses. The median age of participants was 23 years (range 16-44). Among all women, ≥2 SP doses provided Steve M. Taylor1, Alejandro Antonia1, Gaoqian Feng2, Feiko O. no protection from maternal parasitemia [n=435, unadjusted odds ratio ter Kuile3, Steven R. Meshnick1, Stephen J. Rogerson2 (OR) 1.0, (95% confidence interval (CI) 0.5-2.0)], anemia (OR 0.7, 95% 1Gillings School of Global Public Health, University of North Carolina- CI 0.5-1.1), placental parasitemia (OR 0.9, 95% CI 0.4-2.0), and placental Chapel Hill, Chapel Hill, NC, United States, 2University of Melbourne, monocyte infiltration, (OR 0.9, 95% CI 0.4-2.0). We found similar, non- Melbourne, Australia, 3Liverpool School of Tropical Medicine, Liverpool, significant results when evaluating ≥3 SP doses versus <2 doses. However, United Kingdom taking ≥2 SP doses was protective against anemia among primigravidae (n=159, OR 0.4, 95% CI 0.2-0.9). Maternal age, urban living, and use of Pregnancy-associated malaria (PAM) is an important cause of poor birth insecticide-treated nets were not significantly associated with outcomes. outcomes in sub-Saharan Africa, though its prevalence is decreasing. Controlling for age, vaginal delivery, marital status, and living in a house From 1997 to 2005, during which the prevalence of parasitemia at with indoor residual spraying, taking ≥2 SP doses remained protective for delivery decreased from 24% to 5% at a single hospital in Blantyre, anemia among primigravidae (OR 0.4, 95% CI 0.2-0.7). SP for IPTp may Malawi, we analyzed trends in molecular markers of drug resistance confer less protection against malaria than previously observed. Limitations and parasite diversity among 358 Plasmodium falciparum isolates from of this study include strata too small to analyze [e.g. severe anemia (Hb<8 the peripheral blood of delivering women. The average multiplicity of g/dl), and no SP taken], insufficient power to examine birth weight as an infection (MOI), as determined by merozoite surface protein-2 (msp2) outcome, and unverified SP quality. These findings emphasize the urgent genotyping, decreased from 3.8 (standard deviation 1.3) to 1.8 (SD 1.2) need to evaluate alternative medications for IPTp. during the study period. The prevalence of the triple-mutant dihydrofolate reductase (dhfr) haplotype increased from 33% to 100%, and that of the 1489 double-mutant dihydropteroate synthase (dhps) haplotype from 33% to 100%; concomitantly, the prevalence of the combined quintuple-mutant LINKING MALARIA IN PREGNANCY TO POPULATION haplotype increased from 17% to 100%. In contrast, haplotypes of the P. TRANSMISSION DYNAMICS: A MODEL OF THE falciparum multidrug resistance gene (pfmdr1) were unchanged over time, PROGRESSION OF PLACENTAL INFECTION AND THE ROLE OF and mutations at codon 164 of dhfr and codon 581 of dhps failed to PARITY-DEPENDENT IMMUNITY emerge. Accounting for parasite multiplicity, we calculated frequencies of alleles and haplotypes and used these to compute selection coefficients as Patrick G. Walker1, Matthew Cairns2, Feiko Ter Kuile3, Azra a quantitative index of the strength of selection upon mutant alleles and Ghani1 haplotypes. The results demonstrate the molecular correlates of improved 1MRC Centre for Outbreak Analysis and Modelling, London, United control of pregnancy-associated malaria, and serve as a model for the Kingdom, 2Department of Infectious Disease Epidemiology, London School emergence and fixation of drug-resistance in a semi-immune population of Hygiene & Tropical Medicine, London, United Kingdom, 3Liverpool exposed to increasing sulfadoxine-pyrimethamine. Clinical correlation of School of Tropical Medicine, Liverpool, United Kingdom these phenomena will inform drug therapy and resistance surveillance policies. Malaria in pregnancy, which is associated with maternal anaemia, preterm birth, low birthweight and increased mortality for both mother and child, continues to be a major public health issue in many parts of the world. Ensuring that prospective mothers are provided with the most effective treatment or preventative measures is a priority. However, assessment of the effectiveness of any intervention is problematic because the infection status of the placenta can only be assessed at delivery. Thus, for the preceding period of gestation, the prevalence of malaria in pregnancy can only be estimated from peripheral blood which has been shown to

www.astmh.org 450 1491 no association between timing of malaria infection and the likelihood of detecting placental malaria. This study did not have the power to detect GENOMIC DIVERSITY OF VAR2CSA DUFFY-BINDING-LIKE the relationship between these measures of malaria exposure and low DOMAINS IN PREGNANCY-ASSOCIATED MALARIA EXPLORED birth weight or maternal anemia. We will report the relationship between WITH MASSIVELY-PARALLEL PYROSEQUENCING malaria and gestational age at delivery based on ultrasound results. The first half of pregnancy, before women typically seek out antenatal 1 2 2 Steve M. Taylor , Jonathan J. Juliano , Nagesh R. Aragam , Feiko care, is a period of vulnerability to malaria infection. Placental histology 3 4 1 O. ter Kuile , Linda Kalilani-Phiri , Steven R. Meshnick identified approximately half of women who had detectable infection in 1Gillings School of Global Public Health, University of North Carolina- the peripheral blood during pregnancy. Studies with large sample sizes are Chapel Hill, Chapel Hill, NC, United States, 2University of North Carolina needed to determine if histology is an adequate surrogate for intensive School of Medicine, Chapel Hill, NC, United States, 3Liverpool School surveillance in detecting the adverse outcomes associated with malaria of Tropical Medicine, Liverpool, United Kingdom, 4University of Malawi during pregnancy. College of Medicine, Blantyre, Malawi VAR2CSA expression on the surface of erythrocytes infected by 1493 Plasmodium falciparum mediates placental malaria by allowing for the THERAPEUTIC EFFICACIES OF ARTEMISININ-BASED sequestration of infected erythrocytes in the placenta. Because of this COMBINATION THERAPIES IN NIGERIAN CHILDREN WITH specific molecular pathogenesis, VAR2CSA has been advanced as a UNCOMPLICATED FALCIPARUM MALARIA DURING FIVE target for a vaccine against pregnancy-associated malaria. To do so, pathogenic and immunogenic motifs must be identified within var2csa, YEARS OF ADOPTION AS FIRST-LINE TREATMENTS but these efforts have been bedeviled by the sequence diversity of Grace O. Gbotosho1, Akintunde Sowunmi1, Christian H. Happi1, var2csa. Genomic diversity between and within populations of parasites Titilope M. Okuboyejo2 that comprise a single infection is not adequately described by traditional 1Department of Pharmacology and Therapeutics and Institute for Medical genotyping methods because they fail to capture all variants. To this end, Research and Training, University of Ibadan, Ibadan, Nigeria, 2Department we employed massively-parallel pyrosequencing to explore the diversity of of Pharmacology and Therapeutics, University of Ibadan, Ibadan, Nigeria two var2csa Duffy binding-like (DBL) domains in genomic DNA of parasites infecting the placentae and peripheral blood of delivering women in The therapeutic efficacies of 3-day regimens of artesunate-amodiaquine Malawi. After amplifying across hypervariable regions of DBLs 5 and 6, and artemether-lumefantrine, during 5 years of adoption as first-line amplicons were sequenced on a Roche 454 System, which sequences treatments, were evaluated in 811 <12 year-old malarious children. individual amplicons up to 500bp long. We used population genetic Compared with artemether-lumefantrine, amodiaquine-artesunate analysis methods to compare populations of parasites between women, significantly reduced the proportion of children with fever and parasitemia between study sites, and between peripheral and placental blood, and 1d after treatment began (day 1) (P <0.008 for both). The proportion of we computed DBL variant discovery curves. The combination of next- parasitemic children on day 2, and gametocytemia on presentation and generation sequencing and population genetic analysis provides a method carriage reduced significantly over the years (P <0.000001, and P <0.03, by which to capture and exploit the genomic diversity of P. falciparum respectively, test for trend). Overall efficacy was 96.5% (95%CI 94.5- within individual infections. These tools may aid the identification of 98.6) and remained unchanged over the years (P = 0.87, test for trend). motifs that are critical to the pathogenicity and immunogenicity of Kinetics of parasitemias following treatments was estimated by non- pregnancy-associated malaria. compartmental model. Declines of parasitemias were monoexponential with mean elimination half-life of 1.09h (95%CI 1.0-1.16). Parasitemia 1492 half lives and efficacy were similar for both regimens and in all ages. Artesunate-amodiaquine and artemether-lumefantrine remain efficacious INTENSIVE SURVEILLANCE FOR MALARIA DURING treatments of uncomplicated falciparum malaria in Nigerian children 5 PREGNANCY years after adoption. Linda Kalilani-Phiri1, Mwayiwawo Madanitsa1, Patricia Mawindo2, Osward M. Nyirenda2, Phillip C. Thesing3, Blair Wylie4, Miriam K. 1494 3 Laufer EVALUATION OF ADVERSE DRUG REACTIONS TO 1University of Malawi College of Medicine, Blantyre, Malawi, 2Blantyre ARTEMISININ-BASED COMBINATION THERAPY IN A Malaria Project, Blantyre, Malawi, 3University of Maryland, Baltimore, MD, COMMUNITY HEALTH CLINIC IN PAPUA PROVINCE, United States, 4Massachusetts General Hospital, Boston, MA, United States INDONESIA In studies of pregnancy-associated malaria, surveillance is frequently Cut N. Hafifah limited to examination of the placenta for evidence of active or past malaria infection. The sensitivity of placental histology for the detection of Community Health Clinic Medical Emergency and Rescue Committee, malaria during pregnancy is not well studied. We conducted a prospective Jakarta, Indonesia observational study recruiting women in their first or second pregnancy Malaria is one of the main health problem in east of Indonesia. Since who were less than 28 weeks gestation and attending the Ndirande 2004, Ministry of Health Republic of Indonesia has recommended the Antenatal Clinic in Blantyre, Malawi for the first time. Participants had use of orally administered Artemisinin-Based Combination Therapy microscopy and hemoglobin measurement done monthly and whenever (ACT) to treat uncomplicated malaria. However, some patients they were ill. All women received three doses of intermittent preventive experienced uncomfortable adverse reactions due to ACT, leading to its therapy. At delivery, a maternal peripheral, placental and cord blood, and discontinuation. Recognition of these adverse reactions may lead to better placental biopsy samples were collected. A total of 450 women were management. The aim of this study is to determine the prevalence of recruited; 63% (n=285) were primigravidae. Among the 342 women adverse reactions due to orally administered ACT and types of reactions with evaluable birth outcomes, 38 (11%) had malaria at enrollment experienced by uncomplicated malaria patients. A cross sectional study and an additional 22 (6%) had malaria detected on peripheral blood among uncomplicated malaria patients was conducted in Community smear after enrollment. On histological evaluation, 62/342 (18%) had Health Clinic Medical Emergency and Rescue Committe, a non- evidence of placental malaria based on the presence of parasites and/ governmental health organization, in Timika, Papua, Indonesia, between or pigment. Fifty six percent of women with positive malaria smears September 2010 and March 2011. Sample was collected consecutively. during pregnancy had evidence of malaria infection in the placenta and Primary data were collected by interview using a questionnaire to assess 43% without peripheral parasitemia had placental malaria. There was www.astmh.org 451 adverse reactions experienced by the patients, while demographic data 1496 were obtained from medical records. There were 250 uncomplicated malaria patients who fulfilled the inclusion criteria and did not meet any PHARMACOKINETICS, SAFETY AND EFFICACY OF of the exclusion criteria. Adverse reactions of orally administered ACT ARTEMISININ-NAPHTHOQUINE COMBINATION (ARCOTM) were experienced by 48% respondents. Most common adverse reaction THERAPY IN PAPUA NEW GUINEAN CHILDREN WITH was nausea (28%). Approximately 33% respondents who experienced UNCOMPLICATED PLASMODIUM FALCIPARUM AND P. VIVAX adverse reactions considered them intolerable. These findings are MALARIA much higher compared to other studies. Adisa et al (2008) and Cairo et al (2008) found adverse reactions due to ACT in 32.9% and 8.8% Brioni R. Moore1, John Benjamin2, Kevin T. Batty3, Madhu Page- respondents respectively. Nausea, the most common adverse reactions Sharp3, Sam Salman1, Kenneth F. Ilett1, Peter Siba2, Ivo Mueller4, found in this study, was found only in 1,3% patients interviewed by Timothy M. Davis1 Adisa et al and 16% of Cairo et al’s subjects. This difference is probably 1University of Western Australia, Perth, Australia, 2Papua New Guinea due to different types of ACT used and dosage given to the patients. Institute of Medical Research, Madang, Papua New Guinea, 3Curtin Eventhough, only 33% respondents who experienced adverse reactions University, Perth, Australia, 4Walter and Eliza Hall Institute of Medical considered these adverse reactions intolerable, around 22% of these Research, Melbourne, Australia respondents discontinued their ACT against medical advice. In conclusion, prevalence of adverse reactions due to orally administered ACT was quite Artemisinin-naphthoquine (AN) is available as a fixed-dose oral co- frequent. Efforts should be made to minimize the percentage of patients formulation marketed in Papua New Guinea (PNG) and other countries as TM discontinuing their medication. Patients should understand the adverse ARCO (Kunming Pharmaceutical Company). It is currently recommended reactions of ACT that might occur and doctors should carefully manage as single-dose treatment even though the WHO stipulates that artemisinin- these adverse reactions. based combination therapy should be given as a 3-day regimen. To date there have been limited published data on the pharmacokinetics of 1495 naphthoquine, particularly in children. We conducted a randomised study in PNG children aged 5-12 years with uncomplicated falciparum or vivax POPULATION PHARMACOKINETICS OF ARTESUNATE malaria comparing single-dose ARCO (A:N 15:6 mg/kg) given with water AND DIHYDROARTEMISININ IN HEALTHY AND MALARIA- (Group A, n=15), single-dose (A:N 22:9 mg/kg) administered with full- INFECTED SUBJECTS cream milk (Group B, n=17) or as two daily doses (A:N 22:9 mg/kg) given with water (Group C, n=16). Blood samples were collected from each child Carrie Morris1, Stephan Duparc2, Isabelle Borghini-Fuhrer2, at 15 time points after dose, and naphthoquine in plasma was quantified Chang-Sik Shin3, Donald Jung4, Lawrence Fleckenstein1 by liquid chromatography-mass spectrometric assay. Of the 48 children (46 1University of Iowa, Iowa City, IA, United States, 2Medicines for Malaria with falciparum and 2 with vivax malaria), 2 in Group B withdrew because Venture, Geneva, Switzerland, 3Shin Poong Pharmaceuticals, Seoul, of inability to complete study procedures. All regimens were well tolerated Republic of Korea, 4Pharmaceutical Research Services, Cupertino, CA, with no serious adverse events. There were no significant changes in United States pulse, blood pressure, rate-corrected QT interval, or routine biochemistry after treatment, and fever clearance was prompt. The mean 50% ® Pyramax is a pyronaridine/artesunate combination currently under parasite clearance times were 3.9, 3.9 and 4.6 h for Groups A, B and C, evaluation for treatmentof uncomplicated malaria in adult and pediatric respectively. In Group A, 1 patient had Day 23 parasitological failure, and 2 patients. We analyzed the population pharmacokinetics of artesunate children presented with re-infections on each of Days 28 and 42. In Group (AS) and its active metabolite dihydroartemisinin (DHA) in healthy and B and C there were no cases of parasitological failure but 1 Group B child ® malaria-infected subjects participating in eight Pyramax clinical trials. had a re-infection on Day 42. Gametocyte clearance was prolonged with Pharmacokinetic data were available from 166 healthy Caucasian and 20%, 27% and 9% of participants still positive on Day 14 in Groups A, B Korean adults and 631 African and Asian adult and pediatric patients and C, respectively. Using a two-compartment model, the mean terminal with mild to moderate uncomplicated malaria. Plasma AS and DHA elimination half-lives of naphthoquine were 420, 372 and 453 h for Group concentrations were measured using a validated LC-MS method. Non- A, B and C, respectively. The maximum plasma concentration after dose linear mixed effects modeling was used to obtain the pharmacokinetic was similar for all three groups. Both single and two-dose ARCO regimens and variability parameter estimates. A simultaneous parent-metabolite were safe and well tolerated. The long half-life of naphthoquine is similar model was developed consisting of first-order absorption of AS, a to those reported for 4-aminoquinolines and related compounds and one-compartment model for AS, and a one-compartment model protects against recrudescence. Milk co-administration did not appear to for DHA. The population estimates showed that AS absorption was influence naphthoquine pharmacokinetics. rapid, with an absorption rate constant (Ka) of 3.02 h-1. Apparent AS clearance (CL/F) and volume of distribution (V2/F) were 1170 L/h and 1497 1040 L, respectively. Estimates obtained for apparent clearance (CLM/F) and volume of distribution (V3/F) of DHA were 82.0 L/h and 107 L, IN VITRO “PITTING” TO DETECT ARTEMISININ-RESISTANT respectively. Stepwise covariate modeling yielded four significant covariate- PLASMODIUM FALCIPARUM? parameter relationships: weight on CL/F, CLM/F, and V3/F and ritonavir coadministration on CLM/F. Ritonavir coadministration was associated Papa Alioune Ndour with a substantial increase in CLM/F of 30.5 L/hr. Malaria infection did not Inserm-UPMC (Paris 6 University) UMRs945, Paris, France exert a significant influence on any pharmacokinetic parameter. Creatinine Artemisinin-based therapies are currently the reference treatment for clearance, hepatic enzyme elevations, and gender also did not meet the malaria worldwide. Plasmodium falciparum resistant to artemisinin model inclusion criteria. Estimated inter-individual variability was greatest derivatives is spreading in South-East Asia. Tools that would rapidly detect for Ka (159%), followed by V2/F (34.6%). Evaluation of the final model the onset of resistance are urgently needed. Conventional assessment using bootstrapping, visual predictive check, and condition number of P. falciparum drug resistance in vitro is based on the determination of indicated that the model displayed satisfactory robustness, predictive the IC , but IC of artesunate or dihydroartemisinin (DHA) only weakly power, and stability. The final model was utilized to simulate AS and DHA 50 50 correlate with parasite clearance in artemisinin-treated patients. In these concentration-time profiles for representative weights in each of four patients, P. falciparum parasites are expulsed from their host red blood Pyramax® dosing groups. cell by the spleen, a “pitting” process that greatly accelerates parasite clearance, as reported previously. We want to use in vitro pitting as a test to assess parasite resistance in a new, physiologically relevant way. In vitro

www.astmh.org 452 pitting will indeed mimic the major natural parasite clearance mechanism. 1499 Using an ex-vivo human spleen model, we had obtained the pitting of red blood cells containing artemisinin-induced parasite remnants, as reported THE INFLUENCE OF NEVIRAPINE ON ARTESUNATE AND previously. We had also observed that parasite remnants are deposited DIHYDROARTEMISININ EXPOSURE IN HIV-INFECTED along the wall of red pulp sinuses in the spleen of an artemisinin-treated NIGERIAN ADULTS malaria patient. We recently established and validated an in vitro red blood 1 2 3 4 cell filtering device that mimics the mechanical sensing of red blood cells Fatai A. Fehintola , Kimberly Scarsi , Qing Ma , Sunil Parikh , 4 2 5 as they cross the wall of red pulp sinuses in the human spleen, as reported Francesca Aweeka , Babafemi Taiwo , Ibrahim Tope Akinola , Isaac 1 6 6 previously. Using this device we have obtained the pitting of parasitized F. Adewole , Niklas Lindegardh , Aphiradee Phakdeeraj , Oladosu 1 2 1 red blood cells previously exposed to artesunate or DHA. More than 25% Ojengbede , Robert Murphy , Olusegun O. Akinyinka , Gene D. 3 of artesunate-exposed parasitized red blood cells are pitted by a single Morse passage through the filters. This filtering process requires less than 50 1University of Ibadan, Ibadan, Nigeria, 2Northwestern University, Chicago, µl of peripheral blood and less than 5 hours of parasite exposure to the IL, United States, 3University at Buffalo, Buffalo, NY, United States, drug. We are currently identifying the major determinants of the in vitro 4University of California, San Franscisco, CA, United States, 5University pitting process, and will soon determine the pitting rate of parasite isolates College Hospital, Ibadan, Nigeria, 6Mahidol University, Bangkok, Thailand from patients with fast or slow parasite clearance upon treatment with Nevirapine (NVP)-based antiretroviral therapy (ART) may result in important artesunate. drug-interactions with some antimalarials. Artesunate plus amodiaquine (AS-AQ) is a common antimalarial combination used in Nigeria and the 1498 pharmacokinetic (PK) effect of combining AS-AQ with NVP is unknown. DECREASING IN VITRO SUSCEPTIBILITY OF AS and AQ are converted to active metabolites via hydrolysis and FRESH PLASMODIUM FALCIPARUM ISOLATES TO cytochrome p450 pathways, respectively; the latter being most susceptible to drug-drug interactions. We conducted a two-group comparison of DIHIDROARTEMISININ IN COLOMBIA AS-AQ PK in HIV-infected patients receiving NVP-based ART for at least Gustavo Díaz 8 weeks (n=10) to HIV-infected patients not on ART (control; n=11). AS- International Center for Training and Medical Research, Cali, Colombia AQ 200/600mg was given as a daily dose for three days. Blood sample collection for the measurement of AS and AQ was commenced following The main drawback to controlling malaria is the emergence and spread the final dose at 0 (pre-dose), 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 24, 48, of multidrug resistant parasites. Artemether+lumefantrine therapy was 72 and 96h. AS and the active metabolite dihydroartemisinin (DHA) were implemented during 2007 in Colombia for falciparum-malaria, showing quantified with LC-MS/MS. PK parameters [maximum concentration excellent outcomes. Recent reports from Southeast Asia show that (Cmax; mg/L), (volume of distribution (Vd; L), area under the curve (AUC; it is likely that resistance to Artemisinin Combined Therapies-ACTs is mg/L/h), oral clearance (Cl; L/hr), and half-life (t1/2; h)] were determined developing. This phenomenon highlights the need to keep monitoring with non-compartmental analysis (mean ± standard deviation). In the NVP systems for early detection of resistance in other endemic areas. Our group compared to controls, the AS Vd was 1162 ± 856 vs 4525 ± 3535 aim was to determine the spatial/temporal changes in the Plasmodium (p=0.01) and Cl was 1950 ± 543 vs 2995 ± 1180 L/h (p=0.03), while the falciparum phenotypes through the first years of ACTs implementation AUC was 105 ± 31 vs 69 ± 26 (p=0.02). AS t½ was 0.4 ± 0.3 in the NVP in Colombia. From 2008 to 2010, 121 isolates from Quibdó and group and 1.1 ± 0.9h in controls (p=0.06), while the AS Cmax was 108 Tumaco, (North and South Colombian Pacific coast) were evaluated ± 42 vs 71 ± 57 (P> 0.05), respectively. DHA AUC (603 ± 218 vs 883 ± for in vitro susceptibility to mefloquine-MQ, lumefantrine-LUM and 607) and Cmax (298 ± 107 vs 507 ± 429) were also not different between dihidroartemisinin-DHA, through microscopic and ELISA-HRP2 tests. For the NVP and control group (p>0.05). DHA t½ was shorter with NVP (1.6 quality control, the reference strain W2 was evaluated. The IC50s were ± 0.8 vs 3.2 ± 1.4h; P=0.004). AQ data are being analyzed and will be calculated using NH-nonlin software and non-parametric tests were used presented. NVP-containing ART impacted important PK parameters of AS to compare the results. In 2010, DHA showed a significant increased and DHA. These findings will inform future pharmacodynamic and clinical

IC50s (p<0.05) in Tumaco (Geometric Mean-MG in 2008: 2.02nM, outcome studies of AS-AQ in HIV-infected patients receiving NVP-based 2009: 1.2nMand 2010: 4.0nM ). Meanwhile, during 2008-2010, LUM ART. and MQ IC50s were stable in both regions. The parasites from Quibdó were less susceptible to MQ (MG in 2008: 36nM, 2009: 47.8nMand 1500 2010: 30.7nM) compared to those from Tumaco (MG in 2008: 14.3nM, 2009: 25.1nM and 2010: 15.8nM) in this period (p<0.05). Spearman EVALUATION OF A NOVEL MOLECULAR MARKER FOR correlation test between both methodologies was 0.92 (p<0.01). The MONITORING ARTEMISININ RESISTANCE IN PLASMODIUM fact that in vitro DHA IC50s are increasing in Tumaco highlights the need FALCIPARUM MALARIA of continuing with active surveillance. Although all the isolates tested 1 1 1 showed a high susceptibility to DHA and LUM, which is comparable to Gisela Henriques , Khalid Beshir , Teun Bousema , Halidou 2 3 1 4 1 the high efficacy observed in Colombia with ACTs. Because there is cross Tinto , Paul Hunt , Colin Sutherland , Pedro Cravo , Rachel Hallett in vitro susceptibility between LUM and MQ, our findings underline the 1London School of Hygiene and Tropical Medicine, London, United importance of close monitoring of the in vitro response to LUM in the Kingdom, 2Institut de Recherche en Sciences de la Santé, Bobo Dioulasso, North Pacific region. In vivo studies are costly and not sensitive for early Ouagadougou, Burkina Faso, 3Institute for Immunology and Infection detection of resistance (due to the high efficacy of ACTs) and there are no Research, School of Biological Sciences, University of Edinburgh, molecular resistance markers to artemisinin derivates. Therefore in vitro Edinburgh, United Kingdom, 4Instituto de Patologia Tropical e Saúde assays are currently the main strategy for early detection of resistance to Pública, Universidade Federal de Goiás, Goiânia, Brazil ACTs. The human malaria parasite Plasmodium falciparum has evolved resistance to most drugs. There is now evidence of reduced susceptibility to artemisinin derivatives and to Artemisinin Combination Therapy with delayed parasite clearance times. If artemisinin resistance spreads widely, it would threaten global malaria control. We still lack validated molecular markers for monitoring the resistance phenotypes. Using genome-wide strategies in the rodent malaria parasite P. chabaudi our group identified a mutation in a clathrin mu adaptor gene (pccmu) that arose along with

www.astmh.org 453 artemisinin resistance. In order to investigate the possible contribution 1502 of this candidate marker to artemisinin resistance in the human malaria parasite P. falciparum we screened the DNA sequence of the equivalent EFFICACY OF ARTEQUICK (ARTEMISININ-PIPERAQUINE) orthologue, pfcmu, for genetic polymorphisms. We have studied field AND COARSUCAM (ARTESUNATE-AMODIAQUINE) IN THE isolates from an ACT clinical trial in Burkina-Faso that were tested in vitro TREATMENT OF PLASMODIUM FALCIPARUM MALARIA IN for their response to artemisinin derivatives and other antimalarial drugs. VIETNAM We have also evaluated pre- and post- treatment samples from an in vivo ACT clinical trial carried out in Kenya. Genetic polymorphisms in pfcmu Nguyen X. Thanh1, Trieu N. Trung2, Nguyen C. Phong1, Huynh were analysed for association with various measured endpoints in the H. Quang2, Bui Dai1, George D. Shanks3, Marina Chavchich3, two trials that might indicate a drug resistant parasite phenotype. Our Michael D. Edstein3 preliminary results indicate that mutations in this adaptor protein subunit 1Military Institute of Hygiene and Epidemiology, Hanoi, Vietnam, 2Institute may be associated with varying degrees of in vitro and in vivo responses to of Malariology, Parasitology and Entomology, Qui Nhon, Vietnam, artemisinin derivatives, quinine and lumefantrine. We propose this gene 3Australian Army Malaria Institute, Brisbane, Australia should be evaluated further as a potential molecular marker of artemisinin Recent reports of artesunate resistant or tolerant Plasmodium falciparum resistance. malaria in western Cambodia is of immense concern as the fast acting artemisinins combined with the longer elimination half-life drugs such 1501 as mefloquine are now recommended worldwide for first-line treatment PHARMACODYNAMICS OF CURRENT AND NOVEL of uncomplicated P. falciparum malaria. In light of this information ARTEMISININ COMBINATION THERAPIES AGAINST there is an urgent need to evaluate artemisinin combination therapies ARTESUNATE RESISTANT OR SENSITIVE PLASMODIUM (ACTs) in neighbouring countries to Cambodia to determine whether reduced ACT susceptibility has developed with increase failure rates and BERGHEI prolonged parasite clearance times. The objective of the present study Connor O’Brien1, Philipp Henrich1, Daniel Scanfeld1, Dennis was to compare the efficacy of fixed-dosed combinations of artemisinin- Kyle2, David Fidock1 piperaquine (Artequick®) and artesunate-amodiaquine (Coarsucam®) for 1Columbia University, New York, NY, United States, 2University of South the treatment of P. falciparum malaria in south central Vietnam. In an Florida, Tampa, FL, United States open-labelled, randomized clinical pilot study 128 patients (children aged 6-14 years, n=68, adults aged 15-60 years, n=60) were allocated either a Effective treatment of Plasmodium falciparum is threatened by signs of 2-day course of either Artequick® (~2.8 mg/kg artemisinin plus ~16.9 mg/ emerging resistance to artemisinins, the linchpin of Artemisinin-based kg of piperaquine per day) or a 3-day course of Coarsucam® (~4.5 mg/kg Combination Therapies (ACTs) that have been widely adopted throughout of artesunate plus ~12.3 mg/kg of amodiaquine per day), with a follow- malaria-endemic regions. Attempts to define resistance, in isolates from up period of 42 days. Both ACTs were well tolerated, with no obvious patients that showed delayed parasite clearance times following ACT drug associated adverse events. Parasite clearance times ranged between therapy, have yet to replicate an artemisinin-resistant phenotype and no 12 and 60 h for both treatment groups. Of the per-protocol population, definitive genetic target has been identified. Using a P. berghei model, 50 patients were on Artequick® (24 children, 26 adults) and 51 were on we have developed pharmacodynamic profiles of four existing ACTs Coarsucam® (25 children, 26 adults). The PCR genotype corrected cure (dihydroartemisinin-piperaquine, artesunate-amodiaquine, artesunate- rate at day 42 was 98% for both ACTs. This study showed that the two mefloquine and artemether-lumefantrine), as well as the artesunate- ACTs were highly efficacious in the treatment of P. falciparum malaria in pyronaridine ACT under clinical evaluation, generated against drug- adults and children. Further studies of the ACTs are warranted in different sensitive and artesunate-resistant parasites. These profiles are developed regions of Vietnam to determine the nationwide effectiveness of the two from 30-day studies and assess the efficacy of a 3-day dosing regimen ACTs. for each combination as well as efficacy of modifications to treatment frequency and concentration. All regimens are 3-days in duration and 1503 include each ACT component as monotherapy given once daily, ACT [1x] once daily, ACT [1x] split into two daily doses, ACT [1.5x] once MITIGATION OF ARTEMISININ RESISTANCE IN AFRICA: WHAT daily and ACT [1.5x] split into two daily doses. Results to date elucidate SHOULD BE DONE NOW? differences in contribution to treatment between components of ACTs 1 2 3 and suggest pairings and treatment schedules that are maximally effective Ambrose O. Talisuna , Philippe J. Guerin , Carol Sibley , Robert 4 against artesunate-resistant P. berghei. Furthermore, our data suggest W. Snow cross-resistance between artesunate and the partner drugs amodiaquine, 1Worldwide Antimalarial Resistance Network, University of Oxford/KEMRI/ mefloquine and piperaquine, as well as possible antagonism in the Wellcome Trust, Nairobi, Kenya, 2Worldwide Antimalarial Resistance artesunate-amodiaquine and artesunate-mefloquine combinations. Network, University of Oxford, United Kingdom, 3Worldwide Antimalarial These findings imply that some current pairings are at higher risk of Resistance Network, Washington University, St. Louis, MO, United States, treatment failures than others. Integration of these data with ongoing 4University of Oxford/KEMRI/Wellcome Trust Research Programme, Nairobi, pharmacokinetic and genetic studies should shed light on drug mode Kenya of action and mechanisms of resistance in P. berghei, and help guide Artemisinin resistance in falciparum malaria has emerged in Western selection of suitable ACTs in areas where artemisinin resistance is a Cambodia and may have spread westward, an ominous repetition concern. of the spread of resistance to chloroquine over fifty years ago, and later to sulfadoxine-pyrimethamine. This poses a major global public health threat, with the greatest potential effects in sub-Saharan Africa where disease burdens are greatest and systems for drug resistance monitoring and containment are weakest. We do not fully understand the molecular mechanisms underlying artemisinin resistance. The only associated phenotype observed in South East Asia is a decreased parasite clearance rate, not yet seen in Africa. Artemisinin resistance fits the International Health Regulations (2005) definition of a public health emergency of international concern - it is serious, unusual, and has the potential to spread. Most African countries now recommend artemisinin

www.astmh.org 454 combination therapy (ACT) as first line regimen for uncomplicated malaria. 1505 Unfortunately, this policy shift has resulted in the demise of systematic drug resistance surveillance; sub-regional networks have very limited ARTEMISININ RESISTANCE ASSOCIATED WITH Pfmdr1 COPY activities due to lack of funding and changing technical requirements NUMBERS AND ANTI-OXIDANT ACTIVITY IN THE IN VITRO for monitoring ACT efficacy. Well-defined early warning and detection SELECTED PARASITES LINES systems need to be established. We will present strategies that should be urgently adopted by Africa, including: reactivation of regional surveillance Long Cui, Zenlei Wang, Miao Miao, Ramesh Chandra, Jun Miao, networks with clear roles; data sharing to facilitate pooled analyses to Liwang Cui identify rare observations; drug resistance risk factor modeling to bench The Pennsylvania State University, University Park, PA, United States mark baselines; and the development and validation of new tools for To select an artemisinin resistant line in the laboratory, we have subjected monitoring resistance, antimalarial drug pressure, drug quality and Plasmodium falciparum Dd2 strain to dihydroartemisinin (DHA) selection inappropriate drug usage. with step-wise increments of drug concentrations over 13 months. Two

lines (Art1 and Art2) were obtained more than 20-fold increase in IC50 to 1504 DHA. However, the resistance phenotype was unstable and the parasites PILOT OF HOME-BASED MANAGEMENT OF MALARIA could regain susceptibility to DHA after three months of culture in the WITH RAPID DIAGNOSTIC TESTS AND ARTEMISININ-BASED absence of the drug selection pressure. Phenotype analysis showed that the resistant parasite displayed cross-resistance to a number of the COMBINATION THERAPY IN RURAL SENEGAL commonly used antimalarial drugs. No mutations were detected on the Sylla Thiam1, Julie I. Thwing2, Ibrahima Diallo1, Mame Birame reported drug resistant markers, but we identified increased copy number Diouf1, Robert Perry2, Medoune Diop1, Fatou B. Fall1, Mamadou L. of P. falciparum multidrug resistance 1 (pfmdr1) gene associated with Diouf1, Moustapha Cisse1, Mamadou M. Diaw1, Bakary Sambou3, artemisinin resistance. To detect other potential changes in the selected Abderrahmane O. Kharchi4, Tieman Diarrra5, Moussa Thior1 parasites, we performed microarray analysis and detected increased 1National Malaria Control Program, Dakar, Senegal, 2Centers for Disease expression of genes involved in redox metabolism. Collectively, this study Control and Prevention, Atlanta, GA, United States, 3World Health suggests that selection of artemisinin resistance under the laboratory Organization, Dakar, Senegal, 4World Health Organization, Brazzaville, The conditions may be associated with multiple mechanisms. Democratic Republic of the Congo, 5World Health Organization, Harare, Zimbabwe 1506 Malaria remains a major cause of morbidity and mortality in Senegal, ARC3: DETECTING RECENT POSITIVE SELECTION IN and access to care is challenging in part due to the remoteness of many ARTEMISININ RESISTANT MALARIA PARASITES communities. Senegal introduced artemisinin-based combination therapy 1 2 1 (ACT) in 2006, and diagnosis based on rapid diagnostic tests (RDTs) in Christopher Jacob , Olivo Miotto , Shannon Takala Harrison , 3 4 2 5 2007, resulting in 85% of suspected cases in the public health sector Taane Clark , Michael Cummings , Arjen Dondorp , Mark Fukuda , 6 7 8 9 being tested in 2009. To expand access to case management based on Francois Nosten , Harald Noedl , Mallika Imwong , Delia Bethell , 9 9 9 9 parasitologic diagnosis, in 2008 Senegal introduced a pilot for home-based Youry Se , Chanthap Lon , Stuart Tyner , David Saunders , 10 6 7 management with RDTs and ACTs in 20 remote villages >5 km from the Duong Socheat , Aung Pyae Phyo , Peter Starzengruber , Paul 7 11 11 11 nearest health post. Village members chose a volunteer home-based care Swoboda , Gustavo Cerqueira , Joana Silva , Cesar Arze , Stacy 12 12 1 1 provider (HCP) from the community, who received three days of classroom Ricklefs , Stephen Porcella , Matthew Adams , L. Kenefic , S. 13 14 14 13 13 training on malaria case management, followed by a 15 day practical in Campino , S. Auburn , S. Auburn , M. Manske , B. MacInnis , 13 15 2 16 1 the nearest health post. The HCPs were then officially installed and given D. Kwiatkowski , X.Z. Su , N. White , P. Ringwald , C. Plowe a kit including RDTs, ACTs, a case record book, and stock management 1Howard Hughes Medical Institute/Center for Vaccine Development, tools. The chief nurse at the nearest health post was responsible for the University of Maryland School of Medicine, Baltimore, MD, United States, practical training and monthly supervision. Health officials at district, 2Mahidol Oxford Research Unit, Bangkok, Thailand, 3Pathogen Molecular regional, and national levels also performed supervision visits. During the Biology Department, London School of Hygiene & Tropical Medicine, pilot, 869 (93%) of 939 patients were tested by RDT, of whom 290 (33%) London, United Kingdom, 4Center for Bioinformatics and Computational were positive and all reported successfully treated. Of 525 cases referred, Biology, University of Maryland, College Park, MD, United States, 5Armed there were 521 negative tests, two pregnant women, one infant <2 Forces Health Surveillance Center, Silver Spring, MD, United States, months, and one severe case. There were no deaths among these patients. 6Shoklo Malaria Research Unit, Mae Sod, Thailand, 7Institute of Specific A multi-dimensional evaluation was conducted, including a household Prophylaxis and Tropical Medicine, Medical University of Vienna, Vienna, survey and interviews with health officials and community members. Austria, 8Department of Molecular Tropical Medicine and Genetics, Faculty Perceptions were overwhelmingly positive among community members, of Tropical Medicine, Mahidol University, Bangkok, Thailand, 9Armed community and traditional leaders, health post personnel, and district and Forces Research Institute of Medical Sciences, Bangkok, Thailand, 10Center regional health officials. They recommended greater involvement of health for Parasitology, Entomology and Malaria Control, Pnom Penh, Cambodia, post chief nurses in the training, broadening HCP roles to include illnesses 11Institute for Genome Sciences, University of Maryland School of such as diarrhea, pneumonia, TB and malnutrition, reinforcing supervision Medicine, Baltimore, MD, United States, 12Genomics Unit, RML Research and support to the HCP, and extension of this strategy to all unserved, Technologies Section, Research Technologies Branch, Rocky Mountain remote villages in Senegal. Based on these recommendations, the pilot Laboratories, National Institute of Allergy and Infectious Diseases, National was expanded to 408 villages by the end of 2009 and 861 villages by the Institutes of Health, Hamilton, MT, United States, 13Malaria Programme, end of 2010. Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 14Menzies School of Health Research, Darwin, Australia, 15Malaria Functional Genomics Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 16Drug Resistance and Containment Unit, Global Malaria Programme, World Health Organization, Geneva, Switzerland Following reports of poor clinical responses to artemisinin combination therapy (ACT) on the Thailand-Cambodia border, the Artemisinin Resistance Confirmation, Characterization, and Containment (ARC3) pilot www.astmh.org 455 project was initiated to characterize the clinical, in vitro and molecular 1508 basis of artemisinin resistance in Southeast Asia. Four clinical trials of artesunate curative therapy were conducted at two sites in western SAFE AND EFFICACIOUS ARTEMISININ-BASED COMBINATION Cambodia where emerging resistance was suspected; one site on the TREATMENTS FOR AFRICAN PREGNANT WOMEN WITH Thailand-Myanmar border, where prolonged parasite clearance times MALARIA following artesunate-mefloquine treatment had also been reported; and 1 2 3 in Bangladesh, where ACTs have not been used extensively and resistance Umberto d’Alessandro , Halidou Tinto , Linda Kalilani-Phiri , 4 5 6 was not suspected. Parasites collected during these trials were genotyped Harry Tagbor , Michael Nambozi , Theonest Mutabingwa at approximately 8,000 single nucleotide polymorphisms (SNPs) using a 1The Institute of Tropical Medicine, Antwerp, Belgium, 2Institut de molecular inversion probe SNP chip specific to Plasmodium falciparum. Recherche en Sciences de la Sante´ (IRSS)-Center Muraz, Burkina Faso, Genotyping data from 198 samples were used to detect genomic regions Burkina Faso, 3University of Malawi College of Medicine, Blantyre, Malawi, under recent positive selection. Each SNP was scored using three Extended 4Kwame Nkrumah University of Science and Technology, Kumasi, Ghana, Haplotype Homozygosity measures including the Long-Range Haplotype 5Tropical Disease Research Center, Ndola, Zambia, 6Seattle Biomedical test, Integrated Haplotype Score, and Cross-Population Extended Research Institute, Seattle, WA, United States Haplotype Homozygosity. SNPs were also evaluated on their inter- Although malaria is the most important human parasitic disease, few population Fst values using a sliding window approach. The top 5% of loci studies with antimalarial drugs have been carried out in pregnant women. from each statistic were ranked and re-scored. A comprehensive list based Pregnant women are a high-risk group requiring effective antimalarials on the combined rank score revealed a number of novel regions possibly but they are systematically excluded from clinical trials for fear of under recent positive selection, as well as previously described regions teratogenicity and embryotoxicity. This has complicated evidence-based containing resistance genes such as pfcrt on chromosome 7 and dhfr and recommendations for the prevention and treatment of malaria during dhps on chromosomes 4 and 8, respectively. Regions identified as being pregnancy. A multicentre (Burkina Faso, Ghana, Malawi and Zambia), under recent positive selection will be compared to regions identified in a non-inferiority trial on the safety and efficacy of four artemisinin-based genome-wide association study performed using the same sample set, and combinations for the treatment of P. falciparum malaria in pregnant potential candidate genes within those regions will be discussed. women has recently started within the framework of the Malaria in Pregnancy Consortium and the financial support of both the European 1507 and Developing Countries Clinical Trials Partnership (EDCTP) and the COMPARATIVE EFFICACY AND ACCEPTABILITY Gates Foundation. Pregnant women in the second or third trimester OF ARTEMETHER-LUMEFANTRINE VERSUS of gestation and with a confirmed malaria infection are randomised to amodiaquine-artesunate, artemether-lumefantrine, dihydroartemisinin- DIHYDROARTEMISININ-PIPERAQUINE IN KENYAN CHILDREN piperaquine or mefloquine-artesunate. They will be followed up weekly WITH UNCOMPLICATED FALCIPARUM MALARIA until day 63 post-treatment and then monthly until 4-6 weeks and one Kevin O. Onyango1, John M. Ongecha2, Elizabeth Juma3, year post-delivery. Explanatory variables for failure, i.e. drug levels and in Godfrey A. Otieno1, Charles Obonyo2, Lucas Otieno1, Douglas J. vitro resistance of local malaria parasites are also collected. A total of 870 Perkins4, Willis Akhwale5, Bernhards Ogutu1 patients will be recruited to each treatment based on 290 patients in each treatment group in each country (i.e. a total centre sample size of 870 1Centre for Clinical Research, Kenya Medical Research Institute, Kisumu, patients), adding up to a total study sample size of 3480 patients. A 3-arm Kenya, 2Centre for Global Health Research, Kenya Medical Research trial using a “balanced incomplete block design” allow the treatments Institute, Kisumu, Kenya, 3Division of Malaria Control, Ministry of to be distributed in a way to allow a head-to-head comparison and the Public Health and Sanitation, Nairobi, Kenya, 4Centre for Global Health, establishment of relative value of the treatment according to a series of University of New Mexico, Albuquerque, NM, United States, 5Department outcomes. The primary end points are treatment failure (PCR adjusted) of Disease Prevention and Control, Ministry of Public Health and at day 28 and the safety profiles including significant changes in relevant Sanitation, Nairobi, Kenya laboratory values. More than 800 patients have been recruited so far. Artemisinin-based combination therapies (ACTs) have become the Results are expected by 2013. cornerstone for the treatment of uncomplicated falciparum malaria worldwide. In Africa, artemether-lumefantrine (AL) and artesunate- 1509 amodiaquine are the most widely used ACTs. AL has been the first-line treatment for uncomplicated malaria in Kenya since 2006. Despite PARASITE CLEARANCE OF ARTEMISININ-CONTAINING not yet receiving World Health Organization (WHO) prequalification, REGIMENS IN THE TREATMENT OF UNCOMPLICATED dihydroartemisinin-piperaquine (DP) has recently been adopted as a MALARIA: A SYSTEMATIC REVIEW OF PUBLISHED TRIALS second-line treatment in Kenya. This was an open-label, randomized, FROM 2000 TO 2010 comparative trial in children aged 6-59 months with uncomplicated 1 1 2 falciparum malaria conducted in Western Kenya. Parasite clearance rate, Debashish Das , Delia Bethell , Richard Cooksey , Finn 3 1 2 2 sensitivity to and acceptability of AL and DP were monitored. In total, 466 Andersen , Nuch Sapchookul , Patrice Piola , Philippe J. Guerin , 2 2 children were enrolled in the study; they were hospitalized for 3 days for Ric Price , Kasia Stepniewska observed treatment and 72-hour parasite kinetic monitoring, and actively 1WWARN Asia Regional Centre, Bangkok, Thailand, 2WWARN, University followed up at scheduled visits after discharge from hospital on Days of Oxford, Oxford, United Kingdom, 3Menzies School of Health Research, 7, 14, 28 and 42. Hemoglobin levels were assessed on Days 0, 14, 28 Charles Darwin University, Darwin, Northern Territory, Australia and 42. Genotyping for determining treatment outcome was performed The deployment of artemisinin-based combination therapies (ACTs) for on Day 0 and any other day the study participant had a recurrence of the treatment of acute uncomplicated falciparum malaria has become a parasitemia. The study drugs were administered by the parent/guardian key strategy for malaria control. The objective of this study is to review under the observation of a study team member. At discharge from the recent trends in clinical trials of artemisinins and document parasite hospital, a questionnaire on the acceptability of the study drug was clearance times. Publications reported in PubMed, EMBASE, GH and the administered to the parent/guardian. The findings of this study on the Cochrane Libraries between January 2000 and December 2010 were parasite clearance rate, sensitivity to and acceptability of AL and DP in the searched for the following key words: malaria, plasmodium, falciparum, treatment of uncomplicated malaria in Western Kenya will be discussed vivax, ovale, malariae, knowlesi and anti-malarials. 1520 abstracts were following completion of data analysis. indentified and 223 full text articles retrieved. In total 60% (n=96) of 160 clinical trials identified were conducted in Africa and 34% (n=54) in Asia.

www.astmh.org 456 Of the African countries, Nigeria hosted the maximum number of trials 1511 (n=14), whereas in Asia Thailand was the leading country (n=22). In total 138 (86%) studies were randomized and they included 309 treatment THE PHARMACOKINETICS AND PHARMACODYNAMICS arms with an ACT given over 1-7 days. The duration of follow up was at OF A TWO- VERSUS THREE-DAY DOSE REGIMEN OF least 42 days in 44 (28%) studies, 28 days in 107 (67%) studies and less DIHYDROARTEMISININ-PIPERAQUINE IN PATIENTS WITH than 28 days in 8 (5%) studies. A total of 47,855 patients were allocated UNCOMPLICATED MALARIA IN NORTHERN CAMBODIA an artemisinin-containing regimen in 160 trials with samples size ranging from 30 to 1553. The quality of clinical trials (as gauged by randomization, David Saunders1, Pattaraporn Vanachayangkul1, Chanthap Lon2, duration of follow-up, sample size) improved significantly in the second Stuart Tyner1, Raveewan Siripokasupkul1, Mashamon Mitprasat1, half of the decade compared to the first. The proportion of patients Suriya Teopipithaporn1, Youry Se2, Darapiseth Sea3, Sabaithip remaining parasitaemic at 48 and 72 hours could be determined in 125 Sriwichai1, Nillawan Buathong1, Soklyda Chann2, Nou Samon2, and 114 treatment arms, respectively. Following initiation of therapy the Douglas Walsh1, Bryan Smith4, Satharath Prom5, Kevin Leary4, median proportion of patients still parasitaemic at 48 and 72 hours was Duong Socheat3, Delia Bethell1, Paktiya Teja-isavadharm1 6.4% (range 0-73) and 0.5% (range 0-78) respectively. There was no 1Armed Forces Research Institute of Medical Sciences, Bangkok, significant change in the proportion of patients remaining parasitaemic at Thailand, 2Armed Forces Research Institute of Medical Sciences, Phnom 48 and 72 hours in studies conducted between 2000-2005 compared to Penh, Cambodia, 3National Center for Parasitology, Entomology and 2006-2010 (p=0.093 and p=0.215 respectively). Standardised approaches Malaria Control, Phnom Penh, Cambodia, 4U.S. Army Medical Materiel are urgently needed to define with greater precision geographical and Development Activity, Fort Detrick, MD, United States, 5Royal Cambodian temporal trends in the parasite clearance following ACTs; this will require Armed Forces, Phnom Penh, Cambodia analysis of pooled data from individual records. The combination of dihydroartemisinin (DHA) combined with long acting piperaquine (PIP) was recently adopted in Cambodia as the first line agent 1510 against both multi-drug resistant Plasmodium falciparum and P. vivax. IN VITRO CHARACTERIZATION OF THE METABOLISM AND In addition to well-documented safety and efficacy, a post-treatment DISPOSITION OF ARTESUNATE AND DIHYDROARTEMISININ prophylactic effect of DHA-PIP of up to 63 days has been reported, making it potentially valuable in malaria eradication efforts. While a Brandon Pybus, Jason C. Sousa, Xiannu Jin, Chau Vuong, Thulan 3-day course is widely recommended, the Cambodian military currently Luong, Vanessa Collazo, Ai J. Lin, Qigui Li, Michael P. Kozar, Victor employs a 2-day regimen in order to improve compliance. The aim of Melendez this study was to compare the safety, tolerability and pharmacokinetic- Walter Reed Army Institute of Research, Silver Spring, MD, United States pharmacodynamic relationships of a 2 versus 3 day dosing regimen of DHA-PIP using the same cumulative dose to establish the optimal IV Artesunate is currently approved for compassionate use in the US regimen. From September 2010 to February 2011, in an open-label through the CDC, and is seen as a promising alternative to the only FDA- clinical trial, 80 patients with uncomplicated malaria were randomized approved parenteral treatment of severe malaria in the United States, IV 1:1 to receive 9 fixed-dose combination tablets (total dose 320mg of quinidine gluconate. While an effective antimalarial treatment, quinidine DHA and 2880 mg PIP), divided into either a 2 or 3 day course. Plasma has potential serious cardiotoxicity. In support of efforts to obtain FDA piperaquine levels from all volunteers receiving DP were collected at pre- approval for IV artesunate, our lab has performed a battery of in vitro dose (time 0), 4, 24, 48, 72 hr, 7, 14, 21, 28, 35 and 42 days after the metabolism and disposition assays of artesunate and its major metabolite, first dose, and on day of recurrence. Sixteen patients were diagnosed dihydroartemisinin (DHA). Artesunate demonstrated a short (<10 min) with P.f. (20%), 61 with P.v. (76%) and 3 with mixed species infection microsomal half-life in both human and mouse liver microsomes, while (4%). Uncorrected 42-day efficacy rates were not statistically significantly DHA exceeded the assay limits of 60 min. In human hepatocytes, the different between treatment groups by per protocol analysis - 89% for predominant metabolite of artesunate, as expected, was DHA, along with 2 days (95% CI = 76-96%) and 92% for 3 days (95% CI = 80-97%) of its glucoronide. These data were confirmed in vivo using clinical samples DP. Mean parasite clearance times were 11.1 hours for P. vivax, but 72.5 previously analyzed in this lab. In enzyme inhibition studies, artesunate hours for P. falciparum. Piperaquine levels are currently being measured by demonstrated low potential for drug-drug interactions due to enzyme liquid chromatography-mass spectrometry. Pharmacokinetic data will be inhibition with any of the major CYP450s (1A2, 2C9, 2C19, 2D6, and presented, along with pharmacodynamic analysis of parasite clearance and 3A4) while DHA showed only moderate potential for inhibition of 1A2 and PCR-corrected treatment efficacy, with particular attention to drug levels 2C9. Permeability was tested using MDR1-MDCK to assess cell permeation at the time of failure and post-treatment prophylactic effect. and potential as a Pgp substrate. These experiments showed medium/ moderate permeability as compared to standards of known penetrability, and indicate a high potential as a Pgp substrate. The data support observations that AS and DHA are rapidly biotransformed and eliminated as the glucoronide of DHA. Evaluation of other potential metabolites is in progress.

www.astmh.org 457 1512 the effectiveness of DHA + PQ compared to AQ + SP in seasonal IPTc, 1500 children aged 3-59 months were randomized to receive DHA+PQ ARC3: ASSOCIATIONS BETWEEN CANDIDATE GENE or SP+AQ once a month from August to October 2009. A comparator POLYMORPHISMS AND PARASITE CLEARANCE RATE arm with 250 children was also recruited and followed up in parallel. A FOLLOWING TREATMENT WITH ARTEMISININS cross sectional survey was led at the end of transmission season. Primary endpoints were the incidence of clinical malaria attacks. Coverage of all 1 2 Shannon Takala Harrison , Mallika Imwong , Christopher three courses of IPT and compliance of children to daily doses of IPT was 1 3 4 5 Jacob , Cesar Arze , Arjen Dondorp , Mark Fukuda , Francois similar in the two arms. 83% of children in each arm received at least the 6 7 8 8 8 Nosten , Harald Noedl , Delia Bethell , Youry Se , Chanthap Lon , first dose of the 3 courses of IPT and 99% to 100% of them completed 8 8 9 Stuart Tyner , David Saunders , Duong Socheat , Aung Pyae the entire three doses regimen. A total of 328 episodes of clinical malaria 6 7 7 10 Phyo , Peter Starzengruber , Paul Swoboda , Kasia Stepniewska , with any parasitemia (incidence rate (IR) of 0,047(95%IC=[0,042 to 10 3 3 Jennifer Flegg , Gustavo Cerqueira , Joana Silva , Matthew 0,052] episodes per child day at risk (CD)) was observed in the control 1 1 1 1 Adams , Leo Kenefic , Jason Bailey , Amadou Niangaly , N. arm compared to 205 (IR of 0,003(95% IC= [0,002 to 0,003] episodes per 4 11 1 White , P. Ringwald , C. Plowe CD) in DHA+PQ arm and to 147 (IR of 0,002(95% IC= [0,001 to 0,002] 1Howard Hughes Medical Institute/Center for Vaccine Development, episodes per CD) in SP+AQ arm, indicating a protective efficacy (Pe) of University of Maryland School of Medicine, Baltimore, MD, United States, 93% (95% IC= [92 to 94] p<0.0001) for DHA+PQ versus 95% (95% IC 2Department of Molecular Tropical Medicine and Genetics, Faculty of = [94 to 96] p= 0.0001) for SP+AQ. Children in SP+AQ arm were better Tropical Medicine, Mahidol University, Bangkok, Thailand, 3Institute for protected against malaria attacks than those in DHA+PQ arm (Pe = 28% Genome Sciences, University of Maryland School of Medicine, Baltimore, (95% IC=[10 to 42] p= 0.003). At the end of the transmission season, MD, United States, 4Mahidol Oxford Research Unit, Bangkok, Thailand, only SP + AQ retained a substantial efficacy (Pe = 62% (95% CI = [32-78] 5Armed Forces Health Surveillance Center, Silver Spring, MD, United States, p= 0.001) against malaria attacks. The prevalence of parasitaemia was 6Shoklo Malaria Research Unit, Mae Sod, Thailand, 7Institute of Specific reduced by 67%, in the two treated arms. The prevalence of anemia (Hb< Prophylaxis and Tropical Medicine, Medical University of Vienna, Vienna, 10g/dl) was similar in DHA+PQ (40%) and SP+AQ (35.34%) arms with a Austria, 8Armed Forces Research Institute of Medical Sciences, Bangkok, respective Pe of 17% (95% CI = [3 to 29], p = 0.026), and 27% (95% CI Thailand, 9Center for Parasitology, Entomology and Malaria Control, Pnom = [14 to 27], p = 0.001). The mean Hb in SP + AQ arm was significantly Penh, Cambodia, 10World-Wide Antimalarial Resistance Network, Oxford higher than the DHA + PQ group and control group. In conclusion, SP+AQ University, Oxford, United Kingdom, 11Drug Resistance and Containment was the most efficacious for IPTc in our study but its use must be reserved Unit, Global Malaria Programme, World Health Organization, Geneva, to area with low-level SP resistance otherwise DHA+PQ would be a Switzerland suitable alternative. Nearly all malaria-endemic countries have replaced resistance- compromised antimalarial drugs with artemisinin-based combination 1514 therapy, the efficacy of which is also at risk due to the recent emergence A COMPARISON BETWEEN EX VIVO AND IN VITRO of artemisinin-resistant falciparum malaria on the Thailand-Cambodia (CRYOPRESERVED + CULTURE ADAPTED) ARTEMISININ border. Without knowledge of the mechanism of action of the SUSCEPTIBILITIES FOR Plasmodium falciparum ISOLATES artemisinins, genome-wide approaches are being taken to identify loci associated with resistance to this class of drugs. In addition, candidate FROM WESTERN CAMBODIA genes that have been reported in the literature as possibly being Suwanna Chaorattanakawee associated with artemisinin resistance are being evaluated. Full-length Armed Forces Research Insititute of Medical Sciences, Bangkok, Thailand sequences were determined for three candidate genes in Plasmodium falciparum infections in four clinical trials of 7-day curative artesunate Artemisinin derivatives are the most potent first line antimalarial drug therapy conducted as part of the Artemisinin Resistance Confirmation, in use today. However, recent studies along the Thai-Cambodian border Characterization, and Containment (ARC3) pilot project. The sequenced conducted during the past 5 years demonstrate that Plasmodium loci include: pfmdr1, a gene associated with resistance to other falciparum may be becoming resistant to the artemisinin class. Since antimalarial drugs, pfserca, a gene encoding a sarco/endoplasmic clear genetic markers of artemisinin resistance have not been discovered, reticulum calcium-dependent ATPase, and pfubp1, coding for an ortholog in vitro parasite drug susceptibility testing may be helpful to identify of a deubiquitinating enzyme associated with artemisinin resistance in resistance trends. In many in vitro drug sensivity studies, samples are rodent malaria. Polymorphisms in each of these genes, as well as variations collected from untreated subjects in the field and cryopreserved to in copy number estimates of pfmdr1, were evaluated for associations with facilitate sample transport. Because such samples must be thawed and parasite clearance rate and will be presented in the context of results from recultured, the IC50 measured after recovery may diverge from the ex genome-wide analyses from the same studies. Strategies for prioritization vivo IC50. During a dose ranging randomized open label clinical trial of newly identified candidate genes will be discussed. of a 7 day course of mono-therapy artesunate (Artemisinin Resistance in Cambodia - 2; 2008-2009) parasite samples were analyzed ex vivo, 1513 without cryopreservation or culture adaptation, for susceptibility to dihydroartemisinin, artesunate, and other anti-malarial drugs using a

EFFICACY OF DIHYDROARTEMISININ PLUS PIPERAQUIN histidine-rich protein-2 (HRP2) ELISA method. IC50 values obtained for COMPARED TO AMODIAQUIN PLUS SULFADOXIN/ the artemisinins (DHA and AS) did not correlate with treatment outcome. PYRIMETHAMIN IN SEASONAL IPT OF MALARIA IN However, the values were two-fold higher than those obtained in an CHILDREN IN A RURAL AREA OF BOBO-DIOULASSO earlier study (Artemisinin Resistance in Cambodia - 1, 2006-2007) (BURKINA FASO) conducted at the same location. This increase suggests that falciparum parasites in this location are progressively more tolerant to the artemisinin Yves-Daniel Compaoré, Fabrice Some, Issaka Zongo, Noel class of antimalarials. However, we and others have not determined if this Rouamba, Jean Bosco Ouedraogo tolerance is maintained in culture in the absence of drug pressure. We will IRSS/DRO Bobo-Dioulasso, Bobo-Dioulasso, Burkina Faso report the in vitro analysis of cryopreserved parasites and compare them to matching uncryopreserved ex vivo results. Parasite susceptibility to other Intermittent preventive treatment (IPT) is a promising malaria control anti-malarial drugs and change in parasite genetic constituent will also strategy. Optimal regimen and the best option for his delivery remain be described. These results will be helpful to inform future surveillance unclear. The long elimination half-life of Piperaquine (PQ) makes its coformulation with dihydroartemisinin (DHA), suitable for IPT. To assess

www.astmh.org 458 investigating the problem of artemisinin resistance, particularly whether infections. We recruited a total of 610 children aged 6 to 59 months, resistance IC50 phenotypes can be preserved through cryopreservation with uncomplicated P. falciparum malaria and followed them for 28 days and recovery. to assess treatment efficacy. Baseline characteristics were similar in both treatment groups. The analysis revealed no early therapeutic failures (ETF) 1515 in both arms; late clinical failures (LCF) were 1.7% for SP+AS (n=5) vs. 0% SP+AQ (n=0) and late parasitological failures (LPF) were 3.4% SP+AS

INCREASE IN EX VIVO IC50 VALUES TO ARTEMISININS (n=10) vs. 1.4% SP+AQ (n=4) (p>0.05). We observed a rate adequate FROM A SEVEN-DAY ARTEMISININ MONO-THERAPY TRIAL clinical and parasitological response (ACPR) of 94.9% and 98.6% for CONDUCTED IN WESTERN CAMBODIA IS NOT DUE TO SP+AS and SP+AQ respectively (p=0.98). Based on msp2 analysis, the INOCULUM EFFECT rate of re-infection was respectively 4.1% and 1.4% for SP+AS and SP+AQ. After molecular correction, we obtained an ACPR of 99% for 1 2 Stuart D. Tyner , Wiriya Ruttvisutinunt , Kurt Schaecher E. SP+AS, and 100% for SP+AQ (p=0.98). Sulphadoxine-pyrimethamine plus 3 1 1 Schaecher , David L. Saunders , Suwanna Chaorattanakawee , amodiaquine therapy is as efficacious as sulphadoxine-pyrimethamine 1 1 1 Kritsanai Yingyuen , Panjaporn Chaichana , Sittidech Surasri , plus artesunate in the treatment of uncomplicated P. falciparum malaria in 1 4 1 1 Delia Bethell , Duong Socheat , Youry Se , Chanthap Lon , Mark Mali. M. Fukuda5, Douglas S. Walsh1 1Department of Immunology and Medicine, Armed Forces Research 1517 Institute of Medical Sciences, Bangkok, Thailand, 2Department of Retrovirology, Armed Forces Research Institute of Medical Sciences, STUDY OF PLASMODIUM VIVAX DUFFY BINDING PROTEIN Bangkok, Thailand, 39th Area Medical Laboratory, Aberdeen Proving (DBP) OF EASTERN INDONESIAN ISOLATES 4 Ground, MD, United States, National Center for Parasitology, Entomology 1 1 1 5 Andreas Kusuma , Retno Ayu Utami , Leily Trianty , Hidayat and Malaria Control, Phnom Penh, Cambodia, Armed Forces Health 1 2 2 1 Science Center, Silver Spring, MD, United States Trimarsanto , Nicholas Anstey , Richard Price , Rintis Noviyanti , John Reeder3 Studies along the Thai-Cambodian border conducted during the past 5 1Eijkman Institute for Molecular Biology, Jakarta, Indonesia, 2International years demonstrate that P. falciparum may be developing resistance to Health Division, Menzies School of Health Research and Charles Darwin the artemisinin class of antimalarial drugs, the last first-line antimalarial University, Darwin, Northern Territory, Australia, 3Burnet Institute, compounds in use today. Persistence of parasitemia beyond 72 hours Melbourne, Australia has been proposed as a parasitological marker of diminished artemisinin susceptibility. During a dose ranging randomized open label clinical trial Erythrocyte invasion by malaria parasites is a series of complex processes of a 7 day course of mono-therapy artesunate (Artemisinin Resistance and specific interaction between parasite ligands and host receptors in Cambodia - 2; 2008-2009) parasite samples obtained pre-treatment play a key role. Plasmodium vivax uses the critical ligand Duffy Binding (on Day 0) were analyzed ex vivo, without cryopreservation or culture Protein (DBP), to bind its host receptor, Duffy Antigen Receptor for adaptation, for susceptibility to dihydroartemisinin, artesunate, and other Chemokines (DARC), and initiate the formation of tight junction between anti-malarial drugs using a histidine-rich protein-2 (HRP2) ELISA method. merozoite and erythrocyte essential for invasion. The central domain IC50 values obtained for the artemisinins (DHA and AS) did not correlate of Domain II of DBP serves as the critical binding motif to the DARC. with treatment outcome. However, there was a statistically significant However, the genetic diversity of PvDBPII is a major obstacle to vaccine relationship between parasite clearance time greater than 72 hours (PCT development. In order to develop a rational vaccine, more information > 72hrs) and elevated IC50 values for DHA (p<0.003). Previous reports about the sequence polymorphisms of PvDBPII is required, from diverse have implicated an inoculum effect, or the parasitemia of the isolate, as a geographical regions. The current study aims to determine the extent of possible reason for elevated IC50 values against the artemisinins. Here we polymorphisms of PvDBPII from Timika, West Papua, Indonesia and to argue that the association between DHA IC50 value and PCT>72hrs may examine the relationships of different dbp alleles with published sequences not be solely due to inoculum effect, and may represent the early stages through phylogenetic analysis. 95 P. vivax isolates were collected from of in vitro resistance to the artemisinins. the hospital-based study in Timika from 2006-2008. These DNA samples were subjected to PCR-RFLP using Pvmsp-3α to assess its genetic 1516 polymorphisms. Some samples were selected and further subjected to cloning and sequencing to examine the PvDBPII sequence polymorphisms. A RANDOMIZED CLINICAL TRIAL OF ARTEMISININ VERSUS A phylogenetic tree was constructed based on the PvDBPII sequences NON-ARTEMISININ-BASED COMBINATION THERAPY OF obtained in this study and combined with the published sequences using UNCOMPLICATED MALARIA IN MALI Bayesian inference. A total of 101 DBP point-mutations were found from the Timika isolates. Mutations mainly occurred in the critical binding motif 1 2 1 Hamma Maiga , Abdoulhabib Beavogui , Ousmane Toure , of PvDBPII. Most mutations (81%) were non-synonymous; altering amino 1 1 1 Mamadou Tekete , Check Oumar Papa Sangare , Antoine Dara , acids sequence and only about a fifth (23%) were synonymous. R308S, 1 1 1 Zoumana Isaac Traore , Oumar Bila Traore , Souleymane Dama , K371E, D384G, R390H, N417K, L424I, W437R, I503K were the highest 1 1 1 Christelle N’Dong , Hamidou Niangaly , Nouhoum Diallo , Demba frequency polymorphisms seen. In total, about 51 different haplotypes of 1 1 1 Dembele , Ogobara Doumbo , Abdoulaye Djimde PvDBPII were observed. More haplotypes might be found if more clones 1Malaria Research and Training Center, Bamako, Mali, 2Centre de sante de are sequenced. The phylogenetic tree showed that Timika PvDBPII isolates Recherche de maferenya, Conakry, Guinea clustered together with isolates from other geographic regions, including Brazil, Sri Lanka, and Papua New Guinea. Although the PvDBPII sequences Plasmodium falciparum resistance to artemisinin has been reported in of Timika isolates are quite diverse as shown by the number of haplotypes South-East Asia. The potential spread of this resistance is real and makes detected and the topology of the phylogenetic tree, the similarities a search for alternative non-artemisinin-based malaria therapy urgent. We between those sequences are still high. These findings certainly contribute tested the hypothesis that sulphadoxine-pyrimethamine plus artesunate to the development of PvDBPII vaccines against P. vivax infections, and (SP+AS) is as efficacious as sulphadoxine-pyrimethamine plus amodiaquine thus demonstrate its public health significance. (SP+AQ) in the treatment of uncomplicated Plasmodium falciparum malaria. From August to December 2004 and July to December 2005, we conducted a randomized single-blind trial of SP+AS and SP+AQ in two localities in Mali. Parasite genotyping by polymerase chain reaction (PCR) was used to distinguish new from recrudescent P. falciparum

www.astmh.org 459 1518 1520 FREQUENCIES OF SOME HUMAN GENETIC MARKERS AND RETICULOCYTES DERIVED FROM HEMATOPOIETIC STEM RELATIONSHIP WITH PLASMODIUM FALCIPARUM AND P. CELLS CAN BE SUCCESSFULLY CRYOPRESERVED TO BE USED VIVAX MALARIA IN COLOMBIA FOR PLASMODIUM VIVAX INVASION TESTS Lina Gonzalez, Jorge Vega, Jose L. Ramirez, Gabriel Bedoya, Jaime Florian Noulin Carmona-Fonseca, Amanda Maestre Antwerp Institute of Tropical Medicine, Antwerp, Belgium Universidad de Antioquia, Medellin, Colombia The differentiation of hematopoietic stem cells (HCS) into reticulocytes has Malaria in Colombia is highly endemic in the Northwest, Pacific Coast and been previously described as potentially interesting for the development Amazon regions. Despite the high frequencies of Plasmodium falciparum of the Plasmodium vivax in vitro culture. Nevertheless, the need of using infection, severe or fatal malaria cases are rare. Out of 79.909 malaria both freshly derived reticulocytes and fresh P. vivax isolates remained an cases (72% P. vivax - 27% P. falciparum) reported in 2009, only 307 obstacle, particularly for laboratories located in non-endemic countries. were severe (1,4% of falciparum cases) with 0,04% fatality rate. Some We describe a new method for the cryopreservation of reticulocytes proposed that P. falciparum shaped the distribution of ABO blood groups produced after 14 days of culture from HSCs. Invasions assays have been in humans with some groups being protective against severe disease. The carried out with both frozen isolates of P. vivax as well as laboratory Duffy blood group A has marked selective and sickle-cell disease, G-6-PD strains of P. falciparum. Cryopreserved P. falciparum and P. vivax isolates deficiency a more discrete one. Aimed at understanding the blood genetic could equally invade fresh and cryopreserved reticulocytes. This new factors underlying the apparent protection to development of severe technique represents an important advance towards the establishment of disease within a falciparum malaria infected community, we explored the a continuous P. vivax culture as it allows the storage of large quantities of ethnic background, blood types and Plasmodium infection in La Italia on reticulocytes to be later used for the invasion. the Pacific Coast. A descriptive, cross-sectional study in a deprived and mainly rural region was conducted. Sample sizes were Afro-American, 1521 73; Amerindian (Emberá), 74 and Mestizo, 171. Presence of Plasmodium infection was assessed by thick smear, ABO and Rh were determined by BIOMECHANICAL AND NANOSTRUCTUAL CHANGES TO agglutination and Duffy status by PCR and RFLP. For ABO, 69% were O, THE PLASMODIUM VIVAX INFECTED RED BLOOD CELL 21% A, 8% B and 2% AB. A significant association was observed for ABO MEMBRANE status and ethnicity: 100% of Amerindias were group O. Similarly, Duffy 1 2 3 4 genotypes were significantly associated to ethnicity (p=0,003). Expression Ang Li , Bruce Russell , Francois Nosten , Usa Lek-Uthai , 2 5 5 of Rh was confirmed in 97-99% of all subjects, regardless of the ethnic Rossarin Suwanarusk , Mary Ng , Kay En Low , Kanlaya 3 2 2 2 background. For Duffy, the C/C,A/A diplotype carriers were exclusively Sriprawat , Esther G. Koh , Carla Claser , Benoit Malleret , Céline 6 7 2 1 infected by P. falciparum. At locus 131, the frequency of the G allele Borlon , Georges Snounou , Laurent Renia , Lim Chwee Teck was 0,30 in Amerindians and the A allele was 0,69 in Afrocolombians. 1Division of Bioengineering and Department of Mechanical Engineering, Plasmodium infection was confirmed in 17% (falciparum 2,2: 1 vivax), National University of Singapore, Singapore, Singapore, 2Singapore 33% were asymptomatic and 3,1% of falciparum cases developed severe Immunology Network, A*STAR, Singapore, Singapore, 3Shoklo Malaria infection. In conclusion, a very high frequency of group O was confirmed Research Unit, Mae Sod, Thailand, 4Department of Parasitology and in a locality where falciparum infection is frequent. High endogamy and Entomology, Faculty of Public Health, Mahidol University, Bangkok, population differentiation was confirmed after Duffy genotyping. Infection Thailand, 5Electron Microscopy Unit, National University of Singapore, by P. vivax was not detected in CC(FY-/FY-) individuals. Singapore, Singapore, 6Department of Parasitology, Institute of Tropical Medicine, Antwerp, Belgium, 7INSERM UMR S 945, Département de 1519 Parasitologie, Hôpital Pitié-Salpêtrière, Paris, France The infection of reticulocytes by Plasmodium vivax results in significant CRYOPRESERVED PLASMODIUM VIVAX AND changes to the infected red cell membrane (IRBCM). Most importantly, the RETICULOCYTES CAN BE USED FOR INVASION AND SHORT P. vivax IRBCM becomes highly deformable relative to membranes of the TERM CULTURE reticulocyte and mature normocyte. We present a range of data clearly Céline Borlon1, Bruce Russell2, Kanlaya Sriprawat3, Annette demonstrating these biomechanical changes to the IRBCM using single Erhart1, Laurent Renia2, François Nosten3, Umberto D’Alessandro1 cell and (micopipette, microfuidics) and cell population measurements (LORCA). Although the mechanism behind these changes is not fully 1Institute of Tropical Medicine, Antwerp, Belgium, 2Laboratory of Malaria understood, increased deformability would aid P. vivax avoid splenic Immunobiology, Singapore, Singapore, 3Shoklo Malaria Reasearch Unit, clearance. In addition to these biomechanical changes we examined MaeSot, Thailand the development of ‘caveolae’ on the surface of the IRBCM in staged The development of a Plasmodium vivax in vitro culture system is critical ex vivo cultures of P. vivax IRBCs, using TEM, SEM and Atomic Force for the development of new vaccine, drugs and diagnostic tests. Though Microscopy. We show that caveolae-like depressions found in the host short term cultures have been successfully set up, their reproducibility in cells (reticulocytes) and P. vivax IRBCs are morphologically and biologically laboratories without direct access to P. vivax patients has been limited by distinct, thus unrelated. We also show that ‘Schüffner’s dots’, a classical the need of fresh parasites isolates. We have explored the possibility of diagnostic feature of P. vivax, is associated with parasite derived vesicle using both frozen parasite isolates and frozen reticulocytes to perform complexes and not caveolae. invasions and start a short term culture. More than 50 invasion tests were performed. Invasion could be performed with similar efficiency for any of the combination (fresh/frozen reticulocytes and P. vivax isolates) used. This method should be easily replicated in laboratories outside endemic areas and can substantially contribute to the development of a continuous P. vivax culture.

www.astmh.org 460 1522 infected individuals lived in a forested B’laan tribal community in Maasim, Sarangani which was 10-12 km from formal health systems, while 6 of DETECTION OF PLASMODIUM VIVAX PRE-ERYTHROCYTIC the 10 P. vivax-infected individuals lived in a forest fringed T’boli tribe STAGE USING A ROLLING CIRCLE AMPLIFICATION (RCA) community in Kiamba, Sarangani. These infections would have been ASSAY undetected and untreated as no clinical signs of malaria were evident. Our findings to date indicate pockets of ongoing malaria transmission despite Achareeya Korkusol, Sirima Mingmongkolchai, Namtip current control measures, which hamper malaria elimination efforts; Trongnipatt, Ratawan Ubalee, Brian P. Evans, Ratree Takhampunya and the sensitivity of the Pf/Pv combination RDT is insufficient to detect Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand asymptomatic P. vivax infections. On completion of PCR screening, we plan to calculate the comparative performance of the RDT versus PCR in A rolling circle amplification (RCA) assay was developed to detect the pre- diagnosing asymptomatic P. vivax infections in Southern Philippines. We erythrocytic stage in liver cells infected with Plasmodium vivax. The circular will also complete serological assays for exposure to Pf/Pv infections on probe (98bp) in combination with a fluorescence DNA detector probe was the same samples. Together with PCR and questionnaire data we aim to designed to have a complementary sequence (20bp in length) to malaria build a complete epidemiologic profile of malaria in Sarangani and guide 18S rDNA at the 5’ and 3’ends of the probe. The RCA technique was intervention measures to halt transmission. previously developed and applied to detect several pathogens in order to increase detection sensitivity in clinical samples. The target sequence amplification step uses phi 29 or Bst enzymes to enhance the intensity 1524 of the detection signal in the RCA assay. Our results showed that the in A MALARIA DIAGNOSTICS TO BE USED IN THE FIELD: vitro RCA assay for the detection of P. vivax resulted in a fluorescent signal VISUALIZED LOOP-MEDIATED ISOTHERMAL AMPLIFICATION significantly higher relative to the signal resulting from the real-time PCR (LAMP) FOR DETECTION OF PLASMODIUM VIVAX INFECTION method (8.0 versus 0.5 fluorescent intensity). However, the RCA assay required more time and additional steps when compared to the real-time Jun Cao1, Zhi-Yong Tao2, Qi Gao1 PCR assay. In situ RCA was established using an infected blood smear 1Jiangsu Institute of Parasitic Diseases, Wuxi, China, 2Medical College of (thick film) collected from a P. vivax-infected patient and with P. berghei- Soochow University, Suzhou, China infected HepG2 cells collected at 48h post-infection. An infected cell in a blood smear generated a strong RCA signal although the number of Loop-mediated isothermal amplification (LAMP) is a high performance positive cells was lower than that observed with immunofluorescence method for detecting DNA and is of high potential usage in the molecular staining using a heat shock protein 70 (HSP70) monoclonal antibody. In detection of infectious pathogens including Plasmodium spp. However, a liver cell culture infected with P. berghei, the infected cells generated in most malaria endemic areas, which are often resource-limited, current a strong RCA signal and all infected cells were confirmed by an LAMP methods are not feasible for diagnosis due to difficulties in immunofluorescence signal. In conclusion, the RCA technique can detect accurately interpreting results with problems of sensitive visualization P. vivax in infected liver cells even where there are low infection rates and of amplified products, and the high risk of contamination resulting therefore such a technique will allow researchers to better understand the from the high numbers of amplified DNA sequences produced. In this biology of P. vivax in liver cells. study we establish a novel visualized LAMP method in a closed-tube system, and validate it for the diagnosis of malaria in a simulated field 1523 condition. A visualized LAMP method was established by the addition of a microcrystalline wax-dye capsule containing the highly sensitive DNA ASYMPTOMATIC PLASMODIUM VIVAX INFECTIONS IN fluorescence dye SYBR Green I to a normal LAMP reaction prior to the SOUTHERN MINDANAO, PHILIPPINES: A CHALLENGE TO initiation of the reaction. The wax remained intact during isothermal MALARIA ELIMINATION amplification, and released the DNA dye to the reaction mixture only when the temperature was raised to the melting point following amplification 1 1 2 Mary Grace B. Dacuma , Rachel Hallett , Judeline Dimalibot , Soon after cooling down, the solidified wax sealed the reaction mix at the 2 3 4 George Ugaddan , Federico Yadao , Walter Notario bottom of the tube, thus minimizing the risk of aerosol contamination. 1London School of Hygiene and Tropical Medicine, London, United A total of 89 of field blood samples were collected on filter paper and Kingdom, 2University of the Philippines Los Baños, Los Baños, Philippines, processed using a simple boiling method for DNA extraction. This was 3Malaria Control and Prevention, Provincial Health Office, Sarangani then tested by the visualized LAMP method. Compared to microscopy, the Province, Philippines, 4Pilipinas Shell Foundation Sarangani Province, sensitivity and specificity of LAMP were 98.3% (95% CI, 91.1% to 99.7%) Sarangani Province, Philippines and 100%, and were in close agreement with a nested PCR method. In conclusion, this novel, cheap and quick visualized LAMP method is feasible The Philippines is among the 39 countries progressing towards malaria for malaria diagnosis in resource limited field settings. elimination. The country shifted to artemether-lumefantrine in 2009, dispensed insecticide-treated nets, and conducted indoor residual spraying to address malaria in 58 out of 80 endemic provinces. However, in low transmission settings asymptomatic infections especially with Plasmodium vivax can hamper elimination efforts when parasitemia is too low for detection by microscopy in health clinics. In this study, we compared the performance of pLDH/HRP2-based rapid diagnostic test (RDT) for P. falciparum and P. vivax with PCR in detecting asymptomatic infections. Using the RDT in a cross-sectional survey in Sarangani Province to screen 930 participants (aged 12 mos. and above), we detected one P. vivax infection (a prevalence of 0.10%) and 10 P. falciparum infections (1.08%) all of whom were asymptomatic upon presentation. We also collected blood spots on filter paper for molecular and serological assays. To date, we have screened 700 out of 930 blood spots by PCR using species-specific primers for the 18SrRNA gene of Plasmodium. PCR results showed 7 participants (1%) were infected with P. falciparum (4 of whom were RDT negative) while 10 participants (1.4%) were infected with P. vivax, 9 of whom were RDT negative. Four of the 7 P. falciparum-

www.astmh.org 461 1525 with GIS-mapping. Detection of Plasmodium species using microscopy, RDT and Real-time PCR was performed for the active and passive case REAL TIME LOOP-MEDIATED ISOTHERMAL AMPLIFICATION detection. Microscopy, RDT, and real-time PCR in the active surveillance (RealAmp) FOR THE SPECIES-SPECIFIC IDENTIFICATION OF showed approximately 2% positive rates by microscopy or RDT in nearly PLASMODIUM VIVAX 500 individuals. A real-time PCR assay detected the prevalence of 6%. The sensitivity of the RT-PCR in the 96-well format was increased to 10- 1 2 3 Jaymin Patel , Jenna Oberstaller , Mitra Poorak , Maniphet 100 parasites/µl with a glycogen/acetate DNA precipitation at low-speed 1 1 2 Xayavong , Jothikumar Narayanan , Jeremy DeBarry , Ganesh tabletop centrifugation after column extraction. P. vivax and P. malariae 2 4 5 Srinivasamoorthy , Leopoldo Villegas , Ananias A. Escalante , were detected in less than 5% of the malaria-positive patient samples by 1 2 1 Alexandre DaSilva , David S. Peterson , John Barnwell , Jessica RT-PCR. All the P. falciparum isolates were chloroquine-resistant PfCRT 2 1 3 Kissinger , Venkatachalam Udhayakumar , Naomi W. Lucchi K76T genotype and atovaquone-sensitive PfCYTb 268Y by fluorescent 1Division of Parasitic Diseases and Malaria, Center for Global Health, TAQman probe analysis. A reverse transcriptase real-time PCR assay from Centers for Disease Control and Prevention, Atlanta, GA, United States, dried blood on filter papers was able to detect gametocytes. HRP2 antigen 2University of Georgia, Athens, GA, United States, 3Atlanta Research detection by RDT persisted up to 28 days in more than 20% of malaria and Education Foundation, Atlanta, GA, United States, 4Asociación Civil cases in a density-dependent fashion. The geometric mean parasitaemia Impacto Social, Tumeremo, Bolivarian Republic of Venezuela, 5Arizona was 227; 1,342, 5,412, and 10,716 for persistent RDT-positives through State University, Tempe, AZ, United States Day 0; Day 2; Day 7, and Day 28 respectively. Both microscopy and PCR detection diminished in a few days. Studies on malaria seropositivity rates Plasmodium vivax infections remain a major source of malaria-related are in progress. Most of the malaria-positive cases clustered in less than morbidity and mortality. Early and accurate diagnosis is an integral half of the population. Significant asymptomatic populations PCR-positive, component of effective malaria control programs. Conventional molecular RDT and microscopy negative malaria exist. The role of this sub-population diagnostic methods provide accurate results but are often resource- in contributing to continuing transmission is being evaluated. Passive intensive, expensive, have a long turn around time and are beyond surveillance detected less than half of the malaria infections. the capacity of most malaria-endemic countries. Our laboratory has recently developed a new platform called RealAmp, which combines loop-mediated isothermal amplification (LAMP) with a portable tube 1527 scanner real-time isothermal instrument for the rapid detection of malaria EFFICACY OF CHLOROQUINE FOR TREATMENT OF VIVAX parasites. This method was initially tested using Plasmodium genus- MALARIA IN CENTRAL CHINA specific primers, but species-specific diagnosis was not accomplished as there were no primer sets that gave consistent results. Here we describe a Guoding Zhu1, Qi Gao1, Huayun Zhou1, Jun Cao1, Yaobao Liu1, new primer set for the detection of P. vivax. The LAMP assay was designed Yuee Tang2 using three pairs of amplification primers targeting a conserved DNA 1Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for sequence unique to the P. vivax genome using an algorithm we have Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Key developed for genome mining. The amplification was carried out at 64ºC Lab on Molecular Biology of Parasites, Wuxi, China, 2Center for Diseases using SYBR Green intercalating dye for 90 minutes with the tube scanner Control and Prevention, Suining County, Xuzhou, China set to collect fluorescence signals at 1-minute intervals. Clinical samples of P. vivax and other human-infecting malaria parasite species were used Therapeutic efficacy studies allow measurement of the clinical and retrospectively to determine the sensitivity and specificity of the primers parasitological efficacy of medicines and the detection of subtle changes using the 18s ribosomal DNA based nested PCR as the gold standard. in treatment outcome when monitored consistently over time, which The new set of primers showed promising results in detecting laboratory- was considered the gold standard for determining antimalarial drug maintained isolates of P. vivax from different parts of the world. The time efficacy, and their results are the primary data used by national malaria to amplification ranged from 18 to 49 minutes from the start of the control programmes to make treatment policy decisions. This study reaction. The primers detected P. vivax in the clinical samples with 89.29% was to measure the efficacy of Chloroquine (CQ) for treatment of vivax sensitivity and 100% specificity compared to the gold standard nested malaria, and evaluate the incidence of adverse events.From June to PCR method. The new primers also proved to be more sensitive than the October of 2008 and 2009, thirty-eight patients, who met the inclusion published species-specific primers in detecting P. vivax. Further validation criteria, and infected with uncomplicated vivax malaria, confirmed by of this test using prospective testing in endemic countries will help deploy two qualified microscopists in Suining county of central China, were this tool for future field use. enrolled. CQ was administered at the dose of 25 mg base/kg body weight over 3 days(day 0,1,2), and after 28 days follow-up, primaquine was 1526 administered of 0.25mg/kg body weight, taken with food once daily for 14days.The follow-up consisted of a fixed schedule of check-up visits and DETECTION OF ASYMPTOMATIC PLASMODIUM corresponding clinical and laboratory examinations including thick and FALCIPARUM, P. VIVAX AND P. MALARIAE DURING thin blood films for parasite count, auxiliary temperature and adverse LOW-TRANSMISSION SEASON IN THE HILL TRACTS OF events. Patients were classified as therapeutic failures or treatment success BANGLADESH based on the assessments. It was observed that thirty-seven patients were treatment success over time, no fever and parasitemia were occurred on Jasmin Akter1, Sabeena Ahmed1, Chai Prue1, Wasif A. Khan1, day 2 and only one patient with 200/ml parasitemia on day 3. Except the David A. Sack2, Myaing Nyunt2, Gregory Glass2, Timothy Shields2, normal symptoms including headache, unwell, no severe adverse effect Rashidul Haque1, David Sullivan2 was reported. It was concluded that CQ is still effective and safe efficacy, 1International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, there was no obvious data to support the CQ resistance to Plasmodium Bangladesh, 2Johns Hopkins Malaria Research Institute, Baltimore, MD, vivax at present. Nevertheless, according to the discovering in this study, United States the initiative diagnosis technique and method should be developed to detect the possible changing pattern and stage of CQ to Plasmodium During the low transmission season many malaria infections are vivax, to formulate recommendations and to enable the Ministry of Health asymptomatic. To investigate how best to target asymptomatic individuals, and to make informed decisions about the possible need for updating an active randomized, population-based malaria surveillance was initiated of the current national antimalarial treatment guidelines. This study by the JHMRI and ICDDR,B in two unions near hypoendemic Bandabarn in was approved by the national ethical committee, and all the patients the Chittagong Hill Tracts of Bangladesh. The population of 20,000 with signed the informed consent, all information regarding the patients was approximately 4,500 households was enumerated in a baseline census remaining confidential within the study team.

www.astmh.org 462 1528 malaria of which resistance against CQ has never been reported so far. However, all the Philippine isolates (collected in Palawan island in 2009) STAGE SPECIFIC DRUG ACTIVITY OF CHLOROQUINE IN acquired the Y976F mutation. This finding suggests that CQ resistant PLASMODIUM VIVAX MALARIA: IMPLICATIONS FOR EX VIVO vivax malaria will be emerging in the endemic area in the near future or DRUG RESISTANCE TESTING that this mutation may not be critical to CQ resistance, at least in the Philippine population. On the other hand, in South Korea, all the 4 South 1 2 3 4 Douglas H. Kerlin , Jutta Marfurt , Rintis Noviyanti , Qin Cheng , Korean isolates of ours collected in 1999 showed the wild type (Y976) in 2 1 Ric N. Price , Michelle L. Gatton the gene, but later in 2003 and 2007, 2 cases of CQ resistant vivax malaria 1Queensland Institute of Medical Research, Brisbane QLD, Australia, were reported. The correlation of the Y976F genotype and the phenotype 2Menzies School of Health Research and Charles Darwin University, Darwin of P. vivax to CQ has to be further examined to evaluate the reliability of NT, Australia, 3Eijkman Institute for Molecular Biology, Jakarta, Indonesia, this molecular marker for the genetic epidemiology of vivax malaria. 4Australian Army Malaria Institute, Brisbane QLD, Australia The emergence of chloroquine resistance in Plasmodium vivax has 1530 enhanced the need for greater understanding of the epidemiology of FIELD EX VIVO SENSITIVITY TESTING OF PLASMODIUM the disease, the mechanisms of drug resistance in these parasites, and VIVAX AND P. FALCIPARUM: IMPORTANT CONSIDERATIONS effective case management. Historically the Schizont Maturation Test AND A NEW FIELD BASED CYTOMETRIC METHOD (SMT) has been used to monitor drug resistance in asexual stages of P. falciparum. Modifications are subsequently required when the test is Bruce Russell1, Benoit Mallert1, Rossarin Suwanarusk1, Dennis applied to P. vivax due to the high diversity of life cycle stages present Kyle2, Mallika Imwong3, Aung Pyae Phyo4, Cindy S. Chu4, Marcus in blood samples taken from the peripheral circulation. In this study we J. Rijken4, Usa Lek-Uthai5, Georges Snounou6, Francois Nosten4, analysed the results from 760 isolates assessed by the SMT to investigate Laurent Renia1 how development time of the parasite is related to drug sensitivity. We 1Singapore Immunology Network, A*STAR, Singapore, Singapore, confirm the previous hypothesis that chloroquine has a stage specific 2Department of Global Health, College of Public Health, University of activity against P. vivax, and show that this stage specific activity may South Florida, Tampa, FL, United States, 3Faculty of Tropical Medicine, have profound consequences for the interpretation of the SMT. Using Department of Clinical Tropical Medicine, Mahidol University, Bangkok, threshold models we show that increasing assay duration is associated Thailand, 4Shoklo Malaria Research Unit, Mae Sod, Thailand, 5Department with decreased effective concentration (EC50) values. EC50 values are also of Parasitology and Entomology, Faculty of Public Health, Mahidol shown to be linked to the proportion of ring stages in the initial blood University, Bangkok, Thailand, 6INSERM UMR S 945, Département de sample. We further demonstrate that assays with a duration of less than Parasitologie, Hôpital Pitié-Salpêtrière, Paris, France 34 hours (upper 95% CI 39 hours) should not be used to estimate EC50, nor an assay where the abundance of rings stage parasites in the initial The ex vivo sensitivity testing of antimalarials is an important adjunct sample collected at venipuncture does not exceed 66% (upper 95% CI to in vivo resistance testing. Except for the microscopic schizont 90%) of the total parasites. As late stage P. vivax parasites are insensitive maturation assay (MSMA), there are no standardised protocols for side- to chloroquine, the starting composition of parasites in the SMT can by-side comparison of the sensitivity profiles of Plasmodium vivax and P. dramatically affect the estimated EC50 values for chloroquine; the EC50 falciparum clinical isolates. Here we present a practical cytometry based will be erroneously high if only resistant, late stage parasites are exposed. sensitivity assay that provides a viable field based alternative to the MSMA. Application of this threshold modelling approach suggests that similar We discuss a range of important confounders to any sensitivity assay issues may occur for testing of resistance to amodiaquine and mefloquine. utilising clinical isolates, including the presence of host leukocytes, which

The statistical methodology that has been developed also provides a novel decreases antimalarial IC50s, and the initial developmental stage of the means of detecting stage-specific drug activity for new antimalarials. parasite, which in the case of P. falciparum may result in the failure of H3 Incorporation assays. 1529 1531 Pvmdr1 MUTATION FOR GENETIC EPIDEMIOLOGY OF CHLOROQUINE RESISTANT PLASMODIUM VIVAX FIRST REPORT FROM SOUTHERN PAKISTAN ON ALLELIC VARIANTS OF PLASMODIUM VIVAX CIRCUMSPOROZOITE 1 2 2 Moritoshi Iwagami , Pilarita T. Rivera , Elena A. Villacorte , PROTEIN (Pvcsp) AND MEROZOITE SURFACE PROTEIN1 3 1 Weon-Gyu Kho , Shigeyuki Kano (pvmsp1) 1Department of Tropical Medicine and Malaria, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan, 2Department of Afsheen Raza, Najia Ghanchi, Ali Thaver, Mohammad Asim Beg Parasitology, College of Public Health, University of the Philippines Manila, Aga Khan University Hospital, Karachi, Pakistan Manila, Philippines, 3Department of Parasitology, Inje University, College of Plasmodium vivax is the prevalent malarial specie accounting for 70% Medicine, Busan, Republic of Korea of malaria cases in Pakistan.However,basic data on P.vivax genotypes is A mutation in Plasmodium vivax multidrug resistance 1 gene (pvmdr1) lacking from Pakistan.Studies have shown that for P. vivax, polymorphic in codon 976 (Y976F) is reported to be associated with chloroquine (CQ) genes coding for circumsporozoite protein Pvcsp and merozoite surface resistance and is recently used to monitor the distribution and frequency protein 1 Pvmsp1, can be used as reliable genetic markers for conducting of the CQ resistant vivax malaria. In this study, we determined the molecular epidemiological studies. Pvmsp1 gene is a mosaic organization mutation in codon 976 in the pvmdr1, using 28 P. vivax field isolates from of several variable blocks and its genotyping is based on detection of allelic the Philippines (6 isolates), South Korea (4 isolates), Papua New Guinea variants in its three polymorphic fragments (F1 to F3).Pvcsp genotyping (PNG) (4 isolates), India (2 isolates), Indonesia (1 isolate), Thailand (1 is based on detection of either of the two types of nonapeptide repeat isolate), Bangladesh (1 isolate), Nepal (1 isolates), China (1 isolate), Iran (1 units in its central domain; GDRA (A/D) GPQA, namely VK 210 type and isolate), Rwanda (1 isolate), Sudan (1 isolate), Comoros (1 isolate), Brazil (2 ANGA (G/D) (N/D) QPG, namely VK 247 types.To determine allelic variants isolate) and Ecuador (1 isolate), collected in our National Center for Global of Pvcsp and pvmsp1,a descriptive study was done on two-hundred Health and Medicine, Japan, from 1999 to 2011. The Y976F mutation and thirty blood samples of P. vivax collected from Sind and Baluchistan was observed in the 13 isolates out of the 28 isolates (46%): 6 from the during 2008-2009.Pvcsp and pvmsp1were amplified using nested PCR Philippines, 2 from PNG, 1 form India, 1 from Thailand, 1 form Rwanda, 1 methodology.PCR-RFLP was performed for genotyping of Pvcsp while form Sudan and 1 form Comoros. The other 15 isolates possessed the wild different allelic forms of Pvmsp1 were detected by analysis of fragment type (Y976). In the Philippines, CQ has been used for treatment of vivax www.astmh.org 463 size.Overall number of genotypes and their prevalence were defined the infected human, the period of transmission from malaria patients (via arbitrarily by binning 20 base-pair (bp) intervals together. For Pvmsp1,it the vector mosquito) to manifestation as early primary attack cases would was found that in F1 fragment, 12 allelic variants were observed (bp size be 3-4 weeks. If P. vivax malaria is transmitted actively, subsequent minor variation350-550), in F2 fragment 17 allelic variants were observed (950- peaks are expected to be seen after the annual highest peak by 3-4 week 1270 bp) and in F3 fragment 8 allelic variants were observed (250-390 intervals. The obvious minor peak had not been seen until the early 2000s, bp).Thus, a total of 17 genotypes corresponding to pvmsp1 gene were however, it began to be seen in the mid-2000s. During 2006-2009, three found circulating in Southern Pakistan. Pvcsp genotyping in Pakistani or four subsequent minor peaks after the highest peak were observed isolates showed that VK210 variants were predominant(79%, 182/230) by 20-day or 30-day intervals. This phenomenon shows that the length while percent positivity of VK 247 was 13%( 29/230).Respective bp size of transmission period of P. vivax malaria in the ROK has been gradually variation were 600-870bp for VK 210 and 650-820bp for VK 247.We extending. This result may be ascribed to a climate change-mediated conclude that this is,to our knowledge, the first study from Southern temperature rise. Malaria and climate data should be integrated to analyze Pakistan on genetic diversity in pvcs and pvmsp1 gene. Data from this and predict the influence of climate change on malaria occurrence in ROK. study indicates that both pvcs and pvmsp1 can be used as reliable markers for conducting genotyping of P. vivax.Thus, this study may serve as a 1534 baseline data for future research on P. vivax diversity from Pakistan. MORTALITY ATTRIBUTABLE TO PLASMODIUM VIVAX 1532 MALARIA 1 2 2 PLASMODIUM VIVAX INFECTION IN DUFFY-NEGATIVE Nick M. Douglas , Gysje J. Pontororing , Daniel A. Lampah , 1 2 2 INDIVIDUALS IN ETHIOPIA: INDICATIONS AGAINST AN OLD Tsin W. Yeo , Enny Kenangalem , Jeanne R. Poespoprodjo , Anna P. Ralph1, Michael J. Bangs3, Yati Soenarto4, Paulus Sugiarto5, PARADIGM Nicholas M. Anstey1, Ric N. Price1 Tamirat Gebru Woldearegai, Peter G. Kremsner, Jürgen F. J. Kun 1Menzies School of Health Research, Casuarina, Northern Territory, Institute for Tropical Medicine, University of Tuebingen, Tübingen, Australia, 2Papuan Health and Community Development Foundation, Germany Timika, Indonesia, 3Public Health and Malaria Control Department, PT Freeport Indonesia, Timika, Indonesia, 4University of Gadjah Mada, In Ethiopia, the most wide-spread Plasmodium species co-exist and Yogyakarta, Indonesia, 5Mitra Masyarakat Hospital, Timika, Indonesia individuals of variant Duffy blood groups live in this country. A study was conducted to measure the prevalence of malaria and polymorphism Plasmodium vivax causes almost half of all malaria cases in Asia. Although in the Duffy Antigen Receptor for Chemokine (DARC) gene in East and once regarded as benign, studies have highlighted its association with South-Western Ethiopia. Presence of malaria was measured by microscopy severe and fatal malaria. The extent to which P. vivax contributes to and PCR. The polymorphism of DARC was analyzed by DNA sequencing. mortality in endemic regions is not known. We aimed to define the In this study, either P. falciparum or P. vivax infection was detected in all epidemiology of mortality attributable to vivax malaria in southern Papua, examined samples. In the analysis of the Duffy blood group, there were Indonesia by conducting a retrospective clinical records-based audit of 17 (20%) and 24 (22.4%) homozygous Duffy negative individuals in Harar all deaths in patients with vivax malaria at Mitra Masyarakat Hospital. and Jimma study sites. Surprisingly, the data showed that P. vivax infection Between January 2004 and September 2009, hospital surveillance also occurred in three Duffy negative individuals. FYB/FYBnull was found identified 3,495 inpatients with P. vivax monoinfection and 65 (1.9%) to be the dominant genotype in both area. The FYA/FYB and FYB/FYB patients who subsequently died. Charts for 54 of these 65 patients could genotype was associated with susceptibility and FYBnull/FYBnull genotype be reviewed, 40 of whom had pure P. vivax infections on cross-checking. was associated with protection against P. vivax infection. This study Using pre-defined conservative criteria, vivax malaria was the primary documents an emergence of P. vivax infection in Duffy negative individuals cause of death in 5 cases, a major contributor in 17 cases and a minor in the study area. Duffy negative blood group does not provide absolute contributor in a further 13 cases. Extreme anemia was the most common protection of P. vivax infection in the studied population. primary cause of death for patients in the first category. Malnutrition, sepsis with respiratory and gastrointestinal manifestations, and chronic 1533 diseases such as HIV infection were the commonest attributed causes of death for patients in the latter two categories. There were ~293,763 CHANGING TRANSMISSION PATTERN OF PLASMODIUM cases of pure P. vivax infection in the community during the study period VIVAX MALARIA IN THE REPUBLIC OF KOREA: RELATIONSHIP giving an overall minimum case fatality of 0.12 per 1,000 infections. WITH CLIMATE CHANGE The corresponding case fatality in hospitalized patients was 10.0 per 1,000 infections. Although uncommonly directly fatal, vivax malaria is Jae-Won Park an important indirect cause of death in patients with malnutrition, sepsis Graduate School of Medicine, Gachon University of Medicine and Science, syndromes and chronic diseases in southern Papua. Incheon, Republic of Korea The Korean peninsula is an only place where indigenous Plasmodium 1535 vivax malaria has occurred continuously on a large scale in temperate CYTOKINE DYNAMICS AFFECT SUSCEPTIBILITY TO areas. P. vivax malaria was endemic on the Korean peninsula for many PLASMODIUM VIVAX AND P. FALCIPARUM INFECTION AND centuries until the Republic of Korea (ROK; South Korea) was declared as a “malaria elimination” country in 1979. P. vivax malaria re-emerged ANEMIA: STUDIES FROM THE PERUVIAN AMAZON in 1993 in the ROK, and has occurred constantly for more than 15 Jonathan Merola1, Mario Hoenemann1, Jean N. Hernandez2, years after its re-emergence. P. vivax malaria in ROK has been strongly OraLee Branch1 influenced by infected mosquitoes originating from the Democratic 1New York University School of Medicine, New York, NY, United States, People’s Republic of Korea (North Korea). Korean P. vivax malaria has 2Universidad Nacional Amazonia Peruana, Laboratorio de Investigación de shown typical characteristics of unstable malaria transmitted only during Productos Naturales Antiparasitarios de la Amazonía, Iquitos, Peru the summer season, and displays short and long incubation periods. The changing pattern of the transmission period can be predicted by The dynamics of immune factors in Plasmodium falciparum and P. vivax analyzing the seasonal characteristics of early primary attack cases with infection are thought to dictate host symptoms and pathology but have a short incubation period. Such cases began to gradually occur earlier in not been fully elucidated to date. We aimed to describe the cytokine the 1990s after the re-emergence. Considering the sporogony cycle in dynamics of malaria infection and examine malaria species, age and Anopheles mosquito and the asexual cycle via a short incubation period in gender dependent differences. Serum samples were collected by active www.astmh.org 464 weekly blood sampling in the Zungarococha community of Iquitos, Peru 1537 from April 2003-September 2008 from a total of 397 P. falciparum, 515 P. vivax and 39 mixed P. falciparum and P. vivax infections. Cytokine EVALUATION OF NATURALLY ACQUIRED HUMORAL measurements were made using Luminex bead-based assay from sera IMMUNE RESPONSES AGAINST IMMUNOREACTIVE collected from infected individuals one week before, during, one week PROTEINS OF PLASMODIUM VIVAX BY PROTEIN ARRAYS after and one month following infection. Relationships of immune 1 1 1 1 1 markers to strain, age, and gender were assessed. P. falciparum and P. Feng Lu , Jun-Hu Chen , Jian Li , Yang Cheng , Bo Wang , Kwon- 2 3 1 vivax infected males were more prone to febrile malaria infection and Soo Ha , Takafumi Tsuboi , Eun-Taek Han demonstrated higher levels of IFN-γ, TNF-α and IL-10 than infected 1Department of Parasitology, School of Medicine, Kangwon National females (p<0.05). A comparison of cytokine dynamics between the two University, Chuncheon, Republic of Korea, 2Department of Molecular and strains revealed increased IL-4 and IL-6 levels in P. falciparum infected Cellular Biochemistry, School of Medicine, Kangwon National University, individuals one week following treatment (p<0.05). Moreover, IL-10 and Chuncheon, Republic of Korea, 3Cell-free Science Technology and Research IFN-γ levels were markedly decreased one week following treatment of Center, Ehime University, Matsuyama, Japan P. falciparum, differing from the sustained elevation in these cytokines In the previous report, we successfully applied an antibody-based protein observed a week following treatment of P. vivax infection (p<0.05). TNF array for immunoprofiling of Plasmodium vivax infection, and some receptor and IL-10 levels were predictive of febrile infection and parasite highly immunoreactive proteins were identified. To further characterize density in both P. falciparum and P. vivax infection (P<0.005). Independent the antibody reactivity of these immunogenic proteins, we used a Ni-NTA of parasite density, for both malaria species, hematocrit levels were directly surface based protein array to detect the immune responses from sera of correlated to IL-1 (P<0.01) while erythropoietin levels directly correlated vivax malaria parasites. ETRAMP, Pv12, Pv41 and MSP3.9 were studied to with TNF-α (P=0.003). Interestingly, both hematocrit (P=0.02) and compare with the well-characterized vivax vaccine candidate MSP1-19. erythropoietin (P=0.03), were negatively correlated with the co-presence Among the 52 microscopically positive samples, all samples were detected of IL-1 and TNF-α. There was a gender and species related difference P. vivax by the PvMSP1-19 arrays (100% sensitivity), and were 36 (69.2%), in the hematocrit dynamics over time. In conclusion, these findings are 31 (59.6%), 23 (44.2%) and 22 (42.3%) by ETRAMP, Pv12, Pv41 and the first to describe cytokine dynamics of malarial infection in vivo and MSP3.9 arrays respectively. The false positives were obtained 2 (95.0% demonstrate a strong association of inflammatory responses with parasite specificity), 2 (95.0%), 3 (92.5%), 4 (90.0%) and 4 (90.0%) among 40 density, the severity of symptoms, and anemia. The observed differences sera samples from healthy subjects. The ratio (fluorescent intensity of in immune responses between genders and malaria species may explain positive samples/that of negative samples) of antibody response to the distinctions in clinical presentations between males and females as well as PvMSP1-19, ETRAMP, Pv12, Pv41 and MSP3.9 were 17.5, 6.1, 2.6, 3.5 core differences in the pathogenesis of P. vivax and P. falciparum infection. and 4.4 respectively. Although the naturally acquired humoral immune responses against PvMSP1-19 show superior reactivity than others, these 1536 recombinant proteins were also highly recognized by P. vivax infected IMMUNE RESPONSE TO PLASMODIUM VIVAX INFECTION: A patient sera. These results validate the protein arrays for profiling antibody STUDY IN THE CENTRAL CHINA responses to P. vivax infection, and further confirmed that ETRAMP, Pv12, Pv41 and MSP3.9 could also be selected as potential target antigens for Kulachart Jangpatarapongsa1, Hui Xia2, Qiang Fang2, Jetsumon malaria vaccine. Sattabongkot3, Qi Gao4, Liwang Cui5, Baiqing Li6, Rachanee Udomgsangpetch7 1538 1Center for Innovation Research and Technology Transfer, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand, 2Department CHARACTERIZATION AND SEROLOGIC RESPONSES TO of Parasitology, Bengbu Medical College, Anhui, China, 3Department PLASMODIUM VIVAX DUFFY BINDING PROTEIN (DBP) of Entomology, United States Army Military Component, Armed Forces VARIANTS IN RESIDENTS OF PURSAT PROVINCE, CAMBODIA 4 Research Institute of Medical Sciences, Bangkok, Thailand, Jiangsu Samantha J. Barnes1, Francis Ntumngia1, Jesse Schloegel2, 5 Institute of Parasitic Disease, Wuxi, China, Department of Entomology, Chanaki Amaratunga3, Suon Seila4, Duong Socheat4, Hamisu The Pennsylvania State University, State College, PA, United States, Salihu1, Rick M. Fairhurst5, John H. Adams1 6Department of Immunology, Bengbu Medical School, Anhui, China, 1University of South Florida, Tampa, FL, United States, 2La Trobe University, 7Department of Pathobiology, Faculty of Science, Mahidol University, Melbourne, Australia, 3Institute of Allergy and Infectious Diseases, National Bangkok, Thailand Insitutes of Health, Bethesda, MD, United States, 4The National Center for Plasmodium vivax infection possesses a characteristic of relapsing fever Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia, indicating the re-infection by previously hidden parasites in the host. The 5National Institute of Allergy and Infectious Diseases, National Institutes of relapsed infection can lead to activation of memory T cells pool which Health, Bethesda, MD, United States might bring up protective immunity. This study aims to characterize The Plasmodium vivax Duffy Binding Protein (DBP) is the ligand in the natural immune responses in acute P. vivax infected patients living in a major pathway for P. vivax invasion of human reticulocytes, making it sole P. vivax infection cohort in Central China. Lymphocytes were collected an appealing vaccine candidate. Region II of DBP (DBPII) is the minimal from three recruitments: patients infected with P. vivax, malaria-immune portion of the ligand that mediates recognition of the Duffy Antigen and malaria-naive controls. Using flow cytometry, we showed memory T Receptor for Chemokines (DARC) on the reticulocyte surface and cells were elevated in blood during acute infection. The level of γδ T cells constitutes the primary vaccine target. Analysis of natural variation in was two fold higher than that of naïve controls. This suggested that two the coding sequences of DBPII revealed signature evidence for selective populations, memory and γδ T cells, responded specifically to the P. vivax pressure driving variation in the residues of the putative receptor-binding parasites. On contrary, B, NK and NKT cells were decreased during acute site. We hypothesize that anti-DBP immunity in P. vivax infections is infection. In addition, regulatory T cells were reduced, suggesting the non- strain-specific and hindered by polymorphic residues altering sensitivity immune suppressive role of P. vivax parasites. Interestingly, P. falciparum to immune antibody inhibition. To comprehend the human IgG response antigens cross-stimulated T cells obtained from these P. vivax-infected following P. vivax infections we investigated the specificity of serum IgG in patients. These results provided a further insight into interaction between residents living in Pursat Province, Cambodia. Using ELISAs, we quantified P. vivax parasites and host cell-mediated immunity in the exclusive P. the antibody titer against five variant alleles of DBPII. We also sequenced vivax endemic area that could be important for future development of a the DBPII of the field isolates to determine their relationship to the variant successful vaccine designation. alleles used in the ELISAs. When correlating the IgG titer between the DBP

www.astmh.org 465 variants a strain-specific immune response was observed in patients with subjects. Over the next several weeks, we will enroll a total of 20 patients a high antibody titer to DBPII_AH as compared to the other variants. This as the malaria season in Iquitos is now at its busiest. By identifying differed from the correlation of high antibody titers between DBPII_P and termination points in sporogonic development, we aim to provide further DBPII_7.18 (ρ=0.88, p-value<0.0001) and DBPII_P and DBPII_O (ρ=0.87, insight into the biologic mechanisms pertinent to transmission blocking p-value<0.0001). There appeared to be little correlation between specific vaccine strategies and hope the results will help focus our search for polymorphic residues and IgG titer. Understanding the immune response transmission blocking antibodies in simultaneously collected plasma. to the polymorphisms within DBPII will allow further identification of epitopes to enable the production of a more effective P. vivax vaccine. 1541 1539 GENETIC VARIABILITY OF PLASMODIUM VIVAX IN THE NORTH COAST OF PERU AND THE ECUADORIAN AMAZON NOVEL HUMANIZED MOUSE MODELS FOR PLASMODIUM 1 1 1 VIVAX Julio Ventocilla , Jorge Nuñez , L. Lorena Tapia , G. Christian Baldeviano1, Carmen M. Lucas1, Stephen Manock2, Andres G. Rebecca Danner1, Teodor Brumeanu2, Sofia A. Casares1 Lescano1, Kimberly A. Edgel1, Paul C. Graf1 1Naval Medical Research Center, Silver Spring, MD, United States, 1Naval Medical Research Unit Six, Lima, Peru, 2Hospital Vozandes del 2Uniformed Services University of the Health Sciences, Bethesda, MD, Oriente, Shell, Ecuador United States Plasmodium vivax is the most widespread malaria parasite causing Development of humanized animal models able to sustain infection significant morbidity worldwide. In the Peruvian North Coast (PNC), the with Plasmodium vivax is required to increase our understanding of the number of P. vivax malaria cases has steadily increased over the last few biology and pathogenesis of the parasite and to test vaccines and anti- years despite a significant decline in the number of cases in Peru. To malarial drugs. Such models are also expected to allow development of understand the transmission dynamics of P. vivax populations between a convenient mosquito challenge model for vaccination trials. We have the PNC and the neighboring Ecuadorian Amazon (EA), we studied the developed novel humanized mouse strains that were genetically modified genetic diversity and population structure of P. vivax isolated in those to develop a human hematopoietic system upon infusion of stem cells areas. One hundred and twenty blood or serum samples comprising 95 from umbilical cord blood. The humanized mice are immunodeficient, PNC (58 from Piura and 37 from Tumbes, collected from 2008 to 2010) as they are knocked out for Rag and IL2 genes, and at the same time and 25 EA (from Puyo, collected between 2001 and 2004) were assessed express human HLA-DR4. These mice develop human erythrocytes and by 6 polymorphic neutral microsatellite markers. Genetic variability was reticulocytes and our preliminary data indicated their ability to sustain determined by the haplotype frequency and expected heterozygosity P. vivax blood stage infection. The ability of humanized mice to develop (He). Population structure was assessed by Bayesian inference cluster human T cells (both CD4+ and CD8+ subsets), B cells, and significant analysis. We found very low genetic diversity in PNC, with a single allele serum levels of human IgM and IgG, enable them as a unique model to per locus, one haplotype and He=0 in Piura; and 1-3 alleles per locus, test the immunogenicity and protective efficacy of human vaccines. 3 haplotypes and He=0.30-0.32 in Tumbes. In contrast, high genetic diversity was observed in EA, with 4-6 alleles per locus, 15 different 1540 haplotypes and He=0.43-0.70. Population structure analysis revealed three distinct populations correlating with each geographic location. Five out 37 FAILURE TO INFECT: DELINEATING PLASMODIUM VIVAX (14%) isolates from Tumbes had an identical haplotype to that found in DEVELOPMENT CESSATION AMONGST ANOPHELES Piura, suggesting unidirectional gene flow from Piura to Tumbes (100 Km DARLINGI IN THE PERUVIAN AMAZON apart). In addition, one haplotype collected in 2008 in Tumbes showed high similarity to a haplotype found in 2003 in Puyo, which was likely 1 2 2 Megan A. McCaughan , Shira Abeles , Joseph Vinetz , Raul an imported case from the Ecuadorian Amazon. No shared haplotypes 2 3 Chuquiyauri , Carlos Tong Rios between Piura and Puyo was observed (300 Km apart), indicating little 1University of California, San Francisco, San Francisco, CA, United States, gene flow between these two areas. Our study provides important 2University of California, San Diego, San Diego, CA, United States, information on the transmission patterns between the Coastal areas of 3NAMRU, Iquitos, Peru Peru and the Ecuadorian Amazon. Future studies should include isolates Malaria continues to be a top infectious disease burden in tropical and from the South Coastal region of Ecuador in order to identify other routes subtropical areas of the world. Plasmodium vivax is geographically the of parasite dissemination between the North Coast of Peru and Ecuador. most widely distributed cause of malaria, with up to 2.5 billion people at risk and an estimated 80 million to 300 million clinical cases every year. 1542 Recent disease control innovations have included attempts at transmission SIMILAR DIVERSITY OF THE MEROZOITE SURFACE PROTEIN blocking vaccines (TBVs), which target the intra-mosquito part of the complex malaria life cycle. Naturally occurring transmission blocking has 3 ALPHA (MSP-3α) SUBFAMILY IN THAI AND VENEZUELAN been observed such as in a study conducted in the Peruvian Amazon in PLASMODIUM VIVAX POPULATIONS which mosquitoes dissected seven days after parasitemic blood meals Monica Acosta1, Bejamin Rice1, M. Andreina Pacheco1, Leopoldo revealed infection rates of only 50%. To further understand this pattern of Villegas2, Ananias A. Escalante1 incomplete transmission, we are performing membrane-feeding assays in 1Center for Evolutionary Medicine & Informatics, Arizona State University, which vivax-infected blood samples from subjects enrolled in the Peruvian Tempe, AZ, United States, 2Global Fund Malaria Project – Suriname Amazon are fed to first generation lab-reared mosquitoes. Instead (Medical Misión)/PAMAFRO, Paramaribo, Suriname of dissecting at day 7 as typically done to assess for oocysts, we are dissecting mosquitoes at multiple time intervals to detect more specifically Characterizing the polymorphism and identifying possible signatures where transmission drops - is it the failure of microgametes to fertilize of selection continues to be an important task in the study of malarial macrogametes, failure of zygotes to transform into ookinetes or failure parasite antigens. This is of particular importance with regard to of ookinetes to traverse the midgut epithelium and become oocysts. To Plasmodium vivax blood stage antigens, which have not been as account for all developmental stages, dissections begin at 15 minutes post extensively studied as those from the more virulent P. falciparum. Of feed, up to 24 hours, then again at days 4 and 14. Blood samples are then particular significance is the merozoite surface protein 3 (MSP-3) gene stained with Giemsa and examined under microscopy. Of the 6 subjects family, involved in the invasion of the asexual merozoite parasite form into already enrolled, we have found that overall parasitemia was important to the red blood cell. The MSP-3 gene family has undergone an expansion transmission, but that it did not explain the lack of transmission in some in the P. vivax lineage with twelve identified genes, compared to the four www.astmh.org 466 in P. falciparum and two in P. knowlesi. This duplication event has been 1544 posed as being biologically important and could have implications for pathogenicity and parasite interactions with the host immune system. LIMITED GENETIC DIVERSITY AND CLONAL POPULATION Here we describe the genetic diversity and identify possible signatures of STRUCTURE OF PLASMODIUM VIVAX PARASITES FROM selection of three paralogous and syntenic genes of the subfamily MSP-3α A GEOGRAPHICALLY ISOLATED COMMUNITY IN THE (2,559-2,724 base pairs in length), from Venezuelan and Thai population PERUVIAN AMAZON samples. Two of these paralogous genes have only recently been recognized. Despite differences in transmission rates, similar patterns of Christopher Delgado1, Peter Van den Eede2, Veronica Soto1, selection and polymorphism were found for the Thailand and Venezuelan Dionicia Gamboa1, Alejandro Llanos-Cuentas1, Annette Erhart2, samples. Genetic diversity, calculated by the parameter π, revealed that Umberto D’Alessandro2 the majority of sequence diversity was constrained to the N terminal 1Instituto de Medicina Tropical Alexander von Humboldt, Universidad for all three paralogs. This pattern holds when the two populations are Peruana Cayetano Heredia, Lima, Peru, 2Institute of Tropical Medicine compared. In contrast, strong evidence of positive selection was observed Antwerp, Antwerp, Belgium for the N-terminal in the two newly recognized MSP-3α gene copies, Plasmodium vivax has the highest burden of malaria morbidity and is the but only in the Thai population. Finally, evidence of purifying selection at most spread human plasmodium species around the world. However, due the C-terminal for all three genes suggests that it could be of functional to its complex life cycle, little is known about the genetic characteristics importance. If such functional constrain is demonstrated, it may be of these parasites populations and the epidemiology of the disease. In the possible that the MSP-3 family is functionally redundant, a possible benefit present study, we explore the genetic diversity and population structure to the parasite that may be conferred by increased antigenic diversity and dynamics of P.vivax parasites from a community in the Peruvian allowing enhanced evasion of the host immune response. Amazon within the framework of a two years cohort looking for some insights that may explain the epidemiology of this disease within this 1543 area. Thirty-eight patients from San Carlos community, a geographically Epidemic and interepidemic periods structure isolated area, were enrolled and followed up for 2 years after received the Plasmodium vivax population circulating in French radical cure treatment (chloroquine + primaquine). P. vivax infections were Guiana between 2006 and 2010 detected by microscopy and by specie specific PCR. Molecular genotyping of P.vivax parasites using 15 microsatellites was performed. The genetic Lise Musset1, Arielle Salmier1, Eric Legrand1, Benoit de Thoisy2 diversity was determined by calculating the expected heterozygosity (He) 1Laboratoire de parasitologie, Institut Pasteur de la Guyane, Cayenne, and allelic richness. The genetic population structure was determined S 2 French Guiana, Laboratoire de primatologie, Institut Pasteur de la calculating the linkage disequilibrium (pairwise LD and IA ), the probability Guyane, Cayenne, French Guiana of admixture or clonal reproduction (Psex) and looking for clusters of genetically related haplotypes. A limited genetic diversity of P. vivax Since 2005, Plasmodium vivax is the main species circulating in French parasites (He 0.47) and a high prevalence of monoclonal infections (83%) Guiana with an incidence of 51% in 2010. Understanding the genetic -6 S were found. The strong linkage disequilibrium (pairwise LD P<10 and IA structure of this parasite population is essential in order to predict the 0.51), the low probability for admixture (Psex<0.0007) and the finding rapid spread of certain phenotypes of interest (eg resistant parasites) of a few clusters of genetically related haplotypes, described a clonal or for a better understanding of the epidemiology. In this study, 195 population of malaria parasites. The limited genetic diversity and clonal isolates, collected between 2006 and 2010, in five geographical regions of structure of P. vivax population described in this area may represent a French Guiana, were analyzed using six highly polymorphic microsatellite high risk for the rise and spread of drug resistance. Nevertheless, also may markers amplified by a semi-nested polymerase chain reaction method. stimulate positively the development of clinical immunity by the people in We have shown that 28.1% of infections were polyclonal. The parasite the Amazon Basin being reflected in the high proportion of asymptomatic population circulating in this department is likely drawn from a single cases reported in this community. ancestral population. The current population was genetically diverse (He = 0.68), and not structurated in time or space, as shown considering 1545 unique haplotype sampling strategy. In contrast, using all data samples, the population presents higher structuration rates, likely related to CpG-DNA ENCAPSULATED WITH PEPTIDE ANTIGENS OF epidemics, which induce locally, and temporary high levels of inbreeding. PLASMODIUM VIVAX IN MICROPARTICLES ENHANCES THE Demographic history was also investigated, using both exponential SYSTEMIC AND MUCOSAL IMMUNE RESPONSES IN MICE and linear expansion models. Exponential model would consider that USING INTRANASAL MODE OF DELIVERY: AN APPROACH expansion of efficient population size occur mainly during epidemics, TOWARDS MUCOSAL VACCINE FOR MALARIA via clonal expansions. In contrast, linear model relies on genetic diversity acquired during sexual stages of Plasmodium. Exponential model shows Ajaz Bhat1, Vineeta Tanwar1, Jayaprakash Babu2, Riyasat Ali2, Tina a stable efficient population size, suggesting that epidemics do not Mohan2, Sukla Biswas3, Donthamsetty Nageswara Rao2 contribute to long-term Plasmodium expansion. In contrast, linear model 1Vanderbilt University, Nashville, TN, United States, 2All India Institute of shows an expansion of efficient population size since some decades: this Medical Sciences, New Delhi, India, 3National Institute of Malaria Research, is related to a global increase of malaria cases, even when epidemic events New Delhi, India are not considered. We conclude that epidemics may promptly and locally drive genetic information, but do not contribute to a wider, longer-term Due to drug resistance and limitation of growing of Plasmodium vivax and regionally-scaled demographic history of P. vivax. Therefore, a similar parasite in vitro for enough DNA/Protein, we attempted an alternate study encompassing a larger geographical area would be required for a synthetic peptide approach from the deduced amino acid sequences of better understanding of the structure population of P. vivax in the Guiana different antigens of P. vivax constituting all the stages of the life cycle. Shield or even, Amazonia, and to confirm that P. vivax diversity is not For producing efficient and long lasting humoral immune responses, proportional to the transmission level as observed with P. falciparum. PLGA microparticles were used as delivery vehicles and CpG ODN as immunoadjuvants.The adjuvants used were synthetic oligonucleotides containing CpG ( CpG-ODN 1826 and 2006, class B ) motifs. P. vivax peptides viz, MSP 1#1, MSP 1#23, CSP, AMA, and Pvs24 (TBA) possessing B and T cell epitopes were synthesized using Fmoc chemistry, purified to homogeneity by Gel permeation chromatography and HPLC. Peptide purity was confirmed by amino acid acid analysis. These peptide

www.astmh.org 467 antigens were then entrapped in microparticles along with CpG-ODN. 1547 Biodegradable microparticles were prepared from 50:50 PLGA by water-in-oil-in water (w/o/w) solvent evaporation method. Particle size PLASMODIUM VIVAX CIRCUMSPOROZOITE PROTEIN- and size distribution was determined using particle size analyzer.The SPECIFIC CELLULAR IMMUNE RESPONSES AFTER morphology was studied by scanning electron microscopy. Outbred strains IMMUNIZATION WITH THE VMP001/AS01B CANDIDATE of mice were immunized using intranasal route with different peptide MALARIA VACCINE IN MALARIA-NAÏVE INDIVIDUALS formulations. Peptide specific IgG,IgA and SIgA estimation was done by standardized ELISA protocol. Peptides were found to be > 95% pure. Joanne Lumsden1, Saule Nurmukhambetova1, Donna Tosh1, Percentage peptide entrapment was in the range of 60-70%. Percentage Yannick Vanloubbeeck2, Johan Vekemans2, Ripley Ballou2, Joe entrapment for CpG-ODN was in the range of 50-60%. Microparticles Cohen2, Christian Ockenhouse1, Jason Bennett1, Anjali Yadava1 were in the size range of 2-5µm and spherical with smooth surface. 1Walter Reed Army Institute of Research, Silver Spring, MD, United States, Presence of CpG in microparticles along with the peptide antigens 2GlaxoSmithKline Biologicals, Rixensart, Belgium showed serum IgG titre of 51,200-204,800 maintained till 90 days post Plasmodium vivax is the major cause of malaria outside of sub-Saharan immunization.The isotypic profile of the serum IgG revealed IgG2a/2b Africa and inflicts debilitating morbidity and consequent economic impact as the predominant isotypes,maintained till 90 days post immunization. in developing countries. We have developed a novel chimeric recombinant Peptide specific IgA titre in sera ranged between 12,800-25,600 protein VMP001 based on the CSP of P. vivax. The first-in-humans safety, maintained till 90 days post immunization and SIgA titre in washes ranged immunogenicity and efficacy clinical trial of VMP001 formulated in GSK between 800-25,600.Infected mosquitoes fed with high titer Pvs24 (TBA) Biologicals’ adjuvant system AS01 was recently performed in malaria- anti-sera showed signinificant reduction in the oocyst count as revealed B naïve adults. An effective P. vivax vaccine will likely require induction of by membrane feeding assay.This study shows CpG ODN to be a potent both humoral and T cell responses. The aim of this study was to measure mucosal adjuvant to induce immune responses against peptide antigens vaccine-induced T cell responses in PBMCs collected at various time points administered by intranasal inhalation.This is the first reported study with following immunization. Multiparameter flow cytometry was used to mucosal vaccination for malaria. enumerate the frequency and phenotype of T cells producing IL-2, TNF, and IFN-γ after in vitro stimulation of cryopreserved PBMCs with VMP001, 1546 or pools of overlapping peptides corresponding to different regions of + + FINE-SPECIFICITY OF HUMORAL IMMUNE RESPONSES the VMP001 construct. IL-2 CD4 T cell responses were detected in all + + GENERATED IN NAIVE ADULTS FOLLOWING VACCINATION volunteers at one or more time point. TNF CD4 T cell responses were + + WITH VMP001, A PREERYTHROCYTIC VACCINE CANDIDATE detected in 93% of volunteers, albeit at a lower frequency than IL-2 CD4 T cells. In addition, IFN-γ+CD4+ T cell responses were detected in 55% BASED ON THE CIRCUMSPOROZOITE PROTEIN OF of volunteers. TNF+IL-2+ double-producers and IL-2+ single-producers PLASMODIUM VIVAX, FORMULATED WITH GSK were most commonly detected, along with a smaller population of BIOLOGICALS ADJUVANT SYSTEM AS01B cells producing all three cytokines tested. The vaccine-induced CD4+ T Anjali Yadava1, Cysha E. Hall1, Jetsumon Sattabongkot2, Joanne cell responses were strongest and most frequently detected against the M. Lumsden1, Donna M. Tosh1, Johan Vekemans3, W. Ripley N-term region (90% of volunteers) but smaller responses against C-term Ballou3, Joe Cohen3, Yannick F. Vanloubbeeck3, Christian F. and repeat regions were also observed (24% and 21% of volunteers 1 1 respectively). Our results indicate that the VMP001/AS01 vaccine was Ockenhouse , Jason W. Bennett B immunogenic, as indicated by the detection of antigen-specific CD4+ 1Walter Reed Army Institute of Research, Silver Spring, MD, United States, T cell responses in all immunized volunteers. Although the vaccine did 2Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, not induce sterile protection, a small but consistent delay in prepatent 3GlaxoSmithKline, Rixensart, Belgium period was observed in some subjects. Detailed analysis of the induced T We have previously reported on the development of VMP001, a chimeric cell responses may help understand these findings and help improve the circumsporozoite protein based vaccine for P. vivax. In previous studies design of future vaccine constructs. the VMP001 vaccine candidate demonstrated strong immunogenicity, and also demonstrated high efficacy in Aotus monkeys challenged with 1548 infectious P. vivax sporozoites. Based on these data we proceeded to test VMP001 for its safety, immunogenicity and efficacy in humans. This first- A ROBUST PLASMODIUM VIVAX EX VIVO INVASION ASSAY in-human study was designed as a dose-escalating study using three doses Bruce Russell1, Rossarin Suwanarusk1, Céline Borlon2, Fabio T. of VMP001 formulated in GSK Biologicals Adjuvant System AS01 .The B Costa3, Cindy S. Chu4, Marcus J. Rijken4, Kanlaya Sriprawat4, primary study endpoint was to assess vaccine safety. One of the secondary Lucile Warter1, Esther G. Koh1, Benoit Malleret1, Yves Colin5, endpoints of this study was an evaluation of the fine-specificity of humoral Olivier Bertrand5, John H. Adams6, Umberto D’Alessandro2, immune responses generated following vaccination. We evaluated the Georges Snounou7, Francois Nosten4, Laurent Renia1 antibody responses from volunteers to the N- and C-terminal regions 1 as well as the central repeat region of the synthetic VMP001 vaccine Singapore Immunology Network, A*STAR, Singapore, Singapore, 2 construct. The vaccine was immunogenic and all three doses of vaccine Department of Parasitology, Institute of Tropical Medicine, Antwerp, 3 induced antibody responses that spanned the entire length of the vaccine Belgium, Departamento de Genética, Evolução e Bioagentes; Instituto construct. While there were detectable responses to all domains of the de Biologia, Universidade Estadual de Campinas, Campinas - SP, Brazil, 4 5 molecule, there was a distinct hierarchy in terms of the regions recognized Shoklo Malaria Research Unit, Mae Sod, Thailand, INSERM, UMR_S 665, 6 by the vaccinees. The C-terminal region induced the highest magnitude of Paris, France, Global Health Infectious Disease Research, College of Public 7 responses in all three groups, followed by the N-terminal region with the Health, University of South Florida, Tampa, FL, United States, INSERM Repeat domain the lowest. While the magnitude of the titers differed, in UMR S 945, Département de Parasitologie, Hôpital Pitié-Salpêtrière, Paris, general, titers against any single domain correlated to titers against the France others. Individuals having the highest antibody titer to one component We describe a protocol for an ex vivo Plasmodium vivax invasion assay of the vaccine also had high titers to the other domains. Although the that can be easily deployed in laboratories located in endemic countries. vaccine did not induce sterile protection, a small but consistent delay in The assay involves mixing enriched cord blood reticulocytes with matured, prepatent period was observed in some subjects. Detailed analysis of the trypsin-treated P. vivax schizonts concentrated from clinical isolates. immune response may help understand these findings and help improve The invasion efficiencies observed for parasites from 85 isolates were the design of future vaccine constructs. We will discuss vaccine induced highly variable, ranging from 0.1% to 22.3% with a mean of 3.7% antibody avidity and fine-specificity in detail. (95%CI: 2.8%-4.6%). The utility of the protocol for vaccine testing www.astmh.org 468 was demonstrated by using it as an invasion-inhibition assay to assess exists at all institutions including liberal arts colleges. Following the functionally antibodies against DARC, pvMSP1 and pvDBP. This provides experience of teaching a short course in global health in a liberal arts the first biological demonstration that polymorphisms in the pvDBP gene college, student organizations, courses, and officially recognized curricular affect the invasion inhibition efficacy of anti-pvDBP antibody. offerings in global and public health were identified for the 2009-2010 academic year for fifty of the top ranked liberal arts colleges in the United 1549 States. 42% of the colleges had a track, concentration or program; schools that did not have official themes still listed at least one course IS THAT A RODENT IN YOUR LUGGAGE? BUSHMEAT in global or public health. All of the themes were interdisciplinary and CONFISCATIONS REPORTED IN THE CENTERS FOR DISEASE when they were more expansive such as a program, they were organized CONTROL AND PREVENTION’S QUARANTINE ACTIVITY as a multi-college consortium. 48% of them had been developed since REPORTING SYSTEM - UNITED STATES, SEPTEMBER the 2005 academic year. The most number of courses were in the Social 2005-DECEMBER 2010 Sciences (n=9.9±12.2) followed by the Natural Sciences (3.2±5.3). Student organizations in global or public health were present on 30% of campuses Teal R. Bell, Andrew Higgins, Sheryl Shapiro, Heather Bair-Brake, and all but two schools had service, social justice or AIDS organizations. Nina Marano, Noelle-Angelique M. Molinari, Nicole J. Cohen, Gale The values of a liberal education are closely aligned with those of global Galland health: social responsibility, critical thinking, skills in communication, Centers for Disease Control and Prevention, Atlanta, GA, United States analysis and problem-solving, ethical reasoning, and knowledge of the wider world through integrated study in the arts and sciences. Liberal arts Bushmeat, defined as raw or processed meat derived from wild animals, colleges can take several steps to capture this interest in global health and is considered a potential source of infection. The HIV epidemic has been enhance their curriculum. associated with the hunting and processing of bushmeat, and recent studies have found evidence of simian foamy viruses in bushmeat samples confiscated at United States ports of entry. Existing US regulations 1551 prohibit importation of bushmeat from specific animals. However, illegal GETTING THE NUMBERS RIGHT: CONSIDERATIONS FOR importation still occurs, and the exact amount of imported bushmeat is QUANTIFICATION OF MALARIA MEDICINES AND RAPID unknown. This project describes bushmeat confiscation reports in the DIAGNOSTIC TESTS IN RESOURCE LIMITED SETTINGS Centers for Disease Control and Prevention (CDC) Quarantine Activity Reporting System (QARS) and attempts to identify geographic and Naomi Printz seasonal trends. A keyword search was performed in QARS to capture all John Snow Inc., Arlington, VA, United States bushmeat-related reports from September 2005 through December 2010. All relevant reports were reviewed and compiled in an analytic database. Countries continue to scale up interventions for the treatment, diagnosis, All items were categorized by CDC-regulated species, including nonhuman and prevention of malaria. Central to the success of these interventions primate, rodent, bat, bird, unknown, and other. In total, 543 confiscated is ensuring that a consistent supply of products is available whenever and bushmeat items, weighing 2303.6 kilograms (kg), were reported and wherever they are needed. Quantification is the process of estimating the recorded in QARS. The median weight of bushmeat per report was 2 quantities and costs of the products required for a specific health program kg and ranged from 0.1 to 650 kg. Half of confiscated bushmeat was and determining when the products should be delivered to ensure an identified as rodent. The most confiscations were reported in 2008 and uninterrupted supply for the program. Quantification is a critical supply the least in 2006. Africa was the most frequent continent of origin, with chain activity that links information on services and commodities from 68% of all confiscated bushmeat originating from Ghana and Nigeria. the facility level with program policies and plans at the national level, Seasonality was evident, with bushmeat confiscations peaking in late and is then used to inform higher level decision making on the financing spring to early summer after adjustment for travel volume. Four times and procurement of commodities. The results of a quantification can be more bushmeat was confiscated during an enhanced surveillance program used to help maximize the use of available resources for procurement, in June 2010 compared to the same period in previous years, suggesting advocate for mobilization of additional resources when needed, and that items were missed during routine inspections. Even with regulations inform manufacturer production cycles and supplier shipment schedules. in place, bushmeat is smuggled into the United States. Longstanding Malaria presents unique challenges to quantification due to: 1) changing cultural practices make it difficult for persons to accept potential epidemiology due to large scale implementation of effective malaria health risks. Therefore, enforcing penalties associated with bushmeat control interventions in diagnosis (particularly RDTs), net distribution confiscations, along with health education aimed at high-risk groups, may campaigns, and increased use of ACTs; 2) seasonality and geographic be useful to deter import attempts. considerations of disease; historical presumptive treatment of malaria with ACTs, rather than confirmed biological diagnosis; inaccurate records 1550 and weak reporting systems for malaria medicine consumption. The following recommendations are offered to help improve the accuracy TEACHING GLOBAL HEALTH IN THE UNDERGRADUATE of quantifications of malaria medicines: 1) in the short term, the effect LIBERAL ARTS of increased diagnosis and prevention efforts will likely not affect the quantities of ACTs required; 2) when developing a supply plan, program 1 2 2 David R. Hill , Robert M. Ainsworth , Uttara Partap managers should arrange shipments to arrive prior to the peak periods of 1National Travel Health Network and Centre and London School of malaria, to avoid overstocking during the dry season, and understocking Hygiene and Tropical Medicine, London, United Kingdom, 2Williams during the rainy season; 3) investments should be made in strengthening College, Williamstown, MA, United States information systems so that future quantifications can be data driven rather than assumption driven; 4) forecasts should be made based on Teaching public health in the undergraduate curriculum of four-year as many data sources (demographic/morbidity data, services data, and institutions has moved forward over the last five years with the Educated consumption data) as possible. The results of forecasts using different data Citizen and Public Health initiative. It has been driven by leaders in public sources should be analyzed and compared; and 5) quantifications should health, the arts, sciences and humanities, and public health organizations, be updated on a quarterly basis with actual consumption data. as well as by student interest. There has been equal interest in global health with a focus on health equity for all persons and particularly for those in low-income countries. Most undergraduate offerings in global or public health have been aligned with universities with graduate programs in the disciplines. However, is likely that strong interest in global health

www.astmh.org 469 1552 of participant satisfaction indicated that past participation in clinical electives abroad was associated with higher satisfaction scores. Scores URBAN POOR HARD HIT BY CHRONIC CONDITIONS on multiple choice knowledge tests increased following the teaching sessions (p < 0.0001). This finding remained after stratification by site, Ronald O. Ondari1, Sarah N. Orutwa1, Joshua M. Orang’o2, Emily 3 year of post-graduate training, and previous global health experience Nyariki (p<0.01 for all comparisons), suggesting that participants in all subgroups 1Highlands Community Assistance Programme (HICAP), Nairobi, Kenya, demonstrated knowledge gain following participation in all four modules. 2Rescue Hope International, Nairobi, Kenya, 3Mwafrika Institute of Participants with lower pre-test scores demonstrated greater knowledge Development, Nairobi, Kenya gain than those with higher pre-test scores (p<0.0001 for all modules), Hypertension and diabetes are no longer diseases of affluence. High suggesting that participants with low baseline GCH knowledge benefited poverty levels in sub-Sahara Africa have heightened their prevalence.Poor most from the modular curriculum. On the other hand, knowledge gain people are suffering from these conditions because of rapid urbanization, did not differ according to level of residency training or previous global cultural factors, poor health management and general effects of poverty. health experience. Item analysis of the knowledge evaluation instrument The study aimed at understanding diabetes and hypertension among demonstrated that 85% of questions were at an appropriate difficulty urban poor, a total of 5,190 individuals were sampled. Data on their risk level, 35% discriminated well between top and bottom performers, was indicated by tobacco use, alcohol consumption, diet and physical 75% increased significantly after teaching sessions and 78% had a good activity, 12 per cent of adult were current smokers and on average item-to-total score correlation. In summary, satisfaction with the modular smoked eight cigarettes a day. 10 per cent were alcohol users and nearly curriculum was high among participants, and knowledge gain could be a third 32 per cent were heavy drinkers. On physical activity, 15 per cent demonstrated unequivocally. This standardized curriculum could be scaled did not engage in any, while over half (54 per cent) had insufficient up for implementation across North American pediatric training centers. intake of fruits and vegetables, while 37 per cent had high salt intake. The prevalence rates among adult women were slightly higher than men. 1554 Increasing with age, respondents aged 40 and 60 likely having diabetes CLINICAL TRIALS IN RESOURCE-LIMITED SETTINGS and 30 and 60 years having hypertension.Heavy alcohol use leads to damage of pancreases - organ producing insulin regulating how sugars Trudie Anne Lang are metabolized.A damaged pancreas results in less insulin therefore University of Oxford, Oxford, United Kingdom increasing levels circulating sugars (glucose specifically). “Diabetes is therefore a manifestation of imbalance in sugar metabolism.According Clinical trials in developing countries lag far behind wealthier regions to findings, some types of alcohol are a source of “empty calories,” body through a lack of knowledge and skills. More disease management gets a lot of calories without a feeling of satiety.In an ideal situation, trials are needed but there is a lack of access to generic clinical trial one should get calories reflecting feeling of sanctification. More empty tools, guidance and training. Furthermore, researchers are daunted by calories consumed, higher chances gaining too much weight, Interfering regulations and guidelines. Existing capacity development activities are with sugar metabolism and increasing chances of diabetes, Leading linked to specific trials and are disease focused; so limit diversification. damage inner lining of blood vessels which lead to high blood pressure. Other factors such as the cost of travel and remote locations of potential High consumption salt leads to high concentrations salt in blood, which research sites also restrict clinical trial capacity development in these body compensates for by absorbing more water from cells. Resulting an regions. Global Health Clinical Trials (www.globalhealthtrials.org) is a increased blood volume, putting pressure on kidneys and blood vessels, new collaborative web-based platform. It is a free, open access and causing damage manifest as high blood pressure. entirely collaborative platform where anyone working on trials can access guidance, tools resources and share their knowledge, views and 1553 experiences. This resource is evidence-led through integrated participatory action research that enables researchers based in these settings to identify PILOTING AND EVALUATING A GLOBAL CHILD HEALTH the problems in their own context and contribute to solutions themselves. CURRICULUM FOR PEDIATRIC RESIDENCY PROGRAMS The platform was released as a pilot in May 2010 and within a year attracted over 1000 members from 56 developing countries. It is not for 1 2 2 Michael Hawkes , Tobey Audcent , Heather MacDonnell , any one disease or just about product development trials. The aim is to 2 3 4 Katherine Moreau , Jeffrey M. Pernica , Maryanne Crockett , Julie support researchers in running their own trials and diversifying in the types 5 3 2 6 Fisher , Andrea Hunter , Amonpreet Sandhu , Sauve Laura , Tinh- of trial that they conduct. Researchers and their staff use the platform to 7 7 2 2 Nhan Luong , Joanne Liu , David M. Goldfarb , Selim Rashed , seek expert and peer advice on diverse issues such as data management, 2 2 8 Arielle Levy , Anne McCarthy , Jennifer Brenner intend-to-treat-analysis and setting up community advisory boards. There 1University of Toronto, Toronto, ON, Canada, 2University of Ottawa, are free e-learning short courses to take researchers and research staff Ottawa, ON, Canada, 3McMaster University, Hamilton, ON, Canada, pragmatically through all the trial steps and processes. In partnership 4University of Manitoba, Winnipeg, MB, Canada, 5University of Calgary, with WHO/TDR we have created an on-line continuing professional Calgary, AB, Canada, 6University of British Columbia, Vancouver, BC, development scheme. This is the first of its kind and this free opportunity Canada, 7University of Montreal, Montreal, QC, Canada, 8University of to build personal learning and training portfolios whilst guiding Calgary, Calgary, ON, Canada professional development will be highly impactful in creating a cadre of developing country clinical trialists. The Global Health Clinical Trials North American pediatricians care for a growing number of immigrant Programme is an open collaboration and is already gaining widespread and refugee children, and training programs should reflect this changing recognition for being the single point of reference for accessing demographic. A modular global child health curriculum designed for information, experts and support on all aspects of running clinical trials in use during academic half-days was developed and piloted across four resource-limited settings. Canadian post-graduate training centers. Here we present the results of an evaluation of participant satisfaction and knowledge gain using a standardized satisfaction survey and pre/post multiple choice knowledge tests. 125 trainees participated from 4 pediatric training centres. 95% completed 2 or more modules and 42% completed all four. Scores on a standardized satisfaction questionnaire were internally consistent (Cronbach’s alpha=0.88 to 0.95 for the four modules) and indicated a high level of participant satisfaction (mean (SD) satisfaction scores of 3.4 (0.6) to 3.6 (0.6) out of maximum total of 5). Analysis of determinants www.astmh.org 470 1555 transmission). The regression model demonstrated an association between relapse rate and land surface temperature, precipitation, temperature USE OF BIOMETRIC DATA IN LINKING HEALTH suitability index (a measure of temperature conditions which allow for DEMOGRAPHIC SURVEILLANCE SYSTEMS (HDSS) AND sporozoite development), and longest unsuitability period. The map of HEALTH MANAGEMENT INFORMATION SYSTEM (HMIS) the median time to relapse, grouped into three classifications of relapse INFORMATION IN IFAKARA AND RUFIJI HDSSs IN TANZANIA rate (≤60 days, 61-180, and >180 days), showed the occurrence of faster relapse (≤60 days) in regions nearly always suitable for malaria Sadick P. Masomhe, Dr. Rashid Khatib, Dr. Baraka Amuri, transmission, and slow relapse (>180 days) in regions only suitable for Mahmoud Kamusi, Khamis Awadh transmission four to six months of the year. While these results are Ifakara Health Institute, Dar es salaam, United Republic of Tanzania based on a relatively small sample of data, given the limited information published on vivax relapse, the data, from more than 36,000 patients, The objective of the study was provide a mechanism to link demographic offers insight into the poorly understood mechanism of relapse. and social economic information captured within HDSS frameworks and Elimination efforts often leave P. vivax the last parasite standing, due to the health facility-based data. House to house fingerprints and facial its ability to relapse. A better understanding of relapse therefore is an images data collection was done for all HDSS members. Field workers each essential component to its control. equipped with a net book computer, fingerprint scanner, web camera, methylated spirit (for cleaning fingers before fingerprint capture) visited households for 12 months from June 2010 up to May 2011. Field workers 1557 did a daily data backup and field supervisors did a weekly data integration. PREVALENT PARASITEMIA, FEVER, ANEMIA AND INHERITED Once integrated the biometric data were taken to the surrounding health BLOOD DISORDERS IN MESO-ENDEMIC WEST SUMBA, facilities within the Demographic Surveillance Area (DSA) where upon INDONESIA: A RANDOMIZED, CROSS-SECTIONAL ANALYSIS each visit a patient is checked using a combined search and data validation using fingerprints, facial image and the demographic data. Once identified Christian P. Nixon1, Christina E. Nixon2, Dian S. Arsyad2, Krisin the patient’s visitation records including health services attendance, Chand2, Frilasita A. Yudhaputri2, Wajiyo Sumarto2, Suradi diagnosis, service and treatment data were captured and linked to his/ Wangsamuda2, Puji B. Asih2, Sylvia S. Marantina2, Din Syaffrudin2, her HDSS profile. In the two pilot health facilities in Rufiji HDSS from J. Kevin Baird3 September 2010 to May 2011,726 patients have had their HDSS and 1University of California, San Francisco, San Francisco, CA, United States, HMIS information accurately linked using the deployed mechanism. At the 2Eijkman Institute of Molecular Biology, Jakarta, Indonesia, 3Eijkman- moment essential data health system analysis, health planning and policy Oxford Clinical Research Unit, Eijkman Institute of Molecular Biology, formulation is lacking because there is no such a system that links HDSS Jakarta, Indonesia and HMIS information. Upon implementation of the exercise, they were concerns from the public on misuse of the biometric data collected and The expected decline in the intensity of malaria in many areas across use of fingerprint scanner as a means for HIV/AIDS testing and for criminal the globe in response to heightened malaria control efforts has placed tracking. Integrating the HDSS and HMIS data sources would provide both increased emphasis on understanding low to moderate transmission the numerator and the denominator population for computation of both settings such as that typified by Southeast Asia. We conducted a disease incidence rates in the population and health service coverage rates cross-sectional socio-demographic and parasitologic survey in a rarely in the health system. studied region of West Sumba district, East Nusa Tenggara Province, Indonesia from August-November 2010. Blood smear analysis of 960 1556 randomly selected individuals revealed meso-endemic malaria with a point prevalence estimate of 24.7%. Plasmodium falciparum infections A REVIEW OF THE GEOGRAPHICAL VARIATION IN predominated over P. vivax, with a few cases of P. malariae, and a single PLASMODIUM VIVAX RELAPSE RATE case of P. ovale noted. Age specific prevalence rates were highest in children and young adults less than 15 years of age, and geometric 1 2 1 Katherine E. Battle , Thomas Van Boeckel , Peter W. Gething , J. mean parasite densities were found to decrease with increasing age. 3 1 Kevin Baird , Simon I. Hay The prevalence of fever (aural temperature ≥ 37.5 °C) among parasitized 1Spatial Ecology and Epidemiology Group, Department of Zoology, and non-parasitized study participants was 7.2% (17/237) and 5.3% University of Oxford, Oxford, United Kingdom, 2Biological Control and (38/723) respectively, suggesting approximately 30% of fevers were Spatial Ecology, Université Libre de Bruxelles, Brussels, Belgium, 3Eijkman likely to be associated with malaria infection. The inherited red blood Oxford Clinical Research Unit, Jakarta, Indonesia cell (RBC) polymorphisms glucose-6-phosphate dehydrogenase (G6PD) deficiency and Southeast Asian ovalocytosis (SAO) were detected in Plasmodium vivax has the widest global geographic distribution of the 17.5% and 22.6% of study subjects respectively, but were not associated malaria parasites known to affect man. Contrary to past beliefs that with the likelihood of parasitemia (P > 0.2) as detected by conventional it is a “benign” form of malaria, it has been shown to result in severe light microscopy. The prevalence of anemia on the day of survey was disease and mortality. Control of P. vivax is complicated by its ability to 43.4% (403/928), and parasitemic study participants were more likely relapse after treatment of the initial infection. Hypnozoites, a dormant to be anemic than those without parasitemia (odds ratio [OR] = 1.46; liver stage of infection, can initiate a relapse weeks or months following 95% confidence interval [CI] = 1.08-1.96; P = 0.014). These findings initial infection. The widely accepted epidemiological understanding is that are consistent with the few prior studies conducted in the region and strains of P. vivax from various geographical areas exhibit different relapse indicate moderate malaria transmission in an endemic area of East Nusa patterns, such that those found in tropical regions will relapse quickly Tengarra Province, that has been stable enough to induce some anti- (3-6 weeks) and those in temperate regions will relapse more slowly (6-12 parasite and anti-disease immunity. Although we did not observe an months). Support for this belief is provided here, in a systematic review association between either SAO and G6PD deficiency and the likelihood of of published and unpublished reports of P. vivax relapse rates in patients patent parasitemia, the impact of these polymorphisms on other aspects not treated with primaquine, the only drug currently available to treat the of infection such as sub-patent parasitemia, parasite density and severe hypnozoite stage of the infection. Statistical analysis was performed to clinical disease remains to be determined. The relatively small proportion identify the association between the rate of relapse and several climatic of parasitemic study subjects with fever for the given transmission intensity and geographical parameters. The relationship between relapse rate and in this study population merits further attention and will be a focal point environment was illustrated as a global map of 146 P. vivax relapse records of future studies. plotted as georeferenced points over a mask of the longest unsuitability period (the length of time in months an area is unsuitable for malaria

www.astmh.org 471 1558 of over 9000 blood samples were analysed by microscopy. Population parasite prevalence rates ranged from 0 to 57%, with Plasmodium MORBIDITY AND MORTALITY CAUSED BY FALCIPARUM falciparum prevalence between 0 and 29%, P. vivax between 0 and 27% AND VIVAX MALARIA IN HYPO- TO MESO-ENDEMIC and P. falciparum mixed infections between 0 and 9%. P. vivax infections WEST SUMBA, INDONESIA: A TWO-YEAR RETROSPECTIVE dominated in 22% of the villages with detectable parasitaemia. Parasite HOSPITAL-BASED STUDY prevalence decreased significantly with altitude, more so for P. falciparum than for P. vivax. 80% or higher mosquito net usage was independently Siti Nurleila1, Din Syafruddin2, Iqbal Elyazar1, J. Kevin Baird1 correlated with lower parasite prevalence. Microscopy and PCR results are 1Eijkman-Oxford Clinical Research Unit, Jakarta, Indonesia, 2Eijkman compared with historic data and discussed in the context of the large- Institute, Jakarta, Indonesia scale roll out of mosquito nets, change in treatment policy, and observed changes in malaria transmission. Plasmodium vivax causes what has often been referred to as benign tertian malaria. This study aimed to measure the relative contributions of falciparum and vivax malaria to the burdens of hospitalized, severe 1560 and fatal malaria the main referral hospital for West Sumba in eastern THE FORCE OF INFECTION: THE KEY TO UNDERSTANDING Indonesia. We systematically examined records of malaria screening and THE EPIDEMIOLOGY OF PLASMODIUM FALCIPARUM admission at Karitas Hospital (115 beds), which served a community of MALARIA IN PAPUA NEW GUINEAN CHILDREN 280,000 people living with a median prevalence of about 6% of both P. falciparum and P. vivax (ranging from 1:1 to 2:1 in surveys). Conducted Ingrid Felger1, Thomas A. Smith1, Sonja Schoepflin1, Peter in 2010, we limited the survey to calendar years 2008 and 2009. Febrile Zimmerman2, Peter Siba3, Ivo Mueller4 patients seeking treatment at this hospital had routine and reliable 1Swiss Tropical and Public Health Institute, Basel, Switzerland, 2Center for microscopic blood film exams for malaria. Among 18,589 febrile patients Global Health and Diseases, Case Western Reserve University, Cleveland, screened, only 2711 were positive and managed as outpatients. Another OH, United States, 3Papua New Guinea Institute of Medical Research, 3484 found positive for malaria were admitted as inpatients and these Goroka, Papua New Guinea, 4Walter and Eliza Hall Institute, Melbourne, records were screened for completeness, with 35 being excluded, leaving Australia 3449 malaria patient records to evaluate. Relevant content of hospital records were manually transferred into 2 separate case record forms (CRF) Genotyping Plasmodium falciparum parasites in longitudinal studies per patient, which were then each double entered into an electronic provides a robust approach for estimating force of infection (FOI) in the database. Only fully reconciled CRFs were uploaded for analysis. Among presence of superinfections. molFOI, defined as the number of new P. the 3449 admissions for malaria, there were 614 patients classified as falciparum clones acquired over time, is molecular parameter equally severely ill, and 66 patients did not survive. Falciparum malaria accounted suitable for describing basic malaria epidemiology as well as for measuring for 65% and 70% of this morbidity and mortality, and vivax malaria outcomes of clinical trials of antimalarial interventions. We investigated contributed 32% and 27% to these burdens in this community. Whereas the potential of molecular parameters to explain differences in risk of P. infants, children, and adolescents carried 82% of the morbidity and falciparum infections and disease between wet and dry season, among mortality burdens due to P. falciparum (OR=4.6; 3.2-5.8), they carried only different age groups and use versus non-use of insecticide treated 62% of this in P. vivax (OR=1.6; 1.2-2.3). Patients with P. falciparum were bednets (ITN). 264 children 1 - 3 years of age from Papua New Guinea much more likely to be classified as having severe illness (OR=2.7; 2.2-3.3), were followed over 16 months with active detection of infection at but the risk of death with a classification of severe illness was the same 2-monthly intervals and during episodes of febrile illness. PCR for the between falciparum and vivax malarias (OR=1.2; 0.7-2.2). Anemia and highly polymorphic genotyping marker merozoite surface antigen 2 was cerebral syndromes dominated among the severely ill with both falciparum performed in all blood samples. To track individual parasite clones in and vivax malaria (>90% of syndromes for each species). These findings consecutive blood samples with maximal resolution PCR fragments were show P. vivax contributed substantially to the burdens of morbidity and sized by capillary electrophoresis. molFOI was identified as explanatory mortality in West Sumba and seemed to have provoked severe disease variable for precisely describing the risk of P. falciparum illness. molFOI was syndromes essentially identical to falciparum malaria in both character and significantly correlated to incidence of episodes, irrespective of whether risk of death. a parasite density cut off was applied or not. Seasonal variation was

observed in molFOI, and thus in the risk of illness. molFOI was significantly 1559 higher during the rainy season than in the dry season. Our analyses suggest a central role of molFOI for explaining differences in the burden

THE CHANGING EPIDEMIOLOGY OF MALARIA IN PAPUA of clinical P. falciparum malaria in our cohort. molFOI almost completely NEW GUINEA explained spatial variation, age trends and effect of ITN use on incidence. Acquisition of new parasite clones seems to be a major factor for clinical 1 1 1 Manuel W. Hetzel , Celine Barnadas , Gibson Gideon , Nandao illness in these children. This study highlights the suitability of a new Tarongka2, Jonah Iga1, Hector Morris2, Peter M. Siba1, Ivo Mueller3 parameter, molFOI, for understanding the epidemiology of clinical malaria in 1Papua New Guinea Institute of Medical Research, Goroka, Papua New young children. We propose to apply the molecular determined parameter 2 Guinea, Papua New Guinea Institute of Medical Research, Madang, molFOI for monitoring effects of malaria interventions. Papua New Guinea, 3Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia Papua New Guinea (PNG) has the highest malaria transmission outside of sub-Saharan Africa. Moreover, the country’s malaria epidemiology is more complex than many other places with four endemic malaria species and a variety of anopheline vectors filling the diverse ecological niches. Only recently, the National Department of Health has re-activated its malaria control program with the up-scaling of insecticide treated nets supported by a Global Fund to Fight AIDS, Tuberculosis and Malaria grant. A country- wide prevalence survey in 70 randomly selected villages investigated the malaria prevalence and species composition. Follow-up surveys in selected sites assessed changes after the introduction of mosquito nets. Malaria prevalence was assessed by light microscopy and PCR. A total

www.astmh.org 472 1561 1562 PLASMODIUM VIVAX GAMETOCYTE DYNAMICS AND THE WIDESPREAD INFECTION OF WILD-LIVING CHIMPANZEES ROLE OF DRUGS IN REDUCING TRANSMISSION POTENTIAL AND GORILLAS WITH PLASMODIUM VIVAX-LIKE PARASITES Nick M. Douglas1, Julie A. Simpson2, Aung P. Phyo3, Enny Weimin Liu1, Katharina S. Shaw2, Yingying Li1, Sabrina Kenangalem4, Jeanne R. Poespoprodjo4, Pratap Singhasivanon5, Locatelli3, Steve Ahuka-Mundeke4, Bila-Isia Inogwabini5, Gerald Nicholas M. Anstey1, Nicholas J. White5, Francois Nosten3, Ric N. H. Learn1, Jean-Bosco N. Ndjango6, Crickette M. Sanz7, David Price1 B. Morgan8, Mary K. Gonder9, Philip J. Kranzusch10, Peter D. 11 12 13 14 1Menzies School of Health Research, Casuarina, Northern Territory, Walsh , Alexander V. Georgiev , Martin N. Muller , Alex Piel , 15 16 17 Australia, 2Centre for Molecular, Environmental, Genetic and Analytic Fiona Stewart , Michael L. Wilson , Anne E. Pusey , Matthew 18 18 19 Epidemiology, University of Melbourne, Melbourne, Australia, 3Shoklo LeBreton , Nathan D. Wolfe , Eitel Mpoudi-Ngole , Eric 3 1 20 21 Malaria Research Unit, Tak, Thailand, 4Papuan Health and Community Delaporte , George M. Shaw , Julian C. Rayner , Paul M. Sharp , 3 1 Development Foundation, Timika, Indonesia, 5Faculty of Tropical Medicine, Martine Peeters , Beatrice H. Hahn Mahidol University, Bangkok, Thailand 1University of Pennsylvania School of Medicine, Philadelphia, PA, United States, 2Columbia University, New York, NY, United States, 3Institut de Designing interventions that will reduce transmission of vivax malaria Recherche pour le Développement (IRD), University of Montpellier 1, requires a detailed understanding of the dynamics of Plasmodium vivax Montpellier, France, 4University of Montpellier, Montpellier, France, 5Projet gametocytemia. We analyzed data from a large randomized controlled Lac Tumba, World Wildlife Fund, Kinshasa, The Democratic Republic of the trial in Northwestern Thailand and two trials in Papua, Indonesia Congo, 6University of Kisangani, Kisangani, The Democratic Republic of to identify and compare risk factors for P. vivax gametocytemia at the Congo, 7Washington University, St. Louis, MO, United States, 8Lincoln enrolment and during the 6 to 9 weeks following treatment. Overall Park Zoo, Chicago, IL, United States, 9University at Albany, State University 492 patients with P. vivax monoinfections were evaluable from Thailand of New York, Albany, NY, United States, 10Harvard Medical School, Boston, and 476 patients with P. vivax infections (162 of whom had concurrent MA, United States, 11VaccinApe, Bethesda, MD, United States, 12Harvard P. falciparum infections) were evaluable from Papua. In Thailand 84.3% University, Cambridge, MA, United States, 13University of New Mexico, (415/492) of patients with monoinfections were gametocytemic at Albuquerque, NM, United States, 14University of California at San Diego, enrolment (median gametocyte density = 266/µL) compared to 66.6% La Jolla, CA, United States, 15University of Cambridge, Cambridge, United (209/314) in Papua (median gametocyte density = 113/µL; p<0.001 for Kingdom, 16University of Minnesota, Minneapolis, MN, United States, gametocyte prevalence and density comparisons). At both sites there was 17Duke University, Durham, NC, United States, 18Global Viral Forecasting a positive correlation between initial asexual parasitemia and gametocyte Initiative, San Francisco, CA, United States, 19Institut de Recherches density (R=0.53 and R=0.47, p<0.001 for both). High asexual parasitemia Médicales et d’études des Plantes Médicinales Prévention du Sida ou was also associated with an increased risk of gametocytemia during Cameroun, Yaoundé, Cameroon, 20Sanger Institute Malaria Programme, follow-up. In Thailand, the cumulative incidence of gametocytemia The Wellcome Trust Sanger Institute, Cambridge, United Kingdom, between day 7 and 42 following dihydroartemisinin+piperaquine 21University of Edinburgh, Institute of Evolutionary Biology, Edinburgh, (DHA+PIP) was 6.92% versus 29.1% following chloroquine (p<0.001). United Kingdom In Papua, the cumulative incidence of gametocytemia between day 7 and 42 was 33.6% following artesunate+amodiaquine (AS+AQ), Plasmodium vivax accounts for over 50% of malaria cases outside of 7.42% following artemether+lumefantrine and 6.80% following Africa, but is not thought to be transmitted in western and central dihydroartemisinin+piperaquine (p<0.001 for DHA+PIP versus AS+AQ and Africa because of the high prevalence of the Duffy-negative trait in local p=0.4 for DHA+PIP versus AM+LUM). Gametocytemia during follow up populations. Nonetheless, the finding that some individuals in west was associated with concurrent asexual parasitemia in 98.9% (172/174) central Africa harbor antibodies to P. vivax surface proteins, together of cases. Plasmodium vivax gametocyte carriage closely mirrors asexual with reports of P. vivax in some travelers returning from this geographic stage infection. The most important strategy for interrupting P. vivax region, have suggested that there might be an as yet undefined reservoir transmission is prevention of relapses, particularly in those with high of P. vivax. Since we have recently found evidence of multiple species of P. asexual parasitaemia. falciparum-related parasites in apes, we used non-invasive methods to determine whether wild-living chimpanzee and gorilla populations are naturally infected with P. vivax. Using P. vivax specific primers to amplify a diagnostic (~300 bp) mitochondrial DNA (mtDNA) fragment, we screened 3,044 chimpanzee, 1,236 gorilla and 513 bonobo fecal samples from 80 different field sites. Although ape P. vivax was detected at an overall low frequency (~1-2%; possibly because of low fecal parasite loads), we found 45 chimpanzee and 30 gorilla samples from 30 field sites to harbor P. vivax sequences. Ape P. vivax was widely distributed among central (P. t. troglodytes) and eastern (P. t. schweinfurthii) chimpanzees, as well as western (Gorilla gorilla gorilla) and eastern (G. beringei graueri) lowland gorillas, but was absent from bonobos (Pan paniscus). Many of the P. vivax positive specimens also contained Laverania parasites, indicating that ape P. vivax occurs frequently in the context of mixed parasite infections. To confirm that the diagnostic PCR was specific for ape P. vivax, we used single genome amplification to generate larger mtDNA fragments (~3- 4.5kb) from a subset of samples. Phylogenetic analyses of these sequences revealed that the ape parasites were nearly identical to each other as well as to human P. vivax. These findings document widespread infection of wild-living chimpanzees and gorillas with P. vivax-like parasites throughout central Africa.

www.astmh.org 473 1563 higher surrounding viable eggs compared to genital mucosal tissue without infection (p=0.034 and p=0.018, respectively). The density of THE URINARY SCHISTOSOMIASIS-BACTERIURIA epithelial Langerhans cells was not different between women with and CONNECTION: A NEW MODEL TO EXPLORE POTENTIAL without genital schistosomiasis (p=0.25). Tissue containing parasite MECHANISMS eggs was significantly more vascularised (vWF) compared to healthy controls (p=0.017). Immunostain with CD31 identified significantly Yi-Ju Hsieh, Chi-Ling Fu, Anuradha Thathireddy, Michael H. more granulation tissue surrounding viable compared to calcified eggs Hsieh (p=0.032). In conclusion, the findings suggest that S. haematobium Stanford University School of Medicine, Stanford, CA, United States infection may cause changes that could increase HIV susceptibility in the female genital mucosa. Cervicovaginal mucosa with S. haematobium eggs Many studies have posited urinary schistosomiasis as a risk factor for contained more HIV target cells and was significantly more vascularised bacterial urinary tract infections (UTI). Bacteriuria may worsen urinary compared to genital mucosa without eggs. The association with calcified schistosomiasis-linked morbidities such as dysuria and hematuria. One eggs may indicate that S. haematobium infection sustains a long-lasting possible reason for reported high rates of Schistosoma haematobium-UTI increased HIV susceptibility in the female genital mucosa. Further studies co-infection is S. haematobium egg-induced shedding of bacteria-coated are needed to explore the effect of anti-schistosomal treatment on lesions urothelial cells into urine, which could facilitate detection of UTI. Another and cell populations in the genital mucosa. possible mechanism is egg granuloma-related urinary tract obstruction, which could promote urinary stasis and bacterial growth. A third possible mechanism is a skewed immune response to S. haematobium 1565 eggs in the bladder which could preclude an effective local response to DIFFERENTIAL ANTI-MALARIAL IMMUNE RESPONSES IN bacteria. Previously we established a mouse model of S. haematobium SCHISTOSOMA MANSONI AND PLASMODIUM COATNEYI egg-induced, non-obstructive immunopathology. Injection of eggs into CO-INFECTED RHESUS MACAQUES the anterior bladder wall avoids obstruction of the ureters and bladder outlet. We used this model to test the urothelial shedding and immune Amma A. Semenya, JoAnn S. Sullivan, John W. Barnwell, W. skewing hypotheses for the urinary schistosomiasis/UTI association. Evan Secor At 1 and 2 weeks post-injection, 30% and 36% of egg-injected mice Centers for Disease Control and Prevention, Atlanta, GA, United States featured urothelial shedding vs none of the vehicle-injected mice. When mice were transurethrally administered uropathogenic E. coli 7 days Schistosomiasis and malaria are the two leading parasitic diseases after egg injection, bacteriuria occurred at high rates (75%) and titers worldwide. Areas endemic for schistosomiasis and malaria overlap in sub- (median 8x105 cfu/ml) vs vehicle-injected mice and mice infected with Saharan Africa as well as other parts of the world. In a previous study, E. coli only (both groups 33% and median 0 cfu/ml). The bladders of we observed that children harboring schistosomes are twice as likely to egg- vs vehicle-injected mice featured more neutrophils (17.6%±4.25 vs have detectable malaria parasitemia as children who have Plasmodium 5.07%±1.09; p=0.0076) and eosinophils (1.62%±0.26 vs 0.799%±0.799; falciparum infection alone. To test whether a concurrent schistosome p=0.0151), suggesting a role for schistosome-induced immune skewing infection would exacerbate a malaria infection, we used the rhesus in susceptibility to bacteriuria. We are working to further dissect the macaque model. Four macaques were percutaneously exposed to 500 mechanisms by which urothelial shedding and/or immune skewing may cercariae of Schistosoma mansoni. At week eight post-infection, these promote synchronous urinary schistosomiasis and bacteriuria. These macaques plus four additional animals were exposed to the bites of studies will enhance understanding of how bacterial uropathogens may Anopheles dirus mosquitoes that were infected with P. coatneyi. Macaques exploit host immune responses “distracted” by urinary schistosomiasis, with schistosomiasis developed higher parasitemia than macaques and may reveal new approaches to reduce the morbidity of both with malaria alone. In addition, anti-malarial drug treatment was more infections. successful in macaques infected with malaria alone than macaques with schistosome co-infection, suggesting that schistosomiasis impairs the 1564 anti-malaria immune responses that are necessary for effective parasite treatment. To confirm this, we analyzed the malaria specific antibody INCREASED LEVELS OF HIV TARGET CELLS AND responses in both groups of macaques and found that co-infected VASCULARITY IN FEMALE GENITAL MUCOSA WITH macaques displayed a significant impairment in their malaria specific SCHISTOSOMA HAEMATOBIUM INFECTION antibody response. We also conducted studies to understand the role of the route of infection during co-infections. For these studies, we again 1 2 1 Peter M. Jourdan , Gabriele Poggensee , Sigve D. Holmen , Svein exposed four macaques to 500 cercariae of S. mansoni. At eight weeks of 3 4 1 G. Gundersen , Borghild Roald , Eyrun F. Kjetland infection, these macaques plus four additional animals were intravenously 1Centre for Imported and Tropical Diseases, Oslo University Hospital, inoculated with 50,000 P. coatneyi blood stage parasites. These macaques Oslo, Norway, 2Department of Infectious Disease Epidemiology, Robert were also given sub-curative doses of anti-malarial treatment throughout Koch Institute, Berlin, Germany, 3Research Unit, Sorlandet Hospital HF, the infection. Analysis of parasitemia and antibody data demonstrate Kristiansand, Norway, 4Centre for Paediatric and Pregnancy Related no significant differences between co-infected and malaria only infected Pathology, Oslo University Hospital Ulleval, Oslo, Norway macaques, suggesting that the route of malaria exposure might mediate the difference in immunological and pathological outcomes in Schistosoma haematobium frequently causes lesions in the female schistosomiasis and malaria co-infections. genital mucosa. Studies suggest that female genital schistosomiasis may increase the risk of human immunodeficiency virus (HIV) transmission. However, the potential mechanisms for such an association have not yet been explored. The aims of this study are to quantify HIV target cells and blood vessels in female genital mucosa infected with S. haematobium. In a cross-sectional study, cervicovaginal biopsies of Malawian women (n=61) and controls were stained with antibodies to CD3, CD8, CD68 (macrophages), S100 protein (epithelial Langerhans cells), CD31 and vWF (endothelial cell markers). CD4+ T lymphocytes were identified from two consecutive CD3+ and CD8+ 3.5 µm thick sections. The density of CD4+ T lymphocytes was significantly higher surrounding calcified S. haematobium eggs, and the density of macrophages was significantly

www.astmh.org 474 1566 1568 SCHISTOSOME EGG ANTIGENS DIRECTLY ACTIVATE HUMAN THE HUMAN IgE RESPONSE TO TEGUMENTAL-ALLERGEN- TROPHOBLASTS LIKE PROTEINS IN SCHISTOSOMA HAEMATOBIUM Emily A. McDonald1, Sunthorn Pond-Tor1, Ling Cheng1, Remigio Harriet A. Dickinson1, Colin M. Fitzsimmons1, Moussa Sacko2, M. Olveda2, Luz Acosta2, Jennifer F. Friedman1, Jonathan D. Kurtis1 David W. Dunne1 1Center for International Health Research, Rhode Island Hospital, 1University of Cambridge, Cambridge, United Kingdom, 2Institut National Providence, RI, United States, 2Research Institute for Tropical Medicine, De Recherche en Sante Publique, Bamako, Mali Manila, Philippines The exact mechanisms mediating the development of human age- Schistosomiasis infects ~ 40 million women of childbearing age, however, dependent immunity to schistosomiasis have yet to be determined. the impact of schistosome infection on the health of a pregnancy remains However, it is known that IgE levels against SmTAL1 (Sm22.6) correlate poorly understood. Previously, we demonstrated that schistosomiasis with resistance to Schistosoma mansoni reinfection in endemic areas. during pregnancy results in pro-inflammatory mediators in maternal, SmTAL1 is a member of the Tegumental-Allergen-Like protein family. placental and fetal blood. Whether placental tissues themselves respond Members of this family are characterized as having two EF hand domains to schistosome infection remains unclear. In this study, we stimulated (or pseudo EF hand domains) and a dynein light chain. The TAL proteins human trophoblast cells isolated from normal, healthy placentas (n=5) have a high level of sequence similarity, but very different developmental with endotoxin-free schistosome egg antigens (SEA) and assayed culture expression patterns. While the S. mansoni TALs are becoming relatively supernatants for inflammatory and fibrotic mediators. Trophoblasts were well characterized, very little is known about their S. haematobium syncytialized prior to stimulation, as the syncytiotrophoblast is responsible homologues, and the S. haematobium genome has yet to be sequenced. for nutrient, gas, and waste exchange, is directly bathed in maternal Now, using a Sanger EST database, we have employed bioinformatic blood, and therefore the most likely trophoblast cell type to respond to methods to predict the sequence of selected ShTALs and confirmed SEA. Of the analytes examined, secretion of interleukin (IL)-6 and IL-8 transcription with PCR using RNA from S. haematobium egg, cercariae was significantly up-regulated in trophoblasts exposed to SEA compared and worm material. We have used RACE (Rapid Amplification of cDNA to control treated cells (3.74 and 2.49 fold, respectively, both P < 0.05). Ends) to determine their full-length sequences and qPCR to determine the Interestingly, IL-10, commonly associated with reactivity to SEA and critical developmental expression of these ShTALs in within the mammalian host. in pregnancy, was unchanged. In addition, molecules involved in placental We have expressed 3 full-length ShTAL proteins as recombinants- ShTAL1 invasion and growth, including tissue inhibitor of metalloproteinase (predominantly worm), ShTAL2 (egg and worm) and ShTAL8 (egg). These (TIMP)-2 and -3, both showed a trend toward increased expression recombinant proteins were used in isotype-specific ELISA assays with following SEA exposure. Finally, key molecules involved in the availability sera from 403 S. haematobium-infected patients from an endemic area of insulin-like growth factor (IGF), IGF binding protein-1 and -5 showed a of Mali in a cross-sectional treatment and reinfection study. IgG1, IgG4 trend toward increased expression with SEA exposure. These data suggest and IgE antibody responses to these three ShTALs will be discussed in the schistosome antigens directly activate human trophoblasts, resulting in context of the developmental expression of the ShTALs, as it is thought increased production of pro-inflammatory cytokines, decreased availability that the developmental expression pattern of the ShTALs will determine of IGF, and altered remodeling of the placental environment. Enrollment is the qualitative and quantitative isotype responses of the infected ongoing and we are extending our analyses to determine the trophoblast population. The study of the ShTALs provides a unique insight into a much signaling pathways activated by SEA stimulation. This report is the first of understudied parasite species, and the immune responses will help us its kind to examine the effect of SEA directly on the human placenta. understand the mechanisms underlying the age-dependent development of human resistance to schistosomiasis. 1567 1569 A NOVEL ROLE FOR IgE IN HUMAN SCHISTOSOMIASIS CO-ADMINISTRATION OF PRAZIQUANTEL AND Daniel Onguru1, YanMei Liang2, Pauline Mwinzi1, Lisa Ganley- ALBENDAZOLE TO SCHOOL CHILDREN LIVING IN A Leal2 UGANDAN COMMUNITY CO-ENDEMIC FOR SCHISTOSOMA 1 2 Kenya Medical Research Institute, Kisumu, Kenya, Boston University MANSONI AND HOOKWORM: POST-TREATMENT PARASITE- School of Medicine, Boston, MA, United States AND ALLERGEN-SPECIFIC HUMORAL RESPONSES AND Resistance to schistosomiasis is associated with increased levels of serum PATTERNS OF REINFECTION parasite-specific IgE. IgE exerts its functions through its cellular receptors, 1 1 2 FcεRI and FcεRII/CD23; however its functional significance requires further Angela Pinot de Moira , Shona Wilson , Edridah Tukahebwa , 1 1 3 characterization in humans. We previously reported that increased levels Frances M. Jones , Colin M. Fitzsimmons , Joseph K. Mwatha , 4 2 1 of CD23+ B cells correlate with resistance to schistosomiasis in hyper- Jeffrey Bethony , Narcis B. Kabatereine , David W. Dunne exposed populations and sought to define their potential function and 1University of Cambridge, Cambridge, United Kingdom, 2Vector Control relationship with IgE. We found that CD23+ B cells are a heterogeneous Division, Kampala, Uganda, 3The Kenya Medical Research Institute, B cell population with functional and phenotypic differences. Circulating Nairobi, Kenya, 4George Washington University, Washington, DC, United CD23+ B cells are uniquely activated in schistosomiasis and express the States CD23b isoform and CXCR5, the homing receptor for lymphoid follicles. Chemotherapeuatic treatment of schistosomiasis in mono-endemic High CXCR5 expression by CD23+ B cells was associated with the capacity communities is often associated with post-treatment changes in parasite- to home to cognate ligand, CXCL13. CD23-bound IgE cross-linking specific antibody responses. Antibody responses to adult worm antigens increased surface expression of CXCR5 suggesting that CD23b+ B cells generally increase after treatment, whilst those to egg antigens tend to home directly into the lymphoid follicles upon antigen capture. Human decrease or are unchanged. In contrast to schistosomiasis, treatment of schistosomiasis is an intravascular parasitic infection associated with a hookworm is generally associated with sharp declines in specific antibody high antigenic burden in the blood. Thus, circulating CD23+ B cells likely responses. Since these two helminth infections frequently co-exist, WHO capture and shuttle antigens from the blood directly to the splenic follicles now advocates concurrent albendazole and praziquantel treatment. through surface bound IgE, thereby highlighting a new function for both Recent studies have shown that the co-administration of these two drugs IgE and B cells. This process appears to play an important role in the does not significantly alter their safety or efficacy, but little is known about development of protective immunity to schistosomiasis. the effects of combined treatment on post-treatment immune responses

www.astmh.org 475 and subsequent reinfection in co-endemic populations. Since there is also 1571 growing evidence that both parasites may protect against allergy, potential impacts on the incidence of allergic disease is another consideration. SUPPORT FOR INTEGRATED COVERAGE SURVEYS: We investigated changes in total, parasite-specific and allergen-specific EXPERIENCE FROM PLATEAU STATE, NIGERIA IgG , IgG and IgE antibody responses following combined praziquantel 1 4 1 2 2 and albendazole treatment of school-aged children living in a Ugandan Scott McPherson , Jonathan D. King , Elizabeth A. Cromwell , 3 2 2 3 community co-endemic for schistosomiasis mansoni and hookworm. Nimzing Jip , Amy Patterson , Patricia Graves , Emmanuel Miri , 2 2 2 2 Post-treatment changes in schistosome-specific and hookworm-specific Darin Evans , Aryc Mosher , Frank Richards , Paul M. Emerson antibody responses were consistent with observations from mono- 1Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United endemic communities. Antibody responses to adult hookworm or States, 2The Carter Center, Atlanta, GA, United States, 3The Carter Center, Schistosoma mansoni egg antigens either decreased after treatment or Jos, Nigeria were unchanged, whereas those to S. mansoni adult worm antigens In Plateau State, volunteer community drug distributors (CDDs) who increased. Post-treatment increases in IgE to adult worm antigens were provided mass drug administration (MDA) for the established lymphatic associated with reduced susceptibility to S. mansoni reinfection, but there filariasis elimination program were trained to register households for was little evidence for any antibody-mediated resistance to hookworm azithromycin MDA as part of the SAFE strategy to eliminate blinding infection. There was some evidence for a negative association between trachoma. During the same period, LLINs were distributed from central schistosomiasis and allergy. Findings will help predict the public health points as part of an enhanced malaria control campaign. An integrated, effects of combined treatment, as well as providing insight into the cluster randomized survey was implemented in three districts to estimate biology of these two helminth infections and also allergic disease. the true coverage of MDA and LLIN interventions. Randomly selected households within a selected community were visited by a survey team 1570 and all household residents were enumerated. Residents who were APPLICATION OF NEW RECOMMENDATIONS FOR ASSESSING present at the time of the interview were asked to report whether they did or did not take azithromycin for trachoma. Participation was verified WHEN TO STOP MASS DISTRIBUTION OF AZITHROMYCIN using the drug distribution registers where available. Reported nets were FOR TRACHOMA observed and individuals were asked about net use. A total of 364 of Jonathan D. King1, Tesfaye Teferi2, Mulat Zerihun2, Mesele 392 visited households were surveyed. From the surveyed households, Damte2, Firew Ayalew2, Elizabeth A. Cromwell1, Zerihun Tadesse2, responses were recorded for 1 858 out of 2 185 registered persons Teshome Gebre2, Ayelegn Mulualem3, Jeremiah Ngondi1, Paul M. (85.0%). Overall, azithromycin was reported as taken by 56.5% (95%CI Emerson1 42.9-70.2%) of surveyed individuals. Among households reporting to have received the drugs from a CDD, antibiotic coverage was 76.5% (68.0- 1The Carter Center, Atlanta, GA, United States, 2The Carter Center, Addis 84.9%). At least 2 nets were reported owned by 79.7% (69.8-89.7%) Ababa, Ethiopia, 3Amhara National Regional State Health Bureau, Bahir of households and 98.3% (96.9-99.7%) reported receiving the newest Dahr, Ethiopia net from the campaign; 52.0% (39.5-64.5%) of households reported To eliminate blinding trachoma, the World Health Organization that the newest net was being used. The household coverage survey recommends implementing the SAFE strategy which includes annual mass identified households that were not registered and therefore did not drug administration (MDA) with azithromycin. Current impact assessment receive antibiotics. Coverage in future rounds of MDA can be enhanced guidelines suggest MDA may stop after 3 to 5 annual rounds where the by updating treatment registers to cover all households, and identifying prevalence of trachomatous inflammation follicular (TF) among children and training new CDDs in unregistered (and untreated) communities. The 1-9 years of age is below 5%, at the sub-implementation unit (the sub- national target of 2 nets per household was nearly achieved, but future district). We applied the current guidelines in 13/21 districts of South efforts should focus on improving net use. This survey highlights the Wollo zone, Amhara Regional State, Ethiopia after 3 years of annual MDA ease and importance of integrated monitoring of interventions through to determine whether to stop MDA. Ten communities each were selected household surveys. with a probability proportionate to population size in 36 sub-districts in the 13 districts. In each community, one development team (defined 1572 segment of the community) was randomly selected and all residents in all households were registered and those present were screened for MASS DRUG ADMINISTRATION FOR HELMINTHS WITH clinical signs of trachoma. Overall, 38,852 residents were registered from ALBENDAZOLE AND IVERMECTIN IN AN AREA ENDEMIC FOR 9,263 households of whom 33,800 (87.0%) were examined. District- STRONGYLOIDES STERCORALIS, ORAN, ARGENTINA level prevalence of TF in children aged 1-9 years ranged from 0.9% to 1 2 64.4% and sub-district prevalence ranged from 0.8% to 72.9%. A total Alejandro J. Krolewiecki , Eugenia Socías , Silvana Pamela 1 1 3 1 of 6/36 sub-districts and 2/13 districts were below the threshold of 5% Cajal , Marisa Juarez , Carlos Villalpando , Monica Carlos , 1 1 4 1 TF in children aged 1-9 years. Surveyed coverage of persons ever taking Marcela Davila , Ruben Cimino , Karen Palacio , Adriana Di Paolo , 5 6 1 2 antibiotics ranged by district from 74.4% to 95.9% and coverage with Aaron Samuels , Thomas Nutman , Jose Gil , Marcelo Abril , Sonia 2 2 3 5 3 rounds of antibiotics ranged from 23.2% to 91.7%. The experience Tarragona , Silvia Gold , Cesar Jaime , Patrick Lammie in South Wollo demonstrates that impact evaluation designed and 1Instituto de Investigaciones en Enfermedades Tropicales, Orán, Argentina, powered to give a prevalence estimate at the sub-district level are possible. 2Fundacio Mundo Sano, Buenos Aires, Argentina, 3Hospital San Vicente de However, the scale of the work required excruciating attention to detail, Paul, Gerencia Sanitaria, Orán, Argentina, 4Global Network for Neglected was logistically challenging to implement, and produced a demanding Tropical Diseases, Sabin Vaccine Institute, Washington, DC, United States, data-entry load. Future impact assessments would be simplified by the 5Division of Parasitic Diseases and Malaria, Centers for Disease Control and use of electronic data collection. Most importantly, interpretation of the Prevention, Atlanta, GA, United States, 6Laboratory of Parasitic Disease, results is not as simple as stopping MDA in sub-districts below 5% given National Institutes of Allergy and Infectious Diseases, Bethesda, MD, the proximity of hyper-endemic sub-districts. Further analysis of the spatial United States distribution of trachoma between sub-districts is needed. Management of soil transmitted helminths (STH) in highly endemic communities is based on school-based mass drug administration (MDA) programs. Due to its unique characteristics, decisions on the optimal tools needed for evaluating prevalence, monitoring efficacy, selecting appropriate communities for intervention and determining the appropriate medications are further complicated when Strongyloides stercoralis (St www.astmh.org 476 st) is included in the spectrum of targeted STHs. In 2010, a community- 1574 based MDA program was started in Orán, northwestern Argentina, an area highly endemic for STH (including St st) The goal of the program ACCURACY OF COVERAGE SURVEY RECALL FOLLOWING is to assess the performance of single dose combination therapy with AN INTEGRATED MDA FOR LYMPHATIC FILARIASIS, albendazole and ivermectin, and to report the utility of a new recombinant SCHISTOSOMIASIS AND SOIL-TRANSMITTED HELMINTHIASIS antigen based ELISA for St st as a tool for assessing seroprevalence. The 1 2 3 intervention population consisted of approximately 2400 individuals; Philip J. Budge , Kodzo A. Anthony , Edmond Sognikin , 4 4 4 1200 from 3 rural communities, and an additional 1200 from 3 urban/ Amanda Akossa , Els Mathieu , Michael Deming peri-uban communities. Plantations in the rural area and street blocks in 1Epidemic Intelligence Service and Division of Parasitic Diseases and the urban/peri-urban area were used as the unit of randomization; 20% Malaria, Centers for Disease Control and Prevention, Atlanta, GA, United of the population was selected for sampling. The following parameters States, 2Health Development International, Lomé, Togo, 3Ministry of were assessed in each individual: single stool specimen analyzed through Health, Kara, Togo, 4Division of Parasitic Diseases and Malaria, Centers for a comprehensive panel including sedimentation concentration, Harada- Disease Control and Prevention, Atlanta, GA, United States Mori, agar plate and Baermann techniques; St st NIE-ELISA serology Household-based coverage surveys are an important means of ensuring and hemoglobin. We present the preliminary results of the initial pilot that mass drug administrations (MDA) for several neglected tropical intervention in one rural and one urban community. The overall prevalence diseases reach target coverage levels. Such surveys are used for validating of STH by stool examination was found to be 32%, with 12% positive for coverage reported by drug distributors because the two methods St st. Sensitivity increased to 31% for St st when NIE-ELISA was included. are independent of each other; coverage surveys do not depend on A total of 864 individuals from 2 communities with a population of 1127 accurate reporting or denominator estimates. A potential disadvantage persons were treated with ivermectin and albendazole. Active and passive of coverage surveys, however, is recall bias. We tested recall accuracy surveillance revealed no significant (Grade 3 or 4) adverse events. Only 3 in surveys conducted 1, 6, and 12 months after an MDA in Togo, in individuals refused treatment. This initial MDA comprehensively targeting which three drugs (albendazole, ivermectin, and praziquantel) were STH with the added use of St st serology for prevalence calculations is distributed. Independent observers ensured all MDA treatments were a promising approach with an appropriate safety and efficacy proven accurately recorded in registers for comparison with survey responses. regimen. Further inclusion of the remaining communities and rounds of A unique sample of compounds (household groups) was systematically treatment will provide valuable information for defining a strategy for the selected for each survey. All compound residents (mothers answered for management of STH in highly endemic communities. children <10 years of age) were shown examples of pills given during the MDA and asked which they had swallowed. Responses from 506, 1139, 1573 and 963 persons at the 1, 6, and 12 month surveys, respectively, were TWO YEARS EXPERIENCE OF INTEGRATED MASS DRUGS analyzed. Coverage among these persons (defined as having taken at ADMINISTRATION OF NTD CONTROL IN TANZANIA least one MDA drug) was 88%, 87%, and 79%, respectively, according to MDA registers (the lower result at 12 months is likely an artifact due MAINLAND to poor matching of respondents to the MDA register data). Coverage Upendo John Mwingira, A. Nshala, B. Kilembe, M.N. Malecela, estimates based on respondent recall were 88% (95% confidence interval D.W. Mbando [CI] 86-91%), 91% (CI 89-93%), and 90% (CI 87-91%), respectively. Ministry of Health and Social Welfare, Dar Es Salaam, United Republic of Concordance between respondent recall and register data was >93% Tanzania at 1 and 6 months. Respondents generally distinguished between pills similar in appearance; 97% of those taking albendazole (large, white, Tanzania has embarked in an integrated control of Neglected Tropical rectangular, maximum dose 1 tablet) reported taking ½ or 1 tablet, while Disease (NTD) since 2009. Preventive Chemotherapy (PCT) targeted only 32% reported taking only 1 praziquantel tablet (large, white, oval, Diseases endemic in Tanzania are Lymphatic filariasis (LF), Onchocerciasis, dose range 1-5 tablets). Concordance for correct recall of individual Trachoma, schistosomiasis as well as STH. These diseases overlap medications was >80% in all surveys, while concordance for correctly throughout the country. Programme activities include; Social mobilization, remembering pill dosage ranged from 41% to 85%. In this population, CS training, drug delivery, data collection and compilation all in an integrated provided accurate and consistent estimates of overall coverage for up to manner. Implementation Unit (IU) for control activities is districts councils one year following an integrated MDA. These data confirm the usefulness and since there are over 140, a phased scale up is adopted to reach full of CS in monitoring and evaluation of MDA, and suggest that CS might geographical coverage by 2013. Coordination of the NTD control is by be postponed for up to one year without compromising recall accuracy, the government and implementation is in a cascade manner. In the past which might allow for integration into periodic large, multipurpose 2 years the programme has achieved increased therapeutic as well as surveys. geographic coverage towards integrated NTDs control. In 2009, working in 36 IUs (27% geographical coverage) a total of 2500 Health Workers 1575 and 28,000 drug distributors were trained. Advocacy and community mobilization reached 9700 influential people and decision makers. A SPATIAL APPROACH TO QUANTIFYING THE WATER About 10 million at risk population were treated. With increased partner SUPPLY, SANITATION AND HYGIENE (WASH) - HELMINTH involvement and funding in 2011 two phased scale up approach will INFECTION-ANAEMIA CAUSAL PATHWAY IN CHILDREN IN increase the geographical coverage to 75 (56% geographical coverage) SUB-SAHARAN AFRICA and reaching out to 21 million at risk population by the end of year 2011. An integrated training manual, IEC materials and monitoring and Ricardo J. Soares Magalhães, Archie C. Clements evaluation tools have been developed and used in the field. The profile University of Queensland, Herston, Australia of NTDs is scaled up as it is now reflected into the Health Sector Strategic Inadequate water supply, sanitation and hygiene (WASH) are well Plan III, a reference document of the Ministry. With the support of WHO, a known risk factors for infections with schistosomes and soil-transmitted master plan for NTDs is prepared and pharmacovigilance manual has been helminths. Urinary schistosomiasis and hookworm infections are known prepared. Despite the achievements, several challenges were encountered; to cause anaemia which is a severe public health problem in most these include; poor coordination among stakeholders, disease specific countries of sub-Saharan Africa. Using a novel, spatial analytic approach, focus, low urban coverage, lower attention to non PCT NTDs as well as we aimed to quantify for the first time the role of WASH in the risk of non PCT activities like surgeries. Benefits of Integration outweigh the Schistosoma haematobium, S. mansoni and hookworm infection in school challenges and it’s a country’s goal to achieve full and effective integration age children in West Africa; estimate the risk of anaemia in children soonest possible.

www.astmh.org 477 aged 1-4 y (preschool children) attributable to malnutrition, malaria, 1577 and helminth infections; and estimate the number of anaemia cases in preschool children for 2011. We generated predictive maps showing DISSECTING HUMAN ANTIBODY RESPONSE AGAINST probability of absence of WASH. These predictions were then used as DENGUE VIRUS USING REVERSE GENETICS covariates in Bayesian geostatistical models for the three helminth species. 1 2 2 Bayesian geostatistical models were subsequently developed to predict Wahala M. Wahala , Siritorn Butrapet , Claire Y. Huang , 1 the geographical distribution of anaemia of preschool children, adjusting Aravinda M. de Silva for their nutritional status, predicted Plasmodium falciparum parasite 1University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 2Centers for Disease Control and Prevention, Fort Collins, CO, United rate in the 2- to 10-y age group (Pf PR2-10), and predicted prevalence of S. haematobium and hookworm infections. We estimated attributable States fractions of water supply for S. mansoni and S. haematobium to be 47% Human exposed to dengue virus (DENV) develop a type specific and 71% respectively. The attributable fraction of natural floor type was neutralizing antibody (nAb) response against the infecting serotype. 21% for S. haematobium, 16% for S. mansoni and 86% for hookworm. However, the epitopes recognized by these nAbs in polyclonal sera are An estimated 36.8%, 14.9%, 3.7%, 4.2%, and 0.9% of anaemia cases not known. Here, we describe a reverse genetic approach to map the sites could be averted by treating malnutrition, malaria, S. haematobium on the DENV envelope (E) protein that are targeted by nAbs in human infections, hookworm infections, and S. haematobium/hookworm sera. Mouse monoclonal antibodies (mAbs) that strongly neutralize DENV coinfections, respectively. We estimate that in 2011, approximately 6.7 type specifically bind to a unique epitope located on the lateral ridge of million children aged 1-4 y are anaemic in the three study countries. the domain III (EDIII) of the E protein. Sub complex mouse mAbs that This work identified communities in West Africa where preventive neutralize more than one DENV serotype, but not all the four serotypes, chemotherapy integrated with interventions to improve WASH will yield target an epitope located on the A strand of the EDIII. We have used the greatest health benefits. It also identifies the geographical limits of bacterially expressed recombinant EDIII (rEDIII) protein to deplete the anaemia burden and the contribution that malnutrition and parasites EDIII reactive antibodies from polyclonal sera and demonstrated that EDIII make to anaemia. antibodies make only a minor contribution to the neutralization potency of human dengue immune sera. One potential problem with this approach 1576 is that it may not deplete all EDIII reactive antibodies because of folding ESTIMATING COSTS AND FUNDING GAPS OF INTEGRATED differences between the recombinant protein and the native protein on the virus. We used recombinant DENV2 viruses that contain mutations NTD CONTROL PROGRAMS: THE FUNDING GAP ANALYSIS on the lateral ridge (FG loop) and A strand epitopes (AA positions at TOOL (FGAT) 305,307,310), to measure the contribution of human EDIII reactive Brian Chu1, Kathryn Crowley2, Philip Downs2, Achille Kabore2, antibodies in type specific neutralization. The mutant viruses escaped from Jennifer Leopold2, Ruth Yohannes2, Katie Zoerhoff2, Jennifer neutralization by mouse mAbs that bind to lateral ridge and A strand Einberg3, Jennifer Fox3, Molly Mort3 epitopes. However, the 50% neutralization titers of human immune sera were similar for wild type and EDIII mutant viruses. In agreement with 1Task Force for Global Health, Decatur, GA, United States, 2RTI previous EDIII depletion studies, our results indicated that EDIII epitopes International, Washington, DC, United States, 3Independent Consultant, targeted by mouse neutralizing antibodies were not the target of the Seattle, WA, United States human neutralizing response. We hypothesize that nAbs in human sera With neglected tropical disease (NTD) control programs being increasingly mainly target sites located on the domain I /II of the E protein. We have integrated and expanded, it is critical to improve the accuracy of identified putative binding sites on the EDI/II and currently experiments are estimated activity costs and funding gaps for budget and planning in progress to mutate these sites on DENV3 and DENV2 E protein to test purposes. Moreover, there is a growing need to better understand the the hypothesis. cost drivers of implementing integrated control in order to determine an efficient allocation of available resources. To meet these objectives, the 1578 Funding Gap Analysis Tool (FGAT) was developed by the USAID funded NTD Control Program, led by RTI International with WHO endorsement, DENV-3 GENOTYPE SPECIFIC NEUTRALIZATION BY HUMAN to assist national NTD programs to organize and analyze activity costs POLYCLONAL SERUM AND HUMAN MONOCLONAL concordant to their national plans of action. To date, the FGAT has been ANTIBODIES completed by Ministries of Health in over a dozen countries, revealing 1 2 3 several valuable outcomes. The FGAT allows countries to recognize William B. Messer , Scott A. Smith , Jeremy P. Huynh , James E. 2 1 3 cost efficiencies that can be achieved by integrating high cost-driving Crowe , Aravinda M. de Silva , Ralph S. Baric activities such as training, drug delivery, and social mobilization between 1University of North Carolina School of Medicine, Chapel Hill, NC, United disease programs. It also permits countries to calculate the effect of a States, 2Vanderbilt University School of Medicine, Nashville, TN, United significant increase or decrease in key costs, such as fuel or medicines, States, 3University of North Carolina School of Public Health, Chapel Hill, on the program as a whole. Furthermore, the FGAT can direct funding NC, United States into specific program areas by identifying gaps at the sub-activity and Dengue viruses (DENV) are enveloped single-stranded positive-sense RNA district levels that may be undetected at higher levels alone. Such results viruses transmitted by Aedes aegypti and A. albopictus mosquitoes. There ultimately lead to more precise budgets and standardized year-to-year are four genetically distinct serotypes designated DENV-1 through DENV-4. cost estimates. Additionally, the FGAT provides NTD programs data- DENV-3 is further subdivided into four distinct genotypes designated I-IV. related ancillary benefits including a referential database for analysis and The dengue scientific community has long contended that infection with summarized outputs for donor advocacy. In the longer term, the data one serotype confers lifelong protection against subsequent infection with collected by FGAT will allow for in-depth longitudinal and cross-country the same serotype, irrespective of virus genotype. However this hypothesis cost analyses in tandem with program evaluation data. These findings will has never been rigorously tested and the role of DENV genotypic variation be highly beneficial for guiding cost-effective decisions and practical plans in protection from repeated infection is unknown. Recent studies have of action for integrated NTD control programs. shown that in vitro neutralization titers vary substantial by DENV genotype tested and low titers (<1:50) may not be protective. To better understand the role genotypic variation plays in DENV-3 neutralization and protection, we previously designed and constructed a panel of isogenic, recombinant DENV-3 infectious clones, each expressing a representative envelope glycoprotein (E) from a different DENV-3 genotype. When the recombinant www.astmh.org 478 viruses were tested in neutralization assays using human homotypic 1580 primary dengue immune sera, neutralization titers ranged varied by as much as 10-fold, depending on the genotype of the E protein expressed MODULATION OF HUMAN INNATE IMMUNITY BY SRI by the virus. The observed variability in neutralization titers suggests LANKAN DENGUE-3 VIRUSES ASSOCIATED WITH DENGUE that relatively few residue changes in the E glycoprotein may have FEVER AND DENGUE HEMORRHAGIC FEVER significant effects on DENV specific humoral immunity and may influence 1 2 1 antibody mediated protection in the setting of both natural infection Sarah Pagni , Kizzmeka Corbett , Dabeiba Bernal-Rubio , 2 1 and vaccination. To further explore the role of genotypic variation, six Aravinda deSilva , Ana Fernandez-Sesma anti-DENV-3 human mAbs were tested against the clone panel. One 1Mount Sinai School of Medicine, New York, NY, United States, 2University mAb exhibited strong genotype-specific neutralization variability. Using of North Carolina School of Medicine, Chapel Hill, NC, United States recombinant E protein, site-directed mutagenesis in the parent clone Dengue Virus (DENV) is the leading mosquito borne viral threat in the background and the generation of mAb escape mutants against this world with 50 to100 million people infected and 2.5 billion people at mAb the genotype specific amino acid residues were mapped on the E risk annually. Infection by one of the four serotypes of DENV can clinically glycoprotein. The DENV3 recombinant virus panel described here is a present as dengue fever (DF), a febrile illness or the more severe dengue useful tool for mapping antibody responses and assessing the breadth of hemorrhagic fever (DHF) or dengue shock syndrome (DSS), which is natural and vaccine induced immune responses. mostly observed in people experiencing a secondary DENV infection. (DF is very common following primary infections.) There are several potential 1579 reasons as to how these more severe secondary infections arise including ANALYSIS AND IDENTIFICATION OF EPITOPES ON DENGUE antibody dependent enhancement (ADE), T-cell mediated immunity, and VIRUS ENVELOPE PROTEIN RECOGNIZED BY MONOCLONAL differences in viral genotypes. The emergence of a new strain of DENV3 in Sri Lanka in 1989 has been associated with an increase in disease ANTIBODIES AND POLYCLONAL HUMAN SERA BY A HIGH severity, as reported previously. Our project aims to determine if there THROUGHPUT ASSAY are intrinsic differences in the ability of Sri Lankan DENV3 strains isolated Wei-Kung Wang1, Hong-En Lin2, Wen-Yang Tsai1, I-Ju Liu3, Pi- before and after the emergence of DHF to infect human immune cells. Chun Li3, Mei-Ying Liao3, Jih-Jin Tsai4, Yi-Chieh Wu1, Chih-Yun We obtained DENV-3 viruses derived from primary isolates from Sri Lanka Lai1, Chih-Hsuan Lu2, Gwong-Jen Chang5, Han-Chung Wu3 and analyzed their replication and innate immune phenotype in a primary human system. Using monocytes and monocyte derived dendritic cells 1Tropical Medicine, JABSOM, University of Hawaii at Manoa, Honolulu, HI, (DCs) we studied virus replication, infectious particle release and as well United States, 2Institute of Microbiology, National Taiwan University, Taipei, as the innate immune responses in infected cells by plaque assay, qRT-PCR Taiwan, 3Institute of Cellular and Organismic Biology, Academia Sinica, and multiplex ELISA. Our data shows an increase in interferon (IFN) and Taipei, Taiwan, 4Tropical Medicine Center, Kaohsiung Medical University IFN -stimulated gene induction in DCs from multiple donors infected with Hospital, Kaohsiung, Kaohsiung, Taiwan, 5Centers for Disease Control and post-DHF associated DENV-3, compared to the pre-DHF associated DENV- Prevention, Fort Collins, CO, United States 3. Infections of DCs with our DENV-3 isolates in the presence of DENV + The envelope (E) protein of dengue virus (DENV) is major target of sera are ongoing to analyze their phenotype in an ADE context. Future neutralizing antibodies and dengue vaccine development. While previous work will also investigate specific mutations in the DENV NS2B3 and NS5 studies on domain III or domain I/II alone have reported several epitopes genes may correspond to some of the observed differences in virulence of monoclonal antibodies (mAbs) againt DENV E protein, several questions between the viruses. including the possibility of interdomain epitopes, the relationship between epitopes and binding specificity as well as neutralizing potency remains 1581 largely unexplored. We developed a high throughout dot blot assay by using a panel of 67 alanine mutants of predicted surface-exposed ROLE OF ACUTE B CELL RESPONSE IN DENGUE DISEASE E residues as a systemic approach to identify epitopes recognized by SEVERITY AND LONG-TERM IMMUNITY 12 mouse mAbs and polyclonal human sera, followed by confirmation Simona Zompi1, Magelda Montoya2, Marie Pohl1, Angel with capture-ELISA. Three mAbs were found to recognize a novel 2 1 epitope involving residues at domain II central interface, and three Balmaseda , Eva Harris mAbs recognized residues at both domain III and lateral ridge of domain 1Division of Infectious Diseases and Vaccinology, School of Public Health, II, suggesting the presence of interdomain epitopes. Analysis of the University of California, Berkeley, Berkeley, CA, United States, 2Laboratorio conservation index of each epitope residue and conservation score of Nacional de Virología, Centro Nacional de Diagnóstico y Referencia, 91 mouse mAbs reported thus far revealed that the conservation scores Ministerio de Salud, Managua, Nicaragua correlated with the binding specificity. Compared with mAbs generated Dengue, caused by four dengue virus serotypes (DENV-1-4), is the most by traditional protocol, the potent neutralizing mAbs generated by a prevalent mosquito-borne viral disease in humans, causing classic dengue new protocol recognized multiple residues in A strand or residues in C fever (DF) and dengue hemorrhagic fever/dengue shock syndrome strand/CC’ loop of DENV2 and DENV1, as well as multiple residues in (DHF/DSS). While severe disease has been associated with heterotypic BC loop and residues in DE loop, EF loop/F strand or G strand of DENV1. secondary DENV infection mediated by antibody-dependent enhancement These findings have implications for future development of epitope- and stimulation of cross-reactive T cells, the vast majority of secondary specific diagnostics and epitope-based dengue vaccine. Moreover, the infections result in mild or asymptomatic disease, suggesting a protective predominant epitopes of anti-E antibodies in polyclonal sera were found role of the cross-reactive immune response. Little is known about the to include fusion loop residues and some non-fusion residues in the same acute B cell response during DENV infection and its impact on the breadth or adjacent monomer, adding to our understanding of humeral immune and quality of the long-term humoral response. During the 2010 epidemic, responses to DENV at the epitope level. we studied the B cell response in 193 children suspected of DENV infection who were enrolled in a hospital-based study in Managua, Nicaragua, including 127 DENV-positive cases and 66 Other Febrile Illnesses (OFI). The dominant serotype was DENV-3 (82.6%). Sixty-four patients had a primary DENV infection, 54 experienced a secondary infection and 5 cases were undetermined. We measured by flow cytometry the percent of naïve and memory B cells and plasmablasts (PB)/plasma cells (PC) in fresh whole blood and demonstrated a significant increase in PB/PCs in DENV-positive cases when compared to OFI (1.5+/-0.2; n=83 vs. 0.6+/-0.2; www.astmh.org 479 n=40; p=0.003). No significant difference was found among naïve and virus decline seen. In addition, though we are missing data in the early memory B cell compartments. These data correlated with the number of stages of infection, as we are using hospitalised cases, we also look DENV-specific antibody (IgG)-secreting cells measured by ELISpot ex vivo at the determinants of the rate of increase of viraemia. We find that (representing the circulating PC) and after polyclonal in vitro stimulation differences in the modelled immune response growth can recreate the (representing the circulating memory B cells). After a secondary DENV different virus curves of individuals. We see differences in estimated model infection, we found a mean of 2,046 DENV-specific PC/106 PBMCs (14.7% parameters between primary and secondary infection. We identify possible of total IgG) and 870 DENV-specific memory B cells/106 PBMCs (3.8% of differences in the efficiency of virus entry to the cell, initial number total IgG). We are currently measuring the DENV-specific neutralization of antigen recognising immune cells and the efficacy of the immune capacity of the sera, using a flow cytometry-based assay, as well as the response as potential explanations of differences between primary and avidity of the sera in longitudinal samples (3, 6 and 12 months after secondary infections. Incorporating immunological data into our inferential infection) using a competition-based ELISA with recent Nicaraguan framework, we are able to look more in depth at the immunological DENV strains. The breadth of the acute B cell response will be correlated processes governing infection and the magnitude of these processes in with severity of disease and the neutralization capacity and avidity of different people. This suggests differences in pathogenesis which are the serum. This study improves our knowledge of B cell and long-term correlated with primary and secondary disease, and provides more insight humoral responses to DENV infection, which could ultimately impact the into the interplay between virus and immune dynamics. This initial attempt design of safe and effective dengue vaccines. aims to develop a mathematical model of pathogenesis which is tightly coupled to data to describe dengue infection dynamics. As well as helping 1582 in the understanding of the infection process, greater understanding of these processes and their timing could help to understand the RISK FACTORS OF DENGUE HEMORRHAGIC FEVER IN A transmission of the virus and the impact of control measures. We illustrate PREDOMINANT DENGUE SEROTYPE 2-INFECTED CASE- this by considering the potential impact of antiviral drugs on dengue CONTROL STUDY infection and transmission. Junxiong Pang, Agus Salim, Vernon J. Lee, Kee Seng Chia, Yee Sin Leo, David C. Lye 1584 National University of Singapore, Singapore, Singapore DIFFERENCES IN INHERENT SUSCEPTIBILITY TO Dengue, an arthropod-borne virus infection, is of serious concern in PLASMODIUM FALCIPARUM AND RATES OF NATURAL tropical and subtropical regions. Dengue hemorrhagic fever (DHF) is a INFECTION IN POPULATION SUBGROUPS OF ANOPHELES severe form of dengue where several risk factors e.g white race, female GAMBIAE gender, dengue serotype 2, allergy, diabetes and hypertension were Michelle M. Riehle1, Wamdaogo M. Guelbeogo2, Awa Gneme2, reported in different studies. In our 2004 retrospective study where Gregory Snyder1, Karin Eiglmeier3, Inge Holm3, Emmanuel serotype 1 predominated, gender, ethnicity and comobidities were not Bischoff3, Thierry Garnier3, Oumou Niare4, Madjou Sacko4, significantly associated with DHF. Our current retrospective case-control Boubacar Coulibaly4, Sekou F. Traore4, N’Fale Sagnon2, Kenneth D. study explores these risk factors in 2006 and 2007-2008 adult dengue Vernick3 cases in Singapore where serotypes 1 and 2 predominated respectively. 1University of Minnesota, St. Paul/Minneapolis, MN, United States, 2Centre In this study, 149 DHF and 326 DF from the year of 2006 and 669 DHF National de Recherche et de Formation sur le Paludisme, Ouagadougou, and 1141 DF from the year of 2007-2008 were included. We performed Burkina Faso, 3Institut Pasteur, Paris, France, 4Malaria Research and univariate descriptive analysis, and adjusted measures for the association Training Center, University of Bamako, Bamako, Mali were estimated using multivariate logistic regression adjusting for potential confounding variables. Results from 2006 data, similarly in 2004 study, Understanding and characterizing how population subgroups within showed no significant association for gender, comobidities and ethnicity Anopheles gambiae s.s. differ in their susceptibility to P. falciparum with DHF. In contrast, results of 2007-2008 showed age groups 30-39 infection is important for the development of better and more exacting years (adjusted OR= 1.41, p=0.008) and 40-49 years (adjusted OR=1.34, control measures. We combined experimental laboratory infections and p=0.042), female gender (adjusted OR=1.56, p<0.0001), Chinese race capture of naturally infected adult mosquitoes in order to address two (adjusted OR=3.13, p<0.0001), diabetes (adjusted OR=1.82, p=0.016) and important questions (1) differences in the inherent genetic susceptibility diabetes with hypertension (adjusted OR=2.16, p=0.013) as risk factors to malaria parasites, as measured by experimental infection with for DHF. This study also suggested that reported risk factors for DHF may gametocytes, and (2) differences in natural infection rates amongst wild differ between serotype 1 and 2. In conclusion, older Chinese female caught female mosquitoes. The latter approach summarizes natural patients with both diabetes and hypertension may be at greatest risk for variables such as behavior, mosquito age, and bloodmeal source, DHF when serotype 2 predominates. whereas the former approach controls for these variables. Membership in population subgroups was determined by genotyping. 1583 1585 MODELLING DENGUE PATHOGENESIS, CONSIDERING DIFFERENCES BETWEEN PRIMARY AND SECONDARY TRANSPOSON-BASED “FORWARD” GENETICS IN ANOPHELES INFECTION STEPHENSI Hannah Clapham1, Vianney Tricou2, Tien Nguyen Hanh2, Bridget David A. O’Brochta1, Robert A. Harrell2, Channa Aluvihare2, Wills2, Maciej Boni2, Jeremy Farrar2, Cameron Simmons2, Neil Kristina L. Pilitt1, Robert T. Alford1 1 Ferguson 1University of Maryland, College Park, Rockville, MD, United States, 1Imperial College, London, United Kingdom, 2Oxford University Clinical 2University of Maryland Insect Transformation Facility, Rockville, MD, Research Unit, HCMC, Vietnam United States We describe a mathematical model of dengue virus infection within an Transposons can be useful for not only shuttling transgenes into genomes individual, parameterised with sequential viraemia data. In fitting this but, under the right conditions, also for identifying and isolating genes. model to multiple patients’ data, we are able to consider what explains Transposon-based gene- and enhancer-trap technologies are particularly differences seen between people, between primary and secondary powerful because they permit the identification of interesting genes or infection and possibly between different disease severities. We assess enhancers to be recognized based on spatial and temporal patterns of the factors explaining the timing of the peak of virus and the rate of reporter gene expression from the transposon. In the case of transposon- www.astmh.org 480 based gene-traps the transposon insertion site is in the gene responsible isolated subspecies. Linkage disequilibrium analysis of SNPs in cDNAs was for the observed pattern of reporter gene expression, allowing the gene performed on 10 different populations from Senegal, and 10 populations to be easily isolated. Here we report data showing the feasibility of from throughout the world. There was a large amount of disequilibrium performing transposon-based enhancer- and gene-traps in Anopheles among alleles on all three chromosomes. We tested for suppression stephensi using the piggyBac transposon. Transgenic lines with piggyBac of recombination associated with inversions that had been previously elements containing an enhanced cyan fluorescent protein (ECFP) gene detected in Aaf. We crossed “F” males with Higgs White Eye (HWE) Aaa under the regulatory control of the 3xP3 promoter were remobilized after females and then backcrossed male F1 to HWE females. We anticipated crossing to a transgenic line expressing piggyBac transposase. Germ- finding suppressed recombination or that the we locus would be 14cM line transpositions were detected in approximately 1% of the progeny. from the Sex locus as previously reported. Instead the loci were 14cM The activity of the 3xP3 promoter is sensitive to changes in its position apart in some families but unlinked in others suggesting a translocation. within the genome and can detect the presence of local enhancers and All of these results suggest that Aaf is comprised of multiple subspecies promoters. A diverse collection of transgenic lines was made in which in West Africa that can be distinguished based upon chromosomal ECFP expression was in interesting and useful temporal and spatial polymorphisms and molecular genetic markers. patterns demonstrating the potential of this approach. This technology promises to create new opportunities to investigate insect/pathogen 1588 interactions. POTENTIAL GENOTYPIC IDENTIFICATION OF THE BELOW 1586 GROUND PHENOTYPE IN THE CULEX PIPIENS COMPLEX IN CALIFORNIA USING SNP GENOTYPING GENETIC ANALYSIS OF PREFERENCE FOR HUMAN SCENT IN THE DENGUE FEVER MOSQUITO, AEDES AEGYPTI Rebecca T. Trout Fryxell, Stephanie N. Siefert, Yoosook Lee, Gregory Lanzaro, Anthony Cornel 1 2 2 Carolyn S. McBride , Joel Lutomiah , Rosemary Sang , Leslie B. University of California Davis, Davis, CA, United States Vosshall1 The California West Nile virus vector is the Culex pipiens s.l. complex, 1Howard Hughes Medical Institute - Rockefeller University, New York, NY, which displays an array of behavioral adaptations and morphological United States, 2Kenya Medical Research Institute, Nairobi, Kenya differences that make efforts to reduce their numbers challenging. An ancestral, forest form of the Dengue Fever Mosquito, Aedes aegypti, It is unclear if the observed variation is associated with genetically prefers the odor of non-human animals, while a more recently evolved, distinct populations, differing taxa, or if it represents a high degree domestic form strongly prefers human odor. Classic work from the 1970’s of polymorphism within a single panmictic unit of randomly mating and 1980’s showed that these two forms coexist in several places along individuals. The current consensus in California is that the two nominal the coast of East Africa, where they maintain their ecological differences members, “quinquefasciatus” (south of 39°N) and “pipiens” (north 36°N) despite being fully interfertile. This situation provides an unusual and their respective hybrids (between 39° and 36°N), occur in California. opportunity to examine the genetic basis of traits that adapt mosquitoes Because they vector several zoonotic diseases (e.g., dog heartworm, avian to humans. We have verified the continued coexistence of human and malaria, avian pox virus), the Cx. pipiens complex is receiving increasingly animal-adapted mosquito populations in the Rabai region of Kenya, more attention related to defining genetic determinants and variations established laboratory colonies, and documented striking, genetically- in their distinct phenotypes (e.g, blood feeding preference, oviposition based differences in preference for human vs. animal scent. We are now behavior). We hypothesized that a more accurate structure within using high-throughput transcriptome sequencing to identify candidate California can be determined using phenotypes as distinct populations genes in the three mosquito tissues involved in the reception and opposed to geographically distinct populations; consequently, we used processing of odor cues: the antenna, maxillary palp, and brain. single nucleotide polymorphisms (SNPs) to identify the population structure of Cx. pipiens from five counties using above and below ground 1587 collection sites as the structuring phenotype. Our preliminary SNP analysis corroborates other studies; that there is evidence of a single panmictic AEDES AEGYPTI FORMOSUS IS NOT A SINGLE SUBSPECIES IN population with sub-structuring within the state. This may explain the SENEGAL, WEST AFRICA observed variations in distribution and behaviors of this complex. On average, we found a SNP every 7.5bp. Of interest, one genotype was Laura Dickson, Alex Caspary, Michelle Moore, Massamba Sylla, found only below ground suggesting this genotype may be associated William C. Black, IV with oviposition site, breeding site and/or the Cx. molestus sub-species. Colorado State University, Fort Collins, CO, United States Phylogenetic analyses identified this below ground genotype as a Aedes aegypti is the major vector of the Dengue, Yellow Fever, and population distinct from Cx. quinquefasciatus, but genetically similar to Chikungunya viruses. The subspecies Aedes aegypti aegypti (Aaa) has Cx. pipiens. Additional SNP population genetic analyses will further resolve a global distribution while Aedes aegypti formosus (Aaf) is limited to the systematic issues and define the population structure. Analyses based Sub-Saharan Africa. We used McClelland’s scoring of the first abdominal on larger sample sizes and collections from multiple trapping methods tergite to subdivide Aaf collected in Senegal into form “F” (no white and separation based on additional phenotypes will confirm our data scales) and form “G+” (white scales present). Average fecundity was to determine the true nature of Cx. pipiens in California. These results observed within F or G+ forms but few eggs and very few larvae were provide more information on the roles that each potential member or produced in F x G+ crosses. Analysis of the Argo2 gene identified distinct population serves as zoonotic and epizootic vectors. SNPS associated with F or G+ forms. “F” markers predominated in the southeastern, forested regions of Senegal and the “G+”.markers predominated in western and northern Senegal. Analysis of genotype frequencies demonstrated that 29.2% (31/106) of tests departed significantly from HWE and 25 of the 31 significant tests had an excess of homozygotes. Wahlund’s effect is one explanation for this and occurs when a collection is sampled as a single reproductive pool, when there are actually two or more reproductively isolated populations. When the Aaf “G+” were analyzed separately, genotypes fit HWE. However, when Aaf “F” was analyzed, an excess of homozygotes continued to be observed; suggesting that the “F” form may consist of more than one reproductively www.astmh.org 481 1589 1591 IMAGINAL DISCS - A NEW SOURCE OF CHROMOSOMES FOR INTRA-VITAL IMAGING REVEALS MULTIPLE MODES OF GENOME MAPPING OF THE YELLOW FEVER MOSQUITO PLASMODIUM LIVER-STAGE ELIMINATION BY CD8+ T CELLS AEDES AEGYPTI Ian A. Cockburn1, Rogerio Amino2, Fidel Zavala1, Robert Menard2 Maria V. Sharakhova1, Vladimir A. Timoshevskiy1, Fan Yang1, 1Johns Hopkins University, Baltimore, MD, United States, 2Institut Pasteur, Sergei Yu. Demin2, David W. Severson3, Igor V. Sharakhov1 Paris, France 1Virginia Tech, Blacksburg, VA, United States, 2Institute of Cytology, CD8+ cytotoxic T lymphocytes (CTL) can mediate protection against Russian Academy of Sciences, Saint Petersburg, Russian Federation, malaria by eliminating Plasmodium parasites in hepatocytes. To gain new 3University of Notre Dame, Eck Institute, Notre Dame, IN, United States insights into this anti-parasitic activity, we used spinning-disc confocal The mosquito, Aedes aegypti, is the primary global vector for dengue microscopy to visualize the CTL-mediated killing of GFP-labeled parasites and yellow fever transmission. Sequencing of the Ae. aegypti genome in vivo. Killing was indicated by a profound decrease in the fluorescence has stimulated research in vector biology and insect genomics. However, of most of the parasites associated with at least one CTL. Strikingly the current genome assembly is highly fragmented with only ~30% of however, most dying parasites were surrounded by clusters of multiple the genome being assigned to chromosomes. A lack of reliable source CTL. A variety of different modes of parasite death could be distinguished, of chromosomes for physical mapping has been a major impediment ranging from the rapid loss of GFP signal to a progressive attrition of to improving the genome assembly of Ae. aegypti. We propose to parasite fluorescence. CTL-parasite interactions generally lasted for the use mitotic chromosomes from imaginal discs of a 4th instar larva for entire duration of imaging (mean of ~1 death event / 3 h of recording). cytogenetic studies of Ae. aegypti. High numbers of mitotic divisions on These data are consistent with redundant mechanisms being used for the each slide preparation, large sizes and reproducible banding patterns of killing of parasites. In conclusion, the action of CTL on Plasmodium liver the individual chromosomes simplifies cytogenetic procedures. Based on stages is a complex, multi-cellular killing process rather than a rapid and the banding structure of the chromosomes, we have developed ideograms discrete event. for each of the three Ae. aegypti chromosomes and placed 10 BAC clones and a 18S rDNA probe to precise chromosomal positions. The study 1592 identified imaginal discs of a th4 instar larva as a superior source of mitotic chromosomes for Ae. aegypti. The proposed approach allows precise S1P IS ASSOCIATED WITH PROTECTION IN HUMAN AND mapping of DNA probes to the chromosomal positions and can be utilized EXPERIMENTAL CEREBRAL MALARIA for obtaining high-quality genome assembly of the yellow fever mosquito. Constance A. Finney1, Cheryl A. Hawkes1, Dylan C. Kain1, Aggrey Dhabangi2, Charles Musoke2, Christine Cserti-Gazdewich1, 1590 Tamas Oravecz3, W. Conrad Liles1, Kevin C. Kain1 WOLBACHIA-DENGUE INTERACTIONS IN AEDES 1University of Toronto, Toronto, ON, Canada, 2Mulago Hospital, Kampala, MOSQUITOES Uganda, 3Lexicon Pharmaceuticals Inc., The Woodlands, TX, United States Zhiyong Xi, Xiaoling Pan, Peng Lu, Guowu Bian, Robert Parker, Cerebral malaria (CM) is associated with excessive inflammatory responses Deepak Joshi and endothelial activation. Sphingosine 1-phosphate (S1P) is a signaling sphingolipid implicated in regulating vascular integrity, inflammation and T Department of Entomology, Michigan State University, East Lansing, MI, cell migration. We hypothesized that altered S1P signaling during malaria United States contributes to endothelial activation and inflammation, and show that Wolbachia are maternally transmitted Gram-negative intracellular plasma S1P levels were decreased in Ugandan children with CM compared bacteria and infect up to 65% of insect species. The ability of these to children with uncomplicated malaria. Using the Plasmodium berghei alphaproteobacteria endosymbionts to induce the reproductive ANKA model of experimental CM (ECM), we demonstrate that humanized abnormality called Cytoplasmic Incompatibility (CI) has led to a large effort S1PL-/- mice with reduced S1P lyase activity (resulting in increased bio- on developing Wolbachia as a novel genetic strategy for control of vector- available S1P) had improved survival compared to wild type littermates. borne diseases. We recently observed that Wolbachia induce resistances Prophylactic and therapeutic treatment of infected mice with compounds to dengue virus in Aedes aegypti. This provides Wolbachia a “mosquito that modulate the S1P pathway and are in human trials for other vaccine” like feature, which can be introduced, driven through CI, and conditions (FTY720 or LX2931) significantly improved survival in ECM. spread over mosquito population to block transmission of mosquito-borne FTY720 treatment improved vascular integrity as indicated by reduced diseases. We will present our recent works on how Wolbachia-mosquito levels of sICAM, increased Ang1 (regulator of endothelial quiescence) interactions activate Toll pathway and induce expression of anti-dengue levels, and decreased Evans blue dye leakage into brain parenchyma. effectors. The mosquito gene expression profile regulated by Wolbachia Furthermore, treatment with FTY720 decreased IFNγ levels in plasma as will be dissected in the presence and absence of dengue virus, with well as CD4+ and CD8+ T cell infiltration into the brain. Finally, when special focus on immune and redox genes. We will also present evidences administered during infection in combination with artesunate, FTY720 to show why Wolbachia-mediated viral interference does not present treatment resulted in increased survival to ECM. These findings implicate in Ae. albopictus, which naturally carries a Wolbachia superinfection. A dysregulation of the S1P pathway in the pathogenesis of human and Wolbachia-density dependent viral inhibition will be further illustrated murine CM and suggest a novel therapeutic strategy to improve clinical using both mosquitoes and cell lines. We will discuss how our results outcome in severe malaria. will provide novel opportunities to develop environmentally friendly biopesticides or novel strategies for future control of vector-born diseases.

www.astmh.org 482 1593 of Treg IL-6+, IFN-γ+, TNF-α+ and CTLA-4+ cells when compared with IND group. Additionally, Treg cells can suppress the proliferative response in ANTIBODY ENHANCED INTRACELLULAR KILLING OF IND patients, although the mechanism is not IL-10 or CTLA-4 dependent. LEISHMANIA AMAZONENSIS We also demonstrated that effector T cells from IND patients in the presence of Treg cells produce high levels of IFN-γ but also high levels of Christine A. Petersen, Katherine N. Gibson-Corley, Jenny Li, IL-10. On the other hand, cultures of effector T cells in the presence of Bryan Bellaire, Yashdeep Phanse, Douglas E. Jones Treg cells from CARD group produce high levels of IFN-γ and TNF-α, but Iowa State University, Ames, IA, United States no significant amount of IL-10 cytokine. Taken together our data suggest that patients with the IND clinical form of Chagas disease present higher Experimental footpad infection of C3HeB/FeJ mice with the causative percentage of Treg cells. These cells have an important immunoregulatory agent of disseminated cutaneous leishmaniasis, Leishmania amazonensis, role that leads to maintenance of better cardiac function in these patients, leads to a chronic disease with high parasite load and large footpad lesion probably controlling the exacerbated immune response throughout the size that persists even after induction of an antigen-specific CD4+ Th1 modulation of the cytokine environment and/or killing effector cells. host immune response. However, these lesions will heal if the animal is co-infected with L. major. We have used this fact to explore the immune factors that are capable of limiting L. amazonensis during coinfection 1595 and found that B cells and specifically their antibodies play an important ENHANCED PROTECTION OF MICE UNDERGOING role in killing this intracellular protozoan parasite. Using an in vitro TREATMENT FOLLOW BY REINFECTION WITH assay with draining lymph node cells from infected animals we found IFN-γ receptor, iNOS, NADPH oxidase and FcR γ -common chain were TRYPANOSOMA CRUZI required to kill L. amazonensis within infected macrophages. PI3 kinase- Juan M. Bustamante1, Rick L. Tarleton2 dependent superoxide production was detected late during the assay. 1Center for Tropical and Emerging Global Diseases, University of Georgia, We hypothesized that small soluble immune complexes could provide Athens, GA, United States, 2Center for Tropical and Emerging Global a late source of effector molecules for the NADPH oxidase pathway. Diseases and Department of Cellular Biology, University of Georgia, Using soluble cross-linked non-specific IgG2a antibodies we were able to Athens, GA, United States recapitulate parasite killing without immune cells from infected animals. Analysis via Image Stream flow cytometry and confocal microscopy We have previously shown that Trypanosoma cruzi-specific CD8+ T indicated that non-specific antibody did not opsonize the parasite. We cells from mice cured with benznidazole (BZ) expressed high levels of conclude that small soluble IgG2a immune complexes indirectly promoted the memory markers CD62L and CD127 and can transfer a degree of parasite killing via superoxide production within infected macrophages. protection to high dose challenge infection. Here, we evaluated T cell The vast majority of Leishmania infection studies show that during an responses and resistance following repeated treatment and cure. C57BL/6J ineffective immune response antibodies limit macrophage activation and mice were submitted to one or more rounds of infection with T. cruzi and promoted high parasite loads. Our studies add to this knowledge but then cured with BZ followed by a final reinfection with T. cruzi. For mice indicate that during Th1 immunity antibodies can promote an enhanced receiving a tertiary or quaternary challenge, each foot pad was injected macrophage microbicidal response and play a significant role in supporting with T. cruzi expressing the tdTomato protein and the fluorescence T cell-mediated immunity. intensity (photons/cm2/sec) was monitored as a measure of parasite load. All groups were compared to mice with persistent primary infection 1594 with T. cruzi (untreated). Parasite-specific CD8+ T cells from cured mice expanded substantially more upon a rechallenge as compared with the REGULATORY CD4+CD25high T CELLS FROM INDETERMINATE parasite-specific CD8+ T cells from persistently infected/rechallenged mice. PATIENTS WITH CHAGAS DISEASE CAN SUPPRESS THE Additionally, persistently infected mice can clear a secondary challenge EFFECTOR CELLS AND CYTOKINES AND REVEAL AN ALTERED in the site of infection faster than their BZ-treated/cured counterparts. CORRELATION WITH THE DISEASE SEVERITY However, cured mice submitted to two or three rounds of treatment follow by reinfection acquired an enhance protection after a tertiary or 1 2 Juliana A. Gomes , Fernanda Fortes Araujo , Manoel Otavio quaternary challenge as compare with mice receiving only a single round 1 3 3 Rocha , Ana Thereza Chaves , Rafelle Christiane Gomes , Karine of infection and curative treatment. In addition, T. cruzi -specific CD8+ 1 3 4 Silvestre Ferreira , Jacqueline Araujo Fiuza , Tatjana Keesen , T cells from cured mice undergoing three rounds of reinfection and 3 3 Andrea Carvalho Teixeira , Rodrigo Correa Oliveira treatment reexpressed CD127 more rapidly and in higher frequency after 1Federal University of Minas Gerais, Belo Horizonte, Brazil, 2FDA, rechallenge, suggesting that these mice better regulate parasite load upon Washington, DC, United States, 3Centro de Pesquisas Rene Rachou - rechallange, relative to their untreated/persistently infected counterparts. FIOCRUZ, Belo Horizonte, Brazil, 4Federal University of Rio Grande do These results demonstrate that repeated infection and cure fails to induce Norte, Natal, Brazil sterile resistance to infection, calling into question the ability of more conventional vaccination protocols to prevent establishment of infection. Several studies in Chagas disease demonstrate that immunoregulatory mechanisms control the intense immune activity in the chronic phase, preventing a deleterious effect of the excessive immune stimulation. 1596 high Recently, the identification of the human CD4+CD25 FOXP3+T cells DYNAMICS OF LONGITUDINAL KENYAN INFANT (Treg) and its role have been the object of intense studies due to the ANTIMALARIA ANTIBODY RESPONSES putative critical role of these cells in maintaining self tolerance, as well as in the control of immune response. In this study we evaluated the Arlene E. Dent1, Rhonda Kimmel1, Indu Malhotra1, Peter phenotypic profile and the mechanisms by which Treg cells works in Mungai1, Eric Muchiri2, James Kazura1, Christopher L. King1 patients with the indeterminate (IND) and cardiac (CARD) clinical forms 1Case Western Reserve University, Cleveland, OH, United States, 2Division of Chagas disease. Our results showed that patients with the IND clinical of Vector Borne and Neglected Tropical Diseases, Nairobi, Kenya form present significantly higher frequency of Treg cells. Also our data showed that these cells correlate with better heart function (higher LVEF Maternal antibodies transferred to the fetus during pregnancy protect and lower LVDD) in IND patients with chronic Chagas disease. Besides, the infant from malaria infection. These antibodies are thought to wane these patients present significantly higher frequency of Treg IL-10+ and by 6-9 months of age. Infants slowly acquire antimalaria antibodies with IL-17+ cells when compared with non-infected individuals and CARD repeated infections. Antimalaria antibodies were measured in plasma group. Moreover, CARD patients present significantly higher frequency samples collected from a longitudinal cohort of infants born 2007-2009 (Msambweni, Kenya) with blood samples drawn approximately every www.astmh.org 483 6 months from birth to 36 months of age. Plasma samples from 89 1598 infants (~4.3 blood samples/infant) were examined for the presence and magnitude of multiple antimalaria antibodies by a) serology to 13 malaria Continuous in vitro culture of Plasmodium antigens, b) human antibody recognition of Plasmodium falciparum knowlesi in human erythrocytes: a genetically proteins exported to the erythrocyte membrane from 3 different parasite tractable human malarial pathogen strains and c) functional antibody-mediated growth inhibition of cultured 1 2 3 3 parasites. We found that, by serology, antibodies directed against the Robert W. Moon , Arnab Pain , Joanna Hall , Neil Almond , 4 1 1 majority of antigens tested (MSP1, EBA175, EBA181, EBA140, LSA, Graham H. Mitchell , Anthony A. Holder , Michael J. Blackman PfCSP, PfCelTos, Sera5) waned by 4-8 months of age. Antibodies directed 1Division of Parasitology, MRC National Institute for Medical Research, against AMA1, however, did not wane until 16-23 months of age. For London, United Kingdom, 2Pathogen Genomics Group, Computational the majority of the antigens tested, infant antibody levels and prevalence Bioscience Research Center, Chemical Life Sciences and Engineering reached or exceeded birth levels by 28-32 months of age. In contrast, Division, King Abdullah University of Science and Technology, Thuwal, antibodies directed against P. falciparum proteins exported to the Saudi Arabia, 3Division of Retrovirology, National Institute for Biological erythrocyte membrane waned by 4-8 months and stayed at low levels Standards and Control, Potters Bar, United Kingdom, 4Malaria Laboratory, throughout infancy. Growth inhibitory antibodies waned by 4-8 months Department of Immunobiology, King’s College and St Thomas’ Hospitals’ of age and were acquired at low rates throughout infancy. These findings School of Medicine, London, United Kingdom call into question the generally accepted paradigm that antibodies directed Now widely considered the fifth human malaria parasite, P. knowlesi against P. falciparum proteins exported to the erythrocyte membrane has long proven an important model system for malaria, in particular (thought to protect against severe malaria) are acquired rapidly upon in the study of red blood cell invasion. This is due to the relatively large infection whereas antibodies directed against merozoite surface proteins dimensions of P. knowlesi merozoites and their long invasive half- are acquired gradually and after repeated infections. life. Previous work has demonstrated that P. knowlesi cultured in vitro in macaque erythrocytes is highly amenable to genetic modification, 1597 including gene targeting using double-crossover recombination which is Deciphering the moving-junction AMA1-RON2 relatively difficult to achieve in P. falciparum. However, long term culture complex in Apicomplexa parasites of P. knowlesi in human cells has not been possible, limiting its use to laboratories with access to primate facilities. We have now adapted Mauld Lamarque1, Michelle Tonkin2, Brigitte Vulliez-Le a laboratory strain of P. knowlesi to continuous culture in human red Normand3, Graham Bentley3, Martin Boulanger2, Lebrun Maryse1 blood cells. Cultures readily reach high parasitaemia(> 10%), display the 1Universite Montpellier, Montpellier, France, 2Department of Biochemistry expected ~24h replication cycle, and can be synchronised, manipulated & Microbiology, University of Victoria, Victoria, BC, Canada, 3Unité and cloned as easily as culture-adapted P. falciparum. Comparison of the d’Immunologie Structurale, Institut Pasteur, Paris, France original and culture-adapted P. knowlesi lines has shown that without adaptation the original parasites invade and survive very poorly in human The Apicomplexa phylum comprises a wide range of obligate intracellular cells, indicating that genetic or epigenetic changes were required. Current parasites, including Plasmodium spp. and Toxoplasma, responsible for work is focused on comparing adapted and non-adapted lines to identify malaria and toxoplasmosis respectively. These protozoan parasites actively key changes required for growth in human erythrocytes. As well as invade their host cells and this process requires the coordinated secretion facilitating the adaptation of further lines, this knowledge may reveal of specialized secretory organelles, the micronemes and the rhoptries. requirements for infection of humans in the field. The human erythrocyte- At an early stage of invasion, a close contact between the parasite and adapted parasites have been used to produce transgenic P. knowlesi lines the host plasma membranes, referred as the moving-junction (MJ), act as and we will report on progress to further develop genetic methods. We an anchor onto which the parasite relies on to propel itself forward into anticipate that human erythrocyte culture-adapted P. knowlesi will provide the cell. The micronemal protein AMA1 and the rhoptry neck proteins a flexible and genetically amenable model to study many aspects of RON2/4/5 secreted into the host cell are the MJ molecular actors identified malaria parasite biology. so far. We have recently demonstrated a direct interaction between AMA1 displayed on the parasite surface and RON2 embedded in the host plasma 1599 membrane in Toxoplasma and P. falciparum. This binding was shown to be essential for efficient invasion of both parasites. In this study, we have Giardia Flagellar Motility is Not Directly Required refined the RON2 interacting domain to a short segment located between to Maintain Attachment to Surfaces the two predicted C-terminal transmembrane domains. A RON2 synthetic peptide corresponding to this region (RON2sp) was co-crystallized with the Susan A. House, David Richter, Jonathan Pham, Scott C. Dawson AMA1 ectodomain of Toxoplasma and Plasmodium. A structure/function Microbiology, University of California, Davis, Davis, CA, United States analysis led to the identification of key residues essential for the RON2- Giardia trophozoites attach to the intestinal microvilli (or inert surfaces) AMA1 interaction and highlighted the separate co-evolution of the two using an undefined suction-based mechanism, and remain attached partners within the phylum. Overall, our results shed light for rationale during cell division to avoid peristalsis. The ventral flagella, one of four new drug design targeting the invasion process of Apicomplexa parasites. pair of motile flagella, are thought to be responsible for generating a hydrodynamic force that translates to a pressure differential, and hence suction, under the adjacent ventral disc. We defined four distinct stages of attachment using TIRF microscopy, imaging structures of the cell body that contact the substrate during attachment. The lateral crest of the ventral disc forms a continuous perimeter seal with the substrate, a cytological indication that trophozoites are fully attached. We then assessed whether the ventral (or any) flagella are necessary for attachment by using TIRF and biophysical assays to quantify attachment strength. If the hydrodynamic model were correct, both strains with defects in flagellar beating should have been prevented from generating a hydrodynamic current, thereby preventing suction. Following a morpholino-based knockdown of central pair protein PF16, both the beating and morphology of flagella were defective, but trophozoites could still initiate proper surface contacts, resisting detachment under both www.astmh.org 484 normal and shear forces. While trophozoites were able to attach like wild identification of kinetoplasts and nuclei by taking advantage of the type cells, the overall rate of attachment was slower. Over-expression of different sequence binding biases of different fluorescent DNA stains. a dominant negative a2-annexin::GFP (D122A, D275A) resulted in a strain We have successfully used colour deconvolution to separate the signal with defects in the ventral flagellar waveform. This strain was also able to from kinetoplast and nuclear DNA in fluorescence microscopy images. initiate attachment comparable to wild type, with only a slight decrease This produces two new images of the same field of cells, one with only in the ability to withstand normal and shear forces. Thus, when flagellar kinetoplasts and one with only nuclei. These images are amenable to beating is defective, the positioning and orientation of trophozoites are automated analysis and can also simplify manual analysis of complex hindered; however, there is little or no effect on the ability to maintain phenotypes where kinetoplast and nuclear complement and structure attachment as evidenced by a continuous ventral disc seal and continuous are perturbed. Using our approach to staining, image processing and ventrolateral flange contact. automated analysis we can analyse around 20000 cells per hour. We have correlated these automated approaches with manual analysis 1600 of Trypanosoma brucei and Leishmania mexicana revealing distinct advantages in speed and precision. Inroads of calcium: how extracellular calcium enters and enhances invasion-related traits of the 1602 apicomplexan parasite, Toxoplasma gondii Novel drug uptake and potential resistance Douglas Pace, Jing Liu, Samantha Lie Tjauw, Veronica Jimenez, mechanisms in African trypanosomes: Suramin Allysa Smith and Silvia N.J. Moreno 1 1 2 2 Center for Tropical and Emerging Global Diseases and Department of Sam Alsford , Nicola Baker , Ka Fai Leung , Mark Field , David 1 Cellular Biology, University of Georgia, Athens, GA, United States Horn 1Pathogen Molecular Biology Department, London School of Hygiene & The apicomplexan parasite, Toxoplasma gondii, is a highly successful Tropical Medicine, London, United Kingdom, 2Department of Pathology, intracellular parasite capable of invading any nucleated mammalian cell. University of Cambridge, Cambridge, United Kingdom During the lytic cycle, calcium is a critical element required for motility, invasion, and egress. The importance of intracellular calcium for these A better understanding of drug uptake and potential mechanisms of traits has been established. However, little is known regarding the role clinical resistance will facilitate the design, application and assessment of extracellular calcium. Using the fluorescent, cell permeable calcium of more effective therapies. We have carried out loss-of-function screens indicator, Fura2-AM, we show that tachyzoites of T. gondii possess distinct for drug resistance mechanisms in African trypansomes; all five drugs in pathways of extracellular calcium entry. This data was further confirmed clinical use against Human African Trypanosomiasis have been screened with manganese quenching experiments. Specifically, tachyzoites have using genome-scale RNA interference libraries. The approach was a store-operated calcium entry pathway (SOCE), in which the release of validated by the identification of the nitro pro-drug activator, NTR, and calcium from intracellular stores prompts the influx of extracellular calcium the eflornithine transporter, AAT6 (1). Several novel genes have now into the cytosol. Extracellular calcium entry was quantitatively inhibited been identified, and characterization of suramin uptake will be described; by the calcium channel blocker nifedipine. We assessed the influence of although suramin treatment failures have been reported, no molecular extracellular calcium on invasion-related traits and found that extracellular mechanism of clinical or experimental resistance has been documented. calcium had a significant influence on microneme secretion. Conoid Suramin uptake appears to be via receptor mediated endocytosis, and extension and gliding activity (traits associated with invasion efficiency) data will be presented to demonstrate roles for an invariant surface increased significantly in the context of store-operated calcium entry. glycoprotein, an endosomal membrane channel and the ubiquitin Importantly, blockage of extracellular calcium with nifedipine resulted in a pathway. (1) Baker, Alsford & Horn (2011) Genome-wide RNAi screens 75% decrease in invasion. These results support a coordinated entry path in African trypanosomes identify the nifurtimox activator NTR and the of calcium from the extracellular environment by which invasion-linked eflornithine transporter AAT6. Molecular & Biochemical Parasitology 176 traits can be enhanced and intracellular stores can be replenished. We are 55-57. currently characterizing calcium entry pathways using two-electrode cell clamping of Xenopus oocytes expressing T. gondii membrane proteins. 1603 This electrophysiological characterization of T. gondii ionic channels, the first to our knowledge, will be instrumental in a full understanding of The Class II Histone Deacetylase PfHda2 is an calcium permeation pathways in this ubiquitous model parasite. Essential Regulator of Plasmodium falciparum Virulence and Growth 1601 Bradley I. Coleman1, Lindsey Altenhofen2, Manuel Llinas2, Manoj 1 Beyond just counting Ks and Ns; high throughput T. Duraisingh* image analysis of trypanosomatid cell 1Harvard School of Public Health, Boston, MA, United States, 2Princeton organiZation University, Princeton, NJ, United States Richard John Wheeler, Eva Gluenz, Keith Gull Transcriptional control is essential for the survival of the human malaria parasite Plasmodium falciparum. In addition to the unique cascade of Sir William Dunn School of Pathology, University of Oxford, Oxford, United gene expression that characterizes normal asexual growth, parasite Kingdom virulence relies upon regulated expression and silencing within multigene The precise morphology and replication of trypanosomatids facilitates families. This balances the parasite’s needs for antigenic variation and an approach using automated morphometric analysis for investigation immune evasion with the adhesive diversity required for host cell invasion of their cell cycle, life cycle stage differentiation, mutants and chemical and cytoadherence. Despite the importance of this process, the associated insults. High throughput image analysis has the capability to accelerate regulatory enzymes remain incompletely characterized. We have identified morphological measurement and extract more quantitative data from a protein containing a class II histone deacetylase (HDAC) domain, micrographs. However image analysis of trypanosomatid morphology PfHda2, which plays a critical role in P. falciparum transcriptional control. is complicated by the presence of two, often closely apposed, DNA PfHda2 also contains a C-terminal inositol polyphosphate multikinase containing organelles; the kinetoplast and the nucleus. Accurate (IPMK) domain that is conserved among apicomplexan parasites. Protein identification and analysis of these organelles is central to determining cell is detectable in trophozoite and schizont stage parasites and localizes to cycle stage of a cell which is in turn key for understanding the aberrations distinct foci in the nuclear periphery, which in P. falciparum preferentially caused by a drug or mutation. We addressed the difficulties of automated contains clonally variant virulence gene loci. Because multiple attempts www.astmh.org 485 at genetic disruption of the PfHda2 locus were unsuccessful, we targeted vary in their effect on such signaling pathways. We have found that type PfHda2 for inducible degradation using a destabilization domain (DD) I, II and III strains of Toxoplasma differ in their activation of the NF-kappaB tag. We observe a >95% reduction of PfHda2 protein in knockdown pathway, and activation of this pathway is mediated by the polymorphic parasites, leading to elongation of the asexual cell cycle and defective Toxoplasma protein GRA15. Type II strains of Toxoplasma activate both parasite proliferation. As we hypothesized from residence of the protein NF-kappaB p65 nuclear translocation and NF-kappaB p65-mediated within the nuclear periphery, PfHda2 knockdown also leads to loss of transcription, whereas type II GRA15KO strains do not. GRA15 is also var gene transcriptional repression. This effect is completely independent sufficient to activate NF-kappaB p65 when overexpressed in a type I/ from the previously observed cell cycle-related defects. These two distinct III strain or in human cells alone. Studies in knockout cell lines show phenotypes highlight the unique role of PfHda2 as a bi-functional that GRA15 acts downstream of MyD88 and TRIF and upstream or in regulator of parasite growth and virulence. a complex with TRAF6 and the IKKs. Activation of NF-kappaB affects cytokine production, particularly IL-12, a very important cytokine in 1604 Toxoplasma infection, and macrophages infected with a type II GRA15KO strain secrete significantly less IL-12 than macrophages infected with a Genetic analysis of Plasmodium falciparum type II strain. GRA15 also affects cytokine production and parasite growth gametocytogenesis in vivo. GRA15 is a dense granule protein which is secreted into the host cell upon Toxoplasma invasion, representing the first example of a dense Hiromi Ikadai1, Kathryn Shaw Saliba2, Stefan Kanzok3, Kyle granule protein that can modulate a host cell signaling pathway. Jarrod McLean2, Jun Cao2, Kim C. Williamson3, Marcelo Jacobs- 2 Lorena 1606 1Department of Veterinary Medicine, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Japan, 2Department of IL-27-dependent production of IL-10 by Molecular Microbiology and Immunology, Johns Hopkins Bloomberg IFN- + Th1 cells is a critical mechanism for School of Public Health, Johns Hopkins Malaria Research Institute, ү Baltimore, MD, United States, 3Department of Biology, Loyola University protection against severe immunopathology Chicago, Chicago, IL, United States during malaria infection Within the mammalian host, the Plasmodium falciparum parasite has Ana Paula Freitas do Rosario, Anne O’Garra: Jean Langhorne two developmental fates: cyclic asexual replication or terminal sexual MRC National Institute for Medical Research, London, United Kingdom differentiation (gametocytogenesis). The sexual forms of the parasite (gametocytes) are the only form that is able to survive and propagate Infection during malaria is characterized by strong inflammation. in the mosquito vector. Therefore, gametocytes are absolutely essential The establishment of a precise balance between the pro- and anti- for parasite transmission. Very little is known about the mechanisms inflammatory responses may be critical to guarantee control of the involved in the commitment of Plasmodium to sexual differentiation. To parasite and survival of the host. Interleukin-10 (IL)-10, a key regulatory gain insight into these mechanisms, we conducted piggyBac transposon- cytokine, has been shown to protect mice against pathology elicited mediated insertional mutagenesis screened for parasites that were no during acute Plasmodium chabaudi chabaudi AS model of malaria, longer able to form mature gametocytes. Of 736 parasites (clones) however, its crucial cellular source still is a matter of debate. Here, + screened in 3 independent transfection experiments, 29 clones did not we demonstrate that IFN-g Th1 cells are the main producers of IL-10 form gametocytes. We call these clone, insertional-gametocyte mutants throughout acute infection, and as a consequence, mice bearing specific (IGMs). For each IGM, insertion of piggyBac was verified by Southern blot deletion of il-10 in T cells fully reproduce the phenotype observed in -/- + + analysis and the disrupted genes were identified by inverse PCR. This led deficient IL10 mice. The IL-10 IFN-g Th1 cells are highly activated, to the identification of 16 putative gametocytogenesis-disrupting genes. expressing high levels of CD44 and ICOS and low levels of CD127; they Genetic complementation for 4 of the 16 genes was successfully carried also produce more cytokines than the respective single producing cells. + out showing that these genes are essential for gametocytogenesis. To Despite the fact that Foxp3 regulatory CD4 T cells produce IL-10 during + + temporally order the 16 genes, we measured their expression pattern acute infection, highly activated IL-10 IFN-g Th1 cells were shown to be along with the expression pattern of other known gametocyte-specific the essential and sufficient source of IL-10 to guarantee protection against genes in each of the IGMs using RT-PCR. We found a subset of the severe immune-mediated pathology. Finally, in this model of malaria we + + genes that are likely to act in the initial commitment of the parasite demonstrate that the generation of protective IL10 IFN-g Th1 cells is to gametocytogenesis; another subset likely to act during the initial dependent on IL-27 signaling, and independent of IL-21. differentiation after the committed merozoite has invaded a new red blood cell; and a third set likely to act early in gametocyte maturation (transition from a stage I to stage II gametocyte). Thus, we have carried out a comprehensive screen for genes essential to commitment and early differentiation of the P. falciparum gametocyte. This line of investigation may lead to novel strategies to reduce parasite transmission and disease burden. 1605

Strain-specific activation of the NF-κB pathway by GRA15, a novel Toxoplasma gondii dense granule protein Emily Rosowski, D. Lu, L. Julien, L. Rodda, R. Gaiser, K. Jensen, J. Saeij Massachusetts Institute of Technology, Cambridge, MA, United States Toxoplasma gondii is an obligate intracellular pathogen which can modify its environment through the activation and inhibition of host cell signaling pathways. Many different strains of Toxoplasma exist and these strains

www.astmh.org 486

www.astmh.org Author Index A-487

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

Adamani, William 1036, 846 Ahmed, Tahmeed 242 Alhari, David 846 A Adamou, Abdoulaye 1256, 225, Aholoukpe, Bruno 1293 Ali, Ali O. 676 Abaga, Simon 1030, 232, 396, 226, 742 Ahuka-Mundeke, Steve 1562 Ali, Abdullah S. 1161, 1310, 1392, 928 Adamou, Lagare 1405 Aidoo, Michael 632, 989 1437, 673, 675, 676 Abanyie, Francisca A. 627 Adams, A. Paige 295, 941 Aiemamporn, Kanchana 140 Ali, Doreen 1231, 1374, 531, 905 Abbas, Ali K. 675 Adams, Alison P. 234, 1410 Ailes, Elizabeth 110 Ali, Hasmot 562, 940 Abbehusen, Melissa 1057 Adams, A P. 1416 Ainsworth, Robert M. 1550 Ali, Ibne K. M. 1064 Abbott, Mike 849 Adams, David 297 Ajayi, Jerry A. 1144, 128 Ali, Riyasat 1545 Abd El Khalek, Rania N. 77 Adams, John H. 1013, 1242, Ajjampur, Sitara S. R. 1299, 256 Alifrangis, Michael 1180, 170, 389 Abd Elrhman, Laila S. 304 1538, 1548, 33 Ajumobi, Olufemi O. 378 Alim, Beg A. 263 Abd-Alla, Adly 810 Adams, M. 1047 Akala, Hoseah M. 146, 364 Alimi, Fatimah 119 Abdallah, Fadimata W. 1104 Adams, Matthew 1217 Akande, David 119 Alioni, Victor 458 Abdallah, Gumi 1386 Adams, Mohammed 1370 Akanmori, Bartholomew D. 156 Allen, Denise Roth 369, 896 Abdel-Rahman, Bassem 1098 Adams, Matthew 1506, 1512 Akhter, Sadika 1079, 261 Allen, Richards 547 Abdelgadir, Deena M. A. 1250 Adams, Michael 329 Akhvlediani, Tamar 1088, 1301, Almazor, Charles P. 1106 Abdelhamid, Dalia 854 Adams-Forgor, Abudulai 1114 1303 Almeida, Giulliana T. 443 Abdeliev, Z. Z. 418 Adaui, Vanessa 326, 340 Akhwale, Willis 1507 Almeida-de-Oliveira, Natália K. Abdirizak, M. 1291 Adda, Christopher G. 1224 Akinboye, Emmanuel 1175 653 Abdul-Rahman, Bassem 1090 Addae, Michael M. 156 Akinola, Ibrahim T. 1499 Almond, Neil 1598 Abdulla, Salim 369 Addiss, David 1124 Akinosoglou, Karolina-Anthoula Alonso, Alina 579 Abdulla, Salim 426, 890, 956 Adegbola, Richard A. 1077 1453 Alonso, Pedro 1472 Abedin, Jaynal 248, 263 Adegnika, Ayôla A. 499 Akinwale, Olaoluwa 119 Alphey, Luke 395 Abegaz, Berhanu M. 1132 Adelamo, Solomon 846 Akinyemi, Oluwaseun O. 103 Alphonse, Ouedraogo 167 Abeles, Shira 1540 Adeleke, Monsuru 119 Akinyinka, Olusegun O. 1499 Alrefaei, Yousef N. 798 Abi Younes, Jennifer 916 Adelli, Vijender R. 1166, 1171 Akkhavong, Kongsap 796 Alsford, Sam 1602 Abid, Nima 1110 Ademowo, George O. 1168 Ako-Adounvo, Sharon 890 Altamiranda-S, Mariano 807 Abílio, Ana P. 1226 Ademowo, Olusegun G. 802, 987 Akogbeto, Martin 214, 231, 433 Altenhofen, Lindsey 1603 Abiola, Annie W. K. 1192 Adeneye, Adeniyi 119 Akossa, Amanda 1574 Althouse, Benjamin 1350 Abokyi, Livesy 405 Adesina, Olubukola A. 987 Akpan, Henry E. 378 Althouse, Benjamin M. 407, 1051 Abong’o, Bernard O. 1450 Adewole, Isaac F. 1499, 987 Aksoy, Emre 810 Altidort, Myrline 636 Aborigo, Raymond A. 896 Adeyemi, Mitchell 1077 Aksoy, Serap 1061, 1275, 1279, Alto, Barry W. 236 Abot, E. 181 Adiamoh, Majidah 1028, 204 1286, 1288, 458, 800, 810, Aluvihare, Channa 1585 Abot, Esteban 182, 186, 186A, Adjalley, Sophie 1189 861, 976 Alvarado, Luisa I. 1336 954 Adjei, George 1370 Aktar, Amena 72, 960, 963 Alvarado-Salazar, Andrés 1222 Aboubacar, Ben 561 Adoctor, James 896 Akter, Jasmin 1526 Alvarez, Carlos 312, 938, 1341, Aboud, Frances E. 1079 Adoke, Yeka 450 Akter, Shirina 261 1348 Abraham, David 1468, 1479, Adriko, Moses 497, 657 Akweongo, Patricia 1114, 1386 Álvarez, Carolina 1424 1481, 527 Adu-Gyasi, Dennis 1308, 405 Al-Harbi, Naif O. 782, 790 Alvarez, D. 771 Abraham, Eappen 1059 Aebig, Joan 1011, 1218 Al-mafazy, Abdul-wahiyd 675 Álvarez, Natalí 807 Abril, Marcelo 1572, 833 Afebe, Godwin 1357 Al-Mafazy, Abdul-wahid H. 676 Alves, Clarice C. 783 Abu Sayeed, Abdullah 970 Affricot, Emmeline 1117 Al-Saai, Salma 1046 Amador, Manuel 233, 755, 927 Abu-Bakar, Nurhidanatasha 1049 Afilaka, Babatunde 119 Alam, Md. Tauqeer 1428 Amadou, Konaté T. 167 Abu-Elwan, Safa’a 293 Afrane, Yaw A. 215, 1378 Alam, Masud 520 Amajoh, Chioma N. 1144, 128, Abu-Madi, Marawan A. 293 Afroj, Sayma 414 Alam, Mohammad Shafiul 735 401 Abuaku, Ben 655 Agaba, Bosco 630A Alam, Mohammad Murshid 960, Amalfitano, Andrea 1016 Abubakr, Mustafa 304 Agaba, Steven 888 961, 963 Amambua-Ngwa, Alfred 1019, Abudho, Bernard 1035 Agbenohevi, Prince 1090, 1098 Alam, Mahbub U. 263 1188 Abuom, David C. O. 412 Agblewonu, Koffi B. 584 Alam, Uzma 810 Amaral, Murilo S. 443 Acayo, Sarah 1087 Ager, Arba L. 141 Alao, M.J. 1292, 300 Amaratunga, Chanaki 1211, 1538 Achan, Jane 450 Agidi, Perfect S. 1274 Alarcon, Jorge O. 1099 Amayun, Milton 1293 Achanta, Sita S. 173 Agnew, Dalen 477, 581 Alba, Sandra 426, 1080, 1080, Amazigo, Uche V. 584 Acharya, Jyoti 1024 Agnew, Philip 400 1434 Ambasa, Linda 916 Achee, Nicole L. 428, 429, 743, Agomo, Chimere O. 1485 Albarino, Cesar G. 1009 Ambaye, Teklu W. 273 754, 924, 930 Agudelo, Ivony 1339 Albertini, Audrey 986 Amenga-Etego, Seeba 1157, 1308 Achidi, Eric A. 345 Agudelo, Piedad 1340 Albrecht, Daniel 713 Amidou, Diarra 167 Achieng, Loice 766 Aguiar, Joao C. 182, 186, 186A, Alcorta, Yolanda 954 Amin, Mutamid A. 464 Achilla, Racheal A. 45, 765 694, 696 Aldhamen, Yasser A. 1016 Amin, Shyam 109 Achinko, Daniel A. 629 Aguilar, Zoraida 1017 Alefantis, Tim 1015 Amino, Rogerio 1591 Acholonu, Alex D. W. 1153 Aguirre, Sebastian 981 Alencar, Maria Simone de Amman, Brian 1246 Achonduh, Olivia A. 345, 629 Agunyo, Stella 585 Menezes. 354 Amnuaysirikul, Jack 642 Acidri, Rogers 1087 Agunyu, Stella 611 Alera, Maria Theresa P. 315 Amoah, Abena 495 Ackerman, Hans 875 Agyeman-Budu, Akua 174 Alexander, Mathew 1434 Amodu, Audu 164 Ackumey, Mercy 841 Ahdoot, Stella 1223 Alexander, M. 426 Amos, Ben 985 Acosta, Luz 1566 Ahmed, Kwaku B. 495 Alexander, Neal D. 549, 1075 Ampadu, Edwin O. 841 Acosta, Monica 1542 Ahmed, Makdum 1268 Alexander, Paul 849 Amrein, Matthias 356 Acosta, Norma 1467 Ahmed, Md. Atique 351 Alford, Robert T. 1585 Amuri, Baraka 633, 1555 Adam, Ishag 656, 658, 901 Ahmed, Sabeena 1367, 1369, Alfred, Tiono 167 Amuri, Mbaraka 577, 890 Adama, Gansané 167 1526, 911 Alger, Jackeline 1397 Anagnostou, Nicholas 176

www.astmh.org A-488 Author Index The number(s) following author name refers to the abstract number.

Anaseba, Dominic 1114 Aponte, John 1213 Assogba, Benoît S. 189 Ancca Juarez, Jenny 457, 1364, Appawu, Maxwell A. 213 Astete, Helvio 1253, 1259, 1348, B 1365 Appiah, Eric 451 509, 516, 553 Ba, El hadji 362, 893 Andagalu, Ben 146 Appledorn, Daniel M. 1016 Astha, F.N.U. 431 Ba, Yamar 1247 Anderegg, Claudia 676 Arabi, Mouhaman 259 Asuming-Brempong, Elias 487 Ba Fall, Fatou 893 Anders, Robin F. 1, 1224, 1486, Aragam, Nagesh R. 1491 Atabekova, Shoira R. 298 Babakhanyan, Anna 161 683, 695 Arana, Yanina 89 Ataide, Ricardo 690 Baber, Ibrahima 702 Andersen, Finn 1509 Araujo, Fernanda F. 1594 Ataku, Caro E. 727 Babi, Nackson 1087 Andersen, Kristian G. 1407, 1412 Araújo, Juliano M. 783 Ateba Ngoa, Ulysse 499 Babirye, Juliet N. 102 Anderson, Charles 1011, 702 Araz, Engin 253 Athanazio, Daniel A. 1097 Babji, Sudhir 570 Anderson, John F. 1458 Arbe-Barnes, Sarah 997 Athrey, Giridhar 928 Babu, Jayaprakash 1545 Anderson, Jennifer M. 1044, 685, Arboleda, Sair O. 218 Atieli, Harrysone E. 215 Babu, Subash 1482, 23 687, 746, 915 Arca, Phillip 827 Atienza-Herrero, Julián 651 Baby, Mounirou 106 Anderson, Larissa 1271 Archer, Natasha M. 110, 565, 957, Atkins, Graham 1315 Bacellar, Olivia 866 Anderson, Sheri S. 236 959, 622 Atkinson, Jo-An M. 361 Badalova, Nadiya S. 298 Anderson, Tim 1046, 1048, 1429 Arens, Theo 693 Atkinson, Peter 1060 Badolo, Athanase 394 Anderson, Timothy 411 Arensburger, Peter 1060 Atmakuri, Malika 1354 Badu, Kingsley 157 Andersson, Björn 1362 Arevalo, Jorge 324, 326, 334, 340 Atshabar, B. B. 418 Baea, Manasseh 1074 Andersson, Karin 673 Arguello, D. Fermin 507, 1304, Attan-Ayibo, Apussi 1118 Bagny Beilhe, Leila 223 Andonova, Maya 523 1333 Attardo, Geoffrey M. 1061, 1279 Bah, Germanus S. 1001 Andrada, Andrew 1343, 1445 Arguello, Heather E. 631 Atue, Moises 396, 928 Bahashwan, Ahmed A. M. 782 Andrade, Christy C. 1455 Arguello, John 1327, 1446 Atuguba, Frank 1376, 1379, 896 Bahashwan, Saleh A. M. 782, 790 Andrade, Luiza F. 784 Arguin, Paul M. 627 Atuhaire, Aaron 497, 657 Bahelah, Raed 764 Andrade, Zilton A. 498, 799 Arias, Luis M. 1055 Atwal, Sharan 1439 Bahelah, Salwa 764 Andrade-Narvaez, Fernando J. 342 Arifin, S. M. Niaz 377 Atwill, Edward R. 406 Bahia, Ana Cristina 723 Andrae-Marobela, Kerstin 1132 Arifuzzaman, Md. 21, 961 Auburn, S. 1047, 1506 Bahia, Maria Terezhinha 853 André Lin, Ouédraogo 167 Arinaitwe, Emmanuel 453, 535 Auburn, Sarah 1211 Baiden, Frank 1157 Andreadis, Theodore G. 1458 Arinaitwe, Moses 497, 657 Audcent, Tobey 1553 Bailey, Dana 337 Andrews, Elizabeth S. 230 Arkin, Michelle R. 863 Audi, Allan 1474 Bailey, Jason 1512 Andrews, Ross 839 Arlian, Larry 805 Audu, Bala 378 Bailey, Jeffrey 991 Andriamiarimanana, Fehivola 1089 Armah, George E. 1414 Ault, Steven K. 842 Bailey, Robin 549 Andrianaivoarimanana, Voahangy Arman, Shaila 1471 Aumakhan, Bulbulgul 22 Bain, Lisa 152, 907 1089 Armstrong, Adam 77 Auschwitz, Jennifer M. 646 Bair-Brake, Heather 1549 Andriantseheno, Marcellin 60 Armstrong, Philip M. 1458 Avci, Ismail Y. 602 Baird, J. Kevin 383, 672, 1556, Añez, Germán 829, 1342 Armstrong, Stuart D. 1001 Avendaño, Adrian 748 1557, 1558, 449 Ang, Kenny K. H. 863 Armstrong, Tanya 849 Avendaño, Efren 1340 Baitur, D. 449 Angeles, Jose Ma. M. 488 Arnez, Arbrahan M. 928, 1030 Aviles, Ricardo 1397 Baiwog, Francesca 1483 Angelino, Elaine 29 Arnold, Benjamin F. 266 Avilés, William 819 Bakare, Oladapo 1175 Angelwicz, Philip 532 Arnoux, Stéphane 223 Avril, Marion 1486, 161 Baker, Bill J. 605, 1242 Anges, Yadouleton 231 Aroian, Raffi V. 1140 Avula, Bharathi 1167, 367, 995, Baker, Kelly K. 1472 Angov, Evelina 1015, 666, 952, 99 Arole, Shobha 108, 291 996 Bakuza, Jared S. 494 Angulo, Noelia 872 Aronson, Jane 109 Awadh, Khamis 1555 Balabaskaran Nina, Praveen 1040 Angulo-Barturen, Iñigo 136 Arrieta, Germán J. 282 Aweeka, Francesca T. 370, 453, Balasegaram, Manica 318 Annerberg, Anna 901 Arroyo, G. 57, 94 899, 1499 Balbo, Luca 740 Anova, Lalaine 367 Arsanok, Montri 998 Awine, Timothy 1114, 1376, 1379 Baldet, Thierry 189, 214, 432 Ansah, Evelyn K. 1163 Arsyad, Dian S. 1557 Awini, Elizabeth 1308, 1308 Baldeviano, G C. 335, 1541 Ansari, Aftab 979 Artiles, Karen 818 Awino, Justine 413 Baldwin, Albert S. 947 Ansong, Daniel 951 Artis, David 18 Awolola, Samson T. 204 Baleela, Rania M. H. 1356 Anstey, Nicholas 455, 1517, 1534, Artsob, Harvey 1337 Awono-Ambene, Parfait 1453 Balinandi, Stephen 945 1561 Arumugam, Jeeva 23 Ayalew, Firew 1570 Baliraine, Frederick N. 142 Anthony, Kiszewski 931 Arumugam, Sridhar 1481 Ayamba, Samuel 405 Ballance, James 1460 Anthony, Kodzo A. 1574 Arumugam, Thangavelu U. 1, 184 Aydin-Schmidt, Berit 1161 Ballesteros-Rodea, Gilberto 860 Antonia, Alejandro 1490 Arvelo, Wences 1306, 249, 522 Ayeh-Kumi, Patrick F. 1357 Ballo, Amadou 9 Antonio, Martin 1077 Aryeetey, Yvonne 486, 495 Ayele, Berhan 1085 Ballou, W. Ripley 952, 953, 1546, Antunez de Mayolo, E. A. 1466 Arze, Cesar 1047, 1506, 1512 Ayers, Tracy 268, 565, 959 1547 Anuradha, R. 1482 Asad, Mohd 138 Ayi, Kodjo 7 Balmaseda, Angel 1581, 44, 588, Anwary, Mushfique 93 Asante, Kwaku Poku 1308, 405, Ayieko, Cyrus 5 816, 818, 819 Anyanwu, Greg I. 1144, 128 1370, 174 Ayodo, George 163 Balraj, Vinohar 570 Anyirekun Fabrice, Some 1187, Asaolu, Samuel O. A. 448, 788 Ayvar, Viterbo 96 1201 Asgary, Ramin 108, 109, 291 Azabo, Rogers 458 Baltzell, Kimberly A. 1161, 1392, Anyona, Samuel 1091, 1194, Ashley, Elizabeth 1046, 901 Azevedo, Everton C. 1097 1437, 1310 1197, 1208 Ashman, Jill 324 Azim, Tasnim 43, 948 Bamezai, Ramesh N. 767 Anyorigiya, Thomas 1376, 1379 Ashraf, Sania 263 Azman, Andrew 1350 Banania, Glenna 954 Aol, George 1474 Ashton, Michael 901 Azziz-Baumgartner, Eduardo 1268, Banania, J G. 181 Aonuma, Hiroka 394 Asiedu-Asamoah, Mavis 561 1421, 43, 46, 47, 763, 948 Bancone, Germana 988 Apanga, Stephen 1157, 1308 Asih, Puji B. 1249, 1557 Bandeira, Mauricio S. 1097 Apangu, Titus 1087 Asmelash, Aida 24 Bandika, Victor 243 Aparicio, Juan P. 1055 Asoala, Victor 1376, 1379 Bangirana, Paul 28

www.astmh.org Author Index A-489

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

Bangs, Michael J. 1534 Bayard, V. 1355 Bennett, Shannon N. 1344, 592 Biggins, Julia 599 Bangura, James 111 Bayeto, Luis 58 Bennuru, Sasisekhar 539, 542 Bigira, Victor I. 453, 575 Banik, Soma 825 Bayoh, M. N. 925 Benoit, Christine M. 1311 Bigogo, Godfrey 1474 Bansal, Shweta 555 Bayoh, Nabie M. 1252, 1450 Benoit, Joshua B. 1061, 1279 Bijker, Else M. 693, 955 Banu, Sultana S. 414 Bazzone, Lindsey E. 508 Bent, Stephen 38 Billingsley, Katherine M. 766 Barany, Francis 1414 Beach, Raymond 753, 923 Bentley, Graham 1597 Billingsley, Peter F. 1215, 1217, Barasa, Sheila O. 398 Beadell, Jon S. 800 Berg, Elyssa 1463 344, 693 Barban, Veronique 1444, 36, 506 Beare, Nicholas A. V. 970 Berghout, Joanne 1041, 878 Billker, Oliver 643 Barbi, Joseph 66 Beasley, David 1328 Bergmann-Leitner, Elke S. 952, Bilong Bilong, Charles Felix 1136 Barbosa, Lúcio M. 498, 799 Beatty, Mark E. 1327 1015 Bilson, Philip 1308 Barbosa, Philip V. 883 Beatty, P. Robert 589, 590 Bergren, Nicholas 1420 Bin Yunus, Emran 970 Barbu, Corentin M. 457 Beau, Jeremy 605 Bergstrom, Kirk 18 Bingham, Andrea M. 233 Barde, Auwal A. 401 Beau De Rochars, Valery M. 959 Berman, Jonathan 367, 894 Bingham, Georgina V. 920 Baresel, Paul 649 Beavogui, Abdoul Habib 1184, Bern, Caryn 1364, 1365, 872 Binka, Fred N. 890¸ 896 Bari, Arfan 1361 1516 Bernal, M. 79 Binnicker, Matthew J. 631 Baric, Ralph S. 1578 Beck, Hans-Peter 1388 Bernal-Rubio, Dabeiba 1329, Bir Sidhu, Amar 136 Barillas-Mury, Carolina 1025 Beckedorff, Felipe C. Ferrarezi. 443 1580, 981 Birbeck, Gretchen L. 1022 Barker, Christopher 515 Becker, Norbert 939 Berndt, Amanda L. 1462 Bird, Brian H. 1009 Barker, Nicola 1602 Beckers, Pieter 693 Bernhard, Teresa 1233 Bireley, Brandon 1037 Barker Jr, Robert H. 135, 136, 138 Bedir, Orhan 602 Berrie, Eleanor 176 Birkenkamp, Kate E. 1154 Barnadas, Celine 1559 Bedoya, Gabriel 1340, 1518 Berriman, Matthew 1433, 537, Birkett, Ashley 953 Barnes, Kayla G. 29 Beech, Robin N. 447 538 Birlinger, Bertrand-Luc 1298, 288 Barnes, Karen I. 150 Beeghly, Andrew 290 Berthe, Safoura 632 Birrell, Geoffery W. 139 Barnes, Samantha J. 1013, 1538, Beeharry, Yasnee 1403 Bertrand, Olivier 1548 Birren, Bruce W. 1056, 1458, 29, 33 Beer, Netta 675 Berven, Bradley 849 430 Barnes, Tamsin 62 Beers, Megan 65 Beshir, Khalid 1500, 658 Bisanzio, Donal 740 Barnett, Elizabeth D. 1311 Beeson, James G. 1, 1014, 1224, Besigye, Fred 497, 657 Bischoff, Emmanuel 1584 Barnwell, John W. 1428, 1565, 1483, 1486, 184, 683, 690, 691 Bess, Cameron 687 Biswas, Sumi 176, 1220 989, 1525, 637 Beg, Mohammad Asim 1531 Besser, John 75 Biswas, Sukla 1545 Baron, Elinor 269 Beguin, Pierre 1191 Bessong, Pascal O. 775 Biswas, Subas C. 1471 Barr, Kelli L. 1262, 1263 Béhéton, Todoégnon 48 Best, Ivan 334 Bitemo, Michel 662 Barrera, Roberto 1238, 233, 755, Behr, Marcel A. 878, 1041 Best, Wayne 849 Bjorge, Steven 674 927 Bei, Amy K. 30 Bethell, Delia 1047, 1165, 1212, Björkman, Anders 1310, 1392, Barrere, Beatrice 823 Beier, John C. 748 1506, 1509, 1511, 1512, 1515, 673, 675, 1161, 1184 Barreto, Domingos A. 78 Beldjebel, Irad 617, 619 645, 761, 762, 998 Bjornson, Zach 1407 Barrett, Leah J. 775 Belemvire, Allison 1233 Bethony, Jeffrey 1569 Black, Christine 1115 Barron, Eduardo 87 Belén Jiménez-Díaz, María 136 Betson, Martha 1137, 464, 497, Black, Robert E. 1076 Barros, Monica B. L.. 862 Bell, David 986 657 Black, IV, William C. 1587 Barros, Nicolas 1466 Bell, Teal R. 1549 Bett, Andrew J. 1447 Blackman, Michael 1598 Barroso, Lisa 1066 Bellaire, Bryan 1593 Betzel, Christian 1480 Blackstock, Anna 1035 Barry, Alyssa E. 1014 Bellieny, Myrtes S. S. 319, 317 Bhagani, Sanjay 284 Blackwelder, William 1472, 952 Barrysson, Andriamahefazafy 306 Bellini, William J. 416 Bhasin, V K. 147 Blagborough, Andrew M. 1220 Bartelt, Luther A. 775 Belmonte, M. 181 Bhat, Ajaz 1545 Blair, Carol D. 604 Bartholdson, Josefin 30 Belmonte, Maria 182, 954 Bhat, R. M. 147 Blair, Jenny 563 Bartholomay, Lyric C. 1004 Beltramello, Martina 590 Bhatgtacharyya, Mrinal Kanti 173 Blair, Silvia 1210 Bartkovjak, Marian 116, 618 Belur, Giridhar 558 Bhatta, Narendra B. 279 Blake, Isobel M. 549 Bartlett, John A. 435, 828 Ben Hadj Ahmed, Sami 34 Bhatta, Nisha K. 90 Blanas, Demetri 382 Basáñez, Maria-Gloria 549, 669 Benante, John Paul 1281 Bhattacharya, Mihir K. 962 Blanchette, Mathieu 878 Baseer, Imaan 1259 Benavente, Luis E. 632, 1159, Bhattacharya, Shyamal K. 279 Blank, Walter A. 498, 799 Bashir, Ismail Mahat 628 1160 Bhattacharyya, Krishna Nando 124 Blankenship, D’Arbra 143 Basile, Alison J. 942 Benca, Gorge 116, 619 Bhattacharyya, Sunanda 173 Blanton, Ronald E. 498, 799 Basri, Hasan 449 Bendall, Richard P. 1137 Bhattarai, Achuyt 675 Blau, Dianna 1335 Bassene, C. 895 Bender, Thomas 959 Bhattarai, Rachana 91 Blaze, Marie 13 Bassene, Jonas 382 Bendezu, Jorge 171 Bhavnani, Darlene 1273, 1406 Blinkhorn, Richard 1315 Bast, Joshua D. 806 Beneš, Helen 209 Bhokisham, Narendranath 1015 Blitvich, Bradley J. 1004, 943 Bastiaens, Guido J. H. 693, 955 Benedict, Christian 32 Bhonsle, Jayendra 367 Blood, Melinda 1430 Bastos, Cecilia M. 136 Bengaly, Zakaria 322 Bhoomiboonchoo, Piraya 1330, Bloomquist, Jeffrey R. 431 Batchelor, Adrian 695 Bengaly, Zakary 341 1402 Blum, Annalise 1473 Bates, Paul A. 1357, 871 Benjamin, John 1496 Bhuiyan, Md. Saruar 21, 961, 963 Blum, Lauren S. 1112, 296, 82 Bathurst, Ian 136 BenMohamed, Lbachir 623 Bhuiyan, Mejbah U. 1421, 43 Blumberg, Benjamin J. 206 Batonjo, G 1291 Benner, Christine T. 93 Bhupender, Nagpal N. 391 Blumberg, Seth 1010 Battle, Katherine E. 1556, 672 Bennett, Adam F. 1377, 1374, 986 Bi, Yan 190 Boakye, Daniel A. 1129, 213, 495, Batty, Kevin T. 1496, 366 Bennett, Jason W. 1546, 1547, Bian, Guowu 1590 1133, 1463, 486 Batwala, Vincent 25, 27 953 Bichaud, Laurence 1276 Boamah, Ellen 1370 Baus, Esteban G. 1363 Bennett, J. W. 181 Bichha, Ram P. 598 Boaz, Mark 506, 600 Bausch, Daniel G. 111 Bennett, Kristen 287 Bickersmith, Sara A. 199 Bobanga, Thierry 276 Bautista, Christian 1301, 1303 Bennett, Shannon 1258 Bidaisee, Satesh 1102 Bobbili, Naveen 161 Baxi, Sanjiv M. 530 Bennett, Sarah D. 268 Biggerstaff, Bradley 598, 942 Bobogare, Albino 152, 361, 907

www.astmh.org A-490 Author Index The number(s) following author name refers to the abstract number.

Bockarie, Florence 1129 Brackney, Doug E. 52, 55 Buathong, Rome 1053, 1334 Calderon, Flor 324 Bockarie, Moses 802, 1129, 615 Brady, Molly 1124 Buchanan, Ann M. 969 Calderón, Isabelle 1028 Bodhidatta, Ladaporn 1399 Branch, OraLee H. 1389, 886, Bucheton, Bruno 322, 341, 977 Calderon-Arguedas, Olger 748 Boel, Machtel 901 1393, 1535 Buchy, Philippe 1337, 762 Calderwood, Stephen B. 21, 72, Boelaert, Marleen 331 Brando, Clara 684 Buck, Christopher 123 960, 961, 963 Boggiatto, Paola M. 343, 865 Brandy, Nailah 1175 Buckee, Caroline O. 1368, 686 Calvert, Amanda E. 604 Bogyo, Matthew 365 Brandy, Yakini 1175 Buckeridge, David 1385 Calvet, Claudia M. 863 Bohova, Jana 1061 Brantus, Pierre 1124 Buckner, Frederick S. 461, 853 Calzada, J.e. 1355 Bohr, Tiffany M. 357 Bras-Goncalves, Rachel 977 Budge, Philip J. 1574, 582, 1122, Calzada, J. 771 Boivin, Michael J. 28 Brasil, Patricia 1396 1131 Calzetta, Maria 1028 Boldenow, Peter J. 553 Brathwaite, Olivia 309 Budke, Christine M. 91 Cama, Vitaliano 1358, 249 Bolio-González, Manuel E. 328 Brault, Aaron C. 1004, 1455, 54, Buenemann, Michaela 1247 Camara, Mamady 341 Boll, Jessica 211 53 Bui, Cac T. 1150, 1151 Camara, Mama 608 Bolling, Bethany G. 1004, 54 Bree, Mark 135 Bukka, Archana 21 Camara, Makhtar 623 Bolton, Jessica 186, 694 Breiman, Robert F. 1472, 1474, Bukonda, Ngoyi Z. K. 280 Camara, Mamadou 977 Bonaparte, Matthew I. 600 17, 296, 439 Bukonda, Zachaie 630 Camara, Oumou 977 Boncy, Jacques 636, 75, 957 Brelsfoard, Corey 810 Bukrinsky, Michael 795 Camara, Pape 362 Bond, Nell G. 111 Brenner, Jennifer 1553 Bulimo, Wallace D. 45, 765 Camargo, Carolina 867 Bongfen, Silayuv E. 878 Brent, Andrew J. 243 Bull, Pete 690 Cambanis, Alexis 1312 Bongio, Nicholas 1232, 194 Bresee, Joseph 1421, 46, 763 Bulzacchelli, Maria 386 Camiña, M. 79 Bongo, Emile 1293 Bretscher, Michael T. 11, 1388 Bumb, Ram A. 330 Campbell, Carl 531, 905 Bongrand, Pierre 1191 Brewer, Timothy 1385 Bun, Krufinta 1075 Campbell, Corey L. 208 Boni, Maciej 1583, 751 Brewoo, Joseph 1401 Bundy, Donald A. P. 584 Campbell, Karen M. 512 Bonilla, Luis V. 768 Bridges, Daniel 1438, 347, 678 Bunkea, Tol 674 Campbell, Robert A. 880 Bonizzoni, Mariangela 208, 404 Bridges, Michael 1269 Bunn, James 347 Campino, Susanna 1019, 1211, Bonne-Annee, Sandra 527 Brieger, William R. 564 Bunyoeui, Sea 762 1506 Bonney, Joseph H. K. 408 Briët, Olivier 1371 Burdett, Laura A. 6 Campino, S. 1047 Booker, Michael L. 136, 138 Brindley, Paul J. 1000, 794, 798, Burgess, Steven J. 1170, 1173 Campos, Mônica C. Oliveira. 317 Boonplueang, Rapee 808 795 Burke, Donald S. 1052, 316 Can Caeseele, Paul 523 Booton, Gregory C. 250 Britch, Seth C. 806 Burke, Rachel M. 420 Caña, Luisa 1100 Bopp, Cheryl A. 17, 75 Britton, Collette 537 Burkett, Douglas A. 1289 Canestrelli, Daniele 1028 Borchert, Jeff 1087 Broadbent, Kate 882 Burkhard, Peter 684 Cantey, Paul 583 Borghini-Fuhrer, Isabelle 1495, 648 Brodskyn, Cláudia I. 1057 Burkhardt, Martin 1011 Canto-Cavalheiro, Marilene M. Borland, Erin 1246 Brogdon, William 922, 925 Burkot, Thomas R. 1249 319 Borlon, Céline 1519, 1521, 1548 Brooker, Simon 572 Burns, Lorianne C. 706 Canton, Johnathan 64 Borrini-Mayori, Katty 457 Brooks, Alan 1371 Burns, Matthew 1438 Cao, Jun 1524, 1527, 1604, 172 Bosch, Irene 1296, 978 Brooks, Jessica 13 Burrus, Roxanne G. 1237, 929, Cao, Kai 6 Bosco-Lauth, Angela 1005 Brooks, Patricia 613 1243, 387, 930 Capewell, Linda 267, 268 Botero, Adriana 849 Brooks, W. Abdullah 1268, 21, Burstroem, Lotta 643 Cappello, Michael 1133, 1142, Bottazzi, Maria Elena 1479 248 Bushman, Frederic D. 1427 1462, 1463 Bottova, Iveta 1049 Broussard, Cédric 3 Buss, Sarah N. 1065 Capron, André 1038 Bouare, Mamadou 9 Brown, Ammon 642 Bustamante, Juan M. 1595 Caputo, Beniamino 1028, 921 Bouckenooghe, Alain 600 Brown, Catherine 269 Bustamante, Leyla W. 30 Carabin, Hélène 91, 93 Boufana, Belgees 87 Brown, Charles A. 174 Bustinduy, Amaya L. 1380, 500, Caragata, Eric P. 724 Bougoudogo, Flagou 632 Brown, Jocelyn 967 491 Carapetis, Jonathan 839 Bougouma, Edith C. 154, 155 Brown, Julia E. 1248 Buszin, Justin 1469 Carbajal, Luz 20 Boulanger, Martin 1597 Brown, Mark R. 719 Butrapet, Siritorn 1577 Cardenas, Carolina 1347, 747 Boulware, David R. 1119 Brown, Nick J. 713 Butterworth, Alice S. 625 Cardenas, Jenny Carolina 1251, Bourguinat, Catherine 470 Brown, Patrick 1283 Buyungo, Peter 1081 732 Bourke, Claire D. 529 Brown, William M. 749 Byakika-Kibwika, Pauline 274 Cardenas, Lucio Daniel 1251, Bourtzis, Kostas 810 Brownstein, John 1385 Byamukama, Edson 585, 611 1347, 747, 220, 732 Bousari, Olaidé 210 Bruce, Jane 12, 221, 714 Byarugaba, Justus 883 Cardenas Granados, Jenny C. 220 Bousema, J. T. 990 Bruder, Joseph T. 696, 954, 186, Bygbjerg, Ib C. 389, 895 Carey, Cristiam 516 Bousema, Teun 10, 1158, 1439, 181 Caridha, Diana 132, 169 1500, 9, 914 Brumeanu, Teodor 1539 Carlier, Paul R. 431 Bouslough, David B. 1352, 769 Brun, Reto 329 Carlos, Monica 1467, 1572 Boussari, Olayidé 433 Brunetti, Enrico 61, 86 C Carlow, Clotilde K. 473, 471, 472 Boussinesq, Michel 470, 580 Bruno, Travi 332 Carlton, Jane M. 166, 147 Bouyer, Donald 548 Brutus, Laurent 856 Cacciola, Jennifer A. 252, 254 Carmoli, Marya Bowen, Richard A 295, 1005, Bruxvoort, Katia 634, 902 Caccone, Adalgisa 1027, 1030, Carmolli, Marya P. 1343, 1445 1416, 941 Bryan, Cassie M. 643 1285, 1286, 232, 396, 800, 928 Carmona-Fonseca, Jaime 1518 Bowie, Aleah C. 742 Bryant, Clifford 320 Cai, Jenny 137 Carpenter, L. Rand 548 Bowman, Natalie 991 Bryant, Juliet E. 751 Cai, Sumin 1328 Carrasco, L. Roman 1443 Boyd, Alexis 1125 Buakhasit, Darouny 796 Cairns, Matthew 1489, 574, 669, Carrasco, Wilder 1235 Boyd, Scott 818 Bualombai, Pongwit 140 893 Carreño, Numirin 1100, 81 Boyer, Sebastien 736 Bualombai, Pongwit 314 Cairo, Hedley 1113, 1307 Carrillo, Lina 1459 Boyom, Fabrice F. 129 Buathong, Nillawan 1511, 998, Cajal, Silvana P. 1572, 833, 1467 Carrington, Lauren B. 1050 Bozick, Brook 1219 645 Calampia, C. 79 Carter, Jane 1160 Calderón, Alfonso 282 www.astmh.org Author Index A-491

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

Carter, Jennifer A. 1219 Chan, Ernest R. 1430, 1254 Chen, Lin H. 1311 Chitadze, Nazibrola 1088 Carter, Richard 380 Chan, Jo-Anne 690 Chen, Ping 186 Chitnis, Chetan E. 1 Cartozian, Elizabeth 1337 Chan, Miranda 548 Chen, Po-Chi 307 Chitnis, Nakul 1371, 758, 759 Cartwright, Emily 110, 565 Chan Woon, Lu 351 Chen, Ping 696 Chizhikov, Vladimir 514 Carvajal, Daysi 220 Chanarat, Nitima 1165, 762 Chen, Qiao-Hong 431 Chlikadze, Rusudan 1303 Carvalho, Antonio B. 1063 Chancey, Caren 514, 829 Chen, Rubing 1006, 1247 Choi, Jun Yong 863 Carvalho, Augusto M. 866 Chand, Krisin 1557 Chen, Shuqi 978 Choi, Young-Jun 481, 538 Carvalho, Edgar M. 1409, 866, Chand, Meera A. 13 Chen, Tien-Huang 1331 Choi, Young Ho 988 864 Chanda, Emmanuel 434, 922 Chen, Wei-June 1331 Chokejindachai, Watcharee 1014 Carvalho, Leonardo J. M. 641, 881 Chanda, Javan 744, 757, 922 Chen, Xia 1146 Chokheli, Maiko 1301 Carvalho, Lucas P. 866 Chandiwana, Precious 626 Chen, Yun-Chi 1198 Chola, Melissa K. 1220 Casals-Pascual, Climent 875 Chandler, Clare 621 Chen, Yao-Shen 285 Chomel, Bruno B. 406 Casandra, Debora R. 847 Chandler, Clare I. 1162 Chen, Yi-Ying 517 Chong, Chia Yin 824 Casapia, M. 79 Chandler, Clare M. 1109 Chen, Zi 1146 Chongwe, Gershom 195 Casares, Sofia A. 1539 Chandra, Richa 137 Cheng, Allen 839 Chopra, Rohit 558 Casey, Joanne L. 33 Chandra, Ramesh 1505, 203, 717 Cheng, Gong 49 Chou Chu, Lily 1365 Cashman, Kathleen 599 Chandramohan, Daniel 10, 1439, Cheng, Ling 1566 Chow, Angelia 435, 828 Caspary, Alex 1587 451 Cheng, Po-Yung 1421 Chowdhury, Dhuly 844 Cass, Cynthia L. 469 Chandre, Fabrice 214, 400, 433, Cheng, Qin 141, 152, 1528, 169, Chowdhury, Fahima 960, 963 Casseb, Samir M. M. 1349 711 907 Chowdhury, Fazlul K. 1471 Castilho, Tiago 868 Chang, Gwong-Jen 1579, 517 Cheng, Yang 1537 Chreonviriyaphap, Theeraphap Castillo Neyra, Ricardo 1365 Chang, Hsiao-Han 1207, 1431, Chennamaneni, Naveen K. 853 428 Castillo-Salgado, Carlos 388 175 Chentouffi, Aziz 623 Christensen, Bruce M. 481, 538 Castor, Naderge 636 Chang, Jinhong 1442 Chenyambuga, Sebastian 92 Christensen, Jesica A. 1126 Castro, Yagahira E. 1364, 872 Chang, Jung-San 504 Chepchieng, Clifford 806 Christian, Elizabeth 310, 825, 827 Castro-Barcena, Alejandro 328 Chang, Ko 504 Chepkorir, Edith 413 Christian, Parul 115, 562 Castro-Faria-Neto, Hugo Caire Chang, Sandra 1105, 1175 Cherif, Mariama K. 155 Christofferson, Rebecca C. 1346, 880, 881 Chang-Cojulun, Alicia 500 Cherif, Mahamoud S. 179 1347, 227, 305, 586, 587, 732 Castro-Llanos, Fanny 743, 929, Chann, Soklyda 1511, 645, 998 Cherni, Saifedine 1276, 34 Christophides, George 1453 930 Chao, C. C. 1086 Cheruiyot, Agnes 146 Christy, Nathaniel C. V. 1065, 251 Castro-Pinto, Denise B. 317 Chaorattanakawee, Suwanna Cheung, Kong 192 Chu, Brian 1073, 1576, 610 Cavalcante, Paloma A. 80 1514, 1515, 998 Chevillon, Christine 205 Chu, Cindy S. 1530, 1548 Cavanaugh, Kerry 1115 Chaplin, Jason 849 Chewe, Patrick 678 Chu, Yui Yin 1259 CDC Infectious Diseases Pathology Chapman, Jennifer 1423 Chhoeu, Austin H. 640, 367 Chua, Aileen 642 Branch 1335 Chareonviriyaphap, Theeraphap V. Chia, Kee Seng 1582 Chuang, Ilin 954, 181 Ceesay, Alhaji 547 743, 429, 734, 808, 809 Chibale, Kelly 1132 Chuang, Ting-Wu 1255, 513 Celatka, Cassandra 135, 136 Charernmak, Boonsri 1400 Chibunda, Raphael 92 Chubinidze, Marina 1088 Censky, Kaitlin 1066 Charlebois, Patrick 818 Chichester, Jessica A. 703 Chumachenko, Tetyana 1272 Cerami, Anthony 1169 Charles, Mackenzie 477 Chico, Martha E. 937, 1464 Chuquiray, N. 79 Cercone, Emily 1268 Charles, Richelle C. 21, 961 Chigusa, Yuichi 488 Chuquiyauri, Raul 1540 Cerqueira, Gustavo 1047, 1506, Charlwood, Jacques D. 1027 Chijioke-Nwauche, Ifeyinwa N. Churcher, Thomas S. 669, 9 1512 Charman, Susan A. 638, 647, 849 1190, 149 Chusri, Sarunyou 1400 Cesar, Cesar K. M. 1318 Charrel, Remi 1276 Chiko-Like, Matimba 1438 Chwee Teck, Lim 1521 Cesar, Cesar L. M. 1309 Chartrain, Nicole 1328 Chile, Nancy 89 Chy, Sophy 1204 Cespedes, Manuel J. 1099 Charunwatthana, Prakaykaew 970 Chilima, Benson 268, 82 Cienfuegos, Astrid V. 228 Cevallos, William 1273, 1406 Chatelain, Eric 849 Chim, Pheaktra 1430 Cimino, Rubén O. 833, 1572, Ceylan, Ali 253 Chatio, Samuel 896 Chimalizeni, Yamikani 126 1467 Cha, Sung-Jae 1202 Chattaway, Marie A. 69 CHIME MUKWAKWA, Catherine Ciota, Alexander T. 1456 Chaba, Linda 1270 Chaurasiya, Narayan D. 1166, 630A Cirimotich, Chris M. 201, 717, Chabi, Joseph 920 1167, 995 Chimnaronk, Sarin 303 1058 Chacon, Juan C. 1340 Chaussabel, Damien 1125 Ching, Wei-Mei 1086, 1095 Cisneros, Gloria 1359 Chadsuthi, Sudarat 407 Chavchich, Maria 141 Chingoli, Felix 268, 82 Cisper, Neil J. 697 Chafey, Philippe 3 Chavchich, Marina 1502, 169, 139 Chinkhumba, Jobiba 125 Cissé, Badara 362, 382, 893 Chagas-Junior, Adenizar D. 1097 Chaves, Ana T. 1594 Chinnawirotpisan, Piyawan 1322, Cisse, Moustapha 1504, 573 Chagomerana, Maganizo 126 Chaves, Luis F. 749, 750 1325 Ciuoderis, Karl A. 1459 Chai, Jong-Yil 780 Chawan, Ajeeta 386 Chinula, Dingani 744, 757, 922 Claite, Manuela 1381 Chaichana, Panjaporn 1515 Cheasty, Tom 69 Chiodini, Peter L. 13 Clapham, Hannah 1583 Chaisson, Richard E. 1154 Checkley, William 1076 Chiosis, Gabriela 474, 32 Clark, Danielle 1301 Chaki, Prosper 1225 Cheeseman, Ian 1048, 1429 Chiou, Shyan-Song 517 Clark, Daniel 1424 Chakir, Ismael 60 Chelbi, Ifhem 1276, 34 Chiphwanya, John 1231, 531 Clark, David C. 53¸ 54 Chakma, Sumit 735 Chelimo, Kiprotich 688, 944 Chipman, Paul 1441 Clark, Gary G. 399, 403 Chakraborty, Apurba 414 Chemey, Elly 1265 Chippaux, Jean-Philippe 856 Clark, Kristina B. 979, 982 Chakravarty, Sumana 1217 Chen, Beth 1011 Chirabandhu, Tundorn 1402 Clark, Martha 1169 Chaluluka, Ebbie 1179 Chen, Ching-I 53 Chirwa, Brian 678 Clark, Taane 1047, 1506 Chalwe, Victor 1488 Chen, Hua-Wei 1095 Chirwa, Jacob 1438 Clasen, Thomas 265 Chamisa, Andrew 1360 Chen, I-Tzu 285 Chisala, Ngawina V. 353 Claser, Carla 1521 Chams, Linda M. 1134 Chen, Jun-Hu 1537 Chisenhall, Daniel M. 1346, 587 Clayton, April M. 1058 Chan, Adeline 1226 Chen, Jo-Mei 517 Chisha, Zunda 1438, 663, 986 Clayton, Christine 339

www.astmh.org A-492 Author Index The number(s) following author name refers to the abstract number.

Cleaveland, Sarah 41 Cook, Alex R. 1443 Crabtree Ide, Christina R. 248 da Silva, Inês M. 725 Clem, Rollie J. 726 Cook, Jackie 12 Craig, Alister G. 353, 1433, 347, Da Silva-Voorham, Julia 302, 501 Clemens, Emily 917 Cooke, Malcolm 1230 348 Dabiré, Roch K. 189, 1454, 1453, Clemens, John D. 962 Cooke, Mary 1451 Craig, Allen S. 1438, 1488, 195 432 Clemens, Michaela 1090 Cooksey, Richard 1509, 651, 654 Craig, Philip S. 62, 87 Dachraoui, Khalil 1276 Clements, Archie 62, 667, 821 Cooney, Laura A. 1196 Craig, Scott B. 545 Dacuma, Mary Grace B. 1523 Clements, Archie C. A. 1575, 545 Cooper, Philip J. 1464, 937 Craik, Rachel 345 Dadi, Zewdu H. Gondar. 318 Clements, David E. 1447 Cooper, Roland A. 1176, 1172 Cramer, Joanna 1008, 417 Dadzie, Samuel 213 Clemmer, William 1112 Copa, Noemí G. 1055 Crandall, Ian 32 Dagoro, Henry 1074, 933 Clifton, Dana C. 969 Copeland, John 957 Crary, Monica J. 1086, 250 Dagucon, Micaela 104 Clifton, Nicole 111 Coppens, Isabelle 1205 Cravioto, Alejandro 21, 567, 708 Dagur, Pradeep K. 338 Close, David 459 Coppola, Brittni 472 Cravo, Pedro 1500 Dahlström, Sabina 660 Closter, Linda 38 Corbel, Vincent 210, 214, 392, Creek, Darren J. 453 Dahourou, Georges A. 565, 75, Cnossen, Sonya 113, 1313 400, 433, 711 Crespo, Benigno 368, 373 110, 957, 959 Coberly, Jacqueline S. 315 Corbett, Kizzmeka 1580 Crespo-Llado, Keila N. 135 Dai, Bui 1502 Cockburn, Ian A. 1591 Cordaux, Richard 1001 Crespo-Ortiz, Maria del Pilar 1049 Dai, Minxian 359 Coelho, Paulo Marcos Z. 1033 Cordoba, Freddy 289 Crevat, Denis 506, 822, 824 Dai, Weili 700, 701 Coen, Pietro G. 13 Cordova, Jhonny J. 1351, 1235 Crisanti, Andrea 1219, 684 Daily, Harry 471 Coffeng, Luc E. 584 Cornejo del Carpio, Juan G. 1365, Crockett, Maryanne 1553, 968 Daily, Johanna 1023, 1195 Coffey, Lark L. 1007, 1403 457 Crockett, Rebekah J. 811 Dalaba, Maxwell 1114 Cogswell, Alexis A. 1002 Cornel, Anthony J. 1029, 1588 Crompton, Peter D. 358, 4, 689 Dalaperas, Stelios 810 Cohen, Adam L. 1474, 248 Cornelie, Sylvie Cromwell, Elizabeth A. 1570, Dale, Pat 557 Cohen, Joe 1546, 1547, 952, 953 Cornélie, Sylvie 427, 214, 322, 1571, 1383, 846 Dama, Emilie T. H. 322, 341 Cohen, Justin M. 578 341, 393, 711 Crosnier, Cecile 30 Dama, Souleymane 1045, 1183, Cohen, Nicole J. 1549 Corona, Teresa 91 Cross, Nadia 1, 1486, 683 1516 Cohuet, Anna 1453, 1454 Corpuz, June Cantata B. 315 Cross, Nadia J. 691 Damasceno Rego, Carla M. 1349 Coia, Michele 1447 Corrah, Tumani 767 Crowe, James E. 1578 Damdin, Altantuul 597 Coker, Babajide E. 378 Corran, Patrick 910, 914 Crowley, Kathryn 1576 Damdinorj, Tserennorov 597 Coldren, Rodney 1300 Correa, Fabio 554 Crump, John A. 435, 828, 969 Damião, Alcineia 546 Cole, Donald 423 Correa, Margarita M. 199, 228, Cruz, Christopher 516 Damien, Georgia 210, 214 Coleman, Bradley 1603 229, 807 Cruz, María 326 Damon, Inger K. 934 Coleman, Michael 434 Correa, Simon 875 Cruz, Maria 334 Damte, Mesele 1570 Colford, Jr., John M. 266 Correa Oliveira, Rodrigo 1594 Cserti-Gazdewich, Christine 1592 Daneau, Geraldine 623 Colgate, Elizabeth R. 1343 Corredor, Vladimir 654 Cuentas, Andrea 1200 Daniel, Boakye 931 Colin, Yves 1548 Correia, Artur 306 Cui, Liwang 1039, 1505 Daniel, Sylvester 514 Collard, Jean-Marc 1405 Cortada, Pedro 1055 Cui, Long 1505 Daniel-Ribeiro, Claudio T. 1396, Collazo, Vanessa 1510 Cortese, Joseph 29 Cui, Liwang 1536, 639, 652 653, 881 Coller, Beth-Ann 1447 Costa, Anielle d. 1396 Cummings, Derek A. T. 1334, Daniels, Rachel F. 29, 151, 175 Collinet-Adler, Stefan 570 Costa, Fabio T. M. 1548 1350, 502, 1053, 1052, 1051, Danner, Rebecca 1539 Collins, Deirdre 15 Costales, Jaime A. 1362, 869 407, 316 Dao, Adama 1256, 225, 226, 742 Collins, Frank H. 744, 1031, 1249 Costantini, Carlo 1030 Cummings, Hannah E. 66 Dao, Christine 1421 Collins, James J. 444, 1002 Costin, Joshua M. 1441 Cummings, James F. 953 Daou, Modibo 534 Collins, Katharine A. 176 Cot, Michel 48, 8 Cummings, Michael 1047, 1506 Dara, Antoine 1045, 1183, 1516 Colloca, Stefano 176 Cottrell, Gilles 210 Cunningham, Colm 1349 Darbro, Jonathan 219 Colón, Veronica 1336 Coulibaly, Adama 112 Cupp, Ed 614 Darby, Alistair C. 1001 Colpitts, Tonya M. 49, 50 Coulibaly, Boubacar 1584 Currie, Bart 839 Dardick, Ken 38 Comach, Guillermo 302, 501 Coulibaly, Cheick A. 746 Curriero, Frank 316 Darko, Margaret O. 1101 Comer, Andy 414 Coulibaly, Drissa 534, 6, 952 Cursino-Santos, Jeny R. 355 Darmstadt, Gary 558 Commander, Nicola J. 1088 Coulibaly, Flanon 112, 270 Curti, Elena 186, 696 Darriet, Frederic 400 Compaoré, Yves-Daniel 1187, Coulibaly, Mamadou B. 402, 702 Curtis, Kurt C. 781 Das, Ashish 1024 1201¸1513 Coulibaly, Michel E. 1476, 607, Curtis, Maurice 75 Das, Bimal K. 263 Conceiçao-Silva, Fatima 862 608 For the Cysticercosis Working Das, Debashish 1046, 1509 Conde, Silvia 387 Coulibaly, S. 707, 709 Group in Peru 58, 88, 57, 96 Das, Manoj K. 148 Congdon, Seth 991 Coulibaly, Siaka Y. 607, 608 For the Cysticercosis Working Das, Suchismita 1058 Congpuong, Kanungnit 1204, Coulibaly, Yaya Ibrahim 1104 Group 94 Das, Sanchita 1414 140, 314 Coulibaly, Yaya I. 1476, 607, 608 Czaicki, Nancy L. 847 Das, Sushismita 206 Conn, Jan E. 199, 228, 229, 733, Cousin, M. 1292, 707, 709 Czajka, Christina 939 Dasgupta, Debleena 684 807 Coutinho-Abreu, Iliano V. 804, Dasgupta, Gargi 623 Connelly, C R. 236 812 Dash, Aditya P 148 Conroy, Andrea L. 308, 817, 983 Cowan, Linda D. 93 Dash, A P. 147 Consortium, Avid 413 Cowden, Jessica 953 D Dashdavaa, Otgonbaatar 597 Conteh, Abdulai 844 Cowman, Alan F. 1, 1014, 184, DaSilva, Alexandre 1525, 1139 Conteh, Lesong 362 29, 690, 691, 187 D’Alessandro, Umberto 1548, 909 Datagni, Michelle 583 Contreras, Carmen A. 20 Cox, Jonathan 12, 1451, 49, 674 D’Ortenzio, Eric 393 Dave, Kirti 1278 Convelbo, N 709 Cox-Singh, Janet 351 d’Alessandro, Umberto 1508, Dave, Sonia 1278 Conway, David J. 1028, 1188, Coyle, Christina 56 1519, 450, 635, 1544 Davenport, Gregory 1091, 1194, 204, 1019 Crabb, Brendan S. 1, 184, 1014 Da Rosa, José 306 1197, 1208 Conway, Michael J. 50 Crabtree, Mary B. 811, 1246 da Silva, Cícera H. 354 Davey, Dylan F. 1142, 1463 da Silva, Eliana V. P. 1349 www.astmh.org Author Index A-493

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

David, Peter 1430 Demas, Sylvera 566 Diarra, Baba 608 Dodean, Rosie 1172 Davies, David H. 770 Dembélé, Ahmadou 534 Diarra, Issa 534 Dodoli, Wilfred 1374 Davies, Stephen J. 67 Dembele, Benoit 607 Diarrra, Tieman 1504, 1104 Dodoo, Alexander 890 Dávila, Francisco 1335 Dembele, Demba 1045, 1183, Dias, Fil 38 Dodoo, Daniel 1370 Davila, Marcela 1572 1516 Diaw, Mamadou M. 1504, 573 Doherty, Tom 176 Davis, Jennifer 1473 Dembele, Massitan 835 Diaw, Oumar T. 493 Dokomajilar, Chris 1201 Davis, Joe 821 Dembélé, Robert 465 Diaw, Papa A. 623 Doll, Tais A. P. F. 684 Davis, Shevaun P. 1039, 356 Demettre, Edith 711 Diawara, Sory Ibrahim 106 Dolo, Amagana 1218, 155 Davis, Timothy M. E. 1496, 366 Demin, Sergei Y. 1589 Diaz, Emilia 848 Dolo, Housseini 1476, 607, 608 Dawson, Scott 1599 Deming, Michael 1122, 1574 Díaz, Gustavo 1498 Dompnier, Jean-Pierre 1038 Day, Jonathan F. 236 Demirci, Berna 1029 Diaz, Yira R. 858 Don, Robert 1123 Day, Nicholas P. J. 670, 970, 901, Démosthène, Myriam 636 Diaz Toro, Yira 323 Dondji, Blaise 121, 1462 1039 Dena, R. 1160 Dickerson, Donald W. 1233 Dondorp, Arjen M. 670, 970, Dayan, Gustavo 1444, 822, 309 Denlinger, David L. 722 Dickerson, Tobin J. 484 1046, 1047, 1048, 1506, 1512 de Alwis, Adamberage R. 313 Denson, Lee 84 Dickinson, Harriet A. 1568 Donelson, John 1014 De Alwis, Ranjith 753, 923 Dent, Arlene E. 1596, 5, 884, Dicko, Alassane 1184, 1218, 1305 Dong, Ran 973 de Araujo, Fernanda 857 1145, 1382, 411 Dickson, Laura 1587 Dong, Shu 1021 de Araujo Lobo, Jaime M. 227 Dent, Joseph A. 447 Dida, Gabriel O. 1229, 1244 Dong, Xingqi 935 de Azevedo, Frederico T. 354 Dentz, Holly 82 Dieng, Yemou 1180, 170 Dong, Yumei 723, 203, 717 De Boer, Christopher 76 Derbali, Mohamed 1276, 34 Dieter, Kathryne L. 742 Dongus, Stefan 421, 745, 813 de Cózar, Cristina 368, 373 Derendorf, Hartmut 1242 Dieye, Tandakha N. 623 Donnelly, Christl A. 1394, 549 De Donato, Marcos 1100, 81 Derjabin, P. N. 418 Diggs, Carter 954, 181, 952 Donnelly, Martin J. 1027, 430 De Doncker, Simonne 340 Derreure, Jacques 121 Dilernia, JoAnn 107 Doolan, Denise L. 696, 181 De Groot, Anne 485 Derrick, Steven C. 1020, 247 Dillingham, Rebecca 775 Dorňáková, Veronika 1277 De Jesus, Cecilia 1227 Deseda, Carmen 1335 Dillip, Angel 1080, 1434 Doranz, Benjamin J. 310, 595, De la Hoz, Fernando 289 Desouvre, Ninon 579 Dimalibot, Judeline 1523 825, 827 de Laat, Bas 347 Desruisseaux, Mahalia 359 Dimatatac, Frederico 1324 Dorkenoo, Monique 583 de Lamballerie, Xavier 1276 Deu, Edgar 365 Dimitrova, Kristina 523 Dorsey, Grant 422, 453, 530, 535, De Los Santos, Maxy B. 335 Devaney, Eileen 474, 537 Dimopoulos, George 1058, 201, 666, 899 de Melo, Tatiane T. 783 Devine, Gregor J. 1241, 921, 745, 203, 206, 715, 716, 717, 723 Dos-Santos, Stéphanie 427 De Pina, Adilson 306 813 Dindial, Karen S. 1338 Doshi, Saumil S. 248 de Santana, Rosane A. Lins. 354 Dewi, Mewahyu 117 Diniz, Cristovam W. P. 1349 Dosoo, David K. 1308, 1157, 174 De Savigny, Don 577, 841 Dey, Ranadhir 338 Diniz, Daniel G. 1349 Dotson, Ellen M. 1366 de Sena, Isabela C. Campos. 783 Deye, Gregory 367, 640 Diniz, José A. P. 1349 Doty, Jeffrey 237 de Silva, Aravinda M. 1577, 1578, Dhabangi, Aggrey 1592 Dinko, Bismarck 1158, 159 Doucoure, Souleymane 214, 393 313, 831, 1580, 503 Dhakal, Subodh S. 279 Dion, Kirstin 396 Doudoumis, Vangelis 810 de Souza, Dziedzom K. 1129 Dhakhwa, Jyoti R. 1399 Diop, Bernard M. 1191 Douglas, Nick M. 1534, 1561 de Souza, Mikhael 1109, 1162 Dhakshinamurthy, Kalaivani 23 Diop, Medoune 1504, 573 Doumbia, Mory 1044, 106, 915 de Thoisy, Benoit 1543 Dhara, Animesh 719 Diop, R. 707 Doumbia, Saibou 1044, 106, 685, de Vlas, Sake J. 584 Dharmarajan, Guha 1257 Diop, Samba 1104 687 de Vries, Hein 620 Dhawan, Gaurav 386 Diop, Sylvie 1191 Doumbia, Seydou 1104, 402, 106, de Vries, Peter J 888 Dhiambo, Ojera 198 Diosque, Patricio 1362 746 de Wit, Emmie 524 Dhillon, Bal 970 Diouf, Ababacar 685, 687, 915 Doumbia, Salif S. 607, 608 Dean, Jacqueline L. 962 Dhorda, Mehul 454 Diouf, Mame Birame 1294, 1504, Doumbo, Ogobara K. 1045, 1183, Deb, Rinki 1129 Di Paolo, Adriana 1467, 1572 573 1184, 1218, 358, 534, 6, 689, DeBarry, Jeremy 1525, 637 Dia, Ibrahima 1028, 1247, 158, Diouf, Mamadou L. 1504, 573 777, 952, 1516, 702, 9 DeBot, Margaret A. 1219 204, 224 Dissanayake, Gunawardana 753, Doumbo, Safiatou 358, 689 Debrabant, Alain 857 Dia Badiane, Méry 1191 923 Doumtabe, Didier 358, 689 Dechavanne, Celia 3 Diabaté, Abdoulaye 432, 189 DiTusa, Charles A. 993 Douradinha, Bruno 1345 Deewatthanawong, Prasit 756 Diagne, Cheikh 1247 Diuk-Wasser, Maria 1238, 39, 554 Downs, Joni 803 DeHaro, Sergio A. 55 Diakite, Mahamadou 1044, 106, Divis, Paul C. S. 351 Downs, Philip 1576 DeHecq, Jean S. 393 345, 685, 687, 915 DiVita, Margaret A. 1268 Doyle, Patricia S. 320 Deitsch, Kirk 1178 Diakite, Seidina A. S.. 1044, 106 Dixit, Saurabh 1214 Drakeley, Chris 10, 11, 12, 1439, Deitz, Kevin C. 1030 Diallo, Abdoulaye 427 Djegbe, Innocent 433 1451, 666, 9, 910, 914, 990 DeJong, Randall 1025 Diallo, Abdallah A. 1476, 607, 608 Djènontin, Armel 210 Drame, Papa Makhtar 214, 427 Dela Cruz, Florante 53 Diallo, Boubacar 112 Djibo, Ali 1405 Dramuke, Patrick 611 Delaporte, Eric 1562 Diallo, Brehima 402 Djigbenou, Daignon R.A. 143 Draper, Simon J. 1220, 176 Delavechia, Thais 866 Diallo, Diawo 1247 Djimde, Abdoulaye 1183, 1184, Drapkin, Mark 269 Delgado, Christopher 1544 Diallo, Dapa A. 1218, 952 1516, 345, 777, 1045 Drebot, Michael A. 523 Delgado, Rosa 1096 Diallo, Ibrahima 1504, 573 Djogbenou, Luc 189 Drew, Damien R. 1224, 691 Delgado-Ayala, Rosa M. 1417 Diallo, Mawlouth 1247 Dlamini, Sabelo 666 Drexler, Anna L. 731 della Torre, Alessandra 1027, Diallo, Moussa 1256, 225, 226, Dlamini, Siyabonga B. 620 Drowos, Joanna 579 1028, 1030, 921 742 Dlugosz, Lisa 695 Drummond, Jennifer 1435, 680 Deloron, Philippe 48, 8 Diallo, M. M. 1184 Dobano, Carlota 1213 Dryden-Peterson, Scott 24 DelVecchio, Vito 1015 Diallo, Nouhoum 1516, 777 Dobler, Gerhard 518, 597 Dua, Virender K. 166, 147 Demachki, Sâmia 1349 Diamond, Michael S. 590 Dobson, Annette J. 545 Duah, Nancy O. 451, 655 DeMarco, Jean 1456 Diao, Hugo 849 Dobson, Stephen L. 230 Dube, Tina J. T. 913 DeMarco, Ricardo 443 Diarra, Amidou 154, 155, 913 Dodd, Kimberly A. 1009 Dubovska, Helena 619

www.astmh.org A-494 Author Index The number(s) following author name refers to the abstract number.

Dubray, Christine 835 Edgel, Kimberly A. 1541, 335, 387 1326, 1322 Fan, Erkang 461 Dueger, Erica 1090 Edillo, Frances 832 Engebretsen, Ingunn M. S. 102 Fan, Yi-Chin 517 Duffield, Giles E. 1031 Edith, Bougouma 167 Engel, Juan C. 320 Fang, Qiang 1536 Duffy, Patrick E. 1011, 1216, Edstein, Michael D. 139, 141, Enger, Kyle S. 569 Fang, Yaping 207 1305, 701, 1021, 1214, 1215, 1502 Engering, A. 761 Fang, Yue-yi 778 700, 702, 703 Edwards, Amy M. 1415 Engle, Ronald E. 940 Farag, Tamer 1319, 1472 Dufort, Elizabeth 1314 Edwards, Laura J. 911 Engwau, Francis 630A Farag, Tamer H. 17 Dufour, Vanessa 447 Edwards, Nick 176 Enria, Delia 1337 Farell, Margaret 1301 Dugoujon, Jean-Michel 3 Ehart, Annette 635 Enyaru, John 339 Farfan-Ale, Jose A. 943 Dujardin, Jean-Claude 326, 340 Ehrbar, Dylan J. 1456 Enyong, Peter 581 Färnert, Anna 683 Duke, Mary 786 Ehrenmann, François 3 Eom, Keeseon S. 780 Farooq, F. 181 Duman-Scheel, Molly 202 Ehrlich, Allison 868 Epstein, Judith E. 1215 Farooq, Muhammad 806 Dumler, John Stephen 917 Eichinger, Daniel 1141 Epstein, Judith E. 954, 956, 181 Farrar, Jeremy 1583 Dumonteil, Eric O. 328, 815 Eigege, Abel 1036, 1383, 463 Erb, Steven M. 1446, 1327 Farrell, Margaret 1303 Duncan, Christopher J. 176 Eiglmeier, Karin 1584 Ercumen, Ayse 266 Faruque, A.S.G. 567, 1472, 1319 Duncan, Mary Anne 579 Einberg, Jennifer 1576 Erhart, Annette 1519, 1544, 909 Faucher, Jean-François 48 Duncan, Robert 338 Eisele, Thomas P. 1377, 532, 986 Erickson, Bobbie Rae 936 Faust, Saul 176 Duncan, Robert C. 975 Eisen, Lars 428 Erickson, Sara M. 481 Fay, Michael P. 1044, 1218, 1305 Duncombe, Jennifer 821 Eisen, Rebecca 39 Escalante, Ananias A. 1203, 1428, Faye, Babacar 1180, 170, 773, Dunkle, Stacie E. 959 Eisenberg, Joseph N. S.. 1273, 1525, 1542, 885 897 Dunn, Augustine W. 208, 1260 1406, 1271, 569 Esch, Kevin J. 865 Faye, B. 895 Dunn, John 548 Eisenstein, Jana 961 Escueta, Aleyla D. S. 251 Faye, Djibril S. 493 Dunne, David W. 1568, 1569 Ekal, Emmanuel 423 Esfandiari, Javan 546 Faye, Ousmane 1247, 224, 547 Duong, Jason 1037 Ekberg, Greg 696 Esham, Heather 599, 1422 Faye, Oumar 1247 Duong, Socheat 1138 Ekoue, Kouevijdin 893 Esham, Keith 599 Faye, Sylvain 382 Duparc, Stephan 1495, 648 Eksi, Saliha 168 Eskin, Eleazar 29 Fearnley, Emily J. 545 Dupim, Eduardo G. 1063 El Alkamy, Sahar 77 Espérance, Ouédraogo 167 Fedders, Charlotte 954 Dupnik, Kathryn M. 456 El-Hariri, Hazem 1073 Espie, Emmanuelle 1186 Fehintola, Fatai A. 1499 Duque, Juliana 1339 El-Sakkary, Nelly 789 Espina, Luz 829 Feiken, Daniel R. 296 Duraisingh, Manoj 1603 El-Sayed, Badria B. 656, 658 Espinoza, Angelica 312 Feikin, Daniel R. 1474 Duraisingh, Manoj T. 30 El-Sayed, Nasr 1073 Essbauer, Sandra 518, 597 Feitosa, Fabiana 49 Durbin, Anna P. 1343, 1445 El-Setouhy, Maged 1073 Essex, Max 24 Feldmann, Heinz 524 Durham, David 554 El-Shehabi, Fouad 789 Essien Baidoo, Samuel 486 Felger, Ingrid 1388, 1560, 904 Durovni, Betina 1154 El-Zaki, Salah-Eldin J. 656 Estevez, Alejandra 522 Felgner, Phillip L. 770, 4, 1223 Durski, Kara 1119 Elahi, A E M Rubayet 735 Estrada-Franco, Jose G. 1416, Feng, Gaoqian 1490 Durvasula, Ravi 460 Elanga N’Dille, Clément Emmanuel 1419, 295 Feng, Zheng 496 Dutra, Walderez 333 214 Etang, Josiane 432 Fennel, Sean 1160 Dutt, Dhiman 248 Elango, Alamelu D. 1203 Etienne, Manuel 400 Ferdausi, Dilara 124 Dutta, Sheetij 1219, 1224, 666, Elder, John 1351, 511, 509, 555 Eum, Jai H. 719 Ferdig, Michael 1178 688, 695, 727, 952 Elfving, Kristina 1161, 1310, 1392, Eustache, Eddy 1117 Ferdig, Michael T. 1048, 1182, Dutta, Samir R. 15 673, 675 Evangelista, Julio A. 516 1429 DuVall, Adam S. 491, 500 Elguero, Carlos 1314 Evans, Brian P. 1522, 1425, 556 Ferguson, Heather 191 Dweik, Bashir 600 Elias, Maribel 556 Evans, Darin 1036, 1571, 463, Ferguson, Neil M. 1440, 669, Dwyer, Dennis M. 870, 871 Elias, Sean C. 176 533, 846 1583 Dyson, Hugh E. 1088 Ellefsen, Barry 186 Evans, Holly 1128 Fernandes, Guilhermina 1226 Dzinjalamala, Fraction K. 1181, Ellekvist, Peter 644 Evans, Jared D. 1345 Fernandes, Paula J. 1204 900 Elliot, Salena R. 691 Evaristo, Roberto 20 Fernandez, Facundo M. 1109 Ellis, Brian 1140 Evehe, M. s. b. 345 Fernandez, Kate M. 1018 Ellis, Magda K. 760 Eversole, Robert 581 Fernández, Olga L. 858 Ellis, Ruth D. 1218, 1305, 702, Ewer, Katie J. 176 Fernandez, Roberto 1237, 335, E 1011 Existe, Alexandre 636 929 Ellis, William Y. 852, 367 Eyase, Fredrick L. 146, 364 Fernandez-Sesma, Ana 1329, Eagle, Nathan 1368 ElSohly, Mahmoud 367, 996 Ezy, Fauzia 449 1580, 981 Eapen, Alex 391, 147 Elsohly, Mahmoud A. 1167, 995 Fernández-Viňa, Marcelo A. 6 Eappen, Abraham G. 1217 Elwood, Dan 1445 Feroldi, Emmanuel 600 Ebel, Gregory D. 1458, 52, 55, Elyazar, Iqbal 1558, 383 Ferrão, Sabrina B. R. L. 1148 1457 Emami, Noushin 191 F Ferraro, Bernadette 697 Eberhard, Mark 1072 Embrey, Sally R. 420 Ferreira, Andre C. 881 Ebong, Omotayo O. 1190 Embury, Paula 5 Facchinelli, Luca 555 Ferreira, Karine S. 1594 Ebonyi, Augustine 875 Emch, Michael 1103 Fairhurst, Rick M. 1044, 1211, Ferreira-da-Cruz, Maria de Fatima Ecchi, Ebaga 581 Emerson, Paul M. 1085, 1570, 685, 687, 915, 1538 653 Echavarria, Esteban 1327 1571, 846 Fairley, Jessica K. 491, 500 Férrer, Alessandra 16 Echebima, Adaku 1234 Emukah, Emmanuel 1234, 1383, Faiz, M Abul 970 Festo, Charles 634, 902 Echebima, A 612 533, 612 Falade, Catherine O. 1168, 987 Fidock, David A. 1205, 704, 1189, Echevarria, Áurea 317, 319 Enama, Mary 1215 Falconi, Francesca 334 1501 Echochard, Rene 392 Endeladze, Marina 1301 Fall, Fatou B. 1504, 573 Fieck, Annabeth 460 Echodu, Richard 458, 800 Enderes, Corinna B. 1206 Fall, Marema 623 Field, Mark 1602 Eckhoff, Phillip 677 Endy, Timonthy P. 1350, 316, Fally, Markus A. 908, 1290 Fievet, Nadine 8 Edelman, Robert 1217 Famanta, Aminata 106 www.astmh.org Author Index A-495

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

Fikrig, Erol 49, 50 Fortes, Filomeno 653 Galarza, Ivonne E. 1336 Geary, James 477 Filice, Carlo 61 Fortin, Sonia 158 Galdikas, Birute M. F. 885 Geary, Timothy G. 1132, 1193, Filler, Scott 422, 530 Foster, Jeremy 471 Galinsky, Kevin 1213, 430 447, 479, 1123, 477 Filler, Scott J. 1488 Fouché, Bernadette 110, 565, Galland, Gale 1549 Gebre, Teshome 1085, 1570 Fillié, Yvonne E. 499 957, 959 Galligan, Meghan 1431, 151 Gedamu, Lashitew 327 Fillinger, Ulrike 217 Fowkes, Freya J. I. 1, 1014, 1486, Gallinsky, Kevin 1431 Geiss, Michelle 1081 Filomatori, Claudia 981 690 Galloway, Renee L. 544 Gelb, Michael H. 853 Filson, Darren 1375 Fox, Anna M. W.. 643 Gallup, Jack M. 343 Gélvez, Margarita 308, 817, 983 Finlay, Alyssa M. 1233 Fox, Jennifer 1576 Galson, Sophie 291 Genarro, Marila 770 Finney, Constance A. M. 1592, 7 Fox, LeAnne M. 1131, 582, 1122 Galvin, Brendan D. 472 Génesté, Herve J. Leon. 636 Finney, Olivia C. 1199 Foy, Brian D. 725 Gama, Bianca E. 653 Genton, B. 426 Fire, Andrew 818 Foyaca-Sibat, Humberto 93 Gamarnik, Andrea V. 981 Gentsch, Jon 1414 Firestone, Cai-Yen 593 Franchard, Thierry 60 Gamboa, Dionicia 1200, 1210, George, Adjei 174 Fischer, Kerstin 540, 543 Francis, Filbert 681 1544, 162, 171 George, Jovvian 1482 Fischer, Katja 814 Francischetti, Ivo 1283 Gamboa, Dina C. 840 George, Miriam T. 1013, 33 Fischer, Peter U. 1071, 540, 543, Franco, Carlos E. 867 Gamboa, Ricardo 387, 88 George, Sarah 1447 781 Frangos, John A. 641 Gamo, F. J. 368, 373 Georgiev, Alexander V. 1562 Fish, Durland 1062, 38, 39 Frank, Matthias 1206 Gamo-Benito, Francisco J. 135 Gerena, Lucia 638 Fisher, Dale A. 826 Franzén, Oscar 1362 Gan, Victor C. 1323, 1324, 826, Gerner-Smidt, Peter 75 Fisher, Julie 1553 Fraser, Jamie 286, 287 820 Gething, Peter W. 117, 1556, 672, Fitch, Christina E. 563 Frearson, Julie A. 474 Ganeshan, Harini 954, 181 906, 383 Fitzpatrick, Kelly 1457 Freeman, Kira 899 Ganley-Leal, Lisa 1567 Getis, Arthur 553 Fitzpatrick, Megan B. 104 Freeman, Molly 75 Gannavaram, Sreenivas 975 Gettayamicin, Montip 649, 993 Fitzpatrick, Sinead 1356 Freeman, Nicole M. 636 Gao, Qi 1524, 1527, 1536, 172 Geukers, Karen 1193 Fitzsimmons, Colin M. 1568, 1569 Freire, Francisco P. 972 Gao, Xiugong 642 Geyer, Mary Jo 1124 Fiuza, Jacqueline A. 1594 Freitas do Rosario, Ana 1606 Garabed, Rebecca 259 Ghanchi, Najia 1531 Flaherty, John J. 1297, 624 Frempong, Kwadwo K. 213 Garba, Amadou 464 Ghani, Azra C. 1440, 574, 669, Flandin, Jean-Frederic 1408 Frenck, Robert 77 Garboczi, David 539 906, 990, 1489, 910 Flannery, Ellen 202 Frentiu, Francesca D. 724 Garcia, Andres 1368 Gharbi, Myriam 660, 661 Fleckenstein, Lawrence 1495, 648 Freudenheim, Jo L. 296 Garcia, André 48 Ghedin, Elodie 1003, 538, 541 Flegg, Jennifer A. 1043, 1047, Freundt, Natalia 1466 Garcia, Elisabeth 392 Ghosh, Anil 1232 1512, 651 Frey, Stefan 518, 597 Garcia, Gregory E. 727 Giang, Tran V. 505 Flisser, Ana 91 Fried, Michal 1021, 1216 Garcia, Hector H. 58, 59, 87, 88, Giba, Lyudmyla 1272 Flores, Guillermo Zea 614 Friedlander, J A. 638 779, 97, 57, 94, 96 Gibbons, Robert V. 1053, 1322, Flores, Karina 1424 Friedman, Jeff 367 Garcia, Jorge 1397 1326, 1330, 1334, 1400, 1402, Flores, Rosana 1055 Friedman, Jennifer F. 1566 Garcia, J.M. 761 315, 1350, 316, 1325, 1052 Flores-Mendoza, Carmen 387 Fritzen, Charissa 1245, 548 Garcia, Lynne S. 252 Gibson, Claire 1196 Flores-Rivera, Jose 91 Frolov, Ilya 1418 Garcia-Rejon, Julian E. 943 Gibson-Corley, Katherine N. 1593, Florez, Janeth 1296 Frosch, Anne E. P. 1181 Gardiner, Donald L. 625 343 Floridia, Noelia 1467 Fry, Alicia 248 Gardner, Francesca 255 Gichoya, Judy W. 560 Floyd, David 136 Fryauff, David J. 1376, 1379 Gardner, Malcolm 1199 Gichuki, Charity 1197 Fobia, W 455 Fu, Chi-Ling 1563 Gardner, Ruth 1300 Gideon, Gibson 1559 Focks, Dana A. 1262, 1263 Fuchs, James 854 Gardon, Jacques 470 Gidwani, Kamlesh 331 Fofana, Bakary 1045, 1183 Fuchs, Jeremy F. 481 Garges, Eric 1397 Gieselman, Anna 566 Fofana, Djeneba Bocar 106 Fuehrer, Hans-Peter 1290, 908 Garner, Amanda L. 473 Gil, José F. 1055, 1572 Fofana, Mahamadou 112 Fuerst, Paul A. 1086, 250 Garnier, Thierry 1584 Gil, Pedro 658 Folarin, Onikepe A. 452 Fuimaono, Saipale D. 545, 610 Garrett, Nancy 75 Gilbert, Sarah C. 176 Foley, Michael 1224, 33 Fukuda, Mark M. 1165, 1515, Garry, Robert F. 1441, 1412 Gilbreath, III, Thomas M. 212, 738 Foley, Mick 695 1047, 1212, 130, 1506, 1512, Garske, Tini 574, 669 Gildengorin, Ginny 1078, 114, Folk, Scott M. 781 761 Garuti, Helen 136 1221, 1295, 528 Folly, Kouakou 897 Fukumoto, Megumi 180 Gasasira, Anne 530, 535 Gill, Balvinder S. 1332 Foma, Munoh K. 122 Fukumoto, Shiniya 394 Gaspard, Amanda 95 Gillan, Victoria 474 Fong, Rachel 310, 595, 827 Fulhorst, Charles 946 Gass, Katherine 609 Gilles, Jeremie 736 Fonseca, Cristina T. 783, 785, Fuller, Douglas O. 748 Gatabazi, John Baptist 1408 Gillespie, John R. 461 1033 Furlotte, Nicholas A. 29 Gatt, Nina 1224 Gillrie, Mark R. 1039, 356 Fontaine, Krystal A. 1441 Futami, Kyoko 1229, 1244 Gatto, Craig 469 Gilman, Robert H. 1076, 1364, Fontenille, Didier 1453, 223 Gatton, Michelle L. 1528 1365, 58, 872, 1358, 59, 89 Fonton, Noel 392 Gauthier, Susan 1041 Gilson, Paul R. 1 Foo, Karen 1035 Gavidia, Cesar M. 87, 779 Gimnig, John 1231, 1252, 1449, Foote, Eric M. 1317 G Gaye, Oumar 1180, 170, 362, 1450, 531, 925, 1448 Ford-Jones, Lee 968 660, 893, 897 Gindin, Mariel 1143 Formenty, Pierre 1010 Gaayeb, Lobna 158, 241 Gaye, Omar 382 Gindler, Jacqueline 82 Foro, Cesar A. R. 1349 Gadalla, Nahla B. 1439, 656, 10, Gaye, O. 895 Gipson, Gregory R. 1169 Forquer, Isaac 1172 658 Gaynor, Bruce D. 1085 Giraldo-Calderon, Gloria I. 1280 Forrester, Naomi A. 1418, 1006 Gadgil, Meghana 1267 Gaywee, Jariyanart 438 Gire, Stephen K. 1407, 1412 Forshey, Brett M. Gaff, Holly 1287, 515 Gbadoẻ, A 1292, 300 Gisemba, Edward 884 Forshey, Brett M. 1348, 312, 509, Gaie, Marie F. 636 Gbaguidi, Gildas 8 Githeko, Andrew K. 1378, 157, 511, 516, 555, 1341 Gaines, Joanna 565, 566 Gbotosho, Grace O. 1493, 452 212, 215, 217, 376, 425, 404, Gaiser, R. 1605 www.astmh.org A-496 Author Index The number(s) following author name refers to the abstract number.

712 Gouagna, Louis Clement 1453 Guambo, Elizabeth 937 Habbema, J. Dik F. 584 Githure, John 200, 207 Gourbière, Sébastien 815 Gubler, Duane J. 1344, 435, 828, Habib, Nazma 1361 Giudice, Angela 866 Govella, Nico 1252 980 Habib, Najibullah 674 Glass, Gregory E. 735, 1369, 1526 Gowda, Channe 1039 Guclu Kilbas, Zeynep 253 Habler, Verena E. 908 Glass, Pam 1404 Gowda, Kalpana 182, 186A Guda, Tom 200 Habomugisha, Peace 585, 611 Glennon, Elizabeth 731 Graeff-Teixeira, Carlos 1139 Guedes, Marjorie 84 Hafifah, Cut N. 1494 Glesby, Marshall 1409 Graf, Paul C. F. 1541, 335 Guedez, Shirly M. 1235 Hagmann, Stefan 294 Glover, Simon 347 Graham, Barney 1215 Guelbeogo, Moussa 394 Hahn, Beatrice H. 1427, 1562 Gluenz, Eva 1601 Graham, James E. 21 Guelbeogo, Wamdaogo M. 1584, Haile, Ashley 1025 Gneme, Awa 1584 Grais, Rebecca F. 1405, 962 197 Hailu, Asrat 318 Go, Chi Jong 1323 Grant, Warwick N. 814 Guerbois, Mathilde 1418 Hailu, Workagegnehu 318 Gobert, Geoffrey 466, 786 Grassly, Nicholas C. 549 Guerena, Fernando B. 1309, 1318, Haji, Khamis A. 1241 Goblirsch, Sam 61, 86 Gratz, Jean 80 1321 Halasa, Yara 1054, 510 Goda, Sayed 475 Graves, Patricia M. 1234, 1383, Guérin, Philippe J. 660, 661, 1043, Hall, Aron J. 522 Godbehere, Sarah 1016 1571, 533, 612 1186, 1503, 454, 651, 1509 Hall, Cysha E. 1546 Godoy, Adrian 396 Gravett, Courtney A. 1320 Guerra Giraldez, C 57, 94 Hall, E. 79 Goethert, Heidi 37 Gravett, Michael G. 1320 Guerrant, Richard L. 1076, 16, 74, Hall, Joanna 1598 Goff, Tami 14 Gray, Darren J. 496, 62 775, 78, 80, 84 Halldin, Cara N. 1074, 1254 Goka, Bamenla Q. 156 Gray, Jennifer 1306 Guevara, Carolina 516, 938 Hallemeier, Anna G. 563 Gold, Silvia 1572 Gray, Karen-Ann 152, 907 Guevara, J. 79 Haller, Aurelia A. 1008, 1327, Goldberg, Daniel 1169 Gray, Meg 725 Gueye, Abdou S. 1293 1446 Golden, Frances V. 399 Green, A. 345 Gueye, Babacar 1381 Hallett, Rachel 1500, 1523, 658, Golden, Sandra A. 399 Green, Carol 996 Gueye, Lamine 1381 1180 Goldfarb, David M. 1553 Green, Gabriel 192 Gueye Sall, Mbayang 1381 Halpenny, C. 771 Goldman, Ira 637 Greenaway, Christina 113, 1313 Guillerm, Martine 1337 Halsey, Eric S. 1348, 312, 509, Goldsmith-Pestana, Karen 868 Greenaway, Chris 1316 Guillonneau, François 3 591, 1253, 511, 516, 555, 1341 Goldstick, Jason E. 1273, 1406 Greenberg, Robert M. 445 Guimarães, Isabel C. 498, 799 Halstead, Fenella D. 176, 1137 Golightly, Linnie M. 1414 Greenhill, Andrew R. 15 Guimarães, Luiz Henrique 864 Halverson, Gregory 355 Gollob, Kenneth 333 Greenhouse, Bryan 1161, 1392, Guimarães, Luana 864 Hamade, Prudence 1155 Golub, Jonathan E. 1154 1437, 666 Guimarães, Maria Cristina S. 354 Hamady Assadou, Mahamadoun Gomes, Giovanni F. 1349 Greenwald, Rena 546 Guindo, Amadou 402 1218 Gomes, Juliana A. S. 1594 Greenwood, Brian M. 1188, 893, Guindo, Ando 6, 952, 534 Hamainza, Busiku 1377, 195, 744, Gomes, Rafelle C. Fares. 1594 10, 174, 451, 574 Guindo, Agnes 702 986 Gomes de Azevedo, Isaclaudia 881 Gregory, Anstead 332 Guindo, Merepen A. 1218, 1305 Hamano, Shinjiro 1064 Gomez, Gerardo 75 Gregory, Christopher 1304 Guindo, Ousmane 1218 Hamapumbu, Harry 384, 385 Gómez, Giovan F. 229 Gregory, Deye 994 Guindo, Soihibou 158 Hamburger, Matthias 329 Gomez, Maria Adelaida 325 Gregory, M. 79 Guis, Hélène 210 Hamed, K. 1292, 300, 707, 709 Gomez-Puerta, Luis A. 779 Greiman, Stephen E. 480 Guitard, Evelyne 3 Hamel, Mary J. 950 Gomis, Jules Francois 893, 362 Grenfell, Bryan T. 1010 Gull, Keith 1601 Hamer, Davidson H. 1311, 559 Gonder, Mary K. 1562 Grenfell, Rafaella 1033 Gunasekera, Anusha 1215 Hamer, Gabriel L. 749, 755, 1460 Gonzales, E. 57, 94 Gresty, Karryn J. 152, 169 Gunatilleke, Shamila S. 863 Hamid, Firdaus 1465, 272 Gonzales, Isidro 58 Grewal, Paul 1015 Gundersen, Svein G. 1564 Hamidzadeh, Amir 727 Gonzalez, Armando E. 58, 59, 96, Grieco, John P. 429, 743, 754, Gunther, Stephan 939 Hamlin, Katy L. 1072, 610 57, 94 930, 556, 924, 428 Guo, Jiagang 496 Han, Eun-Taek 1537 Gonzalez, Armando for the Griffin, Jennifer B. 1487 Guo, Qin 684 Han, Kay Thwe 1204 Cysticercosis Working Group of Griffin, Jamie T. 1440, 669, 906, Guo, Yayi 1264 Han, Yeon Soo 721 Peru 779 990, 9, 910 Gupta, Charu 857 Hanchana, Sarun 1204 González, Elsa 1424 Griffing, Sean M. 1428 Gupta, Megha 1196 Hancock, Kathy 489 González, Lorenzo 1335 Griffith, Kevin S. 1087, 1115 Gurley, Emily S. 1475, 43, 46, 763, Hanczaruk, Bozena 818 Gonzalez, Lina 1518 Grigoryan, Zoya 109 948 Hand, Carla C. 1169 Gonzalez, Laura 442 Grijalva, Mario J. 1362, 1363, Gutiérrez, Lina A. 229 Handali, Sukwan 1135, 59, 95 Gonzalez, Rommell 387 1366 Gutman, Julie R. 627 Handzel, Thomas 566 González, Rodrigo J. 1257 Grillet, Florence 506 Gutteridge, Clare E. 638 Hanh, Tien N. Thi. 1583 Gonzalez, Zoe 1417 Grimberg, Brian T. 143 Guy, Bruno 1444, 36, 519, 823 Hanley, Kathryn A. 1247 Goodman, Catherine 369, 634, Grinev, Andriyan 514 Guyant, Philippe 12 Hannaman, Drew 186 902 Grisolia, Antonella 61 Guzman, Diamelys 302 Hanpithakpong, Warunee 901 Gorchakov, Rodion V. 234, 1008, Grogl, Max Guzmán, Diamelis 501 Hansen, Kristian 1163 1411, 941 Grögl, Max 459, 367, 642, 852, Guzman, Maria 1337 Hanson, Christopher T. 593 Gordon, Aubree 44, 588, 819 994 Gyan, Ben 1370, 1414 Hanson, Kara 424 Gordon, Gilad 1327, 1401 Gromowski, Gregory D. 593 Gyang, Pam 119 Happi, Christian 1412, 29 Gosh, Probir K. 568 Grondin, Danielle 968 Gyapong, John O. 606 Happi, Christian H. 1493 Gosi, Panita 1212, 998 Gros, Philippe 1041, 878 Gyorkos, Theresa W. 842 Happi, Christian T. 452 Gosling, Roly 1439, 9 Gros, Pierre P. 479 Haque, Farhana 414 Gosling, Roly D. 10 Grossman, Sharon R. 29 Haque, Mohammad Enamul 242 Gosnell, William 1105 Grubaugh, Nathan 1425 Haque, Rashidul 1290, 1367, Goswami, Doli 248 Gruenheid, Samantha 878 H 1526, 251, 254, 255, 257, 46, Goto, Yasuyuki 488 Grueninger, Heiner 372 520, 708, 735, 763, 908 Gotuzzo, Eduardo 1424 Guadalupe, Irene 1464, 937 Ha, Do Quang 751 Haraoui, Louis-Patrick 1316 Ha, Kwon-Soo 1537 www.astmh.org Author Index A-497

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

Harcourt, Brian H. 416 Hedeen, David 1460 Hilla, Marinjho 15 Hossain, M. Jahangir 414, 43 Harding, Alexander S. 1470 Hedeen, Meghan 1460 Hills, Susan L. 598 Hossain, Md. Amir 970 Harding, Simon 126 Heinberg, Adina 1178 Hillyer, Julian F. 728 Hossain, M J. 567 Harding-Esch, Emma M. 549 Helegbe, Gideon K. 156 Hinds, Sarah B. 649 Hostetter, Jesse M. 343 Hardy, Diggory 1371 Helling, Stefan 344 Hinrichs, Dave 1172 Hotez, Peter J. 1143, 1481 Harenberg, Anke 1444 Hemingway, Janet 434 Hinsley, Wes 669 Hou, Tim Y. 1031 Harhay, Michael O. 898 Hencke, Janice D. 251, 257 Hirayama, Kenji 156, 505 Houpt, Eric Harms, Jonathan E. 725 Hendriksen, Ilse 985 Hise, Amy G. 1415 Houpt, Eric R. 80, 1064 Harn, Donald A. 1150, 1151, Henebry, Geoffrey M. 1255, 1387 Hittner, James 1091, 1194, 1197, House, Brent 1301, 1303, 694 1033, 278, 496 Heng, Seng 762 1208 House, Susan 1599 Harrell, Robert A. 1585 Heng, Thay Kheang 998 Hiwat, Helene 1384, 1452 Howell, Katherine 690 Harrington, Laura C. 235, 237, Henkel, Andrea 1059 Ho, Keith T. 349 Howell, Paul 99 756 Henn, Matthew R. 588, 818, Ho, May 1039, 356 Howes, Rosalind E. 117, 672 Harris, Caroline 1453, 745, 813 1457, 1056, 1458 Hoa, Le N. M. 281 Hrobowski, Yancey M. 1441 Harris, Eva 1581, 313, 44, 503, Henrich, Philipp 1501 Hoang, Quoc Cuong 216, 216, Hsia, Yen C. 250 588, 589, 590, 816, 818, 819 Henriques, Gisela 1500 216 Hsiang, Michelle S. 1437, 1392, Harris, Erin E. 646, 1172, 132 Henry, Amy 1258 Hochaus, Guenther 1242 666 Harris, Jason B. 21, 72, 960, 961, Henry, Marie-Claire 210, 214 Hochberg, Natasha S. 1311 Hsiao, Hui-Mein 979, 504, 982 963, 550 Henry-Halldin, Cara N. 739 Hochman, Sarah 1147 Hsiao, Shih-Huai 307 Harris, Juliana V. 357 Hensley, Lisa 1423, 599, 1010, Hodder, Anthony N. 1, 691, 187 Hsieh, Michael H. 1563 Harris, Karen S. 1224 1407, 1422 Hodges, Mary H. 844 Hsieh, Yi-Ju 1563 Harrison, Lisa M. 1462, 1463, Hepburn, Matthew J. 1088, 1301, Hodges, Theresa K. 928 Hu, Branda 600 1133, 1142 1303 Hodgson, Abraham 1114, 1376, Hu, Songlin 375 Harrison, Shannon T. 534 Heppner, D. Gray 952 1379 Hu, Wei 466 Hartl, Daniel L. 1207, 1431, 29, Herath, H. M. T. Bandara 1174 Hoenemann, Mario 1535 Hu, Yan 1140 175 Herbein, Joel F. 251, 252, 257, Hoffman, Irving 991 Huang, Claire Y. H. 1577, 1446, Hartley, Ashley N. 336 254, 255 Hoffman, Stephen L. 1215, 1217, 1327, 604, 53 Hartley, David 515 Herman, Joanna S. 13 693, 956, 185 Huang, Fusheng 196 Hartsel, Joshua A. 431 Hermann, Emmanuel 158, 241 Hofmeister, Erik 1459 Huang, Fang 375 Hartwig, Carmony L. 1176 Hermsen, Cornelus Hogan, Simon 84 Huang, Jing 1006 Hartzell, Joshua 953 Hermsen, Cornelus C. 693, 955, Hoglund, Richard 901 Huang, Junjun 1245 Hartzman, Robert J. 6 185 Hohmann, Elizabeth 21 Huang, Junjun 548 Harvey, DeShannon 297 Hermsen, Rob 682 Hoke, J. Matthew 1176 Huang, Jun 954, 181 Harvey, Kira A. G. 583 Hernandez, Jean N. 1389, 1535, Hol, Wim G. 461 Huang, Liusheng 370, 899 Hashim, Ramadhan 10 886, 1393 Holbrook, Michael R. 1441 Huang, Ruili 1404 Hashim, Rizwan 1361 Hernandez, Yanira 1096 Holcroft, Christina 1313 Huang, Sheng-Xiong 1042 Hashmi, Mohammad Z. G. 1110 Hernandez, Yunuen 870, 871 Holdaway, Heather 1441 Huang, Yuefang 476 Hasina, Tsiry 874 Herrera, Beatriz 1223 Holder, Anthony 1598 Huayanay, Anibal 1237 Hassan, Azmal Ibna 124 Herrera, Yeni 387 Holding, Penny 966 Hubbart, C. 345 Hassan, Hassan K. 233 Herricks, Thurston 877 Holeckova, Katarina 116 Hubert, Véronique 660, 661 Hassan, Juma 675 Herring, Amy H. 1487 Holland, Martin J. 549 Hudson, Peter J. 1010 Hassan, Khaled E. 77 Hertz, Julian T. 435, 828 Hollenbach, Jill 6 Hudson, Thomas H. 852, 459 Hassan, Mahtab U. 970 Hess, Jessica A. 1479, 1481, 527 Hollingdale, M. 181 Huebner, Marc 1477 Hassan, Safa A. 304 Hess, Kimberly 397 Hollingsworth, T. Deirdre 1394, Huestis, Diana L. 1256, 225, 226, Hatch, Steven 978 Hetzel, Manuel W. 1559, 933 669, 906 742 Hatz, Christoph 796 Hewson, R. 418 Holloway, Kimberly 523 Hufert, Frank 518 Hauck, Eric 731 Heyderman, Robert S. 353, 126, Holm, Inge 1584 Huggins, John 1309, 1318, 1321, Haugen, Morgan 202 877 Holmen, Sigve D. 1564 1423 Havelock, Tom 176 Hibbert, Jacqueline 164 Holmgren, Gabrielle 673 Hughes, Michael D. 24 Haverford College Department Hice, Robert 1060 Holroyd, Nancy 538 Hugo, Leon 557 of Biology, Laboratory in Hickey, Patrick W. 1300 Holt, Deborah C. 814, 839 Hui, Charles 968 Microbiology and Molecular Hickman, Mark R. 646, 852, 1172, Homaira, Nusrat 46, 763 Hui, George 1017 Biology 73 132, 134, 367, 459, 642, 994 Hommel, Mirja 1486 Hukumu, Emmanuel 17 Havlir, Diane 899 Hicks, Penni 967 Homsy, Jaco 535 Hulverson, Matthew A. 853 Havt, Alex 16, 74, 78, 80 Hien, Tran T. 1039 Hong, Andrew 457 Humphries, Debbie 1463 Hawela, Moonga 1377, 1438, 744 Higazi, Tarig B. 258 Hong, Young S. 721, 730 Humphrys, Michael 75 Hawkes, Cheryl A. 1592 Higgins, Andrew 1549 Hongjun, Li 264, 797 Hunja, Carol W. 380 Hawkes, Michael 1390, 1553 Higgs, Stephen 1006, 1239, 49, Honko, Anna 599, 1422 Hunsperger, Elizabeth A. 1337, Hawley, William A. 1249, 11 519, 730 Hopkins, Adrian 580 1333, 1335, 507 Hay, J. 418 Hightower, Allen 1035 Hoque, M. Gofranul 970 Hunt, Brenda 803 Hay, Simon I. 117, 1556, 672, 383 Hijar, Gisely 1393 Horiuchi, Kalanthe 942 Hunt, Martin 537 Hayden, Mary H. 1114, 1115 Hildebrand, William H. 6 Horn, David 1602 Hunt, Paul 1500 He, Junying 196 Hildenwall, Helena 985 Horstmann, Rolf D. 767 Hunter, Andrea 1553 He, Mei 1146 Hildreth, Michael B. 1255 Hortiwakul, Tanaporn 1400 Huq, Md. Z. 1369 He, Yan 1146 Hill, Adrian V. S. 176, 1220, 767 Horwood, Paul 15 Hurwitz, Ivy 460 Healer, Julie 1, 184, 187, 691 Hill, Catherine A. 1280 Hosie, Heather 1015 Husain, Mushtuq 414 Healy, Sara A. 1320 Hill, David R. 1550 Hossain, Fakir Bellal 124 Hussain, Aashiq 1414 Heang, Vireak 596 Hill, Philip C. 1077 Hossain, Md. Sharif 1367 Hutnick, Natalie E. 697

www.astmh.org A-498 Author Index The number(s) following author name refers to the abstract number.

Huttenhower, Curtis 360 Issac-Marquez, Angelica 854 Jen, Yen-Hua 504 Joseph, Donald R. 843, 845 Huy, Nguyen T. 505 Issiaka, Soulama 167 Jenkins, Claire 69 Joseph, Jasmine 53 Huy, Rekol 100 Issifou, Saadou 499 Jennes, Wim 623 Joseph, Nidhin C. 386 Huynh, Bich-Tram 8 Ittiverakul, Mali 645, 998 Jensen, K. 1605 Joshi, Deepak 1590 Huynh, Jeremy P. 1578 Ivanek, Renata 91 Jensen, Robert 1170 Joshi, H. 147 Huynh, Uyen K. 561 Ivers, Louise C. 1106, 1120, 622 Jensen, Trevor 666 Joshi, Manju B. 870, 871 Huynh, Vu T. 770 Iwagami, Moritoshi 1529 Jensen, Vicki 599 Joslin, Charlotte E. 250 Huyse, Tine 340 Iyer, Hari 559 Jentes, Emily S. 292 Jöst, Hanna 939 Hwa, Shi Hsia 1401 Iyer, Hari S. 1311 Jeong, Soyi 988 Jourdan, Peter M. 1564 Hwang, Jimee 665, 666 Jeronimo, Selma M. B. 456, 972 Joyce, Karen 18 Hyde, Alexandra 1381 Jerzak, Greta V. S. 1456 Joyce, Kevin 75 Hyde, Molly 1034, 1035 Jespersen, Deborah J. 631 Juárez, Marisa 833, 1467, 1572 Hyde, Terri 82 J Jeyaprakasam, Madhumathi 1480 Julander, Justin 1328 Hynes, Wayne 1287 Jezan, S. Omar 345 Juliano, Jonathan J. 1491, 991 Hyseni, Chaz 1285, 1286, 800 Jackson, Brendan R. 957 Jezorwski, John 822 Julien, L. 1605 Jacob, Christopher 1047, 1177, Jiang, Dadjun 471 Juma, Elizabeth 1107, 1507 1506, 1512 Jiang, Hongying 1195 Jung, Donald 1495, 648 Jacobs, David 961 Jiang, Ju 100, 40, 41, 547 Jusot, Viviane 1405 I Jacobs-Lorena, Marcelo 1059, Jiang, Victoria 1302 1202, 1232, 1432, 1604 Jianguo, Xiao 1332 Iamsirithaworn, Sopon 1053, Jacobson, Lara 566 Jianrong, Dai 264, 787, 797 1334, 407 Jacobus, David P. 139, 141 Jibril, Haruna 24 K Ibadova, Gulnara A. 298 Jacobus, Laura R. 139, 141 Jiddawi, Mohamed S. 962 Ibañez, Javier 373 Jacquet, Maxime 736 Jigjav, Battsetseg 597 K, Ravindran J. 391 Ibanez-Valdes, Lourdes d. 93 Jafali, James 531, 905 Jimenez, Belen 373 Kaabi, Belhassen 34 Ibironke, Olufunmilola A. 448 Jahan, Nusrat 115, 115 Jimenez, Juan A. 88 Kaba, Stephen A. 684 Ibrahim, M. 345 Jahangir, Hossain M. 948 Jimenez, Veronica 1600 Kabali, Conrad 1130 Ibrahim, Marwa I. K. 332 Jaichapor, Boonsong 1425 Jin, Xiannu 1510, 891, 892 Kabamba, Joelle 934 Ibworo, Vincent 17 Jaime, César 833, 1055, 1572 Jinabhai, Champak C. 620 Kabanywanyi, A. 426 Ichimori, Kazuyo 1124 Jain, Shaifali 138 Jip, Nimzing 1571, 846 Kabanywanyi, Abdunoor M. 1185 Idoko, John A. 1144 Jain, Surendra K. 321 Jirage, Dayadevi 169 Kabatereine, Narcis B. 1569, 464, Ienco, Annamaria 921 Jalili, Nasir 299 Jittmala, Podjanee 997 497, 657 Iga, Jonah 1559 Jallow, Muminatou 1019 Joan, Gibble 14 Kabir, B. G. 401 Igbukolu, Cynthia O. 1118, 1101 Jambou, Ronan 1089, 1191, 60, Jochim, Ryan C. 1057 Kabir, Mamun 1064 Ignace, Hakizimana 374 874 John, Chandy C. 163, 688, 883, 5 Kabore, Achille 1576 Ikadai, Hiromi 1432, 1604 James, Auto 307 John-Stewart, Grace 1270 Kabore, Jacques 977 Ikeda, Allison 875 James, Anthony A. 200, 208, Johnke, Julia 1389, 1393 Kabore, Youssouf 154 Ikeda, Eriko 1229 1260 Johnson, Ann-Marie F. 587, 1347, Kabyemela, Edward 1021 Ilboudo, Hamidou 977, 341 James, Eric 693 305 Kachur, S. Patrick 369, 426, 634, Ilett, Kenneth F. 1496, 366 James, Stephanie 946 Johnson, Charles A. 29 665, 902 Imerbsin, Rawiwan 650 James, Theodore W. 1257 Johnson, Dawn 565 Kadkhoda, Kamran 523 Imnadze, Paata 1088, 1301 Jamjoom, Manal 1357 Johnson, J.C. 1422 Kaewpa, Cherdchai 140 Imran, Sayeed 1077 Jamonneau, Vincent 341, 977 Johnson, Jacob 146, 459, 364 Kahindi, Samuel 1451 Imrie, Allison 1332, 592 Jan, Vinje 1414 Johnson, Josh 599 Kahle, Kristen 310, 595, 827 Imwong, Mallika 1047, 1204, Janda, Kim D. 473 Johnson, Jacob 646, 994 Kain, Dylan C. 1592 1506, 1512, 1530, 654 Jangpatarapongsa, Kulachart 1536 Johnson, Marla K. 370 Kain, Kevin C. 1592, 308, 349, Incardona, Sandra 986 Janha, Omar 1019 Johnson, Patrick 1242 817, 983, 7 Ing Tien, Wong 351 Janse, Chris 185 Johnson, Ronald L. 1404 Kaisar, Maria M. M. 272 Iñiguez, Volga 420 Jansens, Annemieke 693 Johnson, Syd 590 Kajungu, Dan 536, 577, 633, 890 Inogwabini, Bila-Isia 1562 Janusz, Kristen B. 598 Johnson, Steven M. 643 Kakeeto, Stella 530 Inoue, Noboru 488 Janusz, Nicholas 327 Joice, Regina C. 360 Kakuru, abel 26, 453, 535 Insisiengmay, Bounnaloth 780 Jara, Jorge 768 Jollow, David 367 Kalanda, Gertrude 1179 Inyama, Petrus U. 1144, 128, 401 Jara, Marlene 326 Jones, Caroline 1107 Kalavska, Andrea 617, 619 Iordanskiy, Sergey 795 Jaramillo, Luz M. 229 Jones, Carl 1245 Kalayanarooj, Siripen 1325, 1330 Irena, Abel 1469 Jaramillo, Nicolás 218 Jones, Caroline 1381 Kalayjian, Benjamin C. 1391, 884 Irungu, Beatrice 145 Jaramillo-Gutierrez, Giovanna Jones, Dominique 1011 Kaligirwa, Nadine 888 Irving, Helen 926 1025 Jones, Douglas E. 1593, 865 Kalilani, Linda 1179 Isely, Christian 1227 Jarman, Richard 1330, 1350, 316, Jones, David S. 1011, 700, 701 Kalilani-Phiri, Linda 123, 1491, Isern, Sharon 1441 1052, 1325, 761, 1322, 1326, Jones, F 79 1492, 1508 Ishengoma, Deus R. S. 1185, 389 1334, 1400, 1402, 205, 438, Jones, Frances M. 1569 Kalivoda, Eric J. 963 Ishida, Hiroshi 367, 994 762, 1053 Jones, Jeffrey L. 774 Kalolella, Admirabilis 634, 902 Islam, Manoshi 1268 Jaske, Erin 1245 Jones, R. M. 703 Kalsy, Anuj 21 Islam, Mahfuza 263 Jawara, Musa 1028, 1030, 204 Jones, Stephen 331 Kalule, John 1314 Islam, Md. Saiful 414 Jaworowski, Anthony 346 Jones, Timothy F. 548 Kalumuna, Charles 274 Islam, Mohammad A. 708 Jayabalasingham, Bamini 1205 Jones-Engel, Lisa 885 Kalyanasundaram, Ramaswamy Islam, Mohammad S. 266 Jayeoba, Oluwemimo 24 Jongkaewwattana, A. 761 525 Islam, Rafiqul 242 Jean Baptiste, Yaro 167 Jorgensen, Pernille 768 Kam, Emily 108 Islam, Ziaul 242 Jean-Louis, Reginald 565 Jose, Christo 460 Kamal, Arzeen 568 Issa, Ouédraogo N. 167 Jeffreys, A. 345 Kamal, Sajid 914 www.astmh.org Author Index A-499

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

Kamali, Maryam 1026 Kayungwa, Benjamin 678 Khan, Inamullah 1361 1571, 613, 846, 463 Kamanga, Aniset 385, 388, 664 Kazembe, Peter N. 123 Khan, Ikhlas A. 1167, 995, 996, King, Mary Rose A. 613 Kamara, Wogba 844 Kazemzadeh, Mehdi 727 367 Kingebeni, Placide M. 1321 Kamate, Beh 1218 Kazimirova, Maria 36 Khan, M. Salah Uddin 948 Kinghorn, Alan D. 854 Kamau, Edwin 953 Kazura, James W. 1069, 1074, Khan, Shifat 261 Kingston, Hugh W. F. 970 Kambi, Jeremy 1291 1075, 1415, 163, 483, 739, Khan, Shabana I. 1167, 995, 996, Kinney, Richard 53 Kamgno, Joseph 470, 580 933, 1596, 5 367 Kioko, Elizabeth N. 801 Kamhawi, Shaden 1057 Ke, Hangjun 1040 Khan, Salah U. 414 Kiptoo, Ignatius 22 Kaminstein, Daniel 290 Kearns, Therese M. 839 Khan, Wasif A. 1290, 1367, 1369, Kirera, Ronald 71 Kaminta, Sylvester 1101, 1118 Keating, Elizabeth M. 558 1526, 708, 908, 911, 735 Kirinoki, Masashi 488 Kamm, Kelly B. 1474, 261 Keating, Joseph 671, 986 Khanakwa, Sarah 27 Kirk, Kiaran 625 Kampondeni, Sam 1022 Keele, Brandon F. 1427 Khanam, Farhana 21, 72 Kirkman, Laura 1178 Kamtchouing, Pierre 121 Keenan, Jeremy D. 1085 Kharchi, Abderrahmane O. 1504 Kirkpatrick, Beth D. 1343, 1445 Kamuliwo, Mulakwa 1373, 1377, Keenan, Martine 849 Khatib, AR 426 Kironde, Fred 683 1438, 1488, 434, 678 Keesen, Tatjana 1594 Khatib, Bakar O. 1241 Kirunda, James 753, 923 Kamusi, Mahmoud 1555 Keiser, Jennifer 793 Khatib, Rashid 536, 633, 890, Kisac, Peter 116 Kamwahi, Shaden 746 Keiser, Philip 766 1555 Kissinger, Jessica 1525, 637 Kamwendo, Debbie 991 Keiser, Tracy 66 Khatib, Rashid A. 577 Kiszewski, Anthony E. 232, 396, Kamya, Moses R. 535, 422, 572, Keita, Abdoul Salam 106 Khatibu, Rashid 1386 1030 530, 889 Keita, Adama Mamby 270 Khatun, Fatema 242 Kitajima, João P. 443 Kandula, Deepika 666 Keita, Adama D. 607 Khatun-e-Jannat, Kaniz 1268 Kitau, Jovin 221 Kaneko, Osamu 652 Keita, Abdoul S. 915 Khemawoot, Phisit 650 Kitching, Roger 557 Kang, Gagandeep 1076, 1299, Keita, Mahamadou M. 112 Khemnu, M. 761 Kitron, Uriel 1380, 1460, 491, 1302, 256, 570, 949 Keita, Sekouba 1305 Khieu, Virak 1138 500, 509, 511, 555, 737, 740, Kang, Hyun Min 29 Kelada, Andrew Y. 870, 871 Kho, Weon-Gyu 1529 749, 750 Kang, Seokyoung 721 Keller, Angela 579, 610 Khodiev, Aybek V. 298 Kittayapong, Pattamaporn 1258 Kankaew, Prasan 1425, 556 Kelley, James F. 301 Khomutov, Volodymyr 1272 Kitutu, Fred 274 Kannady, Khadija 421 Kelly, Ann 1225 Khong, Andrea 849 Kityo, Robert 1246 Kano, Shigeyuki 1529, 180 Kelly, Daryl J. 1086 Khore, Madhavi 1224 Kiulia, Nicholas M. 410 Kanuka, Hirotaka 394 Kelly, Gerard C. 667 Khrutsawadchai, Sakchai 901 Kivouele, Thomas S. N. 662 Kanzok, Stefan 1432, 1604 Kelly, Jane X. 1170, 1172, 1173 Khyang, Jacob 1367, 1369, 911 Kiware, Samson S. 758 Kao, Lin-Hsien 1331 Kelly, Paul 265 Kiang, Richard 768 Kjetland, Eyrun F. 1564 Kapan, Durrell 1258 Kelly-Hope, Louise A. 615, 616, Kibendelwa, Tsongo 1390 Klaerke, Dan A. 644 Kaplon, Jérôme 1405 607, 802 Kiboi, Daniel 145 Klalala, Francois 630 Kappe, Stefan 1196, 643 Kelton, Gaylen M. 1108 Kidima, Winifrida B. 160 Klei, Thomas R. 1479, 1481 Karangayire, Placide 888 Kemere, Jordan 1392, 1437, 666 Kigahe, Brown 890 Klein, Marina 1313 Karangwa, Jean Bosco 888 Kemgne, Eugenie A. M. 129 Kightlinger, Lon 513 Klein, Sabra 940 Karanja, Diana 1034, 1035 Kempaiah, Prakasha 1194, 1197, Kigozi, Ruth 530 Klein, Terry A. 556 Karchmer, Adolf W. 1311 1208 Kihonda, Japhet 745 Klemm, Rolf D. W. 562 Kargbo, Kandeh 111 Kenangalem, Enny 1534, 1561 Kikuchi, Mihoko 505 Kliks, Susie 1337 Karim, Mohammad J. 708 Kenangalem, E. 455 Kilembe, B. 1573 Klimov, Alexander I. 1421 Karlsson, Elinor K. 29, 550 Kendjo, Eric 660 Kilgore, Paul E. 1116 Kline, Daniel L. 403 Kasereka MC, Claude 1390 Kenefic, L. 1047, 1506 Kilima, Peter 1130 Klinger, Jeffrey D. 135, 136, 138 Kasinathan, Ravi S. 445 Kenefic, Leo 1512 Killeen, Gerry 1225, 1252, 421, Klion, Amy D. 607, 608, 1476, Kaslow, Sarah R. 1375 Kenge, Pierre 1030 557, 758, 759, 744, 757 580 Kasper, Matthew R. 596, 79 Kengluecha, Ampornpan 1425 Killeen, Stephanie Z. 1410 Klonis, Nectarios 1049 Kassogue, Yaya 1256, 225, 226, Kennedy, Erin 1035 Kilpatrick, A. Marm 1261 Kloos, Zachary A. 482, 483, 739 742 Kennedy, Malcolm 1481 Kim, Heung-Chul 556 Kloprogge, Frank L. 277 Kast, Kristopher 202 Kennedy, Mark 643 Kim, Hyeonsuk 988 Klungthong, Chonticha 1322, Katabarwa, Moses N. 585, 611 Kenney, Joan L. 234 Kim, Jessica 461 1325, 1326, 1330, 1400 Katalenich, Bonnie L. 1488 Kenny, Shannon 1049 Kim, Kami 1147 Kmit, Igor 116 Kataraihya, Johannes 1185 Kent Crockett, Rebekah J. 1246 Kim, Ryung 1195 Kmush, Brittany 940 Katebe, Cecilia 195 Kenta-Bibi, Emmanuel 122 Kim, Soon Ae 1116 Knobel, Darryn L. 41 Katele, Jacques 1112 Kerdahi, Khalil 776 Kim, Youngsun 1328 Knox, Tessa B. 919, 920 Katikiti, Samson 1435 Kerdpanich, Phirangkul 1402 Kim, Young Woo 988 Knust, Barbara 936 Kato, Agapitus B. 1286 Kerepesi, Laura A. 527 Kima, Peter E. 64 Ko, Albert I. 546 Kattula, Deepthi 1299, 1302, 256, Kerfoot, Maria 849 Kimani, F. 345 Kobayashi, Seiki 1064 570 Kerlin, Douglas H. 1528 Kimani, Naomi 1270 Kobayashi, Tamaki 384, 385 Kaufman, Michael G. 755 Kesse, Nestor 920 Kimmel, Rhonda 1069, 1596 Koch, Oliver 1211 Kaufusi, Pakieli H. 301 Kessy, Flora 1080, 1434 Kinabo, Grace D. 435, 828, 969 Kochar, Abhishek 1024 Kaul, Rupert 7 Kestens, Luc 623 King, Charles H. 1380, 491, 500, Kochar, Dhanpat K. 1024 Kavishe, Reginald A. 681 Keven, John B. 933 737, 1075, 1415 Kochar, Sanjay K. 1024 Kawada, Hitoshi 1229 Kewka, Eliningaya J. 738 King, Christopher L. 1, 1078, Kochel, Tadeusz 511, 516, 1341, Kawatoko, Kenji 1229 Keystone, Jay S. 964, 851, 968 1145, 1221, 1295, 1382, 1391, 830, 1253, 1348, 312, 509, 555 Kawazu, Shin-ichiro 488 Khairnar, Krishna 1067, 1164, 851 143, 1483, 1596, 33, 884, 966, Kochetkova, Olena 1272 Kay, Brian H. 557 Khan, Ashraful I. 960, 963 528 Koepfli, Cristian 904 Kayan, Kingsley 1308 Khan, Amir S. 697 King, Jonas G. 728 Koerich, Leonardo B. 1063 Kayentao, Kassoum 358, 689 Khan, B.K. 345 King, Jonathan D. 1036, 1570, Kogelman, Laura 1311

www.astmh.org A-500 Author Index The number(s) following author name refers to the abstract number.

Koh, Cho Yeow 461 Krcmery, Vladimir 116, 299, 617, Kweku, Margaret A. 451 Laoboonchai, Anintita 1165 Koh, Esther G. L. 1521, 1548 618, 619 Kwiatkowski, Dominic 1019, Laouini, Dhafer 34 Koimbu, Gussy 933 Krecek, Rosina C. 211 1047, 1211, 1506, 345 Larbi, Irene A. 486, 495 Koita, Ousmane A. 671 Kremsner, Peter G. 1532, 499 Kyabayinze, Daniel J. 1155 Larbi, John 1158 Kok, Gerdalize 1435, 680 Krespo Llado, Keila N. 136 Kyelem, Dominique 609, 610 Lares-Villa, Fernando 250 Koka, Hellen 413 Krishna, Sanjeev 351 Kyioshi, Okado 394 LaRocque, Regina C. 961, 963, 21, Kolibab, Kristopher 247 Krishnan, Nivedita 294 Kyle, Dennis 1242, 1501, 1530 292, 550 Kollo, Pierre 121 Kroetz, Deanna L. 370 Larone, Davise 1414 Komisar, Jack L. 954, 953 Krogstad, Donald J. 671 Larsen, David A. 532 Konadu, Dennis 405 Krolewiecki, Alejandro J. 1055, Larson, Annette L. 631 Konah, Stephen 1091, 1194, 1572, 1467, 833 L Larson, Eric T. 643 1197, 1208 Kron, Michael A. 1042 Larson, Peter S. 710 Konate, Amadou T. 913 Kronmann, Karl C. 1090, 1098, LaBadie, Robert R. 137 Larsson, Catherine J. 1343, 1445 Konate, Drissa 1044 655 LaBaer, Josh 961 Laserson, Kayla 17 Konate, Drissa S. 106 Krudsood, Srivicha 1156 LaBeaud, A. Desiree 1078, 1221, Latorre-Barragán, Fernanda 869 Konate, Drissa 685 Kruger, Philip 680 1295, 1415, 944, 114, 528 Lau, Colleen L. 545 Konate, Lassana 1028, 158, 204, Krupka, Malkie 1023, 1195 Labrique, Alain B. 115, 562, 940 Lau, Rachel 1164, 32, 661 224 Kuan, Guillermina 44, 819 Labuda, Lucja 499 Laudsoit, Anne 41 Konate, Siaka 1476, 607, 608 Kubar, Ayhan 602 Labuda, Milan 36 Laufer, Miriam K. 1181, 1492, 900 Kondiek, Nicholas 1091 Kubofcik, Joseph 1069, 1070, Lachica, Ruben 589 Laura, Sauve 1553 Kone, Aminatou 1045 1072 Lackovic, Kurt 1486 Laurence Flevaud, Laurence 1186 Koné, Abdoulaye K. 534, 6, 952 Kuchuloria, Tinatin 1301, 1303 Lafferty, Kevin 493 Laurens, Matthew B. 1217, 534, Kone, Mamady 1218 Kudom, Andreas A. 390 Lafosse, Elsie 957 900, 952 Kone, Younoussou 358, 689 Kuepfer, Irene 577 Laguna-Torres, Alberto 1348 Lauterwasser, Erica M. W.. 365, Kongere, James O. 1244 Kugelman, Jeffrey R. 1413 Lai, Chih-Yun 1579 1176 Kongkaew, Wandee 406 Kuhls, Katrin 1356, 340 Lai, Wen-Ter 307 Laven, Janeen J. 942 Kongmee, Monthathip 809 Kuhn, Susan 968 Laishram, Dolie D. 166 Lavoy, Geoff 422 Koninga, James 111 Kuhn, Walter 290 Lajoie, Gilles 223 Law, Melissa 1125 Koo, Tae-Hee 988 Kukkaro, Petra 1441 Lakwo, Tom 585, 611 Lawrie, Alison M. 176 Kopinathan, Anitha 638 Kulohoma, Benard W. 239 Lal, Amos 1302 Laws, Thomas R. 1088 Koram, Kwadwo 1090, 1098, Kum-Arb, U. 761 Lalaine, Anova 994 Lazarus, Wilfred 1130 1308, 1370, 655, 1376, 1379, Kumar, Aarti 558 Lalani, Tahaniyat 286, 287 Lazebnik, Noam 884 451 Kumar, Jessica A. 1314, 1315 Lalloo, David 1436 Lazo, John S. 459, 852 Koram, Kojo 910 Kumar, Mukesh 603 Lalou, Richard 427 Le, Dao Duc 1204 Korkusol, Achareeya 1522 Kumar, Nirbhay 644 Lam, Polo C. H.. 431 Le, Hoa N. M. 275 Korman, Lawrence 1309 Kumar, Pavan 1482 Lamarque, Mauld 1597 Le Bras, Jacques 651, 660, 661 Koroma, Joseph 1129 Kumar, Rajiv 331 Lambert, Lynn 1011 Le Port, Agnès 3 Korsman, Stephen 93 Kumar, Sanjai 1020, 247 Lambrechts, Louis 1050, 205 Le Xuan, Hung 635 Koru, Ozgur 253, 602 Kumar, T. R. Santha 1205 Lameyre, Valerie 889 Leach, Amanda 1213 Kosoy, Michael 438 Kumar, Vishwajeet 558 Lammie, Patrick 1072, 1467, Leal, Andrea 1263 Kosoy, Olga 942 Kumarasamy, N. 23 1572, 579, 610, 833, 613 Leal, Grace 1006, 1416, 295 Kostyuchenko, Victor 1441 Kumaraswami, V. 1482 Lampah, Daniel A. 1534, 455 Leang, Rithea 674 Kotaki, Akira 1426 Kun, Jürgen F. 1532 Lampe, David 1232, 194 Learn, Gerald H. 1562 Kotloff, Karen 112, 1319, 1077, Kunath, Shajo V. 770 Lamsal, Aashis 1255 Leary, Kevin 1511, 645, 998 1472, 17 Kunene, Simon 666 Lanar, David E. 684, 952, 688 LeBreton, Matthew 1562 Koukounari, Artemis 448, 465, Kuntawungin, Worachet 1165, Lanata, Claudio 1076 Ledeboer, Nathan 1042 549 998 Lanciotti, Robert S. 942 Lederman, Edith 286 Koumare, Sekou 1045, 1183 Kuntz, Karen M. 1119 Lander, Eric S. 29, 430 Ledermann, Jeremy 1246 Kouodjip Nono, Larissa 1136 Kurane, Ichiro 1426 Landis, Sarah H. 1487 Ledgerwood, Julie 1215 Koura, Ghislain 48 Kurewa, Nyaradzai E. 626 Landmann, Frederic 471 Lee, Carla 82 Kouriba, Bourema 534, 952 Kuris, Armand 493 Landmesser, Mary E. 357 Lee, Cynthia 953 Kourouma, Nana 112 Kurscheid, Sebastian 49 Laney, Sandra 1073 Lee, Jiyoung 1274 Kousa, Youssef 1016 Kurtis, Jonathan D. 1216, 1566, Lang, Jean 1444, 36, 506, 519, Lee, John S. 1425 Kovacs, Colin 7 1021 823 Lee, Kristine 1039, 356 Kozar, Michael P. 1510, 638, 646, Kurtzhals, Jurgen A. L. 156 Lang, Trudie A. 1554 Lee, Linda K. 1443, 820 891, 892, 993, 134, 367 Kuru, Teklu 327 Langa, António J. 840 Lee, Ming-Chieh 215, 712 Kozarsky, Phyllis 292 Kurup, Samarchith P. 526 Langat, Bernard 145 Lee, Ming-Na T. 370 Kozlowski, Elyse 1189 Kusi-Appiah, Rita 1118 Langat, Joyce 243 Lee, Patricia J. 646 Kramer, Kenton 1105 Kusriastuti, Rita 383 Langendorf, Céline 1405 Lee, Susan Shin-Jung 285 Kramer, Laura D. 1456 Kusuma, Andreas 1517 Langer, Christine 1 Lee, Sunhee C. 1147 Kramer, Martin L. 136, 138 Kwadwo, Boateng 405 Langhorne, J. 1606 Lee, Vikki 47 Kramer, Vicki 515 Kwadwo, Koram 931 Langlais, David 1041 Lee, Vernon J. 1443, 1582, 826, Kramyu, J. 761 Kwak, Cecilia 1469 Lanteri, Charlotte A. 647, 993, 820 Kranz, Melissa 548 Kwakye-Maclean, Cynthia 841 367 Lee, Yoosook 1029, 1588 Kranzusch, Philip J. 1562 Kwakye-Nuako, Godwin 1357 Lanusse, Carlos 1123 Lee, Yock Ann 1401 Kraus, James M. 853 Kwara, Evans 1308 Lanza, Carlo 997 Lee, Yeuk-Mui 489, 59, 792, 95 Krause, Peter 38 Kwarteng, Anthony 405 Lanzaro, Gregory C. 1029, 1030, Lee Xuan, Hung 909 Kravvariti, Evrydiki 56 Kweka, Eliningaya J. 212 1588 Leed, Susan E. 646 Lanzavecchia, Antonio 590 www.astmh.org Author Index A-501

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

Lees, Rosemary 510 Li, Austin 207 Littrell, Megan 1469, 371 Lopes, Gabriela 194 LeeVan, Elyse 539 Li, Baiqing 1536 Liu, Canhui 1003 Lopez, Anna L. 962 Lefèvre, G 1292 Li, Benwen 478 Liu, Ge 1328 Lopez, Beatriz 249 Lefranc, Marie-Paule 3 Li, Hong 1328 Liu, I-Ju 1579 López, Daniel 1335 Legorreta-Herrera, Martha 1222 Li, Jian 1537 Liu, Joanne 1553 Lopez, Edna M. 315 Legrand, Eric 1543 Li, Jenny 1593 Liu, Jing 1600 López, Giovanni 1424 Leguia, Cristina 1466 Li, Jun 207, 404 Liu, Kit Yeng 1465 Lopez, Liliana 1327, 1446 Lehman, Jennifer 507 Li, Jianyong 431 Liu, Li-Teh 504 Lopez, M. 79 Lehman, Leopold Gustave 1136 Li, Jiyun 461 Liu, Lei 520 López, Roger 44 Lehmann, Tovi 1256, 1257, 225, Li, Lin 541 Liu, Meng 1146 Lopez, Victor 930 226, 742 Li, Meng 720 Liu, Mingli 164 Lopez-Gonzalez, Irene 1416 Leibly, David J. 643 Li, Pi-Chun 1579 Liu, Qingzhen 594 Lopez-Perez, Mary 1210 Leiby, David A. 14 Li, Qigui 1172, 130, 131, 132, Liu, Wenzhe 1328 Lopez-Urbina, Teresa 779 Leimgruber, Stephanie 459 133, 134, 1510, 367, 640, 993 Liu, Weimin 1427, 1562 Lopman, Ben 937 Leite, Álvaro M. 16 Li, Shuhong 978 Liu, Xiangyu 1328 Lord, Cynthia C. 236, 554 Lek, Dysoley 674 Li, Tony 111 Liu, Yi 1255 Lorencz, Erin E. 28 Lek-Uthai, Usa 1521, 1530, 887 Li, Tao 1217 Liu, Yaobao 1527 Lorenzo, Micaela 1359 Leke, Rose G. F. 161, 2 Li, Xiaolian 639 Liu, Yue 520 Loria-Cervera, Nalleli 342 Lemnge, Martha M. 1185, 389, Li, Xinshe 999 Livengood, Jill A. 1008 Loro, Luis M. 1359 681 Li, Yuexin 1172 Liyanage, Tilaka 1071 Loroño-Pino, Maria A. 943 Lemperiere, Guy 736 Li, Yingying 1427, 1562 Lizak, Patricia 370, 899 Loua, Marcel K. 204 Lemusu, Siitia 769 Li, Yuesheng 496 Ljung, Annika 1310 Loutfy, Mona 7 Lengeler, Christian 1080, 1434, Li, Zhiru (Liz) 473 Llanos-Cuentas, Alejandro 1544, LoVerde, Philip T. 446 426, 713 Liang, Guodong 935 324, 340 Low, Kay En 1521 Lenhart, Audrey 1235, 1351 Liang, Michael 32 Llergo, Jose L. 368, 373 Lowe, Rachel 1353 Lennon, Niall J. 1056, 1457, 1458, Liang, Song 259, 778 Llinas, Manuel 1040, 1182, 1189, Lowenberger, Carl A. 885 588, 818 Liang, Xiawu 1199 1603 Lowther, Sara 268 Leno, Mamadou 977 Liang, Xiaowu 770 Lloyd-Smith, James O. 1010 Lozano, Sonia 373 Leo, Yee Sin 1323, 1324, 820, Liang, Xiaodong 941 Lluque, Angela M. 19, 85 Lu, Bao 66 824, 826 Liang, YanMei 1567 Lo, Aminata C. 659 Lu, Chih-Hsuan 1579 Leo, Yee Sin 1582 Liao, Mei-Ying 1579 Lo, Ami C. 897 Lu, D. 1605 Leon, Juan S. 420 Libby, Tanya 1437 Lo, Michael K. 416 Lu, Feng 1537 Leon, Leonor L. P. 319, 317 Lichtner, Franz 1015 Loaiza, Jose R. 199 Lu, Peng 1590 Leon, Mubikayi 1112 Liebman, Kelly 1253 Loan, Huynh Thi Kim 751 Lu, Zhong 514 Leonardo, Lydia R. 488 Liebmann, Katherine 1170 Loayza, Manuel J. 1359 Luambua, Lex 280 Leong, Hoe Nam 824 Lievens, Marc 1213 Lobo, Cheryl A. 355, 183 Luang-Suarkia, Dagwin 1332 Leontsini, Elli 1471 Liles, W. Conrad 308, 817, 983, Lobo, Neil 1451, 744 Luby, Stephen P. 1266, 1268, Leopold, Jennifer 1576 1592 Lobo, Neil F. 11, 1249 1421, 1471, 1475, 248, 261, Leping, Sun 787 Lim, Chang-Kweng 1426 Locatelli, Sabrina 1562 263, 266, 414, 43, 46, 567, Lepore, Tim 38 Lim, Chwee Teck 887 Lochmatter, Corinne 793 568, 763, 948, 1267 Lescano, Andres G. 1541, 335, Lim, Pharath 1211 Lockman, Shahin 24 Lucas, Carmen M. 1541, 335 387 Lima, Angeles 1186 Lofts, Loreen L. 1413 Lucas, John 197 Lessa, Carol 546 Lima, Aldo A. M. 16, 74, 78, 80, Logan, Thomas M. 806 Lucchi, Naomi W. 1525, 637 Lessler, Justin T. 1053, 1334, 316, 84 Logue, Kyle 1254 Luce-Fedrow, Alison 1287 502 Lima, Ila F. 16, 74, 78, 80 Logvinenko, Tanya 21, 961 Luciani, Mark 644 Leung, Daniel T. 72, 960, 963 Lima, Noélia L. 16, 74, 78 Loharikar, Anagha 110, 1265, Luciano, Jacinta 1226 Leung, Ka Fai 1602 Limbach, Keith 186A, 696, 954, 565, 959 Lucke, Colleen E. 697 LeVasseur, James 1459 186, 182, 181 Lok, James B. 1468, 999 Luckhart, Shirley 192, 228, 229, Levens, Joshua 917 Limsalakpetch, A. 761 Lok, Shee-Mei 1441 731, 807 Levert, Keith L. 1135, 1139, 792 Lin, Ai J. 1510 Lokomba, Victor 1487 Ludlow, Louise E. A. 346 Levett, Paul N. 523 Lin, Chang-Yun 1195 Lon, Chanthap 1047, 1165, 1212, Luengchaichaweng, Anicha 140 Levin, Michael 243 Lin, Hong-En 1579 1506, 1511, 1512, 1515, 645, Lugo-Caballero, Cesar 860 Levine, Myron M. 1077, 112, Lin, Yun-Hui (Annie) 695 762, 998 Luhanga, Misheck 1374 1472, 17, 1319 Linares, Nivaldo P. 768 Londono, Berlin L. 747, 1347 Lui, Hong-Di 442 Levine, Rebecca 1460 Lindblade, Kim A. 1306, 522, 249 Londono, Flaminio 1251 Luka, Madalitso 1231, 531, 905 Levy, Arielle 1553 Lindegardh, Niklas 1499, 277, Londono-Renteria, Berlin 220 Luke, Thomas 830 Levy, Karen 1271 453, 901 Londono-Renteria, Berlin L. 1251, Lukens, Amanda K. 1431, 175, Levy, Katelyn H. 1364 Lindh, Magnus 1310 732 29, 151 Levy, Michael Z. 1358, 1364, Lindo, E. 79 Long, Carole A. 1044, 1220, 176, Lulembo, Oliver 1438, 678 1365, 457 Lindow, Janet C. 1343, 1445 184, 685, 687, 915, 1218, 31, Lum, Lucy C. S. 510 Lévy, Prisca 3 Lines, Jo 1449, 424 688 Lumb, Vanshika 148 Lewis, Ian 1040, 1182 Linguissi, Laure Stella Ghoma 662 Long, Kanya C. 312, 591, 1341 Lumbaso, Sam 1291 Lewis, Sheri L. 315 Linse, Sara 683 Long, Lewis 1425 Lumsden, Joanne M. 1546, 1547, Ley, Benedikt 1290 Linthicum, Kenneth J. 806 Longo, Cecilia 1396 953 Ley, Serej D. 240 Liskova, Anna 619 Lopa, Tajnin M. 261 Luna, Tania 1409 Lezama-Davila, Claudio M. 66, Lissandrin, Raffaella 86 Lopera, Juan G. 1459 Luong, Tinh-Nhan 1553 854 Litilit, Dianne 182, 186A Lopera-Mesa, Tatiana M. 1044, Luong, Thulan 892, 1510 Li, Ang 1521, 887 Little, Eliza 1238 685, 687 Lupi, Otilia 1396

www.astmh.org A-502 Author Index The number(s) following author name refers to the abstract number.

Lusingu, John P. A. 681 Mageo, Sharmain 769 Mamun, Abdullah A. 1421 Martin, James 1309, 1318, 1321 Lustigman, Sara 1210, 1479, Magesa, Stephen 221 Mamun, Md. A. 1471 Martin, Joseph G. 592 1481, 525, 541, 542 Magill, Alan 367, 642, 649, 994 Man, W.Y. N. 465 Martin, Julie S. 1141 Luthier, Juan J. 1362 Magnussen, P. 895 Manasatienkij, Wudtichai 1325 Martin, Rowena 625 Lutiali, Peter A. 1244 Magona, Joseph 458 Manceva, Slobodanka D. 703 Martin, Thibaud 433 Lutomiah, Joel 1586, 413 Maguire, Jason D. 286, 287 Mancl, Karen 1274 Martínez, Dianny 1100 Lutucuta-Kosi, Florbela J. I. 653 Mahajan, Sumit S. 1176, 365 Manda, Hortance 930 Martinez, Ofelia 1397 Mahama, Emmanuel 1157, 1308, Mandala, Wilson 348 Martinez-Alfaro, Carlos 1419 Lutwama, Julius J. 811, 1246 1370, 405 Mandia, Victor 985 Martínez-Calvillo, Santiago 860 Luty, Adrian J. F. 1465, 693, 955 Mahanty, S. 57, 94 Mandike, Renata 713 Martinez-Loya, Orlando A. 772 Luyai, Anthony E. 490 Mahapatra, Lily 1478 Manetsch, Roman 1242 Martins, Daniela R. Arantes. 972 Lwetoijera, Dickson W. 188, 745, Maharaj, Payal D. 1455, 54, 53 Manga, Akhenaton 382 Martins, Eduardo 354 813 Maharaj, Rajendra 1435, 150, 668 Mangano, Valentina 154 Martins, Watson 1033 Lwin, Khin Maung 901 Mahieu, Els 579 Mangesho, Peter 621 Martins, Yuri C. 641 Lyashchenko, Konstantin 546 Mahmoud, Eiman 1037 Mangudo, Carolina 1055 Martinson, Francis 991 Lybarfe, Mary Electa 1312 Mahon, Barbara E. 565, 957 Mangwiro, Clement 1360 Maruyama, Eduardo 96 Lyda, Todd A. 871 Mahungu, Tabitha 176 Manianga, Cele 1112 Mary, So 645, 998 Lye, David C. 1323, 1324, 1582, Mai, Absalom 933 Manickam, Nisha 1142 Maryse, Lebrun 1597 820, 826, 1443 Maia, Marta 398 Manix, Catherine 1195 Marzal, M 57, 94 Lyerly, David 1066 Maier, Alexander 690 Manjara, Rutendo 98 Marzan, Florence 899 Lyke, Kirsten E. 1217, 6, 952, 956 Maier, Elizabeth 84 Manjurano, Aphaxard 985 Masaninga, Freddie 1373 Lynde, Carrie 851 Maiga, Amelia W. 1045, 1183 Mankhambo, Limangeni 441 Masanja, Irene 536, 577, 633, 890 Lyon, Mathew 290 Maiga, Boubacar 155 Mann, Henning 643 Masbar, Sofyan 449 Lyth, David 76 Maiga, Hamma 1516 Mann, Victoria 795 Masembe, Charles 1286 Maina, Alice N. 41 Manning-Cela, Rebeca 860 Masomhe, Sadick P. 1555 Maiolatesi, Santina 954 Manock, Stephen 1541 Mason, Carl J. 1399, 438, 761 Maire, Nicolas 1371 Manoff, Susan 1447 Massey, Jr., Holman C. 999 M Maitland, Kirsty 474 Manoj, Gambhir 549 Massougbodji, Achille 8 Majam, Victoria 1020 Manske, M. 1047, 1506, 1019 Masuoka, Penny 556 M, Ilona 597 Majambere, Silas 1241, 675, 745, Mansour, Adel M. M. 77 Matey, Elizabeth 1034 M.V.R., Reddy 1480 758, 813 Mante, Sonny 1124 Mathanga, Don P. 1231, 1372, Ma, Qing 1499 Majanja, Janet 45, 765 Mantel, Nathalie 1444, 36, 506 531, 710, 905, 1181 Mabey, David C. W. 549 Majwala, Albert 440 Manu, Alexander 405 Mather, Michael W. 1040 Mable, Barbara 494 Mak, Tippi 796 Manu, Grace 1370 Mathew, M.A. 1116 Mabuza, Aaron 680 Makame, Makame H. 1392, 1437 Manzi, Fatuma 1386 Mathieu, Els 1574, 583, 835, 837 Macalalad, Alexander R. 588 Makepeace, Benjamin L. 1001 Manzo, Mahamane Laouali 1405 Mathieu-Daude, Françoise 322 MacDonald, Kevin 791 Makharadze, Manana 1088 Marano, Nina 1549 Matias, Abrahan 396 MacDonald, Nicholas 1011 Makhema, Joseph 24 Marantina, Sylvia S. 1557 Matindii, Benjamin Atua 1186 MacDonnell, Heather 1553 Makhviladze, Manana 1301 Maravilla-Moreno, Deyra 1222 Matovu, Enock 339 Mace, Kimberly E. 1488 Makio, Albina 413 Marc, C 300 Matowo, Johnson 221 Macedo, M P. 1149 Makkiya, Adil 475 Marcet, Paula L. 1366, 1460 Matt, Julia 1290 Macedo, Silvia 387 Makoutode, Michel 189 Marchi, Samyra 1097 Mattar, Salim 282 Macedo de Oliveira, Alexandre Makowski, Kai 523 Marcombe, Sebastien 400 Matthias, James 916 1428 Maksoud, Mohamed A. 1301 Marconi, Thiago 866 Mattia, Kimberly-Anne 310, 595, Machado, Alcyone A. 1148, 1149 Makusa, Dayton 678 Marcos, Luis 781 825, 827 Machado, Gerzia M. 319 Malaga, Edith 1364 Mardy, Sek 762 Matu, M. 1160 Machado, Paulo Roberto 864 Málaga Chavez, Fernando S. 457, Marfurt, Jutta 1528 Matzhanova, A. M. 418 Machado, Ricardo L. 1210 1365 Margolis, Harold S. 1333, 544, Maude, Richard J. 670, 970 Machault, Vanessa 427 MalariaGEN Consortium 154 1335, 1337, 507 Maude, Rapeephan R. 1083 Machen, Heather 967 Malecela, Mwele 1130 Marie, Alexandra 711 Maurer, Marcus 121 MacInnis, B. 1047, 1506 Malecela, M.N. 1573 Marín, Juan C. 807 Mauricio, Isabel 1356 MacKay, Andrew 755, 927 Malhotra, Indu 1078, 114, 1145, Marinho, Fábio A. Vitarelli. 785 Maves, R. 79 Mackenzie, Charles 1123, 1124, 1221, 1295, 1391, 1596, 491, Marinotti, Osvaldo 1260, 200, 208 Mawindo, Patricia 1492 1130, 580, 581 528, 966, 500 Marins Povoa, Marinete 1428 Maxwell, Appawu 931 Mackosso, Carole 651 Malila, A 426 Markey, Michael 805 May, Linda 272 MacLeod, William B. 1311 Malima, Robert 221 Markey, Nicole 595 Mayer, Sandra A. 591 Macmullin, Greg 32 Malinowski, Claire 823 Maro, Venance P. 435, 828 Mayeux, Patrick 3 MacNeil, Adam 521, 936, 945 Malleret, Benoit 1521, 1548, 887 Marques Jr., Ernesto T. A.. 1345 Mayhew, George F. 481, 538 Macrito, Attilio 1037 Mallert, Benoit 1530 Marrenjo, Dulcisária 1226 Maynard, Sean K. 67 Madanitsa, Mwayiwawo 1179, Mallewa, Macpherson 126, 876 Marsden, Clare D. 1029 Mayta, H. 57 1492 Mallick, P K. 147 Marsh, Kevin 690 Mayxay, Mayfong 1048 Madrid, Yenny 81 Malmberg, Maja 658 Marshall, John M. 551, 669 Mazier, Dominique 887 Madrill, Nicole 1123, 477 Malpica, Luis 1466 Marshall, Lee 113 Mba-Jonas, Adamma 959 Maecker, Holden T. 358 Maltz, Michele 1275, 1288 Mårtensson, Andreas 1161, 1310, Mbacham, Wilfred F. 129, 629, Maes, Ilse 340 Malukiewicz, Joanna 1203 1392, 1437, 673, 675 345 Maes, Louis 340 Maly, Dustin J. 643 Marti, Hanspeter 1138 Mbala, Placide K. 1318 Maestre, Amanda 1518 Mamadou, Diallo A. 1146 Marti, Matthias 360 Mbalabu, Jean-Claude 276 Magalhães, Marina C. 1349 Mamaty, Abdul-Aziz 1405 Martin, Akogbeto C. 222 Mbale, Emmie W. 126 Magbity, Edward 844 www.astmh.org Author Index A-503

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

Mbando, D.W. 1573 McMahon-Pratt, Diane 868 Metenou, Simon 1476, 608 Mirembe, Justine 25, 27 Mbange, Aristid Herve E. 629 McManus, Donald P. 496, 62, 760, Methaneethorn, Janthima 648 Miri, Emmanuel 1036, 1383, Mbaya, Bridon 347 466, 786 Metta, Emmy 369 1571, 463, 612 Mberikunashe, Joseph 668 McManus, Joshua 849 Metz, Kyle 343 Mirsalis, Jon 996 Mbidde, Edward K. 1087 McMillan, David 367 Meya, David 1119 Miruka, Argwings 22 Mboup, Souleymane 1431, 151, McMorrow, Meredith 636 Meyer, Hermann 1010 Misau, Adamu A. 401 175, 29, 623, 1207 McMullan, Laura 945 Meyer-Weitz, Anna 620 Misczak, John 1328 Mbugi, Patrick J. 217 McNelly, James R. 399 Meyers, Jacob I. 725 Misiani, Eunice 1435, 680 Mbwili, Clara 1438 McNutt, Sarah E. 1462 Meyerson, Howard 143 Misse, Dorothée 214 McAuliffe, Isabel T. 59 McPherson, Scott 1571 Meza, R 79 Mita-Mendoza, Neida 687 McBride, Alan 1097 McQuillan, Jacqueline 538 Mharakurwa, Sungano 384, 385, Mitchell, Adeyemi 1319 McBride, Carolyn S. 1586 Mduluza, Takafira 529 664, 679 Mitchell, David 75 McBride, Flavia 1097 Mead, Daniel 1460 Miao, Jun 1505, 639 Mitchell, Graham 1598 Mcburney, Sean P. 1345 Mead, Edward A. 720 Miao, Miao 1505, 652 Mitprasat, Mashamon 1511, 645, McCall, Matthew B. B. 955 Mead, Paul S. 1087, 1115, 39 Miao, Qianqian (Doreen) 337 650 McCall, Philip 1235, 1351 Meciakova, Michaela 299 Michael, Obaro S. 452 Mitre, Edward 1127, 1128, 1464, McCalmont, William F. 646, 993 Medeiros, Marco A. 546 Michael, Scott F. 1441 1477, 539 McCann, Robert S. 1448 Medetov, Zh. B. 936 Michaelis, Annie 1265 Mittelholzer, Christian 684 McCarra, Matthew 163 Medina, Anicia M. 20 Michalakis, Yannis 1454 Miura, Kazutoyo 1218, 1220, 176, McCarthy, Anne 1553 Medina, Belkis 1100 Michalkova, Veronika 1061, 1279 184, 685, 687, 915 McCarthy, James S. 625, 839 Medina, Belquis 81 Michalski, Michelle L. 538 Mixson-Hayden, Tonya 1428 McCarthy, Sarah 1423 Medrano-Mercado, Nora 344 Michel, Guesly 1117 Miyahira, Yasushi 692 McCaughan, Megan A. 1540 Meek, Sylvia 1155, 12, 674, 714 Michele, Mpunga 1112 Mkoba, Clarence 369 McChesney, James 367 Mehra, Sucheta 562 Michelo, Sandford 265 Mkupasi, Ernatus 92 McChesney, James D. 1167, 995 Mehrez, Mohamed 1073 Michon, Pascal 1014 Mkwanda, Square Z. 616, 835 McClellan, Holly 700, 701 Mehta, Raaj S. 1464 Michyari, Aloke 1131 Mlambo, Godfree 644 McClelland, Michael 21 Mehus, Joseph O. 480 Middha, Sheetal 1024 Mmbando, Bruno P. 389 McClure, Elizabeth M. 1382 Meilia, Rizka 449 Midekisa, Alemayehu 1255, 1387 Moch, Kathy 727 McCollum, Andrea M. 1428, 934 Mejia, Rojelio 525 Midzi, Nicholas 529 Modiano, David 154 McCormick, David W. 441 Melanson, Vanessa 1425 Migot-Nabias, Florence 3 Modise, Taolo A. 24 McCormick, Kevin 1345 Melby, Peter 332 Mihretie, Abere 1387 Moebius, Jacqueline 689 McCoy, J. Philip 338 Melendez, Victor 1172, 130, Mikolasova, Gertruda 299 Moehrle, Joerg J. 997 Mccoy, Margaret E. 684 1510, 367, 891, 892 Mikolasova, Petra 299 Moesh, Stephanie M. 1462 McCracken, John 1306 Melendrez, Melanie C. 1322 Mikolon, Andrea 414 Moffett, Daphne B. 959 McCracken, Michael K. 1346, 305, Mellor, Steven 674 Mikolon, Andrea B. 948 Mohamed, Soghaier A. 304 586, 587 Melnikow, Elena 541 Milazzo, Mary Louise 946 Mohammed, Bakari J. 1392 McCulloch, Charles E. 1085 Melo, Leonardo 354 Milenković, Tijana 1182 Mohammed, Hamish 211 McCullough, Andre 75 Menacho, Freddy 20 Miles, Michael 1356 Mohan, Tina 1545 McDonald, Chloe E. R. 349 Menard, Didier 1204, 1430 Milhous, Wilbur K. 1242 Mohan, Venkata 570 McDonald, Emily A. 1566 Menard, Robert 1591 Millen, Amy E. 296 Mohasin, Md. 72, 960, 963 McDonnell, Joseph 839 Mendez, Jairo 554 Miller, Becky 1429, 1048 Mohmmed, Asif 138 McElroy, Kate L. 1056 Mendez, Sabrina 937 Miller, Barry R. 811, 1246 Mohon, Md. Abu Naser 735 McElroy, Peter D. 1437, 676, 713, Meneely, Philip M. 73 Miller, David M. 416 Moi, Meng Ling 1426 675 Meneses, Claudio 746 Miller, John 1374, 1377, 1438, Moiroux, Nicolas 210 McEwen, Lisa M. 1150, 1151, 278 Menezes, Cristiane A. S. 333 757, 1373, 986 Moke, Fenny 17 McFarland, Deborah R. 1111, 533 Meng, Hao 652 Miller, Janice 38 Mølbak, Kåre 1076 McField, Romeo 124 Mengistu, Netsanet W. M. K. 120 Miller, Jane E. 1228 Molez, Jean-François 893 McGaha, Tommy J. 605 Mengistu, Sofie 109 Miller, Jeffrey R. 1087 Molina, Douglas M. 1199, 770 McGee, Charles 519 Menon, Manoj P. 1111 Miller, Louis 1218 Molina-Cruz, Alvaro 1025 McGrath, S 181 Menon, Pradeep 23 Miller, Robin 754 Molinari, Noelle-Angelique M. McGraw, Elizabeth A. 724, 729 Mens, Petra F. 888 Miller, Scott R. 130 1549 McGready, Rose 1046, 1486, 454, Menya, Diana 576 Miller, William 198 Mollik, Md. Ariful Haque 124 901 Mercado, Juan Carlos 819 Milligan, Paul Molteni, Fabrizo 676 Mcha, Juma H. 1241 Mercereau-Puijalon, Odile 1430 Milligan, Paul J. 1188, 362, 382, Molyneux, Elizabeth M. 441 McHenry, Amy M. 1013 Merci, Claude 600 893 Molyneux, Malcolm 1433, 347, McInnis, Bronwyn 1019 Meremikwu, M. 1292, 300 Mills-Robertson, Felix 1101, 1118 348, 877, 1022 McKenzie, Ellis F. 802 Merkushina, Tatyana A. 298 Milner, Erin E. 646 Mombo-Ngoma, Ghyslain 499 McKeon, Sascha N. 733 Merola, Jonathan 1535 Milner, Jr., Danny A. 29, 353, Monaghan, Andrew 1115 McKerrow, James 320 Merritt, Ethan A. 643 1023, 1147, 360, 1433, 175 Monatrakul, Preeyaporn 153 McKibben, Elisabeth K. 114, 528, Meshnick, Steven R. 1382, 1487, Min, Duk-Young 780 Moncada, Jeanne 1085 1078, 1221, 1295 1490, 1491, 381, 688, 991, Minakawa, Noboru 1229, 1244 Moncayo, Abelardo C. 1245, 548 McKibben, Maxim J. 114, 528, 1103, 1169, 1179 Mingmongkolchai, Sirima 1522 Mondal, Dinesh 1064, 520 966, 1078, 1221, 1295 Mesirov, Jill 1023 Minh, Ngo N. Q. 238 Monestier, Marc 1039 McLaws, Mary-Louise 465 Messenger, Ali M. 1262, 1263 Minja, Happiness D. 379 Mongkolsirichaikul, D. 761, 761, McLean, Kyle 1432 Messer, William B. 1578 Mintz, Eric D. 1472, 17, 268, 565, 761 McLean, Kyle Jarrod 1604 Messina, Jane 1103 82, 957 Mongodin, Emmanuel F. 201 McLellan, Susan 958 Messina, Joseph P. 1448 Miotto, Olivo 1047, 1506 Montague, Mark 1155 McMahan, Lanakila 571 Mestra, Alberto 1134 Mir, Fatima 1110 Monteiro, Glória R. Góis. 972

www.astmh.org A-504 Author Index The number(s) following author name refers to the abstract number.

Monteiro, Robson 1283 Morrissey, Anne B. 435 Mukabayire, Odette 1408, 1408 Mutuku, Francis M. 737 Montes, Martin 1466 Morrow, Matthew P. 697 Mukaka, Mavuto 1231 Mutunga, James M. 431 Montgomery, Jacqui 1433, 348, Morse, Gene D. 1499 Mukasa, Susan M. 1081 Muvdi, Sandra 858 360 Mort, Molly 1576 Mukhopadhyay, Chiranjay 903 Muvdi Arenas, Sandra 323 Montgomery, Joel 937 Mortazavi, Susanne 673 Mukunzi, Silvanos 45, 765 Muwonge, Francis 25 Montgomery, Susan 1034, 1035 Mosca, Andrea 740 Muleba, Mbanga 195 Muyembe‐Tamfum, Jean‐Jacques Montoya, Humberto R. 309 Moses, Lina 1412 Mulele, Chikuli 1488 1010, 1321, 1309, 1318 Montoya, Magelda 1581, 819 Moses, Lina M. 111 Mulembakani, Prime M. 1010 Muzoora, Conrad 440 Moon, Robert 1598 Mosha, Franklin W. 221, 212 Mulenga, Musapa 1058 Mvalo, Tisungane 991 Moonasar, Devanand 1435, 668, Mosha, Jacklin 1439 Mulholland, Eddie 839 Mwakalinga, Victoria 421 680 Mosha, Jacklin F. 10 Mulindwa, Julius 339 Mwakitalu, Esther 1130 Moonen, Bruno 666 Mosher, Aryc 1571, 533 Muliyil, Jayaprakash 1299, 1302, Mwalimu, Stephen 916 Moonga, Hawela 986 Mosher, A. 612 256 Mwandagalirwa, Kashamuka 1103 Moono, Petros 388, 664 Mosquito, Susan 70 Muller, Ivo 691 Mwandama, Dyson 531, 905 Moore, Brioni R. 1496 Moss, Delynn 1072, 59, 610 Muller, Martin N. 1562 Mwangome, Francis K. 966 Moore, Christopher C. 284, 440 Moss, William J. 384, 385, 46, 763 Mullin, Kathleen 343 Mwania, Mercy S. 1244 Moore, Jason 398 Mostafa, Osama N. M. 68 Mulualem, Ayelegn 1570 Mwanza, Mercie 1438 Moore, Julie 916 Mott, Joshua A. 768 Mulunda, Caroline 243 Mwanza Ingwe, Mercy 1373 Moore, Michelle 1587 Motta, Amarilis 1306 Mulwa, Francis 413 Mwanzo, Isaac 215 Moore, Sarah J. 759, 398 Mouchet, François 214, 393 Mumba, Peter 1438, 434, 678, Mwatha, Joseph K. 1569 Moore, Sean R. 84 Mouhamadou, Chouaibou S. 920 922 Mwebaza, Norah 274 Moore, Vanessa M. 1478 Mounsey, Kate E. 814 Mumm, Lawrence 108 Mwesigwa, Gerald 497, 657 Moormann, Ann M. 163, 688, Mount, Howard T. 349 Mundal, Kirk D. 335 Mwingira, Upendo 1130 944 Mounts, Anthony 1421 Muneer, Akhtar 1361 Mwingira, Upendo John 1573 Mor, Siobhan M. 415 Mounts, Anthony W. 47 Mungai, Peter 1078, 114, 1145, Mwinzi, Pauline 1034, 1035, 1567 Mora-Pineda, Geovanny 748 Mourão, Marina d. 784 1221, 1295, 1391, 1596, 528, Myers, Bennett 696 Morais, Ádila L. 972 Moureau, Gregory 1276 500, 1380, 491, 737 Myint, Hla Y. 894, 367 Morales, Alba Lucia 614 Moussa, Dada W. 1104 Mungai, Peter L. Mzilahowa, Themba 1231, 531 Morales, Ingrid 20 Moustafa, Manal 1073 Mungall, Bruce A. 416 Morales-Betoulle, Maria E. 1342 Moustaki, Irini 549 Munir, Mah-e 1421 Moran, Elizabeth 1015 Moxon, Christopher A. 353 Munishi, O. Michael 435, 828 Morassutti, Alessandra L. 1139 Moyano, Luz M. 58, 88 Muniz, André Luiz 1409 N Moratz, Chantal 357 Moyo, Sikhulile 24 Munoz, Benito 136 Moreau, Katherine 1553 Moza, Patricia 1396 Munoz, Beatrice 917 N’Dong, Christelle 1516 Moree, Melinda 843 Mphande, Fingani 1433 Muñoz, Jorge 1335 N’Dri, Louis 920 Moreira, Antonio 306 Mpimbaza, Arthur 274 Muñoz-Jordan, Jorge 507, 978, N’Fale, Sagnon 197 Moreira, Luciano A. 724 Mpoudi-Ngole, Eitel 1562 1056, 1333 N’gonga, Daniel 806 Moreno, Marta 1027, 199 Mrema, Hedwiga 985 Munro, James B. 1177 N’Guessan, Raphael 231 Moreno, Silvia 1600 Msellem, Mwinyi I. 1161, 1310, Munster, Vincent 524 Nabasumba, Carolyn 454 Moreno, Yovany 479 1437, 673, 675 Muratova, Olga 702, 703 Nabity, Scott A. 546 Moreno-Fierros, Leticia 1222 Mshana, Christopher 1080 Murdoch, Michele E. 584 Nabiwemba, Elizabeth 25 Mores, Christopher N. 1346, Msoma, Aulive 1265 Muro, Florida J. 985 Nagajyothi, Fnu 63, 971 1347, 220, 227, 305, 586, 587, Msukwa, Martin 1265 Murphy, Darby 1459 Nagarkatti, Rana 857 732, 747 Msuya, Levina J. 969 Murphy, Jittawadee 176, 181, Nagendar, Pendem 853 Moretz, Samuel E. 685, 687 Mtasiwa, Deo 421 953, 99, 1219, 954 Nahum, Laila A. 784 Morgado, Fernanda N. 862 Mtove, George 917, 985 Murphy, Robert 1499 Naidoo, Inbarani 654 Morgan, David B. 1562 Mu, Jianbing 1214 Murphy, Ryan C. 643 Nair, M.K.C. 1116 Morgan, Juliette 1226 Mubikayi, Luamba 1488 Murugi, Julie 561 Nair, Meera G. 18 Morgan, Marjorie 805 Muchiri, Eric M. 1380, 491, 500, Muruve, Daniel A. 356 Nair, Shalini 1046, 1048, 1429 Morgan, William M. 445 737, 1145, 1391, 1596, 884 Musapa, Mulenga 434, 922 Najnin, Nusrat 568 Morissey, Anne B. 828 Muchiri, Geoffrey 1034 Musicha, Crispin 123 Nakasujja, Noeline 28 Moritz, Mark 259 Mucker, Eric 1423, 599, 1422 Musila, Lillian 413 Nakazawa, Stephen R. 643 Morkel, Cheri 98 Mudiz, Maria Luisa 1227 Musiychuk, Konstantin 703 Nakhasi, Hira L. 338, 975 Morlais, Isabelle 1453, 711 Muehlenbachs, Atis 454 Muskavitch, Marc A. T. 204, 430 Nalusaji, Aisha 454 Morrill, Westin H. 1170, 1173 Mueller, Ellen C. 1477, 539 Musoke, Charles 1592 Namagembe, Allen 274, 630A Morris, Carrie 1495 Mueller, Ivo 1, 1014, 1199, 1483, Musonda, Kasapo 385 Namamba, Jabir 1185 Morris, Glenn 1242 1496, 1559, 1560, 683, 690, Mussa, Abdul 668 Nambozi, Michael 1488, 1508 Morris, Hector 1559 904, 933 Musselwhite, Laura W. 838 Namekar, Madhuri 980 Morris, Paul 1127, 1128, 1477 Mueller, Natascha R. 643 Musset, Lise 1543 Namountougou, Moussa 432 Morris, Sheldon L. 1020, 247 Mues, Katherine 1122, 582 Mustata Wilson, Gabriela 459 Nana-Djeunga, Hugues Clotaire Morris, Ulrika 1161, 673 Muff, Katya 150 Musyimi, Robert 243 470 Morrisey, Joanne M. 1040 Mufti, Zubair 1110 Mutabingwa, Theonest 1508 Nanayakarra, Dhammika 367 Morrison, Amy C. 1235, 1259, Mugerwa, Robert 1081 Mutai, Beth K. 40 Nanayakkara, N. P. Dhammika 1348, 1351, 312, 509, 553, Mugo, Daisy 243 Mutapi, Francisca 529 1171, 1174, 650, 1166, 1167, 555, 591, 750, 511, 930, 1341, Muhindo, Rose 284 Mutebi, John-Paul 1246, 811 649, 995, 996 1253 Muhle, Michael 1189 Muth, Sinuon 1138 Nankabirwa, Joaniter I. 572 Morrison, Thomas E. 947 Muianga, Pedro 150 Mutiso, Agnes 243 Naquira, Cesar 1365 Morrison., Amy C. 516 Muise, Victoria 1132 Mutombo, Polydor N. 465 Náquira, César 457 Naranjo, Nelson J. 228 www.astmh.org Author Index A-505

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

Narasimhan, Vagheesh 360 Nemeth, Nicole 1005 Njie, Madi 875 Nwankwo, L. 612 Narayanan, Jothikumar 1525 Nemser, Bennett 561 Njiiri, James 45 Nwauche, Chijioke A. 1190 Nardone, Glenn 875 Nerurkar, Vivek R. 301, 603, 980, Njiokou, Flobert 470 Nyaaba, Gertrude 1114 Narrod, Clare 556 1105 Njiri, James 765 Nyakundi, Ruth K. 879 Narum, David L. 1, 1011, 163, Nery, Susana V. 840 Njomo, Doris W. 834 Nyangiri, Oscar A. 117, 672 1283, 1486, 701 Netongo, Palmer M. 629 Njoroge, Rose N. 439 Nyariki, Emily 1552 Nascimento, Ana B. 715 Neves, Joshua 186A Njozi, Mustafa H. 577, 536, 890 Nyarko, Edward 1090, 1098 Nascimento, Danielle O. 881 Nevolko, Oleg 1094 Nkhoma, Standwell 1046, 1048, Nyataya, Josphat N. 165 Nascimento, Eduardo 1345 Newbold, Chris 31 1429 Nyehangane, Dan 454 Nascimento, Marcia 1409 Newman, Ruchi M. 1458, 1457 Nkonwa, Innocent 618 Nygren, Benjamin 268, 957 Nascimento, Paulo R. P. 972 Newmark, Phillip A. 1002, 444 Nkrumah, Francis 1376, 1379 Nyirenda, Osward M. 1492, 900 Nascimento, Renata T. 1148, 1149 Newton, Charles R. 101 Nkwengulila, Gamba 494 Nyunt, Myaing M. 1367, 1526, Naser, Abu M. 266 Newton, Paul 1048 Nkwocha, Omeni 1234 1369 Nash, T. 57, 94 Newton, Samuel 1463 Nkwocha, O. 612 Nyuyfomo Tukov, Donatus 580 Nasr, Sussann 422, 530 Neyra, Victor 1200, 171 Noble, Agustina 1134 Nzila, Alexis 145 Nasreen, Sharifa 1421, 261, 948 Ng, Chiu W. 510 Noblecillas, Cesar 387 Nzunza, Rosemary 45 Nasrin, Dilruba 1077, 1319, 1472, Ng, Lee Ching 1443 Noblet, Raymond 605 17 Ng, Mary 1521 Noden, Bruce H. 98 Nassali, Mercy 1155 Ng, Muan Hong 651 Noedl, Harald 1047, 1290, 1506, Nasser, Julio R. 1055, 1467 Ng, Yih Yng 1051 1512, 908 O Nataro, James P. 1472, 17 Ngan, Chantha 596 Noesil, Elysee 1117 Nathan, Rose 1434, 426, 713 Ngando, Raphael 966 Noh, John 489, 59, 792, 97, 95 O’Brien, Connor 1501 Nato, Farida 1191 Nganou-Makamdop, Krystelle Noh, Mi Young 721 O’Brochta, David A. 1585 Naulikha, Jacqueline M. 1270 693, 682 Noisakran, Sansanee 979, 982 O’Connell, Kate 371 Naumova, Elena N. 415, 570 Ngasara, Billy 1185 Nolan, Thomas J. 1468, 999 O’Connor, Katherine A. 565 Nausch, Norman 529 Ngo Duc, Thang 635, 909 Nolen, Tracy 1376 O’Donoghue, Peter 169 Navarro-Lopez, Roberto 1416, Ngondi, Jeremiah 1570, 533 Noonpaksee, Wilai 808 O’Garra, A. 1606 1419 Ngonyani, Hassan 398 Norris, Douglas E. 735, 741 O’Guinn, Monica 1425 Navas, Adriana 325 Ngor, Pengby 674 Norris, Laura C. 741 O’Hanlon, Simon 669 Nayiga, Susan 274 Ngouateu K., Omer Bébé 121 Nosten, François 1046, 1047, O’Meara, Wendy P. 423, 576 Naz, Anila 267 Ngowi, Helena A. 92 1048, 1486, 1506, 1512, 1519, O’Neal, Seth 97 Naz, Abhinav 558 Nguete, Beata 934 1521, 1530, 1548, 1561, 453, O’Neil, Michael T. 638, 646, 993, Nazib, Forida 251 Ngugi, Anthony K. 101 454, 887, 901, 988, 997 1172, 132 Nchimbi, Happy 634, 902 Nguku, Patrick M. 378 Nosten, Francois Notario, Walter O’Neill, Brighid 1361 Ndaiye, Daoda 1159, 1160 Nguon, Chea 1109, 1162, 670 1523 O’Neill, Kerry 474 Ndao, Momar 337 Nguyen, Megan 14 Nou, Samon 762 O’Neill, Katelyn 726 Ndege, Samson 423 Nguyen, Vu 1011 Noulin, Florian 1520 O’Neill, Scott L. 552, 724, 729 Ndekhedehe, Eno 564 Nguyen, Vinh Chau 1337 Novelli, Jacopo 471 O’Reilly, Ciara Ndenga, Bryson A. 217 Nguyen Van, Hong 635 Noviyanti, Rintis 1517, 1528 O’Reilly, Ciara E. 17, 957, 1472 Ndiath, Mansour 382 Ngwa, Alfred A. 1209 Novotny, Joseph 666 O’Shea, Thomas 136 Ndiaye, Daouda 1044, 1207, Nhlema Simwaka, Bertha M. 1374 Nozaki, Tomoyoshi 251 Oakley, Miranda S. 1020 1431, 151, 175, 29, 632, 773 Niane, Fatou K. 893 Nsanzabana, Christian 144 Oaks, Edwin 71 Ndiaye, Jean Louis 1180, 170, Niangaly, Amadou 1512, 534 Nshala, A. 1573 Obala, Andrew 576 362, 382, 773, 893 Niangaly, Hamidou 1516 Ntadom, Godwin 378 Obeng-Adjei, George 156 Ndiaye, Jean L. 897, 895 Niare, Oumou 1184, 1584 Ntep, Marcelline 580 Obeng-Adjei, Nyamekye 697 Ndiaye, Magatte 1180, 170 Nichol, Stuart T. 1009, 1246, 945 Ntoumi, Francine 662 Oberstaller, Jenna 1525, 637 Ndiaye, Mouhamed 362, 382 Nicholls, Ruben Santiago 842 Ntumngia, Francis B. 1013, 33, Obonyo, Charles 1507 Ndiaye, Mouhamadou 773 Nicosia, Alfredo 176 1538 Oboth, Paul 630A Ndiaye, Maguatte 897 Nieto, Luisa M. 1092 Nuckois, John 730 Obrist, Brigit 1080 Ndiaye, Mouhamed 897 Nigussie, Kalemelekot N. 120 Nunes, Márcio R. T. 1349 Ocampo, Martha C. 1459 Ndiaye, Youssoupha 382 Nimmannitya, Suchitra 316 Nuñez, Andrea 588 Ocana-Mayorga, Sofia 1362, 1363 Ndir, Omar 1207, 151, 773 Nimo-Paintsil, Shirley C. 1090, Nuñez, Jorge 1541 Ochieng, Benjamin 17 Ndjamen, Blaise 64 1098 Nurleila, Siti 1558, 449 Ochieng, Cliff 1317 Ndjango, Jean-Bosco N. 1562 Nisalak, Ananda 1052, 1325, Nurmakhanov, Talgat I. 418 Ochieng, Orukani G. 585 Ndonky, Alphousseyni 427 1326, 1330, 1350, 1400, 316 Nurmukhambetova, Saule 1547 Ochoa, Theresa J. 19, 20, 85, 70 Ndonye, Janet 71 Nisbett, Richard A. 107 Nuruzzaman, Md. 568, 1266 Ochola, Elizabeth 1035 Ndour, C. T. 895 Niu, Tianchan 515 Nuthulaganti, Tej 1435, 668, 680 Ochomo, Eric O. 925 Ndour, Papa Alioune 1497 Nixon, Christina E. 1557 Nutman, Thomas B. 1069, 1070, Ockenhouse, Christian 1219, Ndula, Miranda 926 Nixon, Christian P. 1216, 1557 1072, 1125, 1468, 1476, 1478, 1547, 953, 99, 1546, 181, 954 Neafsey, Daniel E. 1018, 1207, Nizame, Fosiul A. 1266 1482, 23, 482, 483, 525, 607, Ockenhouse, Christopher 666 1209, 1431, 1213, 29, 430, 204 Nizame, Md. Fosiul A. 568 608, 1467, 1572, 539, 542 Odashima, Newton S. 1148, 1149 Nebié, Issa 155 Njau, Boniface N. 969 Nuwaha, Fred 102 Oden, Maria 967 Needham, Lisa 1178 Njau, Joseph D. 1111 Nwadike, Jones U. 564 Odermatt, Peter 1138, 796 Neesanant, Pimmnapar 1399 Njau, Ritha 1185 Nwagbara, Juliette I. 742 Odidi, Beatrice 296 Nejsum, Peter 1137 Njei, Basile M. 122 Nwakanma, Davis C. 1188, 204, Odukoya-Maije, Funke 1485 Nelson, Kenrad E. 940 Njei, Nelvis A. 122 1028 Odundo, Elizabeth 71 Nelson, Kara L. 569 Njenga, M. Kariuki 439 Nwane, Philippe 432 Oduntan, Helen A. 363 Nembunzu, Dolores 1112 Njenga, Sammy 1229, 1244 Nwankwo, Lawrence 1234 Oduola, Ayoade M. J. 452 Oduro, Abraham R. 1376, 1379 www.astmh.org A-506 Author Index The number(s) following author name refers to the abstract number.

Odusami, Temitope 1219 Oluput, Robert 76 Osuna, Finnley 765 Padilla, Angel M. 65 Offei Owusu, Irene 213, 606 Olveda, Remigio M. 1566 Oswald, Cate 1117 Padmanabha, Harish 554 Ofori, Micheal 1370, 156 Omalu, Innocent C. 1144, 128 Otchere, Joseph 1133, 1463 Padron-Nieves, Maritza 848 Ofori-Yeboah, Yaw 606 Omar, Rahila 673 Oteng, Eugene K. 31 Paes, Cheryl 310, 595 Ofula, Victor O. 462 Omar, Rahila S. 1161, 675 Otieno, Godfrey A. 1507 Pagano, Ian S. 2 Oghumu, Steve 66 Ombok, Maurice 1450, 925 Otieno, Lucas 1507 Page, Anne-Laure 1405 Ogola, Eric 41 Omollo, Raymond 318 Otieno, Michael 1194 Page, Kristen 76 Ogolla, Sidney 688 Omondi, Angela 146 Otieno, Walter 130, 699 Page, Wendy 839 Ogoussan, Kisito 580 Omore, Richard 17 Otolorin, Emmanuel 564 Page-Sharp, Madhu 1496, 366 Oguike, Mary C. 1158, 1190, 658 Omoregie, Philomena I. 262 Ototo, Edna N. 425 Pagès, Frédéric 427 Ogula, Godfrey 1091 Ondari, Ronald O. 1552 Otozi, Rita 1234, 612 Pagni, Sarah 1580 Ogulla, Godfrey 1194 Ondeng, Alex 17 Otsuki, Hitoshi 184 Pain, Arnab 1598 Oguttu, David 585 Ong’echa, John Michael 1091, Otsyula, Barasa 576 Paintain, Lucy S. 424 Ogutu, Bernhards 130, 1507, 1194, 1197, 1208, 157, 1507 Otsyula, Nekoye 628 Pakpour, Nazzy 192 1292 Ongoiba, Aissata 358, 689 Otten, Mac 1373 Palacio, Karen 1572, 833 Ogwal, Alex 630A Onguru, Daniel 1567 Ottesen, Eric A. 1070, 609 Palacios, Maximiliano 1055 Ogwang, Caroline A. 177 Onikpo, Faustin 1082 Ouafo, Jonas 581 Palma, Sandra 89 Oh, Helen 824 Ono, Takeshi 692 Ouangraoua, Soumeya A. S. 1093 Palma Urrutia, Pedro Pablo 1186 OhAinle, Molly 588, 818, 819 Onwuchekwa, Uma 112 Ouedraogo, Alphonse 154, 155, Pan, Bo 778 Ohrt, Colin 1166, 367, 640, 642, Onwujekwe, Dan 119 707, 709, 913 Pan, Li 854 647, 649, 650, 891, 892, 894, Onyango, Jacinta J. 435 Ouedraogo, Amathe 913 Pan, Xiaoling 1284, 1590 994, 998, 993 Onyango, Jecinta J. 828 Ouedraogo, Andre Lin 990, 155 Pan-Ngum, Wirichada 670 Ojaku, Alex 630A Onyango, Kevin O. 1507 Ouedraogo, Esperance 913 Panchalingham, Sandra 1472, 17 Ojengbede, Oladosu 1499 Onyia, Mgbodichi 1234 Ouedraogo, Issa N. 154 Pandharkar, Trupti 855 Ojo, Kayode K. 643 Onyia, M. 612 Ouédraogo, Jean-Bosco 1187, Pandya, Rajul 1114 Ojo, Tolulope A. 429, 428, 743 Ooi, Eng Eong 1443, 435, 828 1201 Panella, Amanda J. 942 Okatcha, Tunika I. 798 Ooi, Winnie W. 1311 Ouedraogo, Jean Bosco 1513 Pang, Junxiong V. 1582 Okebe, Joseph 875 Opara, Gift 1234, 612 Ouedraogo, J.B. 345 Paolino, Krisopher 953 Okedi, Loyce M. 1286, 800, 458 Opare, Christiana 1101 Ouedraogo, Jean Bosco 432, 901 Papastogiannidis, Petros 136 Okeke, Iruka N. 73, 69 Opi, Daniel H. 350 Ouédraogo, Nebie Issa 913 Pape, W. John 1261 Okell, Lucy Okell, Lucy C. 669, Opoka, Robert O. 28, 883 Ouma, Collins 1091, 1194, 1197, Parameswaran, Poornima 818 990, 424 Opoku-Manu, Jones 1370 1208, 1450, 925 Parashar D. B. 391 Okia, Michael 923, 753 Opondo, Vitalice O. 806 Ouma, Johnson 1286, 810 Parashar, Umesh D. 522 Okiro, Emelda 1107 Opot, Benjamin 45, 765 Oundo, Joseph 17 Paredes, A 57, 94 Okitolonda, Emile 1010, 934 Oraka, Chinedu O. 1084 Ouoba, Raïssa T. A. 601 Parent, Jason 1328 Oko, Lauren 947 Orang’o, Joshua M. 1552 Ouologuem, Dinkorma T. 777 Parikh, Sunil 1499, 370, 453, 899 Okorie, Patricia N. 802 Oravecz, Tamas 1592 Ovalle, Clemencia 858, 867 Parizo, Justin 666 Okoroafor, Iheanyichi 1383 Ord, Rosalynn L. 183 Ovalle Bracho, Clemencia 323 Park, Daniel J. 1207, 1209, 1431, Okoth, George 1473 Ordu, Donald A. 401 Overgaard, Hans J. 1030, 232, 928 175, 29, 882 Oksov, Yelena 541 Oreagba, Ibrahim A. 363 Owino, Simon 916 Park, Gregory S. 883 Oku, Hiroyuki 180 Oresanya, Olusola B. 1383 Owoaje, Eme T. 103 Park, Jae-Won 1533 Okuboyejo, Titilope M. 1493, 452 Oria, Prisca A. 296 Owuor, Boaz 178 Park, Kiwon 1202 Okudo, Charles 146 Oriá, Reinaldo B. 16, 74, 78, 84 Owusu, Ruth 1157, 1308 Parker, Robert 1590 Okumu, Fredros O. 759 Oriango, Robin 1451 Owusu-Agyei, Seth 1157, 1308, Parker, Zahra F. 1278 Oladosu, Oladipo O. 1395 Oriero, Eniyou C. 1188 1370, 1388, 174, 405, 698 Parra, Beatriz 1296 Olang, George 1252 Orillo, Beverly 592 Oxborough, Richard M. 221 Parra, Gabriel J. 1339 Olaya, Sandra 96 Orinda, G 1194 Oyegbami, Banji 271 Parra, Marcela 247 Oldfield, Eric 469 Orji, Bright C. 564 Oyibo, Wellington A. 1395, 1485 Parsons, Michele M. B. 75, 17 Oleinikov, Andrew 1021 Orozco, Susana 503, 589 Oyier, Beryl 1473 Partap, Uttara 1550 Olinger, Jr., Gene G. 1404, 1422, Orozco - Fernández, María del Oyinloye, Oladapo E. 1168 Partidos, Charalambos D. 1008, 599 Carmen 171 Oyugi, Jessica 1231, 1374, 1091 1401, 1411, 417 Oliva, Clelia F. 736 Orozco-Mosqueda, Guadalupe Ozaki, Masayo 1234, 1383 Partin, Kathryn M. 725 Olivarez, Nicholas P. 313 E. 772 Ozaki, M. 612 Partono, Felix 272 Oliveira, Aureo 1033 Orshan, Laor 918 Ozbun, Michelle A. 55 Partridge, Robert 1352 Oliveira, Edward 1033 Ortega, Corrie 1025 Ozekinci, Tuncer 253 Parveen, Shahana 261, 414 Oliveira, Fabiano 746 Ortega, Ynes R. 772, 776 Parzy, Daniel 1191 Oliveira, Guilherme 784 Ortega-López, Jaime 328 Pasvol, Geoffrey 1394, 906 Oliveira, Katia C. 446 Orutwa, Sarah N. 1552 Patel, Jaymin 1525, 989 Oliveira, Paulo 1409 Osborne, Lisa 18 P Patel, Kamala D. 356 Oliveira, Renato 1345 Osei-Kwakye, Kingsley 1157, Patel, Priyesh A. 838 Oliveira, Sergio C. 785 1308, 1370 Pa’au, Molisamoa 613 Patel, Sweta M. 72, 960, 963 Olkowski, Sandra M. 1341 Osman, Husham A. 304 Pablo, Jozelyn 770 Paterson, Yvonne 1150, 1151 Olliaro, Piero L. 898 Osorio, Jorge E. 1008, 1327, Pace, Douglas 1600 Pates Jamet, Helen V. 919, 920 Olorunda, Damilola C. 1484 1339, 1401, 1411, 1446, 1459, Pacheco, M. Andreina 1203, Patil, Anand P. 117, 672, 383 Olson, Christine K. 296 417 1542, 885 Patiño, L. 79 Olson, Kenneth E. 235, 237, 208 Osorio, Salua 554 Pacheco, Romina 1200 Patra, Kailash P. 1223 Olszewski, Kellen L. 1182, 1040 Ospanov, K. S. 936 Pacheco-Tucuch, Freddy 815 Patrick, Edouard 636 Oluoch, David 1291 Osuna, Finley 45 Pacífico, Lucila G. G. 785 Patrickson, John 164 Pack, Angela D. 65 www.astmh.org Author Index A-507

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

Patterson, Amy E. 1234, 1383, Pfaff, Jennifer 825 1321 Price, Ric N. 1528, 1534, 1561, 1571 Pfander, Claudia 643 Pitts, Sidney 164 455 Patterson, Janna C. 1320 Pfeffer, Martin 518 Pitzer, Virginia E. 1010 Prichard, Roger K. 470 Patterson, Noelle B. 186, 186A, Phaiphun, Lucy 901 Pizey, Tim 651 Priest, Jeffrey W. 1072, 610 181, 696, 954 Phakdeeraj, Aphiradee 1499 Placide, Mbala-Kingebeni 1309 Printz, Naomi 118, 1551 Patterson, William 440 Pham, Anh Khoa 964 Plante, Kenneth 1008, 1411, 519, Pritt, Bobbi S. 631 Paul, Rina 115 Pham, Jonathan 1599 1418 Privat-Maldonado, Angela 334 Paul, Repon C. 567 Phanse, Yashdeep 1593 Plaspohl, Sara 297 Proaño, Jefferson V. 91 Paulose, Abraham 1116 Phiboonbanakit, Danabhand 1402 Plewes, Katherine 970 Prom, Satharath 1511, 645, 998 Paulsen, Dave 1245 Philippe, Max 116 Ploemen, Ivo 185 Pronyk, Paul 561 Pavlin, Julie 762, 438 Phillips, Anna E. 448 Plowe, Christopher V. 1045, 1181, Prosser, Wendy 1227 Pavlinac, Patricia B. 1270 Phillips, Pamela L. 1419 1183, 1204, 1211, 1217, 534, Prue, Chai S. 1367, 1369, 911, Paz Soldan, Valerie 1235, 1351, Phiri, Ajib 441 6, 900, 952, 1047, 1506, 1512 1526 511, 509, 555 Phiri, Happy 347 Pochet, Nathalie 1023 Psutka, Rebecca 265 Pearce, Edward J. 999 Phiri, Kamija 1436 Podder, Goutam 414 Puddicombe, Babajide J. 271 Pearlman, Eric 250 Phiri, Masiliso 384 Podust, Larissa M. 863, 320 Puddicombe, Tolulope A. 271 Pearson, Richard 456 Phommasack, Bounlay 780 Poespoprodjo, Jr. 455 Puente, Dayyana J. 83 Peatey, Christopher L. 992 Phong, Nguyen C. 1502 Poespoprodjo, Jeanne R. 1534, Pukrittayakamee, Sasithon 997 Peck, Mireille 959 Phongluxa, Khampheng 796 1561 Punguyire, Damien 1157 Pedersen, Babill S. 626 Phothikasikorn, Jinrapa 808 Poggensee, Gabriele 1564, 378 Puplampu, Naiki 1090, 1098 Pedersen, Karl 1066 Phyo, Aung Pyae 1047, 1506, Pohl, Marie 1581, 816 Purcell, Robert H. 940 Pedro, João M. 840 1512, 1561 Poinsignon, Anne 322, 341, 427 Puri, Mahesh 1116 Peeling, Rosanna 1337 Piameu, Michael 432 Poirot, Eugenie 665 Purizaga, Percy 387 Peeples, Mark E. 416 Piazza, Franco 324 Poku-Asante, Kwaku 1157 Purohit, Vivek 137 Peeters, Martine 1427, 1562 Piccoli, Luca 61, 86 Polhemus, Mark 130, 181, 953 Purswani, Murli 294 Peiris, M. 761 Pichyangkul, S. 761 Pollack, Henry 1141 Pusey, Anne E. 1562 Pekow, Penelope 386 Pickering, Amy J. 1473 Pollett, S. 79 Pusic, Kae M. 1017 Peletz, Rachel 265 Pickering, Jason 1373 Polsomboon, Suppaluck 429, 743 Pusok, Robert 258 Pelleau, Stéphane 1191 Picot, Stephane 1184 Pombi, Marco 1028, 921 Pyae Phyo, Aung 1046, 1530 Pelletreau, Sonia 835 Piedrahita, Diana 1339, 1459 Ponce de Leon, Gabriel 1226 Pyaephy, Aung 997 Peniche, Alex 332 Piel, Alex 1562 Pond-tor, Sunthorn 1021, 1566 Pybus, Brandon 1172, 1510, 367, Penny, Melissa A. 1371 Piel, Frédéric B. 672, 117 Ponder, Elizabeth L. 843, 845 640 Pepin, Kimberly 39 Pierce, Kristen K. 1343, 1445 Pondja, Maria 1226 Pybus, Brandon S. 646, 891, 892, Pereira-Carvalho, Guilhermina A. Pierce, Raymond J. 468, 784 Ponlawat, Alongkot 1322, 1425 993 L. 653 Pierre, Pubien 1117 Pons, María Jesús 70 Perelygin, Andrey 1139 Pierre-Louis, Willythssa 360 Ponte-Sucre, Alicia 848 Perera, Liyanage P. 247 Piesman, Joseph 39 Pontororing, Gysje J. 1534 Perera, Samudrika 1071 Pietri, Annette 1096, 1417 Pool, Christopher D. 1219 Q Perez-Cruz, Luis 436 Pike, Andrew 715 Poorak, Mitra 1525, 637 Perez-Guerra, Carmen 1304 Pilingana, Portipher 559 Porcella, Stephen F. 358, 1047, Qadri, Firdausi 21, 550, 567, 72, Perez-Padilla, Janice 1304, 436 Pilitt, Kristina L. 1585 1506 960, 961, 963 Perkins, Douglas J. 1091, 1194, Pillai, Dylan R. 1067, 1164, 32, Porras, Alexandra 289 Qian, Menbao 778 1197, 1208, 1507 661, 851 Porter, Michael D. 1219 Quagraine, Josephine E. 1133, Perkins, Stuart D. 1088 Pilotte, Nils 610 Portugal, Silvia 358 1463 Perng, Guey Chuen 504, 979, 982 Pimenta, Denise N. 354 Porwollik, Steffen 21 Quang, Huynh H. 1502 Pernica, Jeffrey M. 1553 Pimentel, Guillermo 1098, 1301, Posomboon, Suppaluck 428 Quashie, Neils 655 Perretti, Alessandro 86 1303 Postels, Douglas 876 Quasie, Olga 1101 Perry, Robert 1294, 1504, 573 Pinapati, Richard S. 1182 Postigo, Jorge 856 Queiroz, Rafaella F. Grenfell. 278 Perry, Victor H. 1349 Pinçon, Claire 241 Potchen, Michael J. 1022 Quetz, Josiane S. 16, 74, 78, 80 Person, Bobbie 1035, 566 Pinder, Margaret 1188 Pothier, Pierre 1405 Quick, Robert 1265, 267, 565, Persson, Kristina E. M.. 683 Pindolia, Deepa 1368 Poulton, Ian D. 176 566, 959 Perumal, Kaliraj 1480 Pineda, V. 771 Póvoa, Marinete M. 1210, 199, Quijano-Hernández, Israel A. 328 Pesavento, Patricia 53 Pingle, Maneesh 1414 733 Quillery, Elsa 205 Pesko, Kendra 1457, 1458 Pinheiro, Marcos 456 Powell, Jeffrey R. 1248 Quintana, Fernando 387 Peter, Asaga M. 974 Pinho, Brunno G. 1349 Powers, Ann 1246, 1416, 417, Quintana, Miguel 1397 Petersen, Christine A. 1593, 343, Pinho, Rosa T. 319 811 Quiroz, Wendy L. 1235 865 Pinkerton, Relana 284, 440 Prabaripai, Atchariya 428, 809 Quispe, Victor R. 283 Peterson, A. Townsend 218 Pinot de Moira, Angela 1569 Prada, Ana 19, 85 Quispe Machaca, Victor R. 1365, Peterson, Allison 973 Pinsent, Amy 1394, 906 Pradines, Bruno 660, 661 1364, 457 Peterson, David S. 1525, 637 Pinto, Joao 1028, 1030, 1027 Prakash, Aishya 1122, 582, 1131 Petrarca, Vincenzo 1028, 1030 Pinto, Valerian 1015 Prasad, Abhishek N. 52 Petri, Jr., William A. 1065, 251, Piola, Patrice 1509, 454 Prasad, Neha 330 254, 257, 520, 1066, 46, 763, Pion, Sébastien D. 470 Prata, Mara M. G. 80 R 1064, 255 Piper, Ben K. 1270 Pratt, W.D. 1422 R. Vijayaraghavan 391 Petrucell, Chris 1160 Piriou, Erwan 688 Premakumara, W. D. Y. 1071 Raballah, Evans 1091, 1194, 1197, Petz, Lawrence 1425 Pisarcik, Sarah 1425 Premji, Zul 1185 1208 Petzold, Max 1310, 675 Pitt, Catherine 362 Price, Ric 1509 Racherla, Kishore K. 853 Peyton, David H. 1170, 1173 Pittman, Phillip R. 1309, 1318, Price, Richard 1517 Rachid Viana, Giselle 1428

www.astmh.org A-508 Author Index The number(s) following author name refers to the abstract number.

Rack, Ralph 1233 Rao, Ramakrishna U. 1071, 476, Reville, Patrick K. 66 Riu-Rovira, Therese 1321 Raczniak, Gregory A. 1098 1073 Reyburn, Hugh 985 Rivard, Robert G. 1088, 1301, Rada, Ouk 1109, 1162 Rao, Sowmya R. 292 Reyburn, Rita 962 1303 Radhi, Aamal 475 Rascoe, Lisa N. 1135 Reyes, Anatalio 954 Riveau, Gilles 1038, 158, 241 Radtke, Andrea 1059 Rashed, Selim 1553 Reyes, Lissette 1306, 249 Rivera, Aidsa 1333, 1335, 507, Radzioch, Danuta 66 Rashidul, Haque 1064 Reyes, Maria 87 544 Rahaingosoamamitiana, Corinne Rasmussen, Zeba A. 1076 Reyes, Sean 134 Rivera, Fulton P. 20 1089 Raso, Giovanna 912 Reyes, Sharina 954 Rivera, Irma 1335 Rahalison, Lila 1089 Rasoloharimanana, Lova 874 Reyes-Uribe, Patricia 334 Rivera, Pilarita T. 1529, 488 Rahelimalala, Marthe 1233 Raspone, Oroth 796 Reynolds, Joanna L. 621 Rivera-Dipini, Carmen 436 Rahman, Ahmed Shafiqur 242 Raswiswi, Eric 680 Reynolds, Mary G. 934 Riveron, Jacob 926 Rahman, Atiqur 550 Rath, Bruno 1364 Reznik, Sandra 359 Riveros, Maribel 70 Rahman, Mizanur 1475 Rathod, Pradipsinh K. 877 Rhee, Amy 703 Riyadh, Md. Asrafuzzaman 963, Rahman, Mahmudur 414, 43 Ratsimbasoa, Arsene 1430 Ribacke, Ulf 882 960 Rahman, Mustafizur 43, 948 Rausch, Kelly 1011, 1218 Ribeiro, Grazielle A. 317 Roald, Borghild 1564 Rahman, Mohammad Arif 72, 963 Ravel, Christophe 121 Ribeiro, Guilherme S. 546 Rob, Quick 110 Rahman, Mohammad A. 960 Raviprakash, Kanakatte 830 Ribeiro, Jose 1283 Robb, Katharine 1271 Rahman, Muhammad W. 266 Raviv, Yossef 830 Ribeiro, Paula 447, 789, 791 Robb, Merlin 22 Rahman, M. Z. 948 Rawiwan, Imerbsin 993 Ribeiro-Gomes, Flavia 539 Roberts, Donald 397 Rainwater Lovett, Kaitlin 502 Rawson, Ian 958 Rice, Bejamin 1542 Roberts, Jacquelin M. 774 Raj, Amrita 917 Ray, Debalina 467 Rice, Janet 764 Roberts, Willie 111 Raj, Dipak K. 1216 Ray, Prabhati 367, 640, 642 Richard, Stephanie A. 1076, 583 Robertson, Stephanie 320 Rajaiah Prabhu, Prince 1480 Raynaud, Sandrine 36 Richards, Allen L. 100, 1287, 40, Robinson, James E. 508 Rajan, Latha 764 Rayner, Julian C. 1427, 1562, 30 41 Robles-Murguia, Maricela 812 Rajasingham, Anangu 17 Raza, Afsheen 1531 Richards, Frank 1036, 1571, 463, Roca-Feltrer, Arantxa 1436, 574 Rajerison, Minoarisoa 1089 Razafimahefa, Julien 60 533, 846, 1234, 1383, 585, Rocchi, Vanna 1037 Rajski, Scott 1042 Razakandrainibe, Romy 60, 874 611, 612, 614 Rocha, Claudio 1348, 79 Rajwans, Nimerta 308, 817, 983 Read, Jonathan M. 906 Richards, Jean 1364 Rocha, Manoel Rakers, Lindsay 533 Reams, Melissa 297 Richards, Jack S. 1, 1014, 1486 Rocha, Manoel O. Costa 333, Rakotomanga, Avotiana 1233 Rebholz, Casey 958 Richards, Stephanie L. 236 1594 Rakotondrazaka, Mahenintsoa 60 Rebolledo, Paulina A. 420 Richards-Kortum, Rebecca 967 Rocheleau, Thomas A. 481 Rakotovao, Domohina 874 Rebour, Gilles 1233 Richardson, Jason 1281, 99 Rochford, Rosemary 367, 5, 647, Ralevski, Filip 1067, 851 Recker, Mario 686 Richardson, Jason H. 1322 649 Ralph, Anna P. 1534 Reddy, Michael R. 1030, 232, 396, Richardson, Jason R. 205 Rockett, K. 345 Ram, Malathi 1367, 1369 928 Richardus, Jan Hendrik 584 Rodda, L. 1605 Ram, Pavani Reddy, Sreenivasulu B. 683 Richie, Thomas 186A, 694, 953, Rodenhuis-Zijbert, Izabella 302, Ram, Pavani K. 1266, 1268, 1471, Reddy, Vamsi P. 232 99, 186, 1215, 1376, 181, 182, 501 1473, 1474, 1267, 1475, 248, Reed, Zachary 521 696, 954 Rodnak, Ditthakorn 314 261, 296, 46, 763 Reeder, John 1517 Richman, Adam 1217 Rodpradit, Prinyada 1325 Ramadan, Mohamed A. 782 Refae, Samir 77 Richter, David 1599 Rodrigo, Leighton G. 638 Ramadhani, Habib O. 969 Regev, Aviv 1023 Ricklefs, Stacy 1047, 1506 Rodrigue-Gervais, Ian-Gael 878 Ramal-Asayag, Cesar 1348 Regules, Jason 953 Rico, Alejandro 289 Rodrigues, Amabelia 1028, 204 Ramalho-Ortigao, Marcelo 804, Reich, Nicholas 1052 Ricotta, Emily E. 388 Rodrigues, Janneth 1025 812 Reid, C.R. 1422 Riddle, Mark 77 Rodrigues, Julio 306 Raman, Jaishree 150 Reid, Molly C. 643 Rider, Mark A. 730 Rodriguez, Alejandro 1464 Ramanathan, Roshan 1468 Reid, Michael J. C. 885 Riedel, Nadine 912 Rodriguez, Jairo 978 Ramasamy, Gowthaman 1199 Reiling, Linda 1, 1014, 690 Riehle, Michelle M. 1584 Rodriguez, Joseph A. 1418 Ramesh, Akshaya 482, 483, 739 Reimer, Lisa J. 1069, 1074, 933, Rietveld, Hans 372 Rodríguez, Lucy 1100 Ramineni, Bhanu 1214 1075, 1254, 739 Rigouin, Coraline 467 Rodriguez, Marilis A. 355, 183 Ramirez, Jose L. 1518, 716 Reintz, David M. 1008, 1411 Rijal, Suman 279 Rodríguez, Maria E. 81 Ramirez, Martin 1397 Reis, Mitermayer G. 1097, 498, Rijken, Marcus J. 1530, 1548, 901 Rodríguez, Rosa 1335 Ramirez, Ruth 1339, 1340 546, 799 Rikhsitullaeva, Shakhnoza A. 298 Rodriguez, Silvia 58, 88 Ramirez-Aguilar, Francisco J. 1416, Reis, Patricia A. 881 Riley, Eleanor 910 Rodríguez, Virginia C. 282 1419 Reisen, William K. 515, 1240, Rim, Han-Jong 780 Rodriguez-Acosta, Rosa 1304, Ramirez-Sierra, Maria Jesus 815 1455, 51 Rimoin, Anne W. 1010 1333, 544, 437 Ramos, Juliana P. 1349 Reiserova, Lucia 1328 Rinaldi, Gabriel 1000, 794, 795, Rodríguez-Barraquer, Isabel 1053, Ranade, Ranae M. 461, 853 Reiter, Karine 1011 798 1334, 1350, 502 Ranaivoharimina, Harilala 1233 Remich, Shon 130 Riner, Diana K. 67 Rodriguez-Madoz, Juan R. 981 Rancès, Edwige Remoue, Franck 157, 158, 214, Ringwald, P. 1047, 1506, 1512 Rodriguez-Morales, Ana L. 1222 Rances, Edwige 724, 729 241, 322, 341, 393, 427, 711 Rinn, John L. 882 Rodriguez-Sánchez, Luis M. 1222 Randrianarison, Mickael 60 Rendoo, Davaadorj 597 Rios, Dalila 614 Rodriquez, Silvia 59 Ranford-Cartwright, Lisa C. 191, Renggli, Sabine 713 Rios, Esther J. 1235 Rodulfo, Hectorina 1100, 81 345 Renia, Laurent 1519, 1521, 1530, Rios, Maria 1342, 514, 829 Roeffen, Will 703 Ranjalahy, Michel 1089 1548, 887 Rios, Zonia 312 Roehrig, John T. 604 Ranson, Hilary 433 Renslo, Adam R. 1176, 365, 320 Riscoe, Mike 1172 Roellig, Dawn M. 1358, 249 Rao, D.N. 1480 Restrepo, Berta N. 1339, 1340, Rissa, Youssouf A. 1104 Roels, Thierry 110, 566, 959 Rao, Donthamsetty Nageswara 978 Ritchie, Scott 821 Roestenberg, Meta 185, 693, 955 1545 Revan, Floyd 211 Rith, Sareth 762 Roger, Tine 895

www.astmh.org Author Index A-509

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

Rogerie, François 158 Rubens, Craig E. 1320 Saire-Mendoza, Mardeli 87 Sangare, Moussa Brema 1104 Rogers, Christine 386 Ruckle, Thomas 66 Saiyasombat, Rungrat 1004, 943 Sangare, Samba Adama 270 Rogerson, Stephen 1179, 1433, Rudra, Carole B. 248 Sakande, Souleymane 1187 Sanghvi, Tina 568 1483, 1490, 346, 690 Ruehlen, Nevada 1269 Sakhira, Sonia 1276 Sangoro, Peter 752 Rogier, Christophe 427 Ruiz, Joaquim 19, 20, 70, 85 Salahuddin, Gazi 568 Sanogo, Doh 270 Rojas, Diana 289 Ruiz, Marilyn O. 749 Salanti, Ali 161, 2 Sanogo, Dramane D. 607 Rojas, Laura Jimena 325 Ruiz-Lancheros, Elizabeth 479 Salas, Carola 387 Sanogo, Kassim 1045, 1183 Rojas, Winston 1340 Rukunga, Geoffrey 145 Salas Clavijo, Alejandra 856 Sanogo, Sintry 1305 Rojo-Domínguez, Arturo 860 Rulisa, Stephen 888 Salazaar, Ferdinand V. 743 Sanon, Antoine 322 Roldan, Aleida 1306 Rund, Samuel S. C. 1031 Salazar, Elsa 81 Sanon, Souleymane 913 Rollend, Lindsay 1062, 38 Rundell, Mark S. 1414 Salazar, Ferdinand V. 428 Sanprasert, Vivornpun 1478, 1125 Roller, Ladislav 1061 Rush, Amy C. 478 Saldana, A. 1355 Santalla, Jose 856 Rollin, Pierre E. 416, 521, 414, 945 Russell, Bruce 1519, 1521, 1530, Saldaña, A 771 Santamaria, Cynthia 337 Rollins, Sean 21, 961 1548, 887 Saleh, Maya 878 Santana, Rodrigo C. 1148, 1149 Rolon, Jenniffer 747 Russell, Tanya 757 Salerno, Stacy 1437 Santangelo, Joseph D. 1401, Romani, Franco R. 1099 Ruth, Laird 1317 Sales, Carla P. 354 1446, 1327 Romero, Carolina 1396 Rutstein, Shea 571 Salgado, Doris 1296, 978 Santiago, Cláudio R. Wanderley. Rommel, Christian 66 Rutta, Acleus S. M. 681 Salgueiro, Patrícia 1027 972 Romo, Hannah E. 1420 Ruttvisutinunt, Wiriya 1515 Salihu, Hamisu 1538 Santiago, Gilberto A. 1056 Rook, Kimberly 1268 Rwakimari, John Bosco 753, 923 Salim, Agus 1582 Santiago, Helton C. 539 Roos, David S. 777 Ryan, Elizabeth 1407 Salim, Abdul Hamid 242 Santiago, Jose I. 1269 Roper, Cally 654 Ryan, Eileen 638 Salje, Henrik 1350, 316, 46, 763 Santiago, Luis M. 1333, 437, 507 Roque, Jr., Vito G. 315 Ryan, Edward T. 21, 292, 72, 960, Sall, Amadou Alpha 547, 1247 Santivanez, Saul J. 87, 96 Rosário, Virgílio E. do. 653 961, 963 Sallau, Adamu 1383 Santolamazza, Federica 1027 Rosario-Casablanca, Rafael 436 Ryan, Ray 38 Sallusto, Federica 590 Santos, Antonio C. 1148 Rose, Joan B. 569 Salman, Sam 1496 Santos, Cleiton S. 1097 Rosen, David M. 29 Salmier, Arielle 1543 Santos, Silvane B. 1409 Rosenblatt, Robin A. 1037 Salnot, Virginie 3 Sanz, Crickette M. 1562 Rosenthal, Philip J. 1049, 129, S Salotra, Poonam 975 Sanz, Laura 368, 373 1392, 142, 144, 653, 899, Saly, Kong 645, 998 Sanza, Megan 913 1201, 901 Sa, Juliana M. 1044 Salzer, Johanna 1068 Sao Kry, Nong 12 Rosero, Doris A. 228, 229 Sabat, Greg 442 Sam El, Buth 762 Sapchookul, Nuch 1509 Rosowski, Emily 1605 Sabeti, Pardis C. 1207, 1431, 29, Samai, Mohamed 844 Sapozhnikov, V. I. 418 Ross, Amanda 1371, 904 550, 882, 1407, 1412 Samaké, Yacouba 534 Sapthu, Sunny E.E.N. 499 Ross, Allen G. 496 Sabindy, Samira 1226 Samal, Babru 358 Saraceni, Valeria 1154 Rossi, Carmine 113, 1313 Sabitu, Kabir 378 Samb, Badara 158, 224 Saralamba, Sompob 670 Rossi, Shannan L. 1420 Saborio, Saira 44, 588 Sambol, Nancy 453 Saravia, Nancy 325, 858 Rossignol, Marie 711 Sabot, Kate 1265 Sambou, Bakary 1504 Saravu, Kavitha 903 Rossmann, Michael G. 1441 Sacarlal, Jahit 1213 Sambunny, Uk 1211 Sardesai, Niranjan Y. 697 Rota, Paul A. 416, 414 Sachs, Sonia 561 Sambuta, Athanas 92 Sarkar, Anasuya 66 Roth, Joseph 613 Sack, David A. 1367, 1369, 1526, Samdi, Lazarus M. 1144, 128 Sarkar, Rajiv 1299, 256, 570 Rothman, Alan L. 1322, 1326, 978 708, 735, 911 Same Ekobo, Albert Le Grand 121 Sarpong, Bernard K. 1118 Rothstein, Yarrow 367 Sack, R. Bradley 1076 Samie, Amidou 775 Sarr, Jean Biram 158, 241 Rouamba, Noël 1187, 1201, 1513, Sackey, Sammy 1133 Samon, Nou 1511, 645, 998 Sarr, Ousmane 29 901 Sackey, Samuel O. 606 Sampath, Aruna 367, 650, 894 Sarr, Salemata 623 Roush, William R. 863 Sacko, Madjou 1584 Samuel, Darling 931 Sartono, Erliyani 1465, 272 Rout, Jonathan 1122, 1131, 582 Sacko, Moussa 464, 465, 632, Samuel, Dadzie K. 931 Sathipar, Reshma 620 Routh, Janell 110, 1265, 565 1568 Samuel, Prasanna 1302 Satimai, Wichai 1204, 140, 314 Rowcliffe, Kerryn 169 Sadi, Johari Y. 681 Samuels, Aaron M. 1034, 1572, Satoskar, Abhay R. 330, 66, 973, Rowe, Alexander K. 1082, 1293 Sadowski, Brett W. 638 833 854 Rowe, Christopher G. 700, 701, Saeij, J. 1605 San Martin, Jose L. 309 Sattabongkot, Jetsumon 1214, 1011 Saganda, Wilbrod 435, 828 Sanabria, Darío 1335 1536, 1546, 184, 953 Rowe, Dawne K. 1441 Sagar, Sangeetha L. 1447 Sanchez, Dimarys 1336 Satyanarayana, Neha 1030 Rowell, Candia 497 Sagara, Issaka 1045, 1183, 1184, Sánchez, Jennifer 807 Sauerbrey, Mauricio 614 Rowland, Meghan 548 1218, 1305, 702 Sanchez, Juan F. 387 Sauerwein, Robert W. 693, 703, Rowland, Mark W. 221 Sagatova, M. E. 418 Sánchez-Casco, Myriam 328 955, 185, 682, 956 Rowland, Tobin 1278 Sagerman, Dawn D. 1266 Sanchez-Vargas, Irma 235, 237 Saunders, David 1506, 1511, Rowlands, K. 345 Saggioro, Fabiano P. 1149 Sandeman, Richard M. 814 1512, 367, 645, 650, 1047, Roy, Chad J. 1410 Sagna, Andre Barambey 158 Sanders, John W. 77 1212, 761, 762, 953, 998, Roy, Sharon 110 Sagnon, N’Fale 1584, 394 Sandhu, Amonpreet 1553 1165, 1515 Roy, Smriti 1471 Sago, Laïla 3 Sandiford, Simone L. 715 Savage, Amy F. 861, 976 Roy, Sumitro 568 Saha, Debasish 1077, 1319, 1472 Sandison, Taylor G. 453, 535 Savage, Timothy J. 1329 For the RTS,S Clinical Trials Sahiner, Fatih 602 Sandoval, Carlos 1464 Sawe, Fredrick 22 Investigators 698 Sahu, Bikash 1020 Sanford, Michelle R. 1029 Saxton-Shaw, Kali D. 811 For the RTS,S Clinical Trials Sahu, Rajnish R. 1171, 1166, Sang, Rosemary 1278, 1586, 413 Sayasone, Somphou 796 Partnership 699, 951, 950 1167, 321, 995 Sangare, Check Oumar Papa 1516 Saye, Renion 632 Rúa-Uribe, Guillermo L. 228 Saijo, Masayuki 1426 Sangare, Ibrahim 1454 Sayo, R. 181 Rubahika, Denis 422 Sailaubekuly, R. 418 Sangaré, Laura 1270 Sayson, Stephanie 832 Saingam, Piyaporn 762 www.astmh.org A-510 Author Index The number(s) following author name refers to the abstract number.

Sazzad, Hossain M. S.. 414 Sea, Darapiseth 1511, 645, 998 Shafique, Muhammad 714 Shih, Chen-Chang 517 Scalmanini, Jenna 1473 Sebeny, Peter 77 Shah, Melisa 1474 Shikani, Henry 359 Scandale, Ivan 849 Seckova, Silvia 619 Shah, Naman K. 381 Shikuku, Kibet 22 Scanfeld, Daniel 1189, 1501 Secor, W. Evan 1035, 490, 1034, Shah, Pankhil 754 Shimokawa, Chikako 1064 Scaraffia, Patricia Y. 718 1565 Shah, Syed Akram 1361 Shimp, Jr., Richard 1011, 701 Scarsi, Kimberly 1499 Seda, Hilda 1304 Shaheen, Hind I. 77 Shin, Chang-Sik 1495, 648 Schacht, Anne-Marie 1038, 158, Sedai, Tika R. 598 Shahinas, Dea 1164, 32 Shinondo, Cecilia J. 434 241 Sedegah, Martha 181, 182, 186, Shahnewaz, Md 1319 Shiqin, Howe 435, 828 Schachter, Julius 1085 186A, 696, 954 Shaia, C.I. 1422 Shirley, Rhett 766 Schachterle, Stephen E. 917 Seder, Robert Shakarishvili, Roman 1303 Shlyakhter, Ilya 29, 550 Schaecher, Kurt E. 1212, 1165, Seder, Robert A. 1215, 956, 99 Shakely, Deler 1161, 1310, 1392, Shoemaker, Trevor 945 1515 Segeja, Method D. 389, 681 673, 675 Shongo, Robert 934 Schaffner, Stephen F. 1209, 29 Seguins, Wellington S. 319 Shakya, Geeta 598 Shott, Joseph P. 1305 Schär, Fabian 1138 Segura, Mariela 479 Shalu, M.V. 173 Showalter, Christopher 1460 Schaub, Günter A. 344 Seibou, Michele 1293 Shamblin, J. 1422 Shrestha, Binob 438 Schelinder, Shannon 1269 Seidahmed, Osama 304 Shamblin, Joshua 1423, 599 Shrestha, Bhola R. 1399 Schellenberg, David 382 Seidenberg, Phil 559 Shamloul, Moneim 703 Shrestha, Murari M. 598 Schendle, Robert 843 Seifert, Stephanie N. 1050 Shanakar, Hari 291 Shrestha, Sanjaya K. 1399, 438 Scherer, Philipp E. 971 Seila, Suon 1538 Shandukani, Mbavhalelo 1435 Shun, Tong Ying 459 Schieffelin, John S. 508 Seixas, Magda O. 1097 Shane, Hillary 65 Siakaala, Chadwick 678 Schiehser, Guy A. 139, 141 Sejvar, James 82 Shanks, G. Dennis 244 Sialeeze, Kazungu 559 Schifferli, Jürg A. 793 Sekaran, Shamala Devi 1337 Shanks, George D. 139, 141, 1502 Siam, Hanna A. M. 304 Schilaci, Michael A. 885 Selemani, Majige 536, 577, 633, Shao, Hongguang 999 Siame, Mwiche 679 Schilkey, Faye 52 890 Shapiro, Roger L. 24 Siangla, Joram 157 Schiller, Hilmar 1298, 288 Selvarajah, Suganya 595 Shapiro, Sheryl 1549 Siavi’I, Dottie 610 Schlaudecker, Elizabeth P. 42 Sem, Rithy 1204 Sharakhov, Igor V. 1026, 1589 Siba, Peter M. 1, 1014, 1069, Schlichting, Carl D. 733 Sembène, Mbacké 427 Sharakhova, Maria V. 1589 1074, 1559, 691, 739, 933, Schlienger, R. 426 Sembuche, Samwel 681 Sharker, M. Abu Yushuf 1267, 1199, 1254, 1483, 1496, 15, Schloegel, Jesse L. 1013, 33, 1538 Semenya, Amma A. 1565 1421 1560, 690 Schluter, William 598 Semnani, Roshanak Tolouei 1125, Sharlow, Elizabeth 459, 852 Sibiry, Samake 746 Schmaedick, Mark 610 1478 Sharma, Jabin 23 Sibley, Carol 1503 Schmid, Michael 589 Semrau, Katherine 559 Sharma, Narinder K. 804, 812 Sickerman, Stephen 803 Schmidt-Chanasit, Jonas 939 Semuneguse, Paulos 1387 Sharma, Satish 703 Siddique, Abdullah 254 Schmiedel, Yvonne 499 Senay, Gabriel 1255, 1387 Sharma, Subodh 90 Siddique, Abu B. 562 Schnabel, David C. 765, 146, 45 Senba, Masataka 1064 Sharma, S. K. 147 Sidibe, Bakary 1045, 1183 Schneider, Christof L. 793 Sene, Papa Diogoye 151, 773, Sharma, Suryakant K. 166 Sidick, Aboubacar 433 Schneider, Dominique 856 1431, 175 Sharma, Surya K. 381 Siduda, Sam 630A Schneider, Petra 628 Sener, Kenan 602 Sharma, Vijay L. 166 Siefert, Stephanie N. 1588 Schoepflin, Sonja 1560, 904 Seng-Duong, Lina 619 Sharma, Yagya D 148 Sierra, Gloria M. 501, 302 Scholzen, Anja 693, 955 Senghor, Simon 158 Sharmeen, Nusrat 21 Siethoff, Christoph 997 Schönian, Gabriele 1356, 340 Senn, Nicholas 691 Sharp, Paul M. 1427, 1562 Sihuincha, Moises 1253, 1348, Schountz, Tony 342, 946 Sensalire, Simon 1081 Sharp, Tyler M. 1333, 544, 1335 312, 509, 511, 553, 555, 1341 Schriefer, Albert 864 Seregin, Sergey S. 1016 Sharpe, Arlene 973 Sikaala, Chadwick H. 744, 757 Schriefer, Martin E. 1087 Serghides, Lena 7 Sharrock, Wesley W. 152, 907 Sikder, Shegufta 115 Schubach, Armando O. 862 Serrano, Adelfa E. 135, 136 Shastry, Ananthakrishna 903 Sikulu, Maggy 557 Schuck, Peter 1283 Serrano, Javier 1335 Shaukat, Ayesha 900 Silengo, Shawn J. 1446 Schuldt, Nathan J. T. 1016 Serre, David 1254, 1430 Shaum, Andrea 619 Silk, Benjamin J. 248 Schulze, Alexander 1434, 1080 Sesay, Santigie 1129 Shaw, George M. 1562 Silpapojakul, Kachornsak 1400 Schulze, Kerry 940 Seth, Misago 681 Shaw, Joseph 599 Silva, Cíntia M. Gonçalves. 785 Schwab, Kellogg J. 1264, 1470 Setiabudi, Rianto 449 Shaw, Katharina S. 1562 Silva, Joana 1047, 1506, 1512, Schwartz, Alanna 1437 Seto, Edmund 1437 Shaw, Rachel 1160 1177 Schwartzman, Kevin 113, 1313, Seto, Karen 1238 Shaw Saliba, Kathryn 1432, 1604 Silva, Thiago Luiz A. 861 1316 Settinayake, Sunil 1071 Sheath, Prameet 7 Silva, Virginia P. M. 456 Schwarz, Alexandra 1277, 1364, Seveau, Stephanie 66, 66 Sheehy, Susanne H. 176 Silva-do-Nascimento, Teresa F. 199 344 Seveno, Martial 711 Sheikh, Alaullah 21, 961 Silva-Moraes, Vanessa 1033 Sciotti, Richard J. 646, 852, 459 Severo, Norma P. Franco. 246 Shek, Lynette 824 Silver, Karlee L. 349 Scobie, Heather 1369 Severson, David W. 1589, 202 Shellam, Geoffrey 1332 Silvia, Claudia 886 Scott, Anthony 243 Sevilla Martir, Javier F. 1108 Shen, Ben 1042 Sim, B. Kim Lee 1217, 693 Scott, Jaclyn C. 1446 Sevilleja, Jesus E. A. D. 775 Shepard, Donald S. 1054, 510 Sim, Cheolho 722 Scott, Paul 22 Seydel, Karl B. 1022, 360, 877, Sherrill-Mix, Scott A. 1427 Sim, Kheng 12 Scott, Thomas W. 1050, 1253, 1023, 1195, 347 Sheu, Sheng-Hsiung 307 Sim, Kim Lee 1215 1259, 1322, 1326, 1348, 205, Seye, Mouhamadane M. 493 Shi, Lirong 917 Sim, Shuzhen 716 312, 425, 509, 511, 512, 553, Seymour, Robert 1008, 1410, Shi, Ya Ping 916 Simard, Frederic 1027, 432 555, 750, 1341 1411, 417, 1420 Shibata, Sayaka 461 Simasathien, Sriluck 1402 Scully, Corinne 1404 Seyoum, Aklilu 434, 744, 757 Shield, Jenny 839 Simbauni, Jemimah A. 217 Se, Youry 1047, 1165, 1212, Shackelford, Jacob 1230 Shields, Timothy 1369, 1526 Simen, Birgitte 818 1506, 1511, 1512, 1515, 645, Shaffer, D. 1343 Shiferaw, Zekarias 705 Simmons, Cameron 1583 762, 998 Shaffer, Douglas 22, 71 Shiff, Clive 448, 664, 388 Simmons, Graham 595

www.astmh.org Author Index A-511

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

Simms, Benjamin T. 1142, 1463 Smith, Scott A. 1578 Souleymane, Abdoul A. 1104 Stewart, Fiona 1562 Simpson, Andrew J. H. 1088 Smith, Stephen C. 195 Souleymane, Sanon 167 Stijnberg, Deborah 1152 Simpson, Julie 1486 Smith, Thomas A. 1371, 1388, Soumaoro, Lamine 1476, 607, 608 Stiles, Jonathan 164 Simpson, Julie A. 1561 1560, 904 Soumare, Harouna M. 402 Stinchcomb, Dan T. 1008, 1327, Simpson, Steven 776 Smith, Valerie 13, 906 Sousa, Jason C. 1510, 891, 892, 1401, 1411, 1446, 417 Sims, Jennifer S. 882 Smith Paintain, Lucy 1381 993, 367, 640 Stoddard, Steven T. 1253, 1259, Simwinga, Sandie 267 Smithers, Hannah 192 Sousa, Rosana 864 1348, 312, 509, 555, 1341, 511 Sinden, Robert E. 1220, 176 . Sousa-Figueiredo, José Carlos 464, Stoermer, Kristina A. 947 Sinei, Samuel 22 Smits, Hermelijn H. 1465, 499 497, 1137, 657, 840 Stohard, John R. 840 Singa, Benson 1270 Smucker, Joanne 1355 Souza, Anselmo 1409 Stolk, Wilma A. 584 Singh, Balbir 351 Smythe, Lee D. 545 Souza, Samaly S. 498, 799 Stoller, Nicole E. 1085 Singh, Neeloo 850 Sneed, Kevin 1242 Souza-Neto, Jayme 1058 Stone, Leslie M. 725 Singh, Rupa R. 90 Snider, Cynthia B. 520 Sovann, Ly 762 Stoney, Rhett 292 Singh, Sanjay 33 Snider, Cynthia J. 688 Sovannaroth, Siv 674 Storms, Aaron D. 47 Singh, Shiv S. 148 Snounou, Georges 1521, 1530, Sow, Cheikh 158 Stossel, Andrea 599 Singhasivanon, Pratap 1561, 901 1548, 887 Sow, Doudou 897 Stothard, Russell 1137, 497, 657, Sinha, Sarita K. 279 Snow, Robert W. 1107, 1503 Sow, Samba O. 112, 1472 464 Sintasath, David 674, 714 Snowden, Karen F. 91 Sowunmi, Akintunde 1493, 452 Stott, Blake R. 255 Siqueira, Isadora 1409 Snyder, Gregory 1584 Spaccapelo, Roberta 1219, 684 Stoute, José A. 357, 1355 Sirima, Sodiomon B. 154, 155, Soares, Alberto M. 16, 74, 78, 80 Speare, Richard 839 Stoyanov, Cristina 99 913, 707 Soares, Luciane Marieta 1097 Speck, Stephanie 518, 597 Stoyanov, Danail 1440 Siripaitoon, Pisud 1400 Soares, Renata O. A. 319 Spitzer, Eric D. 1414 Strand, Michael R. 719 Siripokasupkul, Raveewan 1511, Soares Magalhães, Ricardo J. 1575 Spring, Michele D. 953, 181, 628, Streckfuss, Judith 1298, 288 645, 650 Sobsey, Mark 571 954 Stresman, Gillian H. 664 Sissoko, Ibrahim M. 746 Sobti, Ranbir C. 166 Sridaran, Sankar 1428 Strickman, Daniel A. 1289 Sissoko, Mahamadou 9 Socheat, Duong 1046, 1047, Srinivasamoorthy, Ganesh 1525, Stridiron, Andrew 1017 Sissoko, Mahamadou S. 1218, 952 1048, 1165, 12, 1204, 1211, 637 Stroika, Steven 75 Sissoko, Seydou 112 1212, 1506, 1511, 1512, 1515, Srinivasan, Ramalingam 23 Stubbs, Janine 1014 Sisson, Scott A. 465 1538, 645, 670, 714, 998 Sriprawat, Kanlaya 1519, 1521, Studenski, Liora 918 Siu, Edwin 1178 Socías, Eugenia 1572, 1467 1548 Su, Xin-Zhuan 1214, 1047, 1506, Sivarathinasamy, Prabhu 1299, Socías, María E. 833 Sriwichai, Sabaithip 1212, 1511, 652 256 Sodahlon, Yao 580, 583 645, 998 Suarez-Davalos, Maria V. 1363 Siwo, Geoffrey H. 1182 Sodiomon, Sirima B. 167 Ssekabira, Umaru B. 630A Suarez-Ognio, Luis 591 Skarbinski, Jacek 1181, 1231, 531, Soebiyanto, Radina P. 768 Ssekitoleko, Richard 284 Suchdev, Parminder S. 1317 536, 633, 665, 905 Soenarto, Yati 1534 Sserwanga, Asadu 422, 530 Sudoyo, Hera 449 Skelly, Chris 545 Sognikin, Edmond 1574 St. Laurent, Brandyce 1249, 1451 Suen, Shih-Hui 1331 Skerlj, Renato 136 Sohel, Badrul Munir 1268 St. Louis, Mackinley 622 Suffrin, Dmitri 1106 Skinner, Danielle 794 Sohn, Woon-Mok 780 Staedke, Sarah 274, 572 Sugiarto, Paulus 1534, 455 Skinner-Adams, Tina S. 625 Soisson, Lorraine 952, 954, 181 Stancil, Jeffrey D. 553 Sugiharto, Victor 754 Skovmand, Ole 198 Sokhna, Cheikh S. 893, 362 Stanford, Donald 1167, 367, 649, Sugimoto, Jonathan 562 Skrypnyk, Artem 1094 Sokolova, Jaroslava 116, 617, 618, 995 Suh, Kathryn N. 964 Skrypnyk, Valerii 1094 619 Stanisic, Danielle I. 1483, 691, Sukhbaatar, Munkhzul 592 Skyes, Alma 985 Sokolow, Susanne 493 1199, 690 Sukowati, Supratman 1249 Sladeckova, Veronika 617 Solanki, Nehal R. 1020 Stapleford, Kenneth A. 1007 Sulaiman, Irshad M. 776 Sladek, Robert 878 Solomon, Anthony W. 549 Stark, Lillian 803 Sullivan, David 1169, 1367, 1369, Slatko, Barton E. 471 Solorzano, Nelson 929 Starzengruber, Peter 1047, 1290, 1526, 708, 911, 917, 735 Sled, John G. 349 Soltis, Bryony 1309, 1318 1506, 1512, 908 Sullivan, JoAnn S. 1565 Slotman, Michel A. 1030, 396, Soma, Viju 1320 Stauber, Christine 571 Sullivan, Kevin M. 1317 232, 928 Somda, Martin B. 322, 341 Stauffer, William 97 Sultana, Nadira K. 1475 Slovak, Mirko 36 Some, Fabrice 1513, 901 Stear, Mike 494 Suma, T.K. 1124 Slutsker, Laurence 665 Somers, Marie-Andree 561 Stebbings, Kevin 1232 Sumarto, Wajiyo 1557 Small, Dylan S. 457 Sompwe, Eric Mukomen 1186 Steel, Argon 1344, 592 Sumba, Peter O. 163 Smartt, Chelsea T. 236 Sonawane, Sharvari A. 186, 186A Steel, Cathy 1070 Sunahara, Toshihiko 1229 Smit, Jolanda M. 501, 302 Sonenshine, Daniel 1287 Stefanakis, Rianna 845 Sundar, Shyam 331 Smit, Johannes W. A. 272 Song, Langzhou 1328 Stefaniak, Maureen 186A Sundararaman, Sesh A. 1427 Smith, Allysa 1600 Soni, Minty 98 Steiner, Kevin L. 1145 Sung, Jui-Min 984 Smith, Alison 531 Sonnie, Mustapha 844 Steinhardt, Laura 1082, 422 Sunil, Parikh 901 Smith, Bryan 130, 1511, 998 Sonye, George 1229, 1244 Steinhoff, Mark C. 419 Supali, Taniawati 1465, 272 Smith, David 1332 Sosa-Bibiano, Erika 342 Steit, Thomas 1072 Supargiyono, Supargiyono 11 Smith, Darlyne B. 95 Sosthenes, Marcia C. K. 1349 Steketee, Rick 1374, 1377 Sur, Dipika 1319, 1472, 962 Smith, David L. 1368 Sotelo, Evelyn 20 Stepniewska, Kasia 1043, 1044, Surasri, Sittidech 1165, 1212, Smith, Joseph D. 161, 1486 Sotir, Mark J. 292 1046, 1047, 1509, 1512, 651, 1515 Smith, Joshua D. 211 Soto, Veronica 1544 901 Suryadevara, Praveen K. 853 Smith, Jeffery J. 1204 Soto-Gomez, Eunice 1304 Sternberg, Jeremy M. 344 Suryani, Dewi 1332 Smith, Linda C. 193 Soukhathammavong, Phonepasong Steurer, Francis J. 847 Sutanto, Inge 449 Smith, Nathan 423, 576 796 Stevenson, Jennifer 1451 Sutherland, Colin J. 1188, 1439, Smith, Nathaniel 766 Soukou, Koffi B. G. 392 Stevenson, Mary M. 479 656, 657, 10, 1158, 1190, Smith, Ryan C. 1059 Soulama, Issiaka 154 Stewart, Anna 1353 1361, 149, 1500, 658

www.astmh.org A-512 Author Index The number(s) following author name refers to the abstract number.

Sutherland, Laura J. 1078, 1221, Tan, Xiaolin 4 Temyarasilp, Pipatpong 303 Tipmontree, Rungrawee 1236 1380, 500, 944, 491 Tanajura, Davi 1409 Tendongfor, Nicholas 581 Tisch, Daniel J. 1069, 1074, 1075 Suttiprapa, Sutas 1000, 794, 795 Tang, Linhua 375 Tenneth, Dalipanda 307 Titus, Apangu 1115 Sutton, Patrick L. 1389, 1393, 886 Tang, Yuee 1527 Teopipithaporn, Suriya 1511, 998 Tjahjono, Bagus 449 Suvada, Jozef 618 Tangpukdee, Noppadon 1156 ter Kuile, Feiko O. 1490, 1491, Tjauw, Samantha Lie 1600 Suwanarusk, Rossarin 1521, 1530, Taniuchi, Mami 1064 1179, 1489 Tjitra, E. 455 1548, 887 Tanner, Marcel 1371, 421, 956 ter Meulen, Jan H. 1447 Tobin, Ellis 1314, 1315 Suwannachote, Nanatawan 734 Tannich, Egbert 251, 255 Terheggen, Ulrich 691 Todd, Charles W. 847 Suwonkerd, Wannapa 429, 734 Tanowitz, Herbert B. 63, 971, 359 Terlouw, Dianne 1436 Todd, Jim 1107 Suy, Channe 674 Tanwar, Vineeta 1545 Ternier, Ralph 1117 Toet, Hayley M. 814 Swaminathan, Soumya 23 Tanya, Vincent N. 1001 Teshh, Robert 1416 Togo, Amadou 1184 Swierczewski, Brett E. 71 Tanyuksel, Mehmet 253 Tetteh, John K. A. 156 Toh, Cheng-Hock 353 Swoboda, Paul 1047, 1290, 1506, Tao, Zhi-Yong 1524, 172 Tewari, Kavita 99 Tolnay, Airn 1005 1512, 908 Tapia, L. L. 1541, 387 Thaisomboonsuk, Butsaya K. 1326, Tolo, Youssouf 534 Sy, Ngayo 204 Tapia, Milagritos 112, 270 1330, 1330, 1400 Tolosa, Michel 400 Syafruddin, Din 1249, 1558, 1557 Tappero, Jordan W. 535, 110, 565, Tham, Wai-Hong 187, 1014 Tomashek, Kay M. 1304, 1333, Sybertz, Edmund 135, 136 566, 959 Thangamani, Saravanan 1239 436, 437, 507, 544, 1335 Syed, Najam R. 701, 700 Taramona, Claudia 87 Thanh, Nguyen X. 1502 Tomayao, Agnes D. 315 Sylla, Khadim 897 Targonska, Irena J. 93 Tharp, Marla 444 Tomchaney, Michael 202 Sylla, Massamba 1587 Tarleton, Rick L. 1595, 526, 65 Thathireddy, Anuradha 1563 Tomedi, Angelo 104 Sylla, Mamadou B. 112 Tarmizi, Siti 383 Thaver, Ali 1531 Tong, Marcus 666 Sztein, Marcelo B. 6 Tarning, Joel 277, 453, 901 Thavrin, Bou Kheng 714 Tong, Ping Ping 1401 Szumlas, Daniel 402 Tarongka, Nandao 1559 Thayer, Leahh 630A Tong Rios, Carlos 1540 Tarr, Cheryl 75 Thein, Tun-Linn 1324, 1323, 826 Tonkin, Michelle 1597 Tarragona, Sonia 1572 Thellier, Marc 661 Torero, Maximo 96 Tatem, Andrew J. 1368 Thera, Mahamadou A. 1045, Torii, Motomi 1, 184 T Tatipaka, Hari Babu 853 1183, 534, 952 Torr, Stephen J. 1360 Tatishvili, Nana 1303 Theron, Michel 30 Torre, Sabrina 878 Tabachnick, Walter J. 236 Tavares, Margarete 1396 Thesing, Phillip C. 1492, 900 Torrealba, Frannerys 81 Tabernero, Patricia 1109, 1162 Tavul, Livingstone 1014, 690, 691 Thiam, Sylla 1504, 573 Torrero, Marina 1127 Tack, Danielle M. 934 Tayag, Enrique A. 315 Thibaud, Martin 432 Torres, Carolina 229 Tadesse, Zerihun 1570 Taylor, Diane W. 161, 2 Thiemann, Tara C. 1240 Torres, Jomil 1335 Tafes, Hailemariam 318 Taylor, Ethel 268 Thien, Heidrun V. 251 Torres, José 1335 Tagbor, Harry 1508 Taylor, Myra 260, 620 Thior, Moussa 1294, 1504, 573 Torres, Katherine 1200, 171 Tagelsir, N. 345 Taylor, Mark 634 Thiptara, Anyarat 406 Torres, Katherine J. 162 Tagoe, Janice 1098, 1090 Taylor, Robert 449 Thitithayanont, A. 761 Torres, Melissa 485 Taiwo, Babafemi 1499 Taylor, Steven 1179, 1490, 1491 Thomas, S. 761 Torres, Vladimir 357 Taka, Nchang 122 Taylor, Terrie 1023, 1147, 1195, Thomas, Stephen J. 1322, 1326 Torrie, Leah S. 474 Takáč, Peter 1061, 810 126, 1433, 175, 347, 348, 353, Thome, Tamara 1352 Tosh, Donna M. 1546, 1547, 953 Takagi, Aki 1199 877, 1022, 360, 900 Thompson, Andrew 849 Totrov, Maxim M. 431 Takahashi, Daniele 546 Tchiengoua Tchogang, Nina 106 Thompson, Jennifer K. 1, 184, 187 Totten, Daniel C. 209 Takala Harrison, Shannon 1047, Tchioffo Tsapi, Majoline 711 Thompson, Michelle 674 Toubali, Emily H. 844 1506, 1512 Tchuem Tchuenté, Louis-Albert Thomsen, Edward K. 1069, 1074, Touch, Sok 762 Takasaki, Tomohiko 1426 836 933, 1075, 739 Tounkara, Anatole 106 Takayanagui, Osvaldo M. 1148, Tchum, Kofi 1308 Thomson, Andrew 1401 Toure, Brahima 1405 1149 Teelen, Karina 693 Thomson, Cynthia 1327, 1401, Toure, Ousmane 1516 Takeo, Satoru 1, 183, 184 Teepruksa, Phanthaneeya 100 1446 Toure, Sekou 1045, 1183, 1184 Takhampunya, Ratree 1522 Teferi, Tesfaye 1570 Thomson, Rebecca 634, 902 Touré Ndouo, Fousseyni S. 873 Takoubo, Simplice 1293 Teirlinck, Anne C. 693, 955 Thongsripong, Panpiim 592, 1258 Towner, Jonathan 1246, 945 Talaat, Kawsar R. 23 Teixeira, Avelino 1329 Thonkukiatkul, Amporn 184 Townes, David 905 Talbott, Kendra T. 697 Teixeira, Andrea C. 1594 Thorp, Jr., John M. 1487 Townes, John 97 Taldone, Tony 32, 474 Teixeira, Bertinellys 81 Thriemer, Kamala 1290 Townes, Lindsay R. 1372 Taleo, George K. 152, 907 Teja-Isavadharm, Paktiya 130, Thu, Ho Thi Viet 751 Tran, Michael V. 623 Talisuna, Ambrose O. 1503, 450, 1511, 645, 650 Thuma, Philip E. 384, 385, 679 Traore, Adama I. 1256, 225, 742 889 Tejada, Romina A. 1099 Thwing, Julie I. 1294, 1504, 573, Traore, Boubacar 358, 689 Talkington, Deborah 75 Tekete, Mamadou 1516 634 Traore, Bourama 746 Talledo, Michael 1424, 340 Tekete, M.M. 345 Tian, Haijun 1328 Traore, Idrissa 952 Tam, Mifong 479 Tekwani, Babu L. 1166, 1167, Tian, Yu 978 Traore, Karim 106, 534, 1044, 952 Taman, Amira 789 1171, 1174, 321, 995, 996, Tibenderana, James 1155, 450 Traoré, Mamadou 465 Tamarozzi, Francesca 61 367, 649, 650 Tilley, D. 79 Traore, Oumar Bila Tambat, Sejal 386 Telarolli Jr., Rodolpho 245, 246 Tilley, Leann 1049 Traore, Oumar B. 1045, 1183, Tamber, Gurdip 1067 Telfer, Sandra 1089 Timbine, Seydou 225, 226, 742, 1516 Tami, Adriana 302, 501 Telford III, Sam 37 1256 Traore, Sékou Fantamady 106, Tamminga, Cindy 181, 954 Tellez, Yolanda 588, 818 Timmermans, Ans 762, 761 702, 1104, 1584, 402, 607 Tamsma, Jouke T. 272 Tello, Luis 87 Timoshevskiy, Vladimir A. 1589 Traore, Zoumana Isaac 1516 Tan, Asako 1048, 1182, 1429 Tembo, Dumizulu L. 348, 1433 Tine, Roger 1180, 170, 773, 897 Trapaidze, Nino 1088 Tan, John C. 1048, 1429 Tembo, Mathias 267 Tinto, Halidou 1500, 1508 Trape, Jean-François 893 Tan, Kathrine R. 1438, 1488, 195 Temperley, William H. 117 Tiono, Alfred 154, 707, 709, 913 Travassos, Mark A. 534 Tan, Kevin S. W. 887 www.astmh.org Author Index A-513

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

Travassos Da Rosa, Amelia 1416 Tylleskär, Thorkild 102 Van der Vliet, Diane 822 Verlinde, Christophe L. 461, 853 Traylor, Zachary P. 1415 Tyner, Stuart 1506, 1511, 1512, Van Geertruyden, Jean-Pierre 1155 Verma, Aman 1385 Treadwell, Cynthia 579 645, 762, 998, 1047, 1212, van Gemert, Geert-Jan 185, 682 Verma, Saguna 603 Treasure, Carolyn L. 838 1165, 1515, 438 van Gengelen, Jeffrey 76 Vermeire, Jon J. 1142 Treat, Benjamin 186A Van Gessel, Yvonne 649 Vernick, Kenneth D. 1584 Trees, Alexander J. 1001 van Giersbergen, Paul 997 Verret, Wendy 535, 453 Trefethen, Jared 703 Van Kerkhove, Maria 47 Versteeg, Krista 1420 Trefney, M. 1355 U van Mourik, Jan 347 Vertus, Claude 618 Tremblay, Meg 195 Van Nguyen, Hong 909 Verweij, Jaco J. 272 Tren, Richard 397 Ubalee, Ratawan 1522 van Olphen, Alberto 1242 Vestergaard, Lasse S. 621 Triampo, Wannapong 407 Ubalijoro, Eliane 1132 Van Sickels, Nicholas J. 958 Vial, Thomas 506 Triana, Paula R. 501, 302 Uboldi, Alex D. 29 Van Tyne, Daria 1431, 151, 175, Vicarromero, Stalin 312 Trianty, Leily 1517 Udhayakumar, Venkatachalam 29, 882 Vicente, José L. 1027, 1028 Tribble, David R. 286 1428, 1525, 637 Van Voorhis, Wesley C. 643 Victor, Asoala 931 Tricou, Vianney 1583 Udomgsangpetch, Rachanee 1536 Van Weyenbergh, Johan 1424 Vicuña, Yosselin 937 Triglia, Tony 187 Ugaddan, George 1523 van Widenfelt, Erik 24 Vidal, Silvia 878 Trimarsanto, Hidayat 1517 Ulrich, Peter 1169 Van Wynsberghe, Nicole R. 342 Videa, Elsa 44 Trimpe, Lori 1269 Umar, Abba 401 Vanachayangkul, Pattaraporn Videira, Claudia F. 840 Trollfors, Birger 1310 Umar, Mary 1383 1511, 645, 650 Vidigal, Ana Claudia 1396 Trongnipatt, Namtip 1522 Umbers, Alexandra J. 1483 Vanden Eng, Jodi 1226 vieira de Abreu, Adriana 880, 881 Trop, Stefanie 206 Umlauf, Scott 1328 Vander Kelen, Patrick 803 Viera, Sara 136 Trout Fryxell, Rebecca T. 1588 Undurraga, Eduardo 1054 Vanlandingham, Dana 49, 519, Vignuzzi, Marco 1007, 1403 Troye-Blomberg, Marita 155 Undurraga, Eduardo A. 510 730 Viladpai-nguen, S.J. 1486 Troyo, Adriana 748 Unicomb, Leanne 1267, 1471, Vanloubbeeck, Yannick F. 1546, Vilar, Fernando C. 1148, 1149 Trueba, Gabriel 1406 263, 266, 568 1547, 953 Vilcarromero, Stalin 1341, 1348, Trung, Trieu N. 1502 Unnasch, Thomas R. 1003, 233, Vannberg, Fredrik O. 767 509, 591 Tsai, Hung-Chin 285 541, 1242, 605, 803 Vareta, Jimmy 175 Villacis, Anita G. 1363, 1366 Tsai, Jih-Jin 1579, 307, 504 Unoba, Dickson 611 Varga, Laszlo 1008 Villacorte, Elena A. 1529, 488 Tsai, Wen-Yang 1579 Upreti, Shyam R. 598 Varma, Sudhir 1468 Villalpando, Carlos 1055, 1467, Tsai, Yen-Chun 307 Urban, Joseph 1140 Vasconcellos, Erica C. 862 1572, 833 Tsamo, Etienne 129 Urban, Joyce 403 Vasconcelos, Pedro F. C. 1349 Villamizar, Paloma 747 Tsanava, Shota 1088 Urban, Joe 539 Vasilakis, Nikolaos 591, 311 Villani, Fernanda 333 Tsang, Victor C. 489, 58, 59 Urbina, Julio A. 859 Vasquez, Carlos 954 Villanueva-Lizama, Liliana 342 Tsao, Kim 1062 Usmani-Brown, Sahar 38 Vaughan, Jefferson A. 480 Villar, Luis A. 1296 Tse, Sze-Wah 203 Utami, Retno A. Setya. 1517 Vaz-Nery, Susana 464 Villar-Centeno, Luis A. 308, 817, Tsertsvadze, Tengiz 1301, 1303 Utesena, Martina 116 Vazquez, Alfredo Dominguez 614 983 Tsetsarkin, Konstantin A. 1006, Uthaimongkol, N. Vazquez, Susana 1337 Villasis, Elizabeth 1210, 162 311, 519, 591 Uthaimongkol, Nichapat 762, 761 Vázquez-Chagoyán, Juan C. 328 Villegas, Leopoldo 1525, 1542 Tshefu, Antoinette K. 1487, 1292, Utzinger, Jürg 912 Vazquez-Pimentel, Neftaly 1419 Villinger, Francois 979 300 Uwimana, Zena 1408 Vazquez-Prokopec, Gonzalo M. Vinetz, Joseph M. 1210, 1223, Tsiamis, George 810 Uyeki, Timothy 1421 511, 555, 740, 509 1540, 162 Tsuboi, Takafumi 1537, 183, 184, Uyoga, Sophie M. 352 Veerman, J. L. 584 Vineyard, Meighan 51 1, 1486 Uzalili, Veronica 1231 Vega, Jorge 1518 Vinjé, Jan 937 Tsuyuoka, Reiko 12 Vega, Marcia R. 978 Virender, Dua K. 391 Tu, Elmer Y. 250 Veiga, Antonio 306 Virnig, Beth 1119 Tu, Zhijian 1026 Vekemans, Johan 1546, 1547, Visvesvara, Govinda 250 Tubbesing, Ulf 98 V 952, 953 Vitte, Joana 1191 Tukahebwa, Edridah 1569 Vaidya, Akhil B. 1040 Veland, Nicolas 324 Vivas, Daniel 302, 501 Tukesiga, Ephraim 585, 611 Vaillant, Michel 898 Velasco, Juan C. 1359 Vivek, Rosario 949 Tuladhar, Rashmi 973 Valanparambil, Rajesh 479 Velasco, John Mark S. 315 Viviani, Simonetta 822, 823 Tullo, Gregory 685, 687 Valderrama, Liliana 858 Velasco-Salas, Zoraida I. 302, 501 Vlachou, Dina 1453 Tumbu, Paul 618 Valdivia, Hugo O. 335 Velasquez, Daniel E. 249, 872 Vodanovic-Jankovic, Sanja 1042 Tumwebaze, Benon 454 Vale, Glyn A. 1360 Velayudhan, Raman 304 Volkman, Sarah K. 1018, 1207, Tumwine, Lynnette K. 1155 Valecha, N 147 Velazquez, Vylma 1096 1209, 1431, 29, 1213, 151, Tun, Linn T. 1443 Valeika, Steven 916 Velez, Ivan D. 1327, 1446 175, 882 Tungtaeng, Anchalee 649 Valencia, A 79 Velmurugan, Soundarapandian Volkova, Eugenia 1418, 591 Tungu, Patrick K. 221 Valencia, Braulio M. 324 1215 Volper, Esther M. 301, 980 Turell, Michael J. 1278, 1413, 35 Valentiner-Branth, Palle 1076 Venkatesan, Meera 1204, 1211, von Geldern, Tom 849 Turiel, Maíra C. P. 1349 Valenzuela, Jesus G. 1057, 1143, 654 von Seidlein, Lorenz 962 Turituri, Feula 769 344, 746 Venkateswaran, Neeraja 942 von Stebut, Esther 121 Turnsek, Maryann 75 Valerio, Laura 555 Ventocilla, Julio 1541 Vonghajack, Youthanavanh 796 Turyakira, Eleanor 454 Valim, Clarissa 1213, 151 Vera, Hubert 929 Vossbrinck, Charles R. 1458 Tussey, Lynda 1328 Vallance, Bruce 18 Verani, Jennifer 1306 Vosshall, Leslie B. 1586 Tutterrow, Yeung L. 161, 2 Van Boeckel, Thomas 1556 Verastegui, Manuela 1364, 872, Vounatsou, Penelope 1377, 912 Twahir, Hemed 243 van Cuong, Nguyen 751 89 Vujcic, Jelena 261 Twenhawfel, Nancy 599 Van den Broeck, Frederik 340 Verdonck, Kristien 1424 Vujcikova, Julia 617, 619 Twumasi, Miecks 1370 Van den Eede, Peter 1544, 635, Verjovski-Almeida, Sergio 443, Vulliez-Le Normand, Brigitte 1597 Tyagi, Prajesh K. 381 909 446 Vulule, John 1091, 1194, 1197,

www.astmh.org A-514 Author Index The number(s) following author name refers to the abstract number.

1208, 1450, 17, 916, 925, 1448 Wang, Zenglei 652 Werbovetz, Karl 855 Willsey, Graham G. 1456 Vuong, Chau 1510, 892 Wang, Zhisen 849 Were, Tom 1091, 1194, 1197, Wilschut, Jan C. 501, 302 Vyankandondera, Joseph 888 Wanga, Joyce 694 1208 Wilson, Anne 574 Vythilingam, Indra 635 Wangsamuda, Suradi 1557 Wesolowski, Amy 1368 Wilson, Danny W. 184, 691, 1014 Wanionek, Kimberli A. 1343, 1445 West, Jr., Keith P. 115, 562 Wilson, Michael 1463, 174 Wanja, Elizabeth W. 1278 Westby, Katie 1245 Wilson, Mark 539 Wanjala, Christine L. 376, 425 Wever, Claudia 447 Wilson, Mary 864 W Wanji, Samuel 581 Wewers, Mark 66 Wilson, Michael D. 1133 Wanzirah, Humphrey 422, 453, Weyrich, Andrew S. 880, 881 Wilson, Mary E. 1311, 456 Wacker, Mark A. 1182 535 Wheeler, Richard J. 1601 Wilson, Michael L. 1562 Wadegu, Meshack 45, 765 Ward, Brian 337 Wheeler, Sarah S. 1455, 51 Wilson, Mark L. 441, 553, 710, Wagman, Joseph 924 Ward, Danielle T. 1481 Whitacre, Caroline 66 1372 Wagner, Thor A. 1320 Ward, Honorine 1299, 256, 570 White, A C. 1466 Wilson, Shona 1569 Wahala, Wahala M. P. B. 1577, Ward, Sarah M. 1031 White, Graham B. 1289 Wilson, Tony 440 503 Ware, Lisa 953 White, Karen 849 Wimberly, Michael C. 1255, 1387, Wahid, Sobia 914 Warhurst, David C. 656, 658 White, Lisa J. 670 513 Wahl, Angela R. 6 Wariero, James 561 White, Mike 1242 Wimonwattrawatee, Theera 762 Wahlgren, Mats 683 Warnas, Randall 632 White, Michael T. 669, 910 Winch, Peter 1471 Wain, John 69 Warne, Thomas 268, 82 White, Nicholas 1046 Wini, Lyndes 361 Waitumbi, John N. 165, 40, 439, Warrell, David A. 1398, 965 White, N. 1047 Winner, Gregory T. 1230 376, 628 Warrell, Mary J. 1398 White, Nicholas J. 1043, 1561, Winter, Alan D. 537 Walcourt, Asikiya 1175 Wartel-Tram, Anh 822, 824 670, 970, 997, 1048, 1506, Winter, Rolf 1172 Walker, Edward D. 1448, 1450, Warter, Lucile 1548 1512, 901, 1066 Winters, Anna M. 1438 737, 755, 925 Wasley, Annemarie 1303 White, Sarah 123 Winters, Benjamin 1373, 1438, Walker, Edward L. 749 Wasmuth, James 7 Whitehead, John 123 678 Walker, Larry 1171, 1174, 367, Wassmer, Samuel C. New York. Whitehead, Stephen S. 1343, Wiria, Aprilianto E. 1465, 272 647, 649, 650, 894, 994, 1166, 353 1445, 593 Wirsiy, Peter 1227 1167, 995, 996 Wastling, Jonathan M. 1001 Whitehouse, Chris A. 1413 Wirth, Dyann F. 1018, 1207, 1209, Walker, Patrick G. T. 1489 Wasunna, Beatice 1107 Whitehurst, Nicole 1159, 1160 135, 136, 1431, 29, 1023, Walker, Todd W. 806 Watanaveeradej, Veerachai 1402 Whittaker, Maxine 361 1213, 151, 175, 882 Wallace, Derek 822 Waterman, Carrie 1242 Whittington, Jessica 1015 Withers, Mark R. 1321, 1309, Walsh, Douglas S. 1212, 146, Waters, Norman C. 146, 152, 169 Whitty, Christopher J. M. 1163, 1318 1511, 645, 761, 762, 998, Watson, Nora 913 985 Wittlin, Sergio 135, 136 1165, 1515, 364 Watts, Douglas 1247 Wiboon-ut, S. 761 Wojcik, Richard A. 315 Walsh, Frank 585, 611 Wearing, Helen J. 1348 Wichterman, Jennifer 1404 Woldearegai, Tamirat Gebru 1532 Walsh, Michael G. 1461 Weatherall, David J. 117 Wickramasinghe, Rajitha 1121 Wolf, Ashley R. 882 Walsh, Peter D. 1562 Weaver, Anne M. 296 Widdowson, Marc-Alain 1421, 47, Wolfe, Nathan D. 1010, 1562 Walson, Judd L. 1270 Weaver, Bruce 1328 768, 948 Wölfel, Roman 597 Walter, Tita N. 1132 Weaver, Scott C. 1006, 1008, Wiegand, Roger C. 1207, 1431, Wolkon, Adam 1226, 531, 905 Walter, V. 1292 1247, 1410, 1411, 1416, 1419, 29, 135, 136 Won, Kimberly 1035, 1073, 579, Walther, Michael 875 1420, 234, 311, 295, 417, 941 Wiersma, Jorien 693 610, 613 Walton, Shelley F. 814 Weaver, Scott S. 1418 Wierzba, Thomas F. 100 Wondji, Charles S. 224, 926 Walusiku, Batuke 195 Webb, Brett 946 Wijesinghe, Rushika S. 1121, 361 Wong, Dawn M. 431 Wamae, Annah W. 296 Weber, Ingrid B. 1087 Wikel, Stephen 1239 Wong, Jacklyn 1259 Wambua, Joshua 243 Webster, Jayne 1107, 1381, 424 Wilairatana, Polrat 1156 Wong, Paolo A. 1099 Wamburu, Kabura 41 Webster, Joanne P. 465 Wilcox, Bruce 1258 Wong, Sunny H. 767 Wammes, Linda J. 272, 1465 Weetman, David 430 Wilder-Smith, Annelies 824 Wongarunkochakorn, Saowaluk Wamwiri, Florence 810 Weger, James D. 1411, 1008, 417 Wilkerson, Richard C. 199 1212 Wan, Cheng 1032 Wei, Hans 367, 994 Wilkins, Patricia P. 1135, 489, 59, Wongstitwilairoong, Tippa 762 Wang, Andrew 1017 Wei, Wang 264, 787, 797 792, 1139, 58, 781, 97 Woodhall, Dana 835 Wang, Alice 571 Weigand, Roger 138 Wille-Reece, Ulrike 697 Woodrow, Charles J. 651 Wang, Bo 1537 Weil, Ana 269 Williams, Andrew 1220 Woods, Leslie 51 Wang, Eryu 1410, 1418, 295 Weil, Gary J. 1071, 476, 478, 540, Williams, Chris 1023 Woodward, Jimmy 52 Wang, Hongxing 134 543, 781 Williams, Daniel 1435, 680 Woolfit, Megan 729 Wang, Hongju 375 Weina, Peter 130 Williams, David L. 1002, 467, 469 Woolsey, Aaron M. 578 Wang, Huanyu 935 Weiner, David B. 697 Williams, Gail 62, 496 Worell, Caitlin 835 Wang, Mei-Hui 200 Weinrobe, Carolyn 958 Williams, Jeffrey F. 1269 Worges, Matt 1159 Wang, Penghua 49 Weinstein, Philip 545 Williams, Katherine L. 313, 503, Worrell, Caitlin M. 837 Wang, Ruobing 1196, 1199 Weir, William 537 590, 819 Worwa, Gabriella 1455 Wang, Sibao 1232 Weirather, Jason 456 Williams, Maya 516, 591 Wright, Chelsea 1287 Wang, Shu-Hui 307 Weiser, Matthew 991 Williams, Steven 485 Wright, Gavin J. 30 Wang, Tianyi 1345 Weiss, Brian L. 1275, 1288 Williams, Thomas N. 117 Wright, Jennifer 1060 Wang, Wei-Kung 1579 Weiss, Louis M. 63, 359 Williamson, Kim 1604 Wright, Linda L. 1010 Wang, Xiaohong 207 Weiss, Mitchell G. 841 Williamson, Kim C. 1432, 360, Wu, Bo 471 Wang, Xingrong 479 Weiss, Walter 99 4, 168 Wu, David 84 Wang, Xiao Ling 760 Wekesa, Dennis 146 Willis, Juanette 1460 Wu, Han-Chung 1579 Wang, Yuanyuan 1125 Weldegebreal, Teklu 318 Willis, Ruth N. 105 Wu, Lee S. F. 115 Wang, Yanyan 196 Welsh, Robin 485 Wills, Bridget 1583 Wu, Xiao-Jun 442 Wang, Zenlei 1505 www.astmh.org Author Index A-515

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

Wu, Yi-Chieh 1579 Yazdanbakhsh, Maria 1465, 272, Zeng, Qiang 132 Wu, Yimin 700, 701, 703, 1011, 499 Zerihun, Mulat 1570 1214, 1218 Ye, Kenny 1195 Zerlotini, Adhemar 784 Wu, Yineng 861 Yebakima, André 306 Zerpa, R. 79 Wu, Yukun 952, 1472 Yebakima, Andre 400 Zervos, Thomas M. 886 Wu-Hsieh, Betty 984 Yeboah-Antwi, Kojo 559 Zettel Nalen, Catherine 403 Wurapa, Eyako K. 1291, 439, Yeda, Redemptah 146 Zhan, Bin 1140, 1479, 1481 765, 45 Yee Sin, Leo 1443 Zhang, Baosen 935 Wutthithanachot, Wiboonsak 314 Yeka, Adoke 422, 889 Zhang, Hailin 935 Wylie, Blair 1492 Yelmene, Michele L. 1447 Zhang, Jing 131, 132, 133, 134 Wynn, Thomas 539 Yemweni, Anicet 1112 Zhang, Jian 196 Wysocki, Vicki H. 718 Yeo, Tsin W. 1534 Zhang, Peng 367, 640, 642 Wyss, Sarah 460 Yeung, Shunmay 1109, 1162, Zhang, Shuliu 1328 1163 Zhang, Veronica M. 169 Yifru, Sisay 318 Zhang, Wenbao 786 Yimin, Wu 702 Zhang, Yanling 1011 X Yin, Zhengliu 935 Zhang, Zhiwen 1095 Yingyuen, Kritsanai 1515 Zhao, Dazhi 359 Xayavong, Maniphet 1525 Yip, Jana 1224 Zhao, Qinying 137 Xi, Zhiyong 1284, 1590 Yiu, Ying Y. 1140 Zheng, Li-wen 1146 Xia, Ai 1026 Yixin, Ye H. 729 Zheng, Yu-huang 1146 Xia, Hui 1536, 172 Yockey, Brook M. 1087 Zhioua, Elyes 1276, 34 Xiao, Ningchuan 259 Yogeswaran, Parimalaranie 93 Zhong, Daibin 200 Xie, Lisa 131, 132, 133 Yohannes, Ruth 1576 Zhou, Guoqian 1146 Xie, Xiaohui 1260 Yoksan, Sutee 1337, 600 Zhou, Goufa 215, 1378, 157, 212, Ximenes, Maria F. F. 456 Yong, Tai-Soon 780 376 Xu, Hengyi 1017 Yongvanitchit, K. 761 Zhou, Hua-ying 1146 Xu, Shulin 541 Yoon, In-Kyu 1322, 1326, 1330, Zhou, Huayun 1527 1402, 315 Zhou, Hong 184 Yoshino, Timothy P. 442 Zhou, Jingling 346 You, Hong 466, 786 Zhou, Shuisen 375 Y Young, Emily E. 579 Zhou, Xiaonong 492 Yadao, Federico 1523 Young, Lauren E. 1018, 1431 Zhou, Yang 831 Yadava, Anjali 1546, 1547, 953 Young, Sylvia 77 Zhu, Daming 1011, 700, 701 Yagoure, Bilkissou 402 Yousef, Nasser 1480 Zhu, Guoding 1527 Yamaguchi, Yoko 692 Yousheng, Liang 264, 787, 797 Zhu, Jinsong 720 Yamasaki, Tsutomu 183, 184 Yp-Tcha, Marie-Michele 400 Zhu, Xiaohua 855 Yamins, Daniel 29 Yu, Jihnhee 1266, 1268, 1474 Zhuoma, Yangjin 375 Yamo, Emmanuel 1160 Yu, Sun N. 1085 Zielinski-Gutierrez, Emily 1115 Yan, Guiyun 1378, 157, 200, 207, Yu, Zhiguo 1042 Zimmer, Trevor 680 212, 215, 404, 712 Yudhaputri, Frilasita A. 1557 Zimmerman, Dunja 693 Yan, Jian 697 Yukich, Joshua O. 986 Zimmerman, Guy A. 880, 881 Yan, Li 453 Yumiseva, Cesar A. 1363 Zimmerman, Peter A. 1069, 1074, Yan, Lim Wen 828, 435 Yuntian, Xing 264 1230, 1254, 1380, 1430, 482, Yang, Amy 247 Yusibov, Vidadi 703 483, 739, 933, 1075, 1560 Yang, Chao-Fu 1331 Zimmermann, Stefanie 329 Yang, Dujuan 935 Ziniel, Peter D. 469 Yang, Fan 1589 Zinszer, Kate 1385 Yang, Guangxiao 1061 Z Ziperstein, Joshua 583 Yang, Kun 492 Zodaly, Noel 60 Zachariah, Finly 1312 Zody, Michael C. 588 Yang, Maria 697 Zaidi, Anita K. M. 1110, 1472 Yang, Shu Kun 760 Zoerhoff, Kathryn 844, 1576 Zaitsev, Eleina M. 1412 Zöller, Lothar 597 Yang, Yongkun 684 Zakhidova, Nodira A. 298 Yang, Yu Rong 760 Zollner, Gabriela 1281, 918 Zaki, Sherif R. 544 Zollo, Paul H. A. 129 Yang, Yu R. 62 Zaman, K. 242 Yang, Zhaoqing 652 Zompi, Simona 1581, 816 Zaman, Rashid U. 1421, 43, 948 Zongo, Issaka 1187, 1201, 1513, Yano, Kazuhiko 180 Zambrano, Betzana 309 Yao, Yun-hai 1146 345, 901 Zambrano, Julio 302, 501 Zorrilla, Victor O. 335, 1237 Yapar, Mehmet 602 Zamora, Javier 505 Yardley, Vanessa 1361 Zou, Xiaoyan 694 Zanatto, Maria P. 1148, 1149 Zouré, Honorat G. 584 Yaro, Alpha S. 1256, 225, 226, Zang Edou, Estelle S. 873 742 Zoya, John 1374, 905 Zanini, Graziela M. 641 Zubaidah, Siti 1249 Yaro, Jean Baptist 913 Zanis, Michael J. 1280 Yasuda, Chadwick Y. 100, 596 Zulu, Sipho 260 Zanmi Lasante Clinical Committee Zurita-Lagos, Alejandra P. 1362 Yasukawa, Kosuke 1119 1120 Yauri, Veronica 872 Zwang, Julien 1337 Zavala, Fidel 1591 Zyzak, Michael 694, 99 Yaw, Afrane 212 Zegers de Beyl, Celine 714 Yawson, Alexander E. 1027 Zeituni, Amir E. 687 www.astmh.org