TABLE OF CONTENT
Page 002 WELCOME NOTE
Page 005 ORAL PROGRAMME
Page 013 INVITED PLENARY SPEAKER BIOGRAPHIES & ABSTRACTS
Page 038 ORAL ABSTRACTS
Page 088 POSTER LIST
Page 093 POSTER ABSTRACTS
Page 158 LIST OF CONFERENCE PARTICIPANTS
Page 166 CONFERENCE SPONSORS
Page 168 CONFERENCE RELATED FUNCTIONS
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WELCOME NOTE
Page 2 of 170 Dear Delegate
As the Director of the University of Pretoria Institute for Sustainable Malaria Control (UP ISMC), one of the South African Medical Research Council (MRC) Collaborating Centres for Malaria Research, I would like to personally welcome each of you to the 2nd South African Malaria Research Conference, 2016. The conference follows on the successful, first conference hosted by the MRC in Durban in 2015. This second national gathering of the malaria community will again provide researchers the opportunity to showcase novel findings, innovation, ground breaking research and ongoing collaborative efforts towards malaria elimination.
With malaria elimination a foreseeable possibility in South Africa an urgent need exists for fundamental- and applied research and surveillance in malaria endemic areas to eliminate the disease through the use of an integrated management approach, including safer and sustainable alternative malaria control methods. A major hindrance in realising the goal of elimination in South Africa is the cross-border impact of malaria from our neighbouring countries. We are therefore very excited to welcome delegates from our neighbouring and other SADC countries as well as international guests doing research in these countries.
The conference will be co-hosted by the UP ISMC and the MRC Office of Malaria Research (MOMR). Over the three day-long conference scheduled from 31 July to 2 August 2016 you can look forward to 61 oral presentations, of which 12 are plenary presentations, and 67 posters focusing on all facets of malaria. This includes human interventions towards malaria control and elimination including malaria education; health promotion and surveillance; novel methods to control the parasite and block transmission; innovative and alternative vector control methods; the use of various technologies to predict potential outbreaks and also the impact of climate change (remote sensing) and even the use of mathematical modelling to determine the effectiveness of new control tools.
The conference will take place at the Auditorium at the University of Pretoria Groenkloof Campus in Pretoria, South Africa. The Groenkloof Campus is the home of the University of Pretoria’s Faculty of Education. The number of registered delegates expected to attend the conference is around 230, providing students the opportunity to share their malaria-related research and at the same time learn from experts in their respective fields. The conference will aim to save on paper usage and therefore the conference booklet will be available as an interactive document on the conference website for downloading. Delegates will receive a complimentary memory stick (flash drive) that will also contain the booklet and delegates will have the opportunity to add additional material including photos to their memory stick.
Some background information on the UP ISMC. The Institute is a fully integrated, multi-disciplinary, interdepartmental and interfaculty initiative. The aim of the Institute is to coordinate and promote collaborative research on safer and sustainable malaria control and management strategies, and to generate new knowledge and support new activities pertaining to safe malaria control in Africa through fundamental and applied research, supported by research collaboration with regional, national and international partners. The diverse group of researchers within the Institute encompasses all aspects of malaria with research clusters focusing on human health, parasite control and vector control. The establishment of the UP ISMC was approved by the University of Pretoria Senate on 2 June 2016. The Institute was 'upgraded' from the existing UP Centre for Sustainable Malaria Control (UP CSMC), and boasts five years of achievements reached. The UP ISMC is also the proud host of the SARChI Chair on Sustainable Malaria Control.
Page 3 of 170 I wish to thank all our sponsors in advance for contributing towards the conference. Without your contribution this conference would not have been possible. To the delegates participating either through an oral presentation or poster, thank you for sharing your research with all attending the conference. To all the attending delegates a word of thanks for your interest and valuable contribution. I believe that the conference will be informative and create a platform for networking. The conference tea/coffee breaks, lunches and, Cocktail and Gala functions will allow ample time to mingle and share ideas.
I trust you will enjoy the conference and the hospitality that the University of Pretoria has to offer.
Sincerely
Tiaan de Jager Director: UP Institute for Sustainable Malaria Control
Page 4 of 170
ORAL PROGRAMME
Page 5 of 170
PROGRAMME
DAY 1 | Sunday 31 July 2016 Time Event / Presentation title Speaker 09:30 - 11:30 Registration (Tea / Coffee) (posters to be put up) Tiaan de Jager 11:30 - 11:40 Welcome and opening address UP ISMC Lucille Blumberg 11:40 - 12:05 Setting the research agenda around malaria elimination NICD Towards malaria elimination in the MOSASWA (Mozambique, Patrick Moonasar 12:05 - 12:30 South Africa and Swaziland) region NDoH 12:30 - 13:30 Lunch (posters to be put up) Human and environmental aspects: Cross-border, regional control, health Session 1 promotion, malaria education and surveillance 13:30 - 17:00 Session Chairs: Tiaan de Jager (UP ISMC) & Patrick Moonasar (NDoH) Sunday Ukpe 13:30 - 13:50 Malaria Case Management in the context of Malaria Elimination: The role of the population at risk DoH Mpumalanga Leo Braack 13:50 - 14:10 Residual Malaria: Changed mind-sets to quell the seeds of failure UP ISMC Could a song about malaria prevention help protect children under Chad Anderson 14:10 - 15:00 five in Venda rural communities? UP ISMC 4 orals June Petersen Focus on malaria in education in South Africa UP ISMC 10 minutes talk, Eunice Misiani 2 minutes Q&A Malaria deaths: Meeting the 2015 near zero target in South Africa and/or NDoH discussion Targeting foci of transmission towards elimination of malaria in Natashia Morris South Africa MOMR / UP ISMC 15:00 - 15:30 Tea / Coffee (posters to be put up)
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The impact of malaria program interventions and climate change Lactatia Motsuku on mortality and morbidity rates between 2009 and 2016 in Limpopo Province: Preliminary findings NDoH / UP ISMC
Travel patterns and demographic characteristics of malaria cases, Deepa Pindolia the case study of Swaziland's active surveillance system, 2010- 2014 CHAI 15:30 - 17:00 Operational factors associated with increased detection of Khayelihle Bhangu 7 orals infections for reactive case detection (RACD) in Swaziland NMCP Swaziland
10 minutes talk, A study to identify high malaria risk groups and the associated risk Erastus Haindongo factors using a programmatic tool: Malaria elimination risk factor 2 minutes Q&A Uni Namibia and/or assessment discussion Imported cases, sporadic outbreaks and resurgence are the Kaka Mudambo highest threats to malaria elimination in the E8 region SARN - RBM The contribution of a sustainable volunteer base on cross-border Constance Njovu malaria elimination: An impact assessment FCBMI Facilitating factors and barriers to malaria research utilization for Chikondi Mwendera policy development in Malawi UP ISMC Cocktail welcoming
17:00 for 17:30 At conference venue Welcoming page 17:30 - 20:00 VIP guest speakers, entertainment, cocktail dinner and networking for programme
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DAY 2 | Monday 1 August 2016 Time Event / Presentation title Speaker 07:30 - 08:30 Registration (Tea / Coffee) Session 2 Biology and epidemiology of the malaria parasite and transmission blocking 08:30 - 12:15 Session Chairs: Karen Barnes (UCT) & Erick Strauss (SU) Lyn-Marie Birkholtz 08:30 - 08:50 Seeing the wood for the trees: Malaria parasite systems biology and how it aids antimalarial drug discovery UP ISMC Mutant Malaria Parasites: An Overview on their Genetics, Origins Jaishree Raman 08:50 - 09:10 and Spread NICD Andrew Leisewitz Comparing canine Babesia rossi with human falciparum malaria UP ISMC 09:10 - 10:15 Starving malaria: Targeting nutrient uptake in the sexual, Jandeli Niemand 5 orals transmissible stages of Plasmodium falciparum parasites UP ISMC Plasmodium falciparum gametocytes show stage-specific, Riëtte van Biljon 10 minutes talk, chemotype specific transcriptional response to drug perturbation UP ISMC 2 minutes Q&A Synthesis and biological evaluation of heteroaromatic inhibitors of Amanda Rousseau and/or Plasmodium falciparum WITS discussion A comparative, transcriptomic analysis of the effects of a lead Jessica Connacher clinical candidate on the asexual and gametocyte stages of Plasmodium falciparum parasites UP ISMC Tea / Coffee 10:15 - 10:45 Poster session (Odd numbered posters) Quantitative profiling of the Plasmodium falciparum parasite’s Nanika Coetzee histone posttranslational modification landscape during sexual UP ISMC differentiation Cross-reactivity of oxidant drugs against asexual and gametocyte Dina Coertzen stages in Plasmodium falciparum parasites UP ISMC
Heterogeneity of Plasmodium falciparum malaria in Matabeleland Gladwin Muchena 10:45 - 12:15 south: Analysis travel history reported by malaria cases reported 7 orals in routine surveillance MHCC
Assessing risk factors associated with Plasmodium falciparum Eunice Agubuzo 10 minutes talk, exposure using serological data in Bushbuckridge municipality, 2 minutes Q&A Mpumalanga Province, South Africa WRIM and/or discussion Prevalence of glucose-6-phosphate dehydrogenase (G6PD) Shehu Awandu deficient phenotypes and genotypes at select point of care health settings in Vhembe District, Limpopo Province, South Africa UP ISMC Heterogeneity of Plasmodium falciparum transmission in north- Joe Biggs eastern South Africa: Implications for malaria elimination NICD Infection of African malaria vector mosquitoes with Plasmodium Annette Bennette falciparum malaria parasites WRIM Group photo followed by Lunch 12:15 - 13:30 Poster session (Odd numbered posters)
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Session 3 Biology and epidemiology of the malaria vector and remote sensing 13:30 - 17:00 Session Chairs: Basil Brooke (WRIM) & Leo Braack (UP ISMC) Basil Brooke 13:30 - 13:50 Perspectives on translating research into malaria vector control WRIM Spatio-temporal dynamic of asymptomatic malaria in a sahelian Jean Gaudart 13:50 - 14:10 environment Aix-Marseille Uni Anopheles arabiensis habitat suitability model spatially predicts Isaiah Gwitira 14:10 - 15:00 malaria hotspots in Zimbabwe Uni Zimbabwe 4 orals Ashley Burke Is Anopheles vaneedeni a vector of malaria in South Africa? NICD 10 minutes talk, Analysis of the esterase enzymes activity in Anopheles funestus Lizette Koekemoer 2 minutes Q&A adults and/or WRIM discussion Modelling the spatio-temporal dynamics of mosquitoes and its Claire Dufourd response to SIT control UP ISMC 15:00 - 15:30 Tea / Coffee and poster session
The effects of agrochemicals on the life history and insecticide Shune Oliver resistance phenotype of the major malaria vector Anopheles arabiensis (Diptera: Culicidae) NICD Mohlominyana 15:30 - 17:00 Monitoring insecticide resistance in the Anopheles gambiae Mofokeng complex from Kwazulu-Natal, South Africa 5 orals WRIM
Selecting most efficient targets for spatially focused interventions: Sallah Kankoé 10 minutes talk, Malaria incidence versus population mobility Aix-Marseille Uni 2 minutes Q&A and/or Evaluating effects of Landsat-derived environmental covariates for Oupa Malahlela discussion predicting malaria distribution in rural villages of Vhembe District, South Africa SANSA / UP ISMC
Analysing trends and forecasting malaria epidemics in Girond Florian Madagascar using a sentinel surveillance network: A web-based application IPM Gala function
17:30 for 18:00 Protea Manor Hotel, Hatfield Welcoming page 18:00 - 21:00 VIP guest speakers, entertainment, buffet dinner and networking for programme
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DAY 3 | Tuesday 2 August 2016 Time Event / Presentation title Speaker 07:30 - 08:30 Registration (Tea / Coffee) Session 4 Alternative methods and novel interventions for malaria vector control 08:30 - 12:00 Session Chairs: Rajendra Maharaj (MOMR) & Lizette Koekemoer (WRIM) Walter Focke 08:30 - 08:50 New approaches towards malaria vector control UP ISMC Impact of the Lubombo spatial development initiative after twelve Rajendra Maharaj 08:50 - 09:10 years in southern Mozambique MOMR
Pyrethroid-impregnated wall linings: Three years post installation Mildred Ramothole durability, user acceptability, perceived effectiveness, and UP ISMC 09:10 - 10:00 laboratory confirmed efficacy 4 orals Sustained release of repellent mixtures through engineering the Homa Izadi molecular interactions UP ISMC 10 minutes talk, Microporous polyolefins as controlled release devices for Mthokozisi Sibanda 2 minutes Q&A mosquito repellents UP ISMC and/or discussion Mathokoza Planning of housing improvement interventions for malaria Sibandze elimination in Swaziland NMCP Swaziland Tea / Coffee 10:00 - 10:30 Poster session (Even numbered posters) Hindrik Bouwman Environmental problems with DDT as used in malaria control NWU The potential use of serious games in malaria control and Marlize Coleman elimination programmes LSTM Nelusha The larvicidal and brine shrimp activities of eugenol and five Shunmoogam- 10:30 - 12:00 derivatives Gounden 7 orals WITS
Field effectiveness of microbial larvicides on mosquito larvae in Mulamuli Mpofu 10 minutes talk, malaria areas of Botswana and Zimbabwe UP ISMC 2 minutes Q&A and/or Givemore A review of the sterile insect technique for the control of malaria Munhenga discussion vectors in South Africa NICD
Attacking the mosquito on multiple fronts: Insights on optimal Samson Kiware combinations of vector control interventions for malaria elimination from a mathematical model IHI Mathematical modelling guiding elimination in the Asia-Pacific Sheetal Silal region and its potential in Africa UCT Lunch & Poster session 12:00 - 13:30 Poster session (Even numbered posters) Elimination 8 meeting (13:00)
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Session 5 Interventions against the malaria parasite 13:30 - 15:30 Session Chairs: Lyn-Marie Birkholtz (UP ISMC) & Jaishree Raman (NICD) Kelly Chibale 13:30 - 13:50 Antimalarial drug discovery at H3D H3D Current state of development of pantothenamides as anti- Erick Strauss 13:50 - 14:10 plasmodial agents Stellenbosch Uni
Dynamic evaluation of potential transmission blocking Janette Reader antiplasmodials: Rate and stage-specific descriptions of
gametocytocidal compounds UP ISMC Digital image analysis for determining malaria parasite density in John Frean thick blood films NICD 14:10 - 15:20 6 orals Using a risk based approach to develop a quality manual for Cordelia Leisegang public health institutions undertaking investigator-led antimalarial
drug clinical research in South Africa UCT 10 minutes talk, 2 minutes Q&A Bioprospecting traditionally used antimalarial plants by means of Johanna Bapela and/or NMR-based metabolomics UP ISMC discussion Jenny-Lee Novel drug scaffolds for the treatment of multi-drug resistant Panayides Plasmodium falciparum CSIR The molecular determinants for trafficking of ama-1, a malaria Alisje Churchyard invasion protein and vaccine candidate WRIM Rajendra Maharaj 15:20 - 15:25 Wrapping up and summary of the conference MOMR Tiaan de Jager 15:25 - 15:30 Closing of conference UP ISMC 15:30 - 16:00 Collection of posters
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Acronym / Initialism Affiliation and Country Aix-Marseille Uni Aix-Marseille University, France CSIR Council for Scientific and Industrial Research, South Africa DoH Mpumalanga Department of Health Mpumalanga, South Africa E8 Elimination 8, Namibia FCBMI Flowers Cross Border Malaria Initiative, United States of America H3D Drug, Discovery and Development Centre, UCT, South Africa IHI Ifakara Health Institute, Tanzania IPM Institute Pasteur de Madagascar, Madagascar LSTM Liverpool School of Tropical Medicine, United Kingdom MHCC Ministry of Health and Child Care, Zimbabwe MOMR Medical Research Council (MRC) Office of Malaria Research, South Africa NDoH National Department of Health, South Africa NICD National Institute for Communicable Diseases, South Africa NMCP Swaziland National Malaria Control Programme, Swaziland NWU North-West University, South Africa SANSA South African National Space Agency, South Africa SARN - RBM Southern Africa Regional Network - Roll Back Malaria, Botswana SU Stellenbosch University, South Africa UCT University of Cape Town, South Africa Uni Namibia University of Namibia, Namibia UP ISMC University of Pretoria Institute for Sustainable Malaria Control, South Africa Uni Zimbabwe University of Zimbabwe, Zimbabwe WITS University of the Witwatersrand, South Africa WRIM WITS Research Institute for Malaria, South Africa
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INVITED PLENARY SPEAKER BIOGRAPHIES & ABSTRACTS
Page 13 of 170 BIOGRAPHICAL SKETCH
Name: Lucille Blumberg
Institution: National Institute for Communicable Diseases (NICD)
Position: Deputy-Director, Head of Division, and Head of Travel Health
Qualifications: MB BCh, DCH, DTM & H, DOH
A. Personal Statement I trained at the University of the Witwatersrand and then worked in Paediatrics and Critical Care at Baragwanath Hospital in Johannesburg until I discovered the wonderful world of microbes, tropical diseases and zoonoses! I worked at the Tropical Diseases Hospital in Johannesburg treating patients with tuberculosis, malaria, trypanosomiasis, Crimean Congo Haemorrhagic fever and endless cases of tick bite fever. I qualified as a microbiologist and later I joined the newly created National Institute for Communicable Diseases.
Currently, I am the Deputy Director of the National Institute for Communicable Diseases, of the National Health Laboratory Service and currently head of the Public Health Surveillance and Response Division. The Division is responsible for the surveillance of disease outbreaks, travel medicine, and surveillance for selective diseases of public health importance. I am a medical consultant to the Emerging Pathogens Centre on rabies and viral haemorrhagic fevers. My special interests are tropical diseases, travel medicine, malaria, the viral haemorrhagic fevers and rabies.
Professional Membership Member: Various South African expert groups on Ebola, Rabies, Malaria Advisory groups: National Advisory Group on Immunisation; WHO on mass gatherings
B. Contribution to Malaria Research 1. Blumberg, L. Recommendations for the treatment and prevention of malaria: Update for the 2015 season in South Africa. S Afr Med J. 2015, 105(3):175-8. 2. Blumberg, L., Frean, J., Moonasar, D., South African Elimination Committee (Asomugha, C., Baker, L., Barnes, K., Brooke, B., Coetzee, M., de Jager, C., Duvenhage, C., Groepe, M., Kleinschmidt, I., Maharaj, R., Morris, N. and Ukpe, I.S.). Successfully controlling Malaria in South Africa. S Afr Med J. 2014, 104(3):224-227. 3. Morris, N., Frean, J., Baker, L., Ukpe, I.S., Barnes, K.I., Kruger, P., Mabuza, A., Raswiswi, E., Maharaj, R., Blumberg, L., Moonasar, D. Re-defining the extent of malaria transmission in South Africa: implications for chemoprophylaxis. S Afr Med J. 2013, 103(11):861-864. 4. Ukpe, I.S., Moonasar, D, Raman, J., Barnes, K.I., Baker, L., Blumberg, L. Case management of malaria – Treatment and chemoprophylaxis. In: Malaria in South Africa: Moving from control to elimination. S Afr Med J 2013, 103 (10 Suppl 2): 793-798. 5. Moonasar, D., Morris, N., Kleinschmidt, I., Maharaj, R., Raman, J, Mayet, N.T., Benson, F,G, Durrheim, D.N. and Blumberg L. What will move malaria control to elimination in South Africa? S Afr Med J. 2013, 103(10):801-806.
Page 14 of 170 SETTING THE RESEARCH AGENDA AROUND MALARIA ELIMINATION
Authors and Affiliations: Blumberg, L. (1)
(1) National Institute for Communicable Diseases
Abstract The move from malaria control to elimination is a quantum leap, will not be easy to achieve and will undoubtedly require sustained, high-level funding and political commitment for many years. The tools for malaria control are well described, reasonably well researched and have been applied with considerable success in the South African Malaria Control Programme. Moving towards the 2018 target for elimination of malaria in South Africa requires a different set of interventions, some of which have been researched and successfully applied, but often in a different setting to the low to medium malaria transmission situation in South Africa. The potential effects of introducing some of these interventions must be balanced against special local needs and best use of resources, and these considerations must also recognise differences between malaria transmission areas globally. The elimination of falciparum malaria will have to be achieved against a background of decreasing funding for malaria control here because of competing health priorities, and is a major geographic challenge, given that South Africa borders Mozambique, a high-transmission malaria country where the recent focus of malaria control has been its northern parts, rather than the areas directly bordering South Africa. The emergence of drug resistance in SE Asia, the origin for more global drug resistance and increasing vector resistance to insecticides are areas of concern. Key areas for both operational research as well as more ‘cutting edge’ scientific research are addressing vector dominance, the role and safety of gametocytocidal drugs in interrupting malaria transmission, the sensitivity and value of active screening for infections using molecular and other methods, identifying the emergence of markers of parasite resistance, and novel methods for mosquito control. This ambitious goal requires an operational research agenda that addresses the dynamic challenges encountered on the path to elimination, which will need to be flexible and developed in close relation with the national and provincial malaria programmes and a relatively small research community.
Page 15 of 170 BIOGRAPHICAL SKETCH
Name: Patrick Moonasar
Institution: National Department of Health (NDoH)
Position: Director: Malaria Unit
Qualifications: PhD in Public Health
A. Personal Statement I hold a Doctorate in Public Health degree from the London School of Hygiene and Tropical Medicine. Currently, I am the Director for Malaria at the National Department of Health in South Africa for the past 5 years. I have been working in the area of malaria for the past 15 years. I am a member and serve on several committees and boards. I am an honorary lecturer in the School of Public Health at the University of Pretoria in South Africa. I have authored and co-authored over several Scientific Articles, Policy documents and Policy briefs.
Professional Membership Member: South African Malaria Elimination Committee (SAMEC); Southern African Development Community: Elimination 8 Committee; Southern African Roll Back Malaria Network Global Malaria Elimination Group - UCSF- California, USA Member of the Research Advisory Board member of the University of Pretoria Institute for Sustainable Malaria Control (UP ISMC) Board member of MACEPA a subsidiary of Path, Seattle USA Several WHO technical working groups
B. Contribution to Malaria Research 1. Moonasar, D., Goga, A.E., Frean, J., Kruger, P. and Chandramohan, D. An exploratory study of factors that affect the performance and usage of rapid diagnostic tests for malaria in the Limpopo Province, South Africa. Malaria J. 2007, 6:74. 2. Moonasar, D., Morris, N., Kleinschmidt, I., Maharaj, R., Raman, J, Mayet, N.T., Benson, F,G,, Durrheim, D.N. and Blumberg L. What will move malaria control to elimination in South Africa? S Afr Med J. 2013, 103(10):801-806. 3. Maharaj, R., Raman, J., Morris, N., Moonasar, D., Durrheim, D.N., Seocharan, I., Kruger, P., Shandukani, B. and Kleinschmidt I. Epidemiology of malaria in South Africa: From control to elimination. S Afr Med J. 2013, 103(10):779-783. 4. Moonasar, D., Nuthulaganti, T., Kruger, P.S., Mabuza, A., Rasiswi, E.S., Benson, F.G., Maharaj, R. Malaria control in South Africa 2000-2010: beyond MDG6. Malaria J. 2012, 11:294. 5. Morris, N., Frean, J., Baker, L., Ukpe, I.S., Barnes, K.I., Kruger, P., Mabuza, A., Raswiswi, E., Maharaj, R., Blumberg, L., Moonasar, D. Re-defining the extent of malaria transmission in South Africa: implications for chemoprophylaxis. S Afr Med J. 2013, 103(11):861-864.
Page 16 of 170 TOWARDS MALARIA ELIMINATION IN THE MOSASWA (MOZAMBIQUE, SOUTH AFRICA AND SWAZILAND) REGION
Authors and Affiliations: Moonasar, D. (1); Kunene, S. (2); Candrinho, B. (3); Saute, F. (4); Ntshalintshali, N. (5); Maharaj, R. (6); Morris, N. (6)
(1) National Department of Health, South Africa; (2) Ministry of Health, Swaziland; (3) Ministry of Health, Mozambique; (4) Manhica Research Centre, Mozambique; (5) Clinton Health Access Initiative, Swaziland; (6) Medical Research Council, South Africa;
Abstract The main goal of the MOSASWA initiative is to achieve zero local malaria transmission in Swaziland, South Africa and Mozambique’s-Maputo Province by 2020 and achieve pre-elimination status in southern Mozambique (Gaza and Inhambane provinces) by 2025. The Initiative aims to build on the successes of the Lubombo Spatial Development Initiative (LSDI). The MOSASWA initiative has received political support from each of the participating countries through the signing of a declaration committing them to tackling malaria from the border areas of each country. The key objectives of the initiative are to: harmonise policies; strengthen sub-national capacities; expand access of malaria elimination interventions; focus on mobile and migrant populations, malaria risk localities and residents to reduce and interrupt malaria transmission; mobilise resources and advocate for increased long-term financing to achieve and sustain malaria elimination across the MOSAWA region. In this paper we present findings of a situational analysis conducted in the MOSASWA region. A situational analysis was conducted in 2015, using data from the three countries Health Management Information Systems (HMIS), national Malaria Information Systems (MIS), and sentinel surveillance systems. Malaria incidence, prevalence and other relevant indicators were extrapolated from these datasets. The situational analysis conducted upon the launch of the MOSASWA initiative revealed an incidence rate of 2.64 cases per 1000 population at risk in South Africa, 2.34 in Swaziland and 219 in Mozambique, in 2014. Incidence rates in Mozambique ranged between 0.01 and 500 at the sub-district level. Comparison of malaria cases recorded after the closure of the LSDI in 2010 and again in 2014 just prior to the establishment of MOSASWA revealed increases of 36%, 42% and 6% in Maputo Province (Mozambique), South Africa and Swaziland respectively between 2011 and 2014. The gains achieved through the LSDI malaria initiative have been eroded subsequent to its cessation in 2011. A renewed cross-border collaboration is required to ensure coordination between the three countries in reducing malaria in the MOSASWA region. The MOSASWA initiative provides a unique opportunity to accelerate the trajectory towards malaria elimination through the implementation of evidence based strategies at country and regional level.
Page 17 of 170 BIOGRAPHICAL SKETCH
Name: Idongesit Sunday Ukpe
Institution: University of Pretoria – UP ISMC
Position: Principal Specialist / Senior Lecturer: Family Medicine, Mpumalanga Department of Health South Africa, and University of Pretoria
Qualifications: MB BCh, DTM&H, MMed (Family Medicine)
A. Personal Statement I am a qualified medical practitioner who also holds a diploma in Tropical Medicine and Hygiene from the London School of Hygiene and Tropical Medicine. I am also a Fellow of the Royal Society of Tropical Medicine and Hygiene; Member of the South African Academy of Family Physicians, as well as Fellow of the Australasian College of Tropical Medicine.
Currently I am the Principal Specialist: Family Medicine, Mpumalanga Department of Health as well as a senior lecturer in the Department of Family Medicine at the University of Pretoria. My current research interests are in communicable disease management and control. He assists with the training of health care workers on malaria case management at the primary health care and district hospital levels.
Professional Membership Fellow, Royal Society of Tropical Medicine and Hygiene Fellow, Australasian College of Tropical Medicine South Africa Malaria Elimination Committee (Chairperson: Sub-committee on Treatment and Chemoprophylaxis) Department of Family Medicine, University of Pretoria Member University of Pretoria Institute for Sustainable Malaria Control (UP ISMC) Member, South African Academy of Family Physicians
B. Contribution to Malaria Research 1. Blumberg, L., Frean, J., Moonasar, D., South African Elimination Committee (Asomugha, C., Baker, L., Barnes, K., Brooke, B., Coetzee, M., de Jager, C., Duvenhage, C., Groepe, M., Kleinschmidt, I., Maharaj, R., Morris, N. and Ukpe, I.S.). Successfully controlling Malaria in South Africa. S Afr Med Jl. 2014, 104(3):224-227. 2. Morris, N., Frean, J., Baker, L., Ukpe, I.S., Barnes, K.I., Kruger, P., Mabuza, A., Raswiswi, E., Maharaj, R., Blumberg, L. and Moonasar, D. Re-defining the extent of malaria transmission in South Africa: Implications for chemoprophylaxis. S Afr Med J 2013, 103 (11): 861-864. 3. Ukpe, I.S., Moonasar, D, Raman, J., Barnes, K.I., Baker, L., Blumberg, L. Case management of malaria – Treatment and chemoprophylaxis. In: Malaria in South Africa: Moving from control to elimination. S Afr Med J 2013, 103 (10 Suppl 2): 793-798. 4. Ukpe, I.S. Plasmodium ovale in South Africa. Trans R Soc Trop Med Hyg. 1998, 92(5):574. 5. Ukpe, I.S. Continuing medical education in unstable malaria areas. Bull WHO. 1999, 77(11):948.
Page 18 of 170 MALARIA CASE MANAGEMENT IN THE CONTEXT OF MALARIA ELIMINATION: THE ROLE OF THE POPULATION AT RISK
Authors and Affiliations: Ukpe, I.S. (1)
(1) Department of Family Medicine, University of Pretoria
Abstract In Malaria Programme, malaria case management intervention mainly entails programmatic measures aimed at diagnosing malaria early and promptly treating it effectively to minimise morbidity and prevent mortality, prevent onward transmission to others, and prevent the emergence of drug resistance. In the context of malaria elimination, defined in terms of zero local transmission of malaria in a specific geographical area, with regards to malaria case management intervention, the role of the population at risk of malaria in the battle to eliminate local transmission of malaria is to present any ‘malaria-like’ illness early, within 24 hours of onset, to formal health care facility for diagnosis and prompt effective treatment to prevent onward transmission. Late presentation of malaria to formal health care facilities negatively impacts on the goals of malaria case management intervention. For malaria elimination therefore, there is need to explore innovative ways of educating the population on malaria to promote early help seeking at formal health care facilities when ill with malaria symptoms.
Page 19 of 170 BIOGRAPHICAL SKETCH
Name: Leo Braack
Institution: University of Pretoria – UP ISMC
Position: Associate Professor in the Dept of Medical Virology, and an Entomologist
Qualifications: BSc, BSc (Hons), PhD in Entomology
A. Personal Statement My early career was as a research scientist in the Kruger National Park of South Africa, but included responsibility for malaria control in this 20,000km2 area now hosting around a million visitors per annum. My broader research responsibilities covered general medical and veterinary entomology but with particular focus on malaria, the role of blowflies in anthrax transmission, and ticks. I also collaborated substantially in research on haemorrhagic fever viruses in small mammals. Later I was promoted to Chief Scientist, and subsequent promotions took me out of an active research role and into international conservation area management. Four years ago I resumed an academic career by joining the University of Pretoria where I am now enjoying a renewed focus on malaria, in particular research into vector biology. I successfully applied for a Gates Foundation “Grand Challenges Exploration Fund” which led to some fascinating research mainly in Uganda on biting behaviour of all the main African malaria vectors. That research on biting behaviour led to a successful application, with Professor Focke at University of Pretoria and others in Mozambique and Germany, for funding from the Deutsche Forschungs Gemeinschaft for development of novel polymer carrier materials to impregnate with combination repellent/insecticidal compounds to reduce outdoor biting by malaria vectors. At the request of a major gas-drilling company in Mozambique, I have assisted in assessing and refining their malaria management programme in a high-intensity transmission area of Mozambique.
Professional Membership Member, WHO Team investigating reservoir hosts for Ebola and Marburg viruses Member, CDC team in Uganda to research potential Ebola/Marburg hosts Management Committee member: UP Institute for Sustainable Malaria Control (UP ISMC)
B. Contribution to Malaria Research 1. Braack, L.E.O. Large contractors in Africa: conundrums with malaria chemoprophylaxis. Malaria J. 2016, 15:207. 2. Brisco, K.K., Cornel, A.J., Lee, Y., Mouatcho, J. and Braack, L.E.O. Comparing efficacy of a sweep net and a dip method for collection of mosquito larvae in large bodies of water in South Africa F1000Res. 2016, 5:713. 3. Braack, L., Hunt, R., Koekemoer, L.L., Gericke, A., Munhenga, G., Haddow, A.D., Becker, P., Okia, M., Kimera, I. and Coetzee, M. Biting behaviour of African malaria vectors: 1. where do the main vector species bite on the human body? Parasit Vectors. 2015, 4(8):76. 4. Munhenga, G., Brooke, B.D., Spillings, B., Essop, L., Hunt, R.H., Midzi, S., Govender, D., Braack, L. and Koekemoer, L.L. Field study site selection, species abundance and monthly distribution of anopheline mosquitoes in the northern Kruger National Park, South Africa. Malar J. 2014, 24(13):27. 5. Govere, J., Braack, L.E., Durrheim, D.N., Hunt, R.H. and Coetzee M. Repellent effects on Anopheles arabiensis biting humans in Kruger Park, South Africa. Med Vet Entomol. 2001; 15(3):287-92.
Page 20 of 170 RESIDUAL MALARIA: CHANGED MIND-SETS TO QUELL THE SEEDS OF FAILURE
Authors and Affiliations: Braack, L. (1); Birkholtz, L. (1); Stoltz, A. (1); Focke, W. (1); Kruger, T. (1); Bornman, R. (1); Mouatcho, J. (1); de Jager, C. (1)
(1) UP Institute for Sustainable Malaria Control
Significant reductions in malaria mortality and morbidity have been achieved over the preceding 15 years, fanning hopes that this time round global eradication can be achieved. The reality is that the easy fruits have been harvested, successes are beginning to taper off and serious challenges are increasingly evident. One frustrating element is the difficulty in dealing with Residual Malaria along the fringes of core malaria distribution. Are we flogging the same old tools a little too doggedly and failing to embrace new sets of thinking and elimination tools? Traditional malaria control had an overwhelming focus on indoor residual spraying (IRS) and long lasting insecticide treated nets (LLINs), both depending on mosquitoes entering homes and submitting to insecticide exposure in a confined area. The battle-ground has shifted and is now in the wide open spaces outside, with an enemy in multiple guises and an ability to attack from different flanks. Residual Malaria is fed at different scales and from widely divergent sources. There are external factors that contribute to the challenge of festering malaria, such as imported malaria cases, imported gametocyte carriers, reduced donor funding etc. that are difficult to deal with, and there are internal factors such as insecticide resistance, shifts in vector behaviour such as increased outdoor biting and early-night biting, the neglected issue of secondary vectors that may allow residual malaria to smoulder, problems with malaria control programme efficiencies, inadequate regulatory foundations to deal with gametocyte carriers etc., and these are the factors that are more within our control and require some honest introspection. As cross-cutting challenges are imperfect knowledge and understanding of the complexities that contribute to residual malaria, lack of availability or adoption of alternative control tools, inadequate resources, and sometimes lack of inadequate political support. Finally, there is the bottom-line reality that malaria is a disease of poverty, and at the end of the day the ultimate factor that will lead to eventual global eradication is sufficient economic upliftment that enables rural people to escape the behavioural factors that expose them to malaria vectors in the first place.
Page 21 of 170 BIOGRAPHICAL SKETCH
Name: Lyn-Marie Birkholtz
Institution: University of Pretoria – UP ISMC
Position: Professor in the Department of Biochemistry & DST/NRF South African Research Chair in Sustainable Malaria Control
Qualifications: BSc, BSc (Hons), MSc, PhD in Biochemistry
A. Personal Statement I obtained my PhD (Biochemistry) in 2003, jointly performed at the University of Pretoria (UP) and at the Bernhard Nocht Institute for Tropical Medicine (Hamburg, Germany). I am currently a Professor of Biochemistry at UP and I established the Malaria Parasite Molecular Laboratory (www.biochem.up.ac.za/M2PL) as part of the UP Institute for Sustainable Malaria Control (UP ISMC). I was awarded a DST/NRF South African Research Chair in Sustainable Malaria Control (July 2013) and hold a NRF B3 rating. My research interest is centred on the physiology, biochemistry and pharmacology of malaria parasites with a specific focus on discovering biochemical distinctions between the malaria parasite and the human host, which are exploitable for novel antimalarial chemotherapeutical design. Principal contributions include 1) elucidation of structure-activity properties of drug-target proteins involved in underexplored metabolic pathways and 2) global analyses of the functional genome of the parasite after perturbation of such metabolic pathways.
Professional Membership Member: Women in Science, SA; South African Society of Biochemistry and Molecular Biology; South African Malaria Initiative; European Virtual Institute for Malaria Research (EviMalaR); International Biochemical Society; American Chemical Society; American Society for Microbiology Chair: SA-French SAFeTI research partnership workshops, SA; SA–Sweden research partnership program bilateral agreement (NRF-SIDA) workshops, SA Management Committee member: UP ISMC
B. Contribution to Malaria Research
1. van Brummelen, A.C., Olszewski, K.L., Wilinski, D., Llinas, M., Louw,A.I. and Birkholtz LM. Co- inhibition of Plasmodium falciparum S-Adenosylmethionine Decarboxylase/Ornithine Decarboxylase Reveals Perturbation-specific Compensatory Mechanisms by Transcriptome, Proteome, and Metabolome Analyses. J Biol Chem. 2009, 284 (7):4635-4646. 2. Verlinden, B.K., Niemand, J., Snyman, J., Sharma, S.K., Beattie, R.J., Ross, J., Woster, P.M and Birkholtz LM. Discovery of Novel Alkylated (bis)Urea and (bis)Thiourea Polyamine Analogues with Potent Antimalarial Activities. J Med Chem. 2011.54(19):6624-6633. 3. Reader, J., Botha, M., Theron, A., Lauterbach, S.B., Rossouw, C., Engelbrecht, D., Wepener, M., Smit, A., Leroy, D., Mancama, D., Coetzer T,L. and Birkholtz LM. (2015) Nowhere to hide: interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination. Malaria J. 2015 14:213. 4. Verlinden, B.K., de Beer, M., Pachaiyappan, B, Besaans, E., Andayi, W.A., Reader, J., Niemand, J., van Biljon, R., Guy, K., Egan, T., Woster, P.M. and Birkholtz LM. Interrogating alkyl and arylalkylpolyamino (bis)urea and (bis)thiourea isosteres as potent antimalarial chemotypes against multiple lifecycle forms of Plasmodium falciparum parasites. Bioorg Med Chem. 2015. 23(16): 5131-5143. 5. Birkholtz, L., Bornman, R., Focke, W., Mutero, C. and de Jager C. Sustainable malaria control: transdisciplinary approaches for translational applications. Malaria J. 2012. 11:431-442.
Page 22 of 170 SEEING THE WOOD FOR THE TREES: MALARIA PARASITE SYSTEMS BIOLOGY AND HOW IT AIDS ANTIMALARIAL DRUG DISCOVERY
Authors and Affiliations: Birkholtz, L. (1)
(1) Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, Private bag x20, Hatfield, Pretoria, South Africa, 0028
Abstract Malaria parasites have an extremely complex life cycle consisting of two distinct developmental phases, asexual proliferation as well as sexual differentiation. The characteristic pathogenesis associated with the disease is largely due to a massive increase in parasite numbers (e.g. Plasmodium falciparum) during their asexual amplification in human erythrocytes. However, a minor portion of these parasites are able to undergo sexual differentiation with every asexual cycle, forming male and female gametocytes that ensure continued transmission by being taken up by female Anopheline mosquitoes. Both asexual and sexual development is tightly controlled and regulated to ensure normal progression, however, the molecular mechanisms governing these processes have not been clarified. This is quite surprising given the impact that this information could have in developing new tools to target the parasite. Systems biology describes the ability to study all molecular components that make up a biological entity. This allows predictions on the dynamics and complexity of organisms, details that could not be obtained from the study of individual parts alone. Drug development is based on the realisation that disease phenotypes are complex and that discovery of new drugs should be driven by insights into biological responses. This paper will present large scale transcriptomics, proteomics and epigenomics data on malaria parasite asexual and sexual stage development and highlight peculiarities therein that could aid drug discovery processes. ‘-Omics’ analysis were performed for all stages of parasite development within its asexual proliferative cycle as well as all developmental stages characterising sexual differentiation. On all parasite samples, transcriptomics (DNA microarray), proteomics (LC-MS/MS) and epigenomics (histone PTM nano- LC MS/MS) were performed. Biological replication enabled quantitative statistical validation of the subsequent datasets in a head-to-head profile. Gene enrichment and interaction networks were applied and the data used to interrogate changes in the biology of the parasite upon drug perturbation, enabling drug mode-of-action deconvolution. We have completed, for the first time, systems level analyses of the functional genome of malaria parasites across their entire life cycle, including asexual and sexual development. Horizontal probing of these datasets allowed descriptors of regulatory processes including regulation of gene expression through key transcription factors (Api-2G), kinases (e.g. NEK, NIMAs and CDKs) and epigenetic marks. The large dataset enabled interrogation of the mode-of-action of lead antimalarials and their derivatives, indicating divergence in action where different metabolic pathways were targeted in a chemotype dependent manner. Systems level analyses of the malaria parasite biology reveals key mechanisms for development of these organisms that could be targeted as the parasites’ Achilles heel. By using such global strategies encompassing all biological processes at once, drug discovery endeavours will be supported by identifying novel druggable processes, describe drug mode-of-action and ensure drug diversity.
Page 23 of 170 BIOGRAPHICAL SKETCH
Name: Jaishree Raman
Institution: National Institute for Communicable Diseases (NICD)
Position: Medical Scientist, Parasitology Reference Laboratory
Qualifications: BSc, Higher Diploma, BSc (Hons), MSc, PhD in Zoology: Molecular Systematics
A. Personal Statement After my post-doc, I joined the Malaria Research Unit of the South African Medical Research Council and during my 10 year tenure I established and led a successful research programme focusing primarily on understanding the emergence and spread of antimalarial drug resistance within southern Africa. Antimalarial resistance marker data generated by the laboratory as part of a robust surveillance programme has been used to inform antimalarial drug policy in Swaziland, South Africa and Mozambique. In 2014 I moved my research programme to the National Institute for Communicable Diseases, where research activities within the programme are now directed to towards assisting South Africa achieve its malaria elimination goal. I am currently assessing the appropriateness of implementing new tools like ultra-sensitive PCR and LAMPs in rural settings as means of facilitating the early detection of sub-patent malaria carriers. Gametocyte detection and the elimination of gametocytes are also major research areas as prolonged low level gametocyte carriage is thought to be one on the major sources of sustained transmission in South Africa.
Professional Membership Member: Case Management and Surveillance Subcommittees of the South African Malaria Elimination Committee (SAMEC); Research Constituency of the Southern African Resistance Network (SARN) of the Roll Back Malaria Initiative; African Plasmodium Diversity Network (PDNA) Technical Advisory Group Member: Diagnostics and Case Management Subcommittee of the E8 Malaria Elimination Initiative; Molecular Module of the Worldwide Antimalarial Resistance Network (WWARN)
B. Contribution to Malaria Research 1. Maharaj, R., Raman, J., Morris, N., Moonasar, D., Durrheim, D., Seocharan, I., Kruger, P., Shandukani, B. and Kleinschmidt, I. Epidemiology of malaria in South Africa: from control to elimination. S Afr Med J. 2013, 103(10):779-783. 2. Raman, J., Mauff, K., Muianga, P., Mussa, A., Maharaj, R. and Barnes, K.I. Five years of antimalarial resistance marker surveillance in Gaza Province, Mozambique, folllowing artemisinin-based combination therapy roll out. PLoS One. 2011, 6(10): e25991. 3. Raman, J., Little, F., Roper, C., Kleinschmidt, I., Cassam, Y., Maharaj, R. and Barnes, K.I. Five years of large scale dhfr and dhps mutation surveillance following the phased implementation of artesunate plus sulphadoxine-pyrimethamine in Maputo Province, Southern Mozambique. Am J Trop Med Hyg. 2010, 82(5):788-794. 4. Allen, E., Little, F., Camba, T., Cassam, Y., Raman, J. Boulle, A. and Barnes, K.I. Efficacy of sulfadoxine-pyrimethamine with and without artesunate for the treatment of uncomplicated Plasmodium falciparum malaria in southern Mozambique: a randomized controlled trial. Malar J. 2009, 8(141):1-10. 5. Raman, J., Sharp, B., Kleinschmidt, I., Roper, C., Streat, E., Kelly, V. and Barnes, K.I. Differential effect of regional drug pressure on dihydrofolate reductase and dihydropteroate synthethase mutations in Southern Mozambique. Am J Trop Med Hyg. 2008, 78(2):256-261.
Page 24 of 170 MUTANT MALARIA PARASITES: AN OVERVIEW ON THEIR GENETICS, ORIGINS AND SPREAD
Author and Affiliations: Raman, J. (1)
(1) National Institute for Communicable Diseases
Abstract Human malaria parasites have ensured their survival over the decades by rapidly evolving mechanisms which negate the intended effects of antimalarials. The genetic basis of resistance to commonly used monotherapies, such as chloroquine, mefloquine and sulfadoxine-pyrimethamine, is surprising simple. It generally involves single nucleotide polymorphisms (SNPs) in essential transporter genes or within the target-site itself. These SNPs either interfere with the movement of the antimalarial to the target site or with the binding of the antimalarial to the target-site. Although the genetic basis for artemisinin resistance has not been fully elucidated, indications are that while SNPs are involved, they are not located in transporter genes. In this overview we will briefly discuss the origin of drug resistant parasites found in southern Africa, while focussing on the genetic mechanisms underpinning antimalarial drug resistance to chloroquine, mefloquine, sulfadoxine- pyrimethamine and artemisinin. We will also speculate on the potential threat artemisinin resistant parasites pose to malaria control/elimination efforts within southern Africa and propose interventions that should be implemented to prevent the establishment of artemisinin resistant parasites within southern Africa.
Page 25 of 170 BIOGRAPHICAL SKETCH
Name: Basil Brooke
Institution: National Institute for Communicable Diseases (NICD) & WITS Research Institute for Malaria (WRIM)
Position: Associate Professor & Head of Laboratory: Vector Control Reference Laboratory
Qualifications: Malaria vector biology and control
A. Personal Statement My research work over the past 15 years has focused on an array of genetic and biometric issues related to the development, inheritance, expression, relative stability and pleiotropic effects of insecticide resistance in African malaria vector species. This work has been applied to the development of strategies designed to maintain effective malaria vector control in the face of insecticide resistance, particularly in the southern African region. I have also been integrally involved in the assessment of new malaria vector control products and alternative methods of vector control.
Detecting and understanding the epidemiological consequences of insecticide resistance and other important phenotypes in malaria vectors forms an important component of the research support needed for South Africa’s drive toward malaria elimination. Boosting control effectiveness towards elimination requires a vector surveillance system that allows for the collation of data on vector species distribution, resting and biting behaviour, and the occurrence of insecticide resistance in malaria affected regions. Colleagues and I are therefore in the process of setting up projects to boost malaria vector surveillance in South Africa by assessing the merits of a series of passive and active vector collection/trapping techniques in those provinces that experience seasonal malaria transmission, and in which indoor residual spraying (IRS) based vector control is conducted annually.
Professional Membership Fellow of the Royal Entomological Society (FRES) Health Professions Council of South Africa (HPCSA) Entomological Society of Southern Africa (ESSA)
B. Contribution to Malaria Research 1. Brooke, B.D., Kloke, G., Hunt, R.H., Temu, E.A., Koekemoer, L.L., Taylor, M.E., Small, G., Hemingway, J. and Coetzee, M. Bioassay and biochemical analyses of insecticide resistance in southern African Anopheles funestus. Bull Entomol Res. 2001, 91: 265-272. 2. Brooke, B.D. and Koekemoer, L.L.. Review. Major effect genes or loose confederations? The development of insecticide resistance in the malaria vector Anopheles gambiae. Parasit Vectors .2010, 3: 74. 3. Brooke, B.D., Koekemoer, L.L., Kruger, P., Urbach, J., Misiani, E. and Coetzee, M. Malaria Vector Control in South Africa. S Afri Med J. 2013, 103(10 Suppl 2): 784-788. 4. Oliver, S.V. and Brooke, B.D. The effect of multiple blood-feeding on the longevity and insecticide resistant phenotype in the major malaria vector Anopheles arabiensis (Diptera: Culicidae). Parasit Vectors .2010, 7: 390. 5. Samuel, M., Oliver, S.V., Wood, O.R., Coetzee, M. and Brooke, B.D. Evaluation of the toxicity and repellence of an organic fatty acids mixture (C8910) against insecticide susceptible and resistant strains of the major malaria vector Anopheles funestus Giles (Diptera: Culicidae). Parasit Vectors .2015, 8: 321.
Page 26 of 170 PERSPECTIVES ON TRANSLATING RESEARCH INTO MALARIA VECTOR CONTROL
Authors and Affiliations: Brooke, B. (1, 2)
(1) Centre for Opportunistic, Tropical & Hospital Infections, National Institute for Communicable Diseases, Johannesburg (2) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg
Abstract Malaria vector control relies principally on insecticide application. Indoor spraying of residual insecticides (IRS) and the distribution of insecticide treated nets (ITNs) are proven control technologies that have been used to good effect over the past 15 years, a period in which malaria incidence has decreased by 37% globally and by 42% in Africa. Nevertheless, applications of these technologies require constant management, refinement and development primarily because of the burgeoning incidence of insecticide resistance in target vector populations coupled with a very limited number of insecticide classes that are currently available for public health use. Additionally, residual (outdoor) malaria transmission is becoming increasingly prevalent in control settings including South Africa’s malaria affected regions, requiring additional control technologies and applications. Insecticide resistance in malaria vectors is now so widespread that it is necessary to apply sound management principles to all malaria affected regions under the auspices of the Global Plan for Insecticide Resistance Management. One of the five pillars of this plan details the critical need for regular vector surveillance linked to operational and translational research. A retrospective analysis covering 15 years of direct research experience in malaria vector ecology, insecticide resistance and alternative methods of vector control illustrates the vital link between surveillance and research, and ultimately shows that one can never ask too many research questions in pursuit of efficient and sustainable malaria control.
Page 27 of 170 BIOGRAPHICAL SKETCH
Name: Jean Gaudart
Institution: Aix-Marseille University, Marseille, France
Position: Assistant Professor
Qualifications: Medical degree, MSc, PhD in Mathematics
A. Personal Statement As a medical doctor, Public Health specialist and Biostatistician, I have the necessary breadth of knowledge and expertise to strengthen the biostatistical/biomathematical core of the current project. My research is focused on the analysis of the spatio-temporal dynamic of Malaria and communicable diseases. As an academic researcher, I have developed biostatistical methods for spatio-temporal cluster analysis, and to assess the dynamic of malaria in collaboration with international teams. Furthermore, I have developed mathematical models to assess relationships between malaria transmission and different factors such as environment, economic behaviour and mobility. Acutely aware of the importance of strong continued international partnership, over 16 years, I have developed a strong collaboration with the Malaria Research and Training Centre in Bamako, Mali (Pr Ogobara Doumbo), strengthening the data-management and biostatistics unit. As an example of my leadership roles, I am currently leading a research team working on Health, Space Environment and development. Within the AMMA consortium (African Monsoon Multidisciplinary Analysis), I’m leading the network Climate and Malaria. Furthermore, strongly aware of building capacity, I am the director of a Master program on Public Health, Society and Development, with around 50 students internationally per year and I am a doctoral supervisor for PhD students from numerous countries (Mali, Burkina Faso, Senegal, Togo), together with partner researchers.
Professional Membership Member: French Statistical Society; International Biometric Society; French Society of Exotic Pathology; Studies Committee, Faculty of Medicine, Marseille; French-speaking Epidemiologist Society; Global Health Committee, French national research agency [ANR] Expert: French ethic committee [CPP Sud-Méditerranée 1], Marseille ; Clinical Investigation Centre’s [CIC] Marseille and La Reunion, France
B. Contribution to Malaria Research
1. Gaudart, J., Touré, O., Dessay, N., Dick,o A.L., Ranque, S., Forest, L., Demongeot, J and, Doumbo, O.K. Modelling malaria incidence with environmental dependency in a locality of Sudanese savannah area, Mali. Malaria J. 2009, 8:61. 2. Gaudart, J., Ghassani, M., Mintsa, J., Rachdi, M., Waku, J. and Demongeot, J. Demography and diffusion in epidemics: Malaria and Black Death spread. Acta Biotheoretica 2010, 58(2-3):277- 305. 3. Demongeot, J., Gaudart, J., Mintsa, J. and Rachdi, M. Demography in epidemics modelling. Commun Pure Appl Anal. 2012, 11(1):61-82. 4. Demongeot, J., Gaudart ,J., Lontos, A., Promayon, E., Mintsa, J. and Rachdi M. Zero-diffusion domains in reaction-diffusion morphogenetic and epidemiologic processes. Int J Bifurcat Chaos. 2012, 22:1250028_1-23. 5. Berthélemy, J.C., Thuillez, J., Doumbo, O.K. and Gaudart, J. Malaria and protective behaviours: is there a malaria trap? Malaria J .2013, 12:200.
Page 28 of 170 SPATIO-TEMPORAL DYNAMIC OF ASYMPTOMATIC MALARIA IN A SAHELIAN ENVIRONMENT
Authors and Affiliations: Coulibaly, D. (1); Piarroux, R. (2); Travassos, M.A. (4); Tolo, Y. (1); Laurens, M.B. (4); Kone, A. K. (1); Traore, K. (1); Sissoko, M. (1); Niangaly, A. (1); Diarra, I. (1); Daou, M. (1); Guindo, B. (1); Kouriba, B. (1); Plowe, C.V. (4); Doumbo, O.K. (1); Thera, M.A. (1); Gaudart, J. (3)
(1) Malaria Research & Training Center, Department of Epidemiology of Parasitic Diseases, Faculty of Medicine and Dentistry, University of Sciences, Techniques &Technologies, Bamako, Mali; (2) Aix-Marseille University, UMR MD3, F-13385 Marseille, France; (3) Aix-Marseille University, IRD, INSERM, SESSTIM UMR_S912, F-13385 Marseille, France; (4) Division of Malaria Research, Institute for Global Health, University of Maryland School of Medicine, Baltimore, MD
Abstract In Mali, despite scaling up of control strategies, there is no clear decreasing trend of malaria incidence. Undetected infected individuals may constitute a reservoir when environmental conditions are unfavourable for transmission. This study aimed at assessing spatio-temporal variations of malaria incidence and asymptomatic carriage. A cohort of 400 children (0-14 years, Bandiagara, Mali, 2009-2014), was followed monthly and when ill to capture malaria infection and illness data. Periodicity and phase shifts between clinical episodes, asymptomatic carriage and rainfall were determined by using wavelet-transform. For low and high transmission periods, symptomatic and asymptomatic malaria were mapped at the household level. High and low risk spatial clusters were identified by using Kulldorff’s approach. Risk factors for symptomatic and asymptomatic malaria were assessed using nested case-control analysis, matched by transmission- period, neighbourhood and bednet use. Co-factors included age, gender, previous infections, hemoglobin-type, and G6PD-deficiency status. Adjusted Odd Ratios (ORa) were estimated using conditional logistic regression. Time-series analysis showed a persistent seasonal pattern, with a 3 month shift between rain and malaria infection. Within high or low transmission periods, clusters in the town center were at low risk for clinical malaria (resp. RR=0.61, RR=0.23). Three significant high risk clusters were identified during the high transmission period, but only one of these during the low transmission period. Asymptomatic carriages were clearly persistent, even during low transmission period. For each high transmission period, the ratio of asymptomatic carriers to clinical cases was around 0.5, but increased to 5 during low transmission periods. Nested case-control analyses showed that experiencing clinical episode was protective against subsequent clinical episodes (ORa=0.61 CI95%[0.38;0.96]) and asymptomatic carriage (ORa=0.26 [0.07;0.93]), but asymptomatic carriage during a previous high transmission period was a risk factor (ORa=2.67 [1.52;4.69]) for asymptomatic carriage. Despite implementation of seemingly appropriate control activities, malaria transmission patterns remained stable. This unexpected stability of malaria transmission was associated with persistent asymptomatic carriage during dry seasons. Parasite carriage is critical for undetected circulation during inter-epidemic periods, where environmental local conditions are unfavourable. This spatio-temporal pattern of parasite carriage could be a window of opportunity to interrupt transmission and to improve control strategies.
Page 29 of 170 BIOGRAPHICAL SKETCH
Name: Walter Focke
Institution: University of Pretoria – UP ISMC
Position: Professor in Department of Chemical Engineering, and Director: Institute of Applied Materials
Qualifications: BEng (Chem), MEng, PhD in Materials Science & Engineering
A. Personal Statement I am a full Professor in Chemical Engineering and Director of the Institute of Applied Materials at the University of Pretoria. Chemical product formulation is my core research competence. It is the science and technology employed to design chemical products in order to satisfy one or other market need. It is a multidisciplinary activity that is built on the scientific principles of phase equilibrium thermodynamics, surface chemistry, colloids, polymers, emulsions and suspensions, mixing technology, etc. In practice it is important to know how these principles are connected to desirable end-use product properties, e.g. texture, shelf life, weathering behaviour, adhesion, corrosion, biodegradability, etc. While this approach can in principle be applied to any arbitrary chemical system, my research groups focus on pyrotechnics for mining applications, clay and graphite nanoparticles as polymer additives and controlled release of actives from polymer matrices. The latter activity seeks applications in malaria vector control. Past and present malaria related research projects include development of polypropylene bed nets, polyethylene wall linings, insecticidal “white wash” and reducing the volatility of mosquito repellents.
Professional Membership Member: American Chemical Society; Polymer Processing Society; International Pyrotechnics Society, South African Institute of Chemical Engineers; Editorial board of the J Vinyl Additive Technology; Editorial board of the International Journal of Adhesion & Adhesives; Editorial board of International Polymer Processing Management Committee member: University of Pretoria Institute for Sustainable Malaria Control (UP ISMC)
B. Contribution to Malaria Research Our research efforts seek to find safer ways to control malaria vector mosquitoes through longer lasting repellent technologies (to protect people outdoors) and long-life insecticide impregnated netting to provide indoor protection. A primary target is to prolong the activity of WHO approved insecticides in order to make them suitable as DDT replacements in IRS.
1. Merckel, R.D., Focke, W.W., Sibanda, M.M., Massinga, P. and Crowther, N.A.S. Co-Intercalation of Insecticides with Hexadecyltrimethylammonium Chloride in Mozambican Bentonite. Mol Cryst Liq Cryst. 2012, 555(1):76-84. 2. Birkholtz, L., Bornman, R., Focke, W., Mutero, C. and Jager C. Sustainable malaria control: transdisciplinary approaches for translational applications. Malar J. 2012, 11:431. 3. Sibanda, M.M. and Focke, W.W. Development of an insecticide impregnated polymer wall lining for malaria vector control. Malar J. 2014, 13(1):80. 4. Akhtar, M.U. and Focke W.W. Trapping citronellal in a microporous polyethylene matrix. Thermochim Acta.2015, 613(1):61–65. 5. Kruger, T., Sibanda, M.M, Focke, W.W., Bornman, M.S. and de Jager C. Acceptability and effectiveness of a monofilament, polyethylene insecticide-treated wall lining for malaria control after six months in dwellings in Vhembe District, Limpopo Province, South Africa. Malar J. 2015, 14:485.
Page 30 of 170 NEW APPROACHES TOWARDS MALARIA VECTOR CONTROL
Authors and Affiliations: Focke, W.W. (1, 2); Androsch, R.T. (3); Braack, L.E.O. (2); De Jager, C. (2); Izadi, H. (1, 2); Kruger, T. (2); Maharaj, R. (2, 4); Sibanda, M.M. (1, 2)
(1) Institute of Applied Materials, Department of Chemical Engineering, University of Pretoria; (2) UP Institute for Sustainable Malaria Control, University of Pretoria; (3) Center of Engineering Sciences, Martin Luther University Halle-Wittenberg; (4) Office of Malaria Research, Medical Research Council, Durban South Africa
Abstract Malaria is a debilitating disease that infects upwards of 200 million people annually. This malaria burden can be reduced dramatically by reducing the probability of mosquito bites. Vector control is a strategy that aims to achieve this by decreasing the number of malaria transmitting mosquitoes as much as possible. Effective malaria vector control techniques, sanctioned by the WHO, include indoor residual spray (IRS) and long-lasting insecticide impregnated nets (LLINs). However, both methods target only endophilic vector populations. While topical applications of repellents such as N,N-diethyl-m-toluamide (DEET) do reduce the risk of infection outdoors, the residual efficacy lasts only a few hours. Malaria elimination will require new approaches to deal more effectively with mosquitoes whether they bite indoors or outdoors. This communication outlines some vector control approaches currently pursued by the University of Pretoria Institute for Sustainable Malaria Control. The target product concepts include low-cost wall linings for indoor use and long-life repellent anklets for outdoor protection. The latter may also benefit from more effective, longer lasting repellents formulations. Multifilament polypropylene yarn, containing 0.38 % β-cyfluthrin, were successfully produced on conventional production-scale fibre-spinning machine. The performance of bed nets, knitted from the yarns, complied with the WHO bioassays requirements even after 25 washes. The WHO specifications are ≥ 95% knockdown after one hour and ≥ 80% mortality after 24 hours. Polyethylene-based mesh impregnated with either alphacypermethrin or deltamethrin was evaluated as wall lining in an extensive field trial. Bioassays showed that the mesh retained its effectiveness (100% mortality in WHO cone tests) beyond three years from the date of installation. Novel topical mosquito formulations were designed that outperformed DEET and IR3535 in arm-in-cage repellence tests conducted at the South African Medical Research Council. Foot-in- cage bioassays indicate that anklets, in which DEET was trapped inside a microporous polymer matrix, retained significant mosquito repellence activity even after 54 days of accelerated ageing. Novel product ideas targeting malaria vector control were shown to hold promise in laboratory tests and even in preliminary field trials. More work is required to optimize the active ingredients formulations and the delivery systems in order to provided effective and long-lasting protection against malaria transmission.
Page 31 of 170 BIOGRAPHICAL SKETCH
Name: Rajendra Maharaj
Institution: Medical Research Council Office of Malaria Research (MOMR)
Position: Director: MOMR & Extraordinary Professor of Entomology
Qualifications: BSc, BSc (Hons), MSc, PhD in Entomology
A. Personal Statement I completed a PhD investigating environmental factors influencing malaria transmission. Thereafter, I was offered a position as the National Malaria Control manager at the national Department of Health, where I was responsible for directing the malaria control activities in the country and advising on policy issues. My 21 years’ experience working in all aspects of malaria research and control, included research and control of both the malaria parasite and mosquito vector, conducting indoor residual house spraying programmes, organising and participating in entomological and parasitological surveys, utilising global positioning system (GIS) technology to identify hot-spots of malaria transmission, direct and manage malaria control interventions and provide a management tool for use by malaria control managers. Regionally, I was the Principal Investigator of the Lubombo Spatial Development Initiative (LSDI). This initiative succeeded in decreasing the prevalence of malaria in all three countries by over 90% in five years. Currently, I am the Director of the MRC Office of Malaria Research.
Professional Membership Technical Advisor: IAEA; WHOPES; WHO on the use of DDT; WHO on writing GFATM proposals Member: SADC Malaria Task; SA Malaria Elimination Committee; Subcommittee for Vector Control; KwaZulu-Natal Malaria Advisory Group; International Atomic Energy Association Expert Group on Sterile Insect Technique; African Network for Vector Resistance; Expert Advisory Group to the Regional Director of the WHO; Malaria Elimination Group; AMFm; SHIP Scientific Advisory Committee; University of Pretoria Institute for Sustainable Malaria Control (UP ISMC) Advisory Board member Co-Chairman, the WHO Working Group on Indoor Residual Spray Steering Committee Member: South African Malaria Initiative; Novel Drug Development Platform; Southern African Roll Back Malaria Network; DDT Expert Advisory Group
B. Contribution to Malaria Research
1. Mavundza, E.J., Maharaj, R., Finnie, J.F., Kabera, G. and Van Staden, J. An ethnobotanical survey of mosquito repellent plants in uMkhanyakude district, KwaZulu-Natal province, South Africa. J Ethnopharmacol. 2011, 137: 1516-1520. 2. Moonen, B., Cohen, J.M., Snow, R.W., Slutsker, L., Drakeley, C., Smith, D.L., Abeyasinghe, R.R., Rodriguez, M.H., Maharaj, R., Tanner, M. and Targett G. Operational Strategies to Achieve and Maintain Malaria Elimination. Lancet. 2010; 6, 376(9752):1592-603. 3. Maharaj, R., Morris, N., Seocharan, I., Kruger, P., Moonasar, D., Mabuza, A., Raswiswi, E. and Raman J. The feasibility of malaria elimination in South Africa. Malar J. 2012, 11:423. 4. Moonasar, D., Morris, N., Kleinschmidt, I., Maharaj, R., Raman, J, Mayet, N.T., Benson, F,G,, Durrheim, D.N. and Blumberg L. What will move malaria control to elimination in South Africa? S Afr Med J. 2013, 103(10):801-806. 5. Maharaj, R., Raman, J., Morris, N., Moonasar, D., Durrheim, D.N., Seocharan, I., Kruger, P., Shandukani, B. and Kleinschmidt I. Epidemiology of malaria in South Africa: From control to elimination. S Afr Med J. 2013, 103(10):779-783.
Page 32 of 170 IMPACT OF THE LUBOMBO SPATIAL DEVELOPMENT INITIATIVE AFTER TWELVE YEARS IN SOUTHERN MOZAMBIQUE
Authors and Affiliations: Maharaj, R. (1); Moonasar, D. (2); Morris, N. (1)
(1) Medical Research Council of South Africa, Durban; (2) National Department of Health, South Africa
Abstract The Lubombo Spatial Development Initiative (LSDI) was a tri-country initiative between Mozambique, South Africa and Swaziland. This initiative established an effective cross-border malaria control programme especially in Mozambique. As part of the LSDI, vector control was implemented using efficacious insecticides and early diagnosis and treatment. The initiative ended in 2011 after 12 years but the success of the LSDI has not been recorded. This study aims to update the paper published in 2007 highlighting the first five years of the initiative. The data collected over the life of the LSDI was analysed. The decrease in prevalence/incidence was used as a proxy indicator of impact. Surveys conducted in Mozambique provided the baseline against which success was measured. Entomological collections were conducted at sentinel sites in Maputo Province whilst prevalence surveys were conducted in the same households in children between the ages of 2-<15. Interventions to control the mosquito vector (spraying with carbamate insecticides) and the roll-out of effective treatment (artemesinin based combination therapy) was implemented. Exit traps fitted to windows of selected households in each sentinel site provided information pertaining to vector species and population numbers prior to and after indoor residual spraying. Anopheles arabiensis and An. funestus were found to be the main vectors in Mozambique. The introduction of indoor residual spraying decimated mosquito population numbers to a point where it was no longer feasible to collect vector population data. From the information generated using different classes of insecticides, it was evident that An. funestus was less affected by pyrethroids than An. arabiensis. The malaria prevalence in the age group under study decreased from an average of 70% to an average of <5% by the time activities were discontinued in Mozambique. The massive reduction in prevalence in southern Mozambique was proof that co-ordinated cross-border malaria control has an enormous impact on disease transmission. Long term control in contiguous border areas can alleviate the disease burden in all countries.
Page 33 of 170 BIOGRAPHICAL SKETCH
Name: Kelly Chibale
Institution: University of Cape Town
Position: DST/NRF SARChI chair in drug discovery, Professor of organic chemistry, Director: of H3D, Director: MRC/UCT Drug Discovery & Development Research Unit
Qualifications: B.Sc. Ed., PhD in Chemistry
A. Personal Statement I am a full Professor of Organic Chemistry at the University of Cape Town (UCT) in South Africa, a Full Member of the UCT Institute of Infectious Disease & Molecular Medicine (IDM), a Tier 1 South Africa Research Chair in Drug Discovery, founding Director of the Medical Research Council (MRC) Drug Discovery and Development Research Unit at UCT and the Founder and Director of the UCT Drug Discovery and Development Centre (H3D). I obtained my PhD in Synthetic Organic Chemistry from the University of Cambridge in the UK (1989-1992). This was followed by postdoctoral stints at the University of Liverpool in the UK (1992-94) and at the Scripps Research Institute in the USA (1994-96). I was a Sandler Sabbatical Fellow at the University of California San Francisco (2002), a US Fulbright Senior Research Scholar at the University of Pennsylvania School of Medicine (2008) and a Visiting Professor at Pfizer in the UK (2008).
My research focus is on drug discovery of potential drugs that fight malaria, tuberculosis, cancer, helminths, cardiovascular and fibrosis diseases
Professional Membership Fellow of the Royal Society of Chemistry (FRSC) Fellow of the Royal Society of South Africa (FRSSAf) Member of the American Chemical Society
B. Contribution to Malaria Research 1. Greenbaum, D., Mackey, Z., Hansell, E., Doyle, P., Gut, J., Caffrey, C.R, Lehman, J., Rosenthal, P.J., McKerrow, J.H. and Chibale K. Synthesis and Structure Activity Relationships of Parasiticidal Thiosemicarbazone Cysteine Protease Inhibitors against P.falciparum, T. brucei and T. cruzi. J. Med. Chem. 2004, 47, 3212-3219. 2. Ndakala, A.J., Gessner, R.K., Gitari, P.W., October, N., White, K.L., Hudson, A., Fakorede, F., Shackleford, D.M., Kaiser, M., Yeates, C., Charman, S.A. and Chibale K. Antimalarial Pyrido[1,2- a]benzimidazoles. J. Med. Chem. 2011, 54, 4581–4589. 3. Younis, Y., Douelle, F., .Feng, T-S., González Cabrera, D., Le Manach, C., Nchinda, A.T., Duffy, S., White, K.L., Shackleford, D.M., Morizzi, J., Mannila, J., Katneni, K., Bhamidipati, R., Zabiulla, K.M., Joseph, J.T., Bashyam, S., Waterson, D., Witty, M.J., Hardick, D., Wittlin, S., Avery, V., Charman, S.A. and Chibale K. 3,5-Diaryl-2-aminopyridines as a Novel Class of Orally Active Antimalarials Demonstrating Single Dose Cure in Mice and Clinical Candidate Potential. J. Med. Chem. 2012, 55, 3479−3487. 4. Younis, Y., Street, L.J., Waterson, D., Witty, M.J. and Chibale K. Cell-based Medicinal Chemistry Optimization of High Throughput Screening (HTS) Hits for Orally Active Antimalarials-Part 2: Hits from SoftFocus Kinase and other Libraries. J. Med. Chem. 2013, 56, 7750. 5. Njoroge, M., Njuguna, N., Mutai, P., Ongarora, D., Smith, P. and Chibale K. Recent Approaches to Chemical Discovery and Development Against Malaria and the Neglected Tropical Diseases Human African Trypanosomiasis and Schistosomiasis. Chem. Rev. 2014, 114, 11138−11163.
Page 34 of 170 ANTIMALARIAL DRUG DISCOVERY AT H3D
Authors and Affiliations: Chilabe, K (1)
(1) Drug Discovery and Development Centre (H3D), Department of Chemistry and Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Rondebosch 7701, South Africa;
Abstract A limited skilled manpower base and poor access to technology platforms and/or enabling technologies are some of the challenges that have historically hampered African-led innovation in antimalarial drug discovery. One of the most effective ways of addressing these challenges is to prosecute projects through networks of partnerships while at the same time building local capacity on the back of these projects. This lecture will introduce the University of Cape Town Drug Discovery and Development Centre (H3D) and describe representative antimalarial drug discovery projects, and how these projects have been critical to build drug discovery infrastructure, experience and expertise.
Page 35 of 170 BIOGRAPHICAL SKETCH
Name: Erick Strauss
Institution: Stellenbosch University
Position: Professor in the Department of Biochemistry
Qualifications: BSc, BSc (Hons), PhD in Chemistry & Chemical Biology
A. Personal Statement My research group is currently one of the leaders in the field of coenzyme A (CoA) biosynthesis and in the development of inhibitors that target this pathway. We were the first identify a gene that encodes the binfunctional CoaBC protein that catalyses the second and third steps of CoA biosynthesis in bacteria. We uncovered the mode of action of the natural product CJ-15,801, and demonstrated its selective inhibition of the PPCS activity of the CoaBC protein of Staphylococcus aureus. My group has also made important contributions to our understanding of bacterial pantothenate kinases (PanKs), the first enzyme of the CoA pathway. We were the first to characterize the type III PanKs (PanKIII) in 2005, publish the first description of a PanKIII structure in 2006, followed by a further characterization of the binding and mechanism of the protein in 2008. I was integral to the rediscovery of the pantothenamide class of CoA antimetabolites in 2002, and have since published several studies on the synthesis and application of these compounds as antibacterial and antiplasmodial agents (with Dr. Kevin Saliba from ANU, Australia). A highlight of this work was the development of a modified pantothenamide which shows inhibition against the proliferation of P. falciparum with a potency similar to chloroquine.
Professional Membership Western Cape Coordination Committee, South African Chemical Institute American Society for Biochemistry and Molecular Biology American Society for Microbiology South African Chemical Institute South African Society for Biochemistry and Molecular Biology American Chemical Society
B. Contribution to Malaria Research 1. Macuamule, C.J., Tjhin, E.T., Jana, C.E., Barnard, L., Koekemoer, L., De Villiers, M., Saliba, K.J. and Strauss, E. A pantetheinase-resistant pantothenamide with potent, on-target and selective antiplasmodial activity. Antimicro. Agents Chemother. 2015, 59, 3666–3668. 2. De Villiers, M., Barnard, L., Koekemoer, L., Snoep, J.L. and Strauss, E. Variation in pantothenate kinase type determines the pantothenamide mode of action and impacts on coenzyme A salvage biosynthesis. FEBS J. 2014. 281, 4731–4753. 3. Moolman, W.J.A., de Villiers, M. and Strauss, E. Recent advances in targeting coenzyme A biosynthesis and utilization for antimicrobial drug development. Biochem Soc Trans. 2014, 42:1080–1086. 4. De Villiers, M., Macuamule, C., Spry, C., Hyun, Y.-M., Strauss, E. and Saliba, K.J. Structural modification of pantothenamides counteracts degradation by pantetheinase and improves antiplasmodial activity. ACS Med Chem Lett. 2013, 4:784−789. 5. Spry, C., Macuamule, C., Lin, Z., Virga, K.G., Lee, R.E., Strauss, E. and Saliba, K.J. Pantothenamides are potent, on-target inhibitors of Plasmodium falciparum growth when serum pantetheinase is inactivated. PLoS ONE. 201,. 8(2):e54974.
Page 36 of 170 CURRENT STATE OF DEVELOPMENT OF PANTOTHENAMIDES AS ANTIPLASMODIAL AGENTS
Authors and Affiliations: Macuamule, C.J. (1); Tjhin, E.T. (2); Jana, C.E. (1); Barnard, L. (1); Koekemoer, L. (1); Spry, C. (2); De Villiers, M. (1); Saliba, K.J. (2,3); Strauss, E. (1)
(1) Department of Biochemistry, Stellenbosch University, Stellenbosch, South Africa; (2) Research School of Biology, The Australian National University, Canberra, ACT, Australia; (3) Medical School, College of Medicine, Biology and Environment, The Australian National University, Canberra, ACT, Australia
Abstract Pantothenate, the precursor of the essential metabolic cofactor coenzyme A (CoA), is the only vitamin that Plasmodium falciparum must obtain from extracellular sources during blood-stage proliferation. Analogues of pantothenate can interfere with the biosynthesis of CoA from pantothenate, or hijack the biosynthetic process to form non-reactive CoA analogues that act as anti-metabolites that have the potential of inhibiting a range of CoA-dependent processes. Recently, a pantothenamide showing potency similar to that of chloroquine (IC50 ~20 nM) was discovered. However, the compounds are also sensitive to degradation by serum enzymes, reducing their half- life and consequently their potency. The development of pantothenamides as antiplasmodial agents with clinical potential therefore becomes a delicate balancing act of target-dependent potency on the one hand, and the introduction of degradation-resistant modifications on the other. We are following a two-pronged approach: first, introducing structural modification that renders pantothenamides resistant to degradation, followed by determination of the resulting compounds’ change in antiplasmodial potency (i.e. an empirical approach). Second, pursuing the discovery of the pantothenamides’ molecular target of inhibition, to allow the introduction of modification that increases degradation resistance while maintaining target-selectivity (i.e. a rational approach). We have been successful in discovering those structural modifications that provides the most protecting against degradation, while maintaining excellent potency. However, our understanding of the molecular target of pantothenamide-mediated inhibition is still at an early stage. The further advancement of the pantothenamides as antiplasmodial agents with clinical potential will largely depend on our ability to pin-point their molecular mechanism of action, and whether this constitutes action at a single or multiple targets. Current strategies whereby this can be achieved is mainly based on reconciling our knowledge of parasite biology with that of CoA-dependent enzymology; this work therefore as much wider implications and will significantly impact on our understanding of central metabolism in apicomplexan parasites.
Page 37 of 170
ORAL ABSTRACTS
Page 38 of 170 COULD A SONG ABOUT MALARIA PREVENTION HELP PROTECT CHILDREN UNDER 5 IN VENDA RURAL COMMUNITIES?
Abstract Theme Education & health promotion
Authors and Affiliations Anderson, C.M. (1); McCrindle, C.M.E. (2); Kruger, T. (1)
(1) University of Pretoria School of Health Systems and Public Health (SHSPH) and University of Pretoria Institute for Sustainable Malaria Control (UP ISMC), University of Pretoria, South Africa; (2) University of Pretoria School of Health Systems and Public Health (SHSPH), University of Pretoria, South Africa
Abstract Malaria kills over half a million individuals each year, mostly children <5 years of age, with 90% of cases occurring in Sub-Saharan Africa. The aim of the study was to develop a song for children < 5 years of age in Venda rural communities in Limpopo, South Africa, that included the five main determinants known to decrease the risk of malaria. Literature review does not indicate that any songs focussing on malaria prevention have previously been developed for this age group of children. A literature review of determinants contributing to malaria in rural children <5 years of age in Thohoyandou, Limpopo, South Africa was conducted for the development of a song, with age appropriate lyrics describing behaviour likely to minimise the risk of malaria. Data was obtained from expert opinion surveys and focus groups using a Likert scale and the Delphi Method, to modify the song according to responses. A total of 8 out of 10 experts suggested revisions based on speed of music and clarity of words; after revision, two focus groups in the study area approved the song after further modifications. The song was edited three times based on the feedback from both focus groups and expert opinions, until recommendations converged for a complete song. A culturally and age appropriate song to help prevent malaria in children >5 years of age has been created. The music was engaging and informative enough that the community, preschool teachers and malaria experts agreed that this song could help with behavioural changes to prevent malaria in children <5 years of age.
Page 39 of 170 FOCUS ON MALARIA IN EDUCATION
Abstract Theme Education & health promotion
Authors and Affiliations Petersen, J. (1); Schubart, O. (1); Adebanji, O. (1); Steyn, M.G. (1); Phatudi, N. (1); Kruger, T. (1)
(1) Faculty of Education, University of Pretoria & University of Pretoria Institute for Sustainable Malaria Control
Abstract Malaria is a major health dilemma with over 90% of deaths occurring in Africa, South of the Sahara, most in children under the age of five years. Annual reports from Limpopo Province indicate that Ha- Makuya in the Vhembe District has the highest incidence rate of malaria. Three separate studies focused on how improved knowledge of malaria might affect the attitudes, awareness, and learning of those affected by malaria. A qualitative research approach was followed and data were collected using interviews, observation and document analysis. Interviews with Primary School principals, teachers and learners, as well as with parents, district officials and health care workers were conducted over the period of a week. Interviews were transcribed and returned to the participants for confirmed accuracy. Comprehensive field notes were kept on observation, and analysed documents included the Curriculum Assessment Policy Statement (CAPS). A lack of curriculum content on malaria in the Foundation Phase was observed. Most of the children and teachers interviewed had limited knowledge about the causes and prevention of malaria. It was also observed that the Department of Health (DoH) was at the forefront of malaria education, with negligible collaboration from the Department of Basic Education (DoBE). Malaria appears not to affect the teaching and learning in the classroom as there are effective measures in place to deal with suspected cases of malaria. As young children are mostly affected by malaria, the integration of health education in the Life Skills subject in the Foundation Phase will provide them with fundamental knowledge of malaria. Parents and teachers are keen on the Department of Education including malaria education in the Foundation Phase curriculum. Improved collaboration between role players will also lead to optimised opportunities regarding the role that schools could play in reducing the impact of malaria through education.
Page 40 of 170 MALARIA DEATHS: MEETING THE 2015 NEAR ZERO TARGET IN SOUTH AFRICA
Abstract Theme Case Management
Authors and Affiliations Misiani, E.A. (1); Motsuku, L.M. (1,2,3); Ukpe, I.S. (3,4); Asomugha, C. (5); Mabuza, A. (4); Mnyaluza, J. (5); Raswiswi, E. (6); Mabunda, E. (7); Shandukani, M.B. (1); Raman, J. (2); Moonasar, D. (1).
(1) National Department of Health, Pretoria; (2) National Institute for Communicable Diseases; (3) University of Pretoria; (4) Mpumalanga Provincial Department of Health; (5) Gauteng Department of Health; (6) KwaZulu-Natal Department of Health; (7) Limpopo Department of Health
Abstract The Global Malaria Action Plan set out national mortality targets of near zero for all preventable malaria deaths. This study sought to determine South Africa’s progress in achieving this goal at various levels and determining factors associated with deaths in hospitals within high reporting districts. A retrospective data review of malaria related deaths in various districts in the four provinces of Gauteng, Mpumalanga, KwaZulu-Natal and Limpopo was carried out from 2011/12 malaria season to 2015/16 season (the seasons run from June to July of the following year). Malaria mortality trends analysis and chi square test were conducted. Frequencies and proportions were generated. The odds ratios and p value for factors associated with deaths in the highest reporting facilities will be estimated using STATA statistical software. The number of malaria deaths fluctuated throughout the study period with a peak observed in 2013/14 season. Between 2013/14 and 2014/15 the number of deaths decreased by 15.4%. During the study period, Tintswalo, Donald Fraser and Ngwelezana hospitals reported the highest numbers of deaths contributing to 18.69% (n=20), 27% (n=30) and 69% (n=18) of the reported malaria related deaths in Mpumalanga, KwaZulu- Natal and Limpopo provinces respectively. In Gauteng and Mpumalanga, late health seeking behaviour of malaria patients led to most deaths. South Africa moved steadily towards the zero malaria deaths target between 2011 and 2014 until the 2013/2014 outbreak. Public hospitals located near migration routes reported the highest numbers of deaths. Appropriate case management and health education are necessary for reduction of deaths and the results of this study will be useful in guiding policy to achieve near zero deaths in future. Malaria mortality, near zero, case management
Page 41 of 170 TARGETING FOCI OF TRANSMISSION TOWARDS ELIMINATION OF MALARIA IN SOUTH AFRICA
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Morris, N. (1); Shandukani, B. (2); Moonasar, P. (2); Kruger, P. (3); Mabuza, A. (4); Raswiswi, E. (5); Raman, R. (6); Brooke, B. (6); Groepe, M.A. (7); Varney, S. (8); Misiani, E. (2) (1) Medical Research Council (2) National Department of Health, Malaria Directorate (3) Malaria Control Programme, Limpopo Province (4) Malaria Control Programme, Mpumalanga Province (5) Malaria Control Programme, KwaZulu-Natal Province (6) National Institute of Communicable Diseases (7) World Health Organisation (8) Clinton Health Access Initiative
Abstract The national strategy for malaria elimination adopted by South Africa in 2012 provides for targeted intervention and response to accelerate towards zero local transmission (NDoH, 2012). World Health Organisation (WHO) operational guidelines for elimination (2012) suggest that malaria during the elimination and pre-elimination phases is heterogeneous and that intervention should be targeted at known foci of transmission for maximum impact in interrupting transmission (Bousema, 2012). The WHO defines a focus as “a defined, circumscribed locality situated in a currently or formerly malarious area with the continuous or intermittent epidemiological factors necessary for malaria transmission.” Seven distinct classes of foci are defined: endemic, residual active, residual non-active, new active, new potential, cleared up and pseudo foci (WHO, 2012). We developed empirical criteria based on the varied epidemiology of the three malaria endemic provinces of South Africa in order to classify foci of transmission in the country and proposed appropriate intervention packages per foci type. Confirmed malaria case data reported into the three provincial Malaria Information Systems (MIS) in KwaZulu-Natal, Mpumalanga and Limpopo Provinces were extracted as aggregate per locality for the 1996/1997 to 2015/2016 malaria seasons, stratified by source of infection (local, imported and unclassified). Empirical criteria were developed in line with WHO foci classification definitions and taking account of provincial, district, municipality and locality level trend over time. Intervention packages were proposed per foci type, including various combinations and intensities of vector surveillance and control, reactive and proactive case detection, case investigation, prompt diagnosis and treatment, community mobilisation, health promotion and advocacy. A preliminary desktop exercise applying empirical criteria to malaria case data at the locality level detected 70 residual active, 110 new potential, 261 endemic, 684 residual non-active, 37 new active and 1095 cleared up foci of malaria transmission across the nine malaria endemic districts of South Africa. An additional 261 localities were classified as endemic and still appropriate for control strategies, while a further 300 localities reported imported cases exclusively, either currently or historically. Results of the desktop classification exercise conducted by an expert working group will be the subject of provincial technical group consultations, discussion and ratification. Adjustments based on programmatic knowledge of case reporting and investigation at the locality level in each province are expected. Phased implementation of intervention packages is planned during the 2016/2017 malaria season. With all nine malarious districts of South Africa now in the elimination phase, a more detailed understanding of heterogeneity at the local level is required in order to target intervention most effectively to interrupt transmission. Large scale foci classification is a crucial next step in the national drive towards elimination, and will serve to inform targeting of specific and appropriate intervention packages.
Page 42 of 170 THE IMPACT OF MALARIA PROGRAM INTERVENTIONS AND CLIMATE CHANGE ON MORTALITY AND MORBIDITY RATES BETWEEN 2009 AND 2016 IN LIMPOPO PROVINCE: PRELIMINARY FINDINGS
Abstract Theme Epidemiology
Authors and Affiliations Motsuku, L.M. (1, 2, 3), Baloyi, R.E. (1, 2, 3), Misiani, E.A. (2, 3), Shandukani, M.B. (2, 3), Moonasar, D.P. (2)
(1) National Institute for Communicable Diseases; (2) National Department of Health; (3) University of Pretoria
Abstract The impact of malaria program interventions and climate change on morbidity and mortality rates between 2009 and 2016 in Limpopo provinces of South Africa. Malaria is a challenging public health problem in Africa. South Africa is embarking on elimination strategy to achieve zero local malaria transmission by 2018. However Limpopo province is one of endemic provinces reporting highest cases and deaths every season despite interventions in place. This study is aimed at exploring the impact of Limpopo province malaria program interventions and climate change on morbidity and mortality rate: 2009-2016. A retrospective data review of malaria cases and death between 2009/10 season and 2015/16 season was carried out. Data on training, health education, malaria treatment, Indoor Residual Spraying (IRS) and climate change were collected. Malaria incidence rates and case fatality rate from 2009/10 season until 2015/16 season was calculated. Time series analysis was used to determine any association between mortality/morbidity (dependent variable) and interventions/climate change (Independent variable). There were 24071 cases and 401 deaths reported in Limpopo province during the study period. There was significant decrease in number of cases from 2009/10 season to 2015/16 season by 86.7% (p<0.005). The highest incident rate and case fatality rate were noted in 2013/14 season outbreak, 84 per 100000 populations and 1.5% respectively. The results for impact of IRS, climate change and training will be reported after the analysis is complete. South Africa had an increase in malaria cases and deaths between 2012/13 season and 2013/14 season. This increase is attributed to Limpopo province outbreak. Case fatality rate increases as incidence rate increases. Time series analysis will be second phase of the study in order to explore impact of case management on deaths and other interventions. Malaria mortality, Incidence trends, Malaria interventions
Page 43 of 170 TRAVEL PATTERNS AND DEMOGRAPHIC CHARACTERISTICS OF MALARIA CASES, THE CASE STUDY OF SWAZILAND’S ACTIVE SURVEILLANCE SYSTEM, 2010-2014
Abstract Theme Regional malaria and crossborder issues affecting elimination
Authors and Affiliations Dlamini, N. (1); Tejedor‐Garavito, N. (2); Pindolia, D. (4); Soble, A. (4); Ruktanonchai, N. (2, 3); Alegana, V. (2, 3); Le Menach, A. (4); Ntshalintshali, N. (4); Dlamini, B. (4); Smith, D.L. (5); Tatem, A.J. (2, 3); Kunene, S. (1)
(1) National Malaria Control Programme, Swaziland; (2) WorldPop project, University of Southampton, UK; (3) Flowminder Foundation, Sweden; (4) Clinton Health Access Initiative; (5) Institute for Health Metrics, US
Abstract As Swaziland makes progress towards its malaria elimination goal, understanding the potential of imported cases to instigate local transmission and sustain local parasite reservoirs becomes critical to understand and evaluate. Travel history (reported travel within the last 8 weeks) from routine surveillance data between January 2010 and June 2014 was analysed to describe and compare travel patterns of passively detected RDT-confirmed index cases to travel patterns of actively-identified RDT negative individuals (contacts living in the vicinity of an index case within receptive areas were tested during re-active case detection). Reported destinations of travel were geo-located to the smallest administrative boundary possible. A total of 67% of confirmed index cases (n=1,517) reported travel history. Of 9,859 contacts screened during reactive case detection, 105 (1%) were RDT positive. 876 index cases travelled outside Swaziland. Travel history reports for index cases increased by 48% annually for travel outside Swaziland and 27% annually for travel within Swaziland. 25% of individuals screened during reactive case detection reported travel outside Swaziland, of which 22% tested positive for malaria. Mozambique was the most likely travel destination of positive individuals, with Maputo City, Inhambane and Gaza being the most likely destinations. 96% percent of RDT positive travellers outside Swaziland were either Swazi (52%) or Mozambican (45%), however Swazis were more likely to test negative. 84% of the 755 males and 60% of the 440 females that travelled outside Swaziland tested positive. Overall, 24 to 45 year olds were most likely to report travel. Additionally, paths of transmission, important border crossings and means of transport were identified. Results from this analysis can be used to direct national and regional targeting of interventions, over space, time and by population groups. Cross-border collaboration is necessary to manage importation of malaria at the regional level for countries targeting malaria elimination.
Page 44 of 170 OPERATIONAL FACTORS ASSOCIATED WITH INCREASED DETECTION OF INFECTIONS FOR REACTIVE CASE DETECTION (RACD) IN SWAZILAND
Abstract Theme Epidemiology
Authors and Affiliations Bhangu, K. (1); Mkhonta, N. (1); Ntshalintshali, N. (2); Schwartz, A. (3); Dlamini, B. (2); Nhlabathi, N. (1); Novotny, J. (2); Sturrock, H. (4); Zulu, Z. (1); Dorsey, G. (4); Gosling, R. (3); Kunene, S. (1); Greenhouse, B. (3); Hsiang, M. (3, 4)
(1) National Malaria Control Programme, Swaziland Ministry of Health; (2) Clinton Health Access Initiative, Swaziland; (4) Global Health Group, University of California, San Francisco, United States of America; (5) Department of Pediatrics, University of Texas South-western Medical Center
Abstract Reactive case detection (RACD) is strategy whereby household members and neighbors of index cases detected through passive surveillance are actively screened for malaria infection. It is recommended for malaria surveillance programs seeking to eliminate malaria, however the work is resource intensive and low yield and there may be ways to optimize operations. Using prospectively collected reactive case detection data in Swaziland all laboratory-confirmed index cases were targeted for a case investigation at their places of residence. If there was potential for local transmission, family and neighbors residing within 500m were targeted for rapid diagnostic testing (RDT), collection of a dried blood spot for LAMP testing, and a survey to assess risk factors. Multivariate models using a generalized estimating equations (GEE) approach were used to test the associations between potential risk factors and LAMP positivity, with models accounting for clustering at the level of the index case. From August 2012 to March 2015, 373 index cases residing in receptive areas received RACD. 29% (108/373) of RACD events resulted in the identification of at least one additional LAMP+ infection, versus 13% of RACD events having at least one RDT positive case. Index cases were more likely to lead to the identification of an additional LAMP+ case if RACD coverage was at least 80% (AOR 2.0, 95% CI 1.2-3.4), RACD occurred within 7 days of the index case presentation (AOR 2.5, 95% CI 1.3-4.6), if the index case was female (AOR 1.8, 95% CI 1.0-3.2), worked in farming (AOR 2.4, 95% CI 1.0-5.6), and was not protected with vector control interventions (AOR 2.1 95% CI 1.2-3.6). Compared to screening everyone within 500m of an index case, a strategy whereby only individuals residing within 200m of an index case were screened, with expansion to 500m if there was an RDT+ in the index case household or the index case was female or farming would enable less screening (76% of individuals) and identification of 93% of LAMP+ infections. Use of highly sensitive diagnostics, as well as analysis of RACD and index case level risk factors for a given setting can inform the design of optimal RACD scenarios that maximize efficiency and yield.
Page 45 of 170 A STUDY TO IDENTIFY HIGH MALARIA RISK GROUPS AND THE ASSOCIATED RISK FACTORS USING A PROGRAMMATIC TOOL: MALARIA ELIMINATION RISK FACTOR ASSESSMENT TOOL (MERFAT)
Abstract Theme Epidemiology
Authors and Affiliations Haindongo, E. (1); Smith, J. (2); Mumbengegwi, D. (1); Sturrock, H. (2); Gosling, R. (2)
(1) Multi-disciplinary Research Centre, University of Namibia, Windhoek, Namibia; (2) Malaria Elimination Initiative, Global Health Group, University of California, San Fransisco, USA
Abstract Namibia has seen considerable reductions in the number of malaria infections in recent years. Following these gains in the reduction, the country is transitioning from low-controlled malaria to elimination or the absence of locally acquired infections. As overall endemicity levels declines, malaria transmission becomes increasingly focal and reservoirs of infection are maintained in geographical areas and populations at higher risk of infection. The study region historically experienced a high burden of malaria due to its tropical climate and porous borders with high endemic areas. The western Zambezi region is a low transmission setting, making it a choice for an elimination risk factors assessment tool. The study aims to 1) identify local risk factors for symptomatic malaria and 2) to develop an easily implementable and cost-effective programmatic tool. The study is being conducted at 6 out of 11 health facilities since January 2015 – May 2016. The Malaria Elimination Risk Factor Assessment Tool (MERFAT) is a health facility based case-control study, in which every incident cases is assigned two control profiles matched by age and gender, profiles were also assigned to a health facility by probability proportional to the size of the health facility catchment population. Preliminary results indicate that frequent at night outdoor activity (> 7 p/ week) in the early evening hours (6-9 pm) may be vulnerable. Environmental factors such as total rainfall a month prior to diagnosis was also associated with a high risk. Travelling individuals were not at higher risk. Coverage in case and control households was low (35%). Preliminary results have found that people with malaria are less likely to use their ITNs. Future interventions should thus be targeted at students, fishermans and children. Furthermore, Indoor Residual Spraying (IRS) can potentially also be strengthened.
Page 46 of 170 IMPORTED CASES, SPORADIC OUTBREAKSAND RESURGENCE ARE THE HIGHEST THREATS TO MALARIA ELIMINATION IN THE E8 REGION
Abstract Theme Regional malaria and crossborder issues affecting elimination
Authors and Affiliations Mudambo, K.T.S.M. (1); Mosweunyane, T. (2); Moonasar, D. P. (3); Kunene, S. (4); Mberikunashe, J. (5)
(1) Roll Back Malaria - East and Southern Africa Secretariat, Gaborone, Botswana; (2) Botswana National Malaria Control Program, Gaborone, Botswana; (3) South Africa National Malaria Program, Pretoria; (4) Swaziland National Malaria Program, Mbabane; (5) Zimbabwe National Malaria Program, Harare
Abstract The successful elimination of malaria and attainment of the 2020 - 2030 Goal in the Elimination Eight (E8) countries (Angola, Botswana, Mozambique, Namibia, South Africa, Swaziland, Zambia and Zimbabwe) will very much depend on successful cross-border collaboration, prevention of import of cases, outbreaks and resurgence. Since 2012, there has been an increase in imported cases, outbreaks and resurgence in several E8 countries. The aim of this paper is to highlight the threat paused by import of cases, outbreaks and resurgence and their impact on elimination. A desk review of imported cases, outbreaks and resurgence (2013 - 2016) was carried out in 5 E8 countries. Telephone conversations were used clarification and elicit for information. A total of 22,006 confirmed imported cases were observed in 3 Front-line countries between 2013 - 16 (6,367, 5,937 and 2,114) respectively for 2013, 2015 and 2016. During the same period, 2282, 5881 and 789 local cases were observed respectively. Imported cases came from as far as South Sudan, Eretria and Ghana. The review also showed an increase in sporadic outbreaks in both front-and-second-line countries with a total of 15,637 cases being recorded in 2014. Not all outbreaks are recorded and resurgence occurred in some malaria free districts. In second-line countries, cases are not disaggregated, making it difficult to quantify the imported burden. There are no standardized indicators/tools for defining imported cases, outbreaks and thresholds. The observed surge in imported cases, outbreaks and resurgence constitutes a threat that could derail the Front-line countries 2020 elimination goal. The E8 needs to develop Regional standardized indicators/tools/thresholds/definitions that will guide the quantification/definition of the burden of imported cases, strengthen regional cross-border surveillance system and sharing of information. A complete E8 review of the status of imported malaria, the design of cross-border strategies, strengthening Port Health Authorities, mobile units and introduction of screening to halt this threat is recommended.E8 scorecard is a toll that can strengthen progress tracking. It was concluded that unless the E8 standardizes indicators/tools/thresholds/definitions/harmonizes policies, the goal to eliminate malaria in the 4 front-line countries by 2020 will not be achieved. Key words: resurgence, import, outbreak, elimination
Page 47 of 170 THE CONTRIBUTION OF A SUSTANABLE VOLUNTEER BASE ON CROSS-BORDER MALARIA ELIMINATION: AN IMPACT ASESSMENT
Abstract Theme Regional malaria and crossborder issues affecting elimination
Authors and Affiliation Njovu, C. (1); Givens, D. (1)
(1) J.C. Flowers Foundation Isdell:Flowers Cross Border Malaria Initiative
Abstract Since 2004 the Isdell:Flowers Cross Border Malaria Initiative (I:FCBMI) has supported extensive malaria prevention work in “last mile” communities, where national health systems are challenged by cross-border disparities in resources and malaria delivery tactics, as well as cost and logistic challenges. The engagement of Malaria Community Volunteers (MCVs) is a core function of I:FCBMI, which serves to strengthen health system capacity to reach remote and vulnerable communities. An impact assessment was carried out in order to measure the contribution of I:FCBMI MCVs to malaria elimination goals within the border regions of Namibia, Angola, Zimbabwe, and Zambia. In 2010, we focused our work on the cross border regions of Namibia, Angola, Zimbabwe and Zambia in support of the E8 strategy. In these communities, national malaria prevention interventions were promoted and carried out by MCVs who live in the area. Once recruited, MCVs were trained to conduct community mobilization, malaria education and sensitization, rapid diagnostic testing, LLIN distribution, and household data collection under the supervision of I:FCBMI country coordinators and focal points based in the cross-border areas. In the 4 countries, 885 unpaid MCVs were trained in partnership with local churches and governments on both sides of each border. This work has been coupled with advocacy for cross border policy and harmonization, as well as operational research, including effective monitoring and evaluation. Quantitative and qualitative data from four case studies reveal the impact of MCV activities. The impact assessment demonstrates that MCVs were able to extend the reach of formal government and partner programs, contributing to measurable impact on malaria control in these hard to reach areas. Innovative partnerships will be critical to further the goal of elimination in challenging cross-border areas. The evaluation underscores the importance of community mobilization and volunteer retention to achieve a sustained decrease of malaria incidence rates in each community.
Page 48 of 170 FACILITATING FACTORS AND BARRIERS TO MALARIA RESEARCH UTILIZATION FOR POLICY DEVELOPMENT IN MALAWI
Abstract Theme Education & health promotion
Authors and Affiliations Mwendera, C.A. (1); de Jager, C. (1); Longwe, H. (2); Phiri, K. (3); Hongoro, C. (1)(4); Mutero, C.M. (1)(5)
(1) University of Pretoria Institute for Sustainable Malaria Control (UP ISMC), School of Health Systems and Public Health, University of Pretoria, Pretoria, South Africa; (2) ICAP at Columbia University, Mailman School of Public Health, Pretoria, South Africa; (3) School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; (4) Population Health, Health Systems and Innovation, Human Sciences Research Council (HSRC), Pretoria, South Africa; (5) International Centre of Insect Physiology and Ecology, Nairobi, Kenya
Abstract Faced with challenges of limited resources, developing countries need to implement interventions with proven track record. Research data is key in providing evidence for policy development leading to successful interventions. However, research utilization in policy development is an intricate process requiring the understanding of contextual factors. There are factors which facilitate while others inhibit research utilization for policy development in a country. This study was conducted to assess factors which facilitate and/or hinder malaria research utilization for policy development in Malawi. We used a qualitative approach through in-depth interviews with 39 key informants that included malaria researchers, policy makers, programme managers, and key stakeholders. Purposive sampling and snowballing technique were used in identifying key informants. Interview transcript were entered in NVIVO 11 software for coding and analysis. The results showed that facilitating factors include global efforts in championing evidence informed policy making through knowledge translation, which has percolated to developing countries. Malawi government's commitment has also shown to be key in this cause. Various government tools (such as guidelines for policy development and analysis, and the National Health Research Agenda), and the establishment of institutions such as the Knowledge Translation Platform, and Knowledge Translation Unit, have been identified to pursue utilization of research in policy development. Other facilitating factors include; the availability of research institutions and their collaboration with the National Malaria Control Programme; stakeholders supporting and funding malaria research. The barriers to research ulitization in policy development included the lack of platforms for researchers to engage with the public; politics; researchers' lack of communication skills to policy makers, collaboration with policy makers and among researchers themselves; funder driven research; unknown World Health Organization policy position; and the lack of a malaria research repository. Overall, the study identified facilitating factors to malaria research utilization for policy development. These factors need to be systematically coordinated to address the identified barriers and improve on research utilization. Hence, the need to develop a framework than can facilitate this process.
Page 49 of 170 COMPARING CANINE BABESIA ROSSI WITH HUMAN FALCIPARUM MALARIA
Abstract Theme Modeling towards elimination
Authors and Affiliations Leisewitz, A. (1); Goddard, A. (1); De Gier, J. (2); Van Engelshoven, J.M.A.J.A. (2); Schoeman, J. (1)
(1) Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, South Africa; (2) Utrecht University, Veterinary Students
Abstract Babesia infection has been compared to human malaria infection for many decades. There are aspects of the dog disease that may well provide a unique animal model useful to the malaria research community. We collected clinical, haematological, biochemical and cytokine data (on a subset) on 320 cases of community acquired B. rossi infection. Data was not complete on all cases. 239/320 (74,69%) were uncomplicated and 81/320 (25,31%) were complicated. 47/320 (14,69%) were collapsed at presentation and 34/320 died (10,63%). 61/317 (19,24%) were hypoglycaemic (<3,3 mmol/L) with the complicated group having significantly lower glucose than the uncomplicated (p<0,01). 182/209 (87,08%) were hyperlactataemic (>1,0 mmol/L) with the complicated group being more so than the uncomplicated (p<0,01). 264/ 317 (83,28%) were anaemic (PCV<37%) and 3 cases were haemoconcentrated (PCV>55%) and all 3 died. There was a significant difference between the haematocrit of complicated and uncomplicated cases (p<0,01). Bilirubin (>25,6 umol/L) was elevated in 57/230 (24,78%) and the degree of elevation was correlated with the disease severity (p<0,001). Acute nephrosis is a serious complication. 31/320 (9,69%) had elevated creatinine (>133 umol/L) and there was a significant difference between those that died and survived (p<0,01). Using accepted criteria for defining systemic inflammatory syndrome, the syndrome was more likely to be present in complicated cases (60/81 complicated cases were SIRS positive). Parasitaemia was predictive of disease severity with significant differences present between complicated and uncomplicated cases (P<0,01). The concentration of GM-CSF, IFNγ, IL-2, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, MCP-1 and TNFα were determined in a subset of cases. IL-6, IL-10, MCP-1 and TNFα were elevated in 26/55 (47,27%), 54/55 (98,18%), 44/55 (80%) and 2/55 (3,64%) of ill dogs respectively (p<0,001). IL-8 was lower in sick dogs (p<0,001). IL-6, MCP-1 and TNFα were associated with disease severity. The criteria used to describe clinical canine babesia here are typically used to describe and classify malaria in humans. There are clear similarities between these two diseases suggesting that canine babesiosis should be investigated for its model potential.
Page 50 of 170 STARVING MALARIA: TARGETING NUTRIENT UPTAKE IN THE SEXUAL, TRANSMISSIBLE STAGES OF PLASMODIUM FALCIPARUM PARASITES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations van Biljon, R.A. (1); Naude, M. (1); Birkholtz, L. (1); Niemand, J. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa.
Abstract The P. falciparum parasite has to contend with a series of environmental changes as the parasite progresses through its life cycle from the sexual stage in the Anopheline vector, through the bloodstream of the human host and various intracellular locations within the hepatocytes and erythrocytes of the human host. This has led to unique evolutionary adaptations that allow survival in these changing environments. It is known that the asexual intraerythrocytic stages of the P. falciparum parasite have developed a range of strategies to obtain essential nutrients from the extracellular environment, as well as to facilitate waste disposal. These processes are effective drug targets since uptake and export mechanisms are either used by antimalarial agents to reach their target site, or are directly targeted to kill the parasite, as is the case with the new spiroindolone class of antimalarials. Furthermore, in P. falciparum parasites, transporters can also be involved in antimalarial drug resistance, as is the case with chloroquine, amongst others. Given the importance of uptake processes, it is therefore surprising that there is extremely limited information available about these processes in the sexual, transmissible form of the parasite. The permeome of sexual stage P. falciparum gametocytes was characterized by in silico analysis of the genome to identify all possible uptake proteins. The presence of the transcripts of these genes during gametocytogenesis was determined using DNA microarray analysis. Global functional characterization of the uptake and subsequent metabolism of a variety of carbon and nitrogen substrates by gametocytes was performed using high-throughput Phenotype MicroArrays™. Channels, transporters and uptake processes present in gametocytes were identified and classified. The presence of these uptake processes during stage I-V of gametocytogenesis was determined and correlated with the cellular energy metabolism pathways of these life cycle stages, thereby clarifying phenotypic differences between stage I-V gametocytes. Nutrient uptake processes in P. falciparum gametocytes were characterized in order to ultimately target these as transmission blocking strategies.
Page 51 of 170 PLASMODIUM FALCIPARUM GAMETOCYTES SHOW STAGE-SPECIFIC, CHEMOTYPE SPECIFIC TRANSCRIPTIONAL RESPONSE TO DRUG PERTURBATION
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Van Biljon, R.A. (1); Altenhofen, L. (2); Van Wyk, R.D.J. (1); Niemand, J. (1); Paquet, T. (3); Chibale, K. (3); Llinás, M. (2); Birkholtz, L.M. (1)
(1) Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, Private bag x20, Hatfield, Pretoria, South Africa, 0028; (2) Pennsylvania State University, University Park, PA 16802, USA; (3) H3D, Drug Discovery and Development Centre, University of Cape Town, South Africa, 7701
Abstract The discovery of transmission blocking drugs that target the sexual stages of Plasmodium falciparum parasites is a key component of an effective malaria elimination strategy. P. falciparum gametocytes undergo an extended, two week maturation process, making it difficult to pinpoint precise times in development where targeting the sexual stage will be effective. While one of the goals set for candidate antimalarials by the Medicines for Malaria Venture (MMV) is targeting multiple stages of the parasite lifecycle, chemical screening of a number of antiplasmodial compounds has illustrated not only differential susceptibility between asexual and sexual stage parasites but also between early and late stage P. falciparum gametocytes. The molecular mechanisms behind this phenomenon as well as the biological response of the metabolically inactive gametocyte under drug pressure are still mostly unexplored. In this study, two potent antiplasmodial compounds identified as part of a transmission-blocking drug screening platform by the South African Malaria Transmission Consortium (SAMTC) were investigated for their effect on the gametocyte transcriptome using microarray analysis. Each of the compounds showed differential potency when tested against early (stage II-III) or late (stage IV-V) gametocytes in vitro. Compound MMV666810 inhibited early gametocyte viability by 50% at 600 ± 88 nM and late-stage gametocyte viability at 180 ± 8 nM, while compound MMV674850 inhibited early gametocyte viability by 50% at 4.5 ± 3.6 nM and late-stage gametocyte viability at 28.7 ± 0.2 nM. A transcriptomic study conducted on gametocytes under treatment conditions (both early and late stage) yielded a unique transcriptional response, for immature and mature stages of gametocyte development as well as for each of the compounds in question. Among the transcripts affected by drug treatment were epigenetic regulators, transcriptional regulators and kinases that respond uniquely depending on the compound and stage of gametocytogenesis. These data describe the “fingerprint” of two potent antiplasmodial compounds on differentiating malaria parasites and demonstrate both the ability and specificity of the gametocyte to respond to drug exposure.
Page 52 of 170 SYNTHESIS AND BIOLOGICAL EVALUATION OF HETEROAROMATIC INHIBITORS OF PLAMSODIUM FALCIPARUM
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Rousseau, A.L. (1); Seanego, T.D. (1); Klein, H.F. (1); Molatsane, T. (1); van Zyl, R.L. (2) and Hoppe, H.C. (3)
(1) Molecular Sciences Institute, School of Chemistry, University of the Witwatersrand, Johannesburg 2000, Gauteng, South Africa; (2) Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, Gauteng, Johannesburg 2000, South Africa; (3) Department of Biochemistry and Microbiology, Rhodes University, Grahamstown 6140, South Africa
Abstract Our research has focused on the design and synthesis of small molecules with the potential to inhibit Plasmodium falciparum. One series of compounds have been designed to target folate metabolism in the parasite. Although drug resistance has limited the clinical usefulness of the most commonly used antifolate drugs, such as pyrimethamine and cycloguanil, antifolates are still attractive as antimalarials as they represent an inexpensive regime for combating chloroquine- resistant malaria, an important consideration for the treatment of malaria in Africa. We designed and synthesised a potent series of flexible cycloguanil analogues, active in the nanomolar range against drug resistant strains of P. falciparum. However, problems with the synthesis of these compounds and the fact that they were isolated as a mixture of enantiomers prompted us to consider pyrimidine analogues of these compounds. Synthetic approaches to these compounds and their antimalarial activity will be described. A second series of compounds we have focused on are designed to target the parasitic kinases PfCDPK1 and PfCDPK4. We hope to design a single compound capable of treating malaria and blocking transmission of malaria to the mosquito vector. Our progress on the synthesis of these compounds will be described.
Page 53 of 170 A COMPARATIVE, TRANSCRIPTOMIC ANALYSIS OF THE EFFECTS OF A LEAD CLINICAL CANDIDATE ON THE ASEXUAL AND GAMETOCYTE STAGES OF PLASMODIUM FALCIPARUM PARASITES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Connacher, J. (1), Niemand, J. (1), Paquet, T. (2), Chibale, K. (2); Birkholtz, L.M (1)
(1) Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, Private bag x20, Hatfield, Pretoria, South Africa; (2) University of Cape Town, Private Bag X3, Rondebosch 7701, South Africa
Abstract Malaria remains a serious health concern in the developing nations of Africa, Asia and South America. The World Health Organisation reported a total of 584 000 malaria related deaths for 2014 and African children under the age of 5 from the principal demographic group to be affected. Furthermore, resistance to the widely-used artemisinin combination therapies (ACTs) is rapidly developing across the globe with a similar trend as previously observed for formerly used antimalarial drugs. For this reason, new drug development is still at the forefront of malaria research. Specifically, the identification of antimalarial compounds using high-throughput screening (HTS) serves as a foundation for this. A novel series of compounds were recently identified through HTS as having antimalarial activity in the low nanomolar range against both the asexual and gametocyte stages of the malaria parasite (Younis et al., 2012). The lead compounds of the series, MMV390048 and MMV642943 are currently undergoing clinical trials. Despite this, no knowledge currently exists concerning the effects induced by these compounds on the P. falciparum parasite at the transcriptome wide level. The use of functional genomics tools such as microarrays can provide an understanding of the changes in gene expression that occur following drug treatment. In order to ascertain such changes, we used a P. falciparum specific microarray to determine transcriptomic changes that occur in asexual parasites as well as early and late gametocytes after treatment with MMV390048. The transcriptome of malaria parasites treated with this antimalarial was then also correlated to those of derivatives and lead backup compounds. This enabled specific indications of signature fingerprints that enable mode-of- action deconvolution of gametocytocidal compounds within this series. Network analysis indicates conserved nodes and neighbour analysis potential derivitisation functionality. Cross-sectional comparisons validate the singular biological action of MMV390048 across all life cycle stages of malaria parasites.
Page 54 of 170 QUANTITATIVE PROFILING OF THE PLASMODIUM FALCIPARUM PARASITE’S HISTONE POSTTRANSLATIONAL MODIFICATION LANDSCAPE DURING SEXUAL DIFFERENTIATION
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Coetzee, N. (1); Sidoli, S. (2); Garcia, B.A. (2); Birkholtz, L. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa; (2) Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA
Abstract The epigenetic gene regulatory mechanisms of the Plasmodium falciparum parasite have become increasingly investigated for chemotherapeutic applications. Histone post-translational modifications (PTMs) and the replacement of the core histones by variant histone proteins have been implicated in regulating gene expression during the intra-erythrocytic development of P. falciparum parasites and are hypothesized to contribute to sexual commitment and antigenic variation in the parasite. Epigenetic control has recently been directly linked to gametocyte commitment (regulated by the AP2-G transcription factor) and gametocyte overproduction (disruption of heterochromatin silencing involving heterochromatin protein 1 and histone deacetylase 2). Previous studies have identified histone PTMs in mixed asexual populations, however, important epigenetic regulatory mechanisms may be present during gametocytogenesis that could be vital for malaria transmission, if not pathogenesis. In this study, we present the quantitative landscape of histone PTMs across the entire asexual and sexual P. falciparum parasite’s life cycle in human blood. Intact histone proteins were acid-extracted from eight representative P. falciparum life cycle stages (ring, trophozoite and schizont asexual stages and stage I-V gametocytes) and propionylated to be analyzed using LC-MS/MS. The dataset serves to identify and quantify novel and previously identified P. falciparum histone PTMs, as the parasites progress through asexual and sexual development. The relative histone yield was found to increase (P<0.001) as the parasites progress through asexual development, to early and late gametocyte development. The histone PTM landscapes of the eight life cycle stages were found to have a highly conserved distribution of known euchromatin and heterochromatin marks (e.g. H3K9ac, H4ac, H3K9me3, etc.). Asexual stage associated histone PTMs include known activation and silencing marks (e.g. H3K14ac and H3K9me1), whereas sexual stage associated histone PTMs include mostly known activation marks (e.g. H3K4me1, H3K18ac and H3K23ac). Only a few histone PTMs were found to be associated with differential expression during life cycle development. This comprehensive characterization of histone PTMs enables deciphering of the degree to which it forms part of the stage-specific epigenetic gene regulation in malaria parasites.
Page 55 of 170 CROSS-REACTIVITY OF OXIDANT DRUGS AGAINST ASEXUAL AND GAMETOCYTE STAGES IN PLASMODIUM FALCIPARUM PARASITES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Coertzen, D. (1); Reader, J. (1); Botha, M.E. (1); de Kock, C. (2); Smith, P. (2); Haynes, R. K. (3); Birkholtz, L. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa; (2) Division of Pharmacology, Department of Medicine, Groote Schuur hospital, Old Main Building K45, University of Cape Town, Observatory, 7925; (3) Centre of Excellence for Pharmaceutical Sciences, Faculty of Health Sciences, North-West University, Potchefstroom, South Africa
Abstract Despite a decline in disease incidence due to the implementation of various control strategies, effective malaria elimination and eradication remains highly dependent on chemotherapeutic intervention through the use of antimalarial drugs. However, the formation of drug resistance against antimalarial combination therapies threatens the use of current treatment strategies, therefore the continuous development of novel antimalarial drugs is required to prevent untreatable infections from occurring. Artemisinin derivatives, the last line of effective antimalarial drugs, have shown to elicit a highly selective cytotoxic effect against Plasmodium parasites due to induction of an oxidative stress response during the intraerythrocytic stage of the parasites life cycle. The aim of this study is to determine the in vitro activity of several lead oxidant drugs as well as their novel derivatives against both the asexual and gametocyte stages of P. falciparum parasites. The identification of lead oxidative compounds that show cross-stage activity aim to be developed into triple-drug antimalarial therapies in combination with redox drugs and quinolone drugs, for which their respective activities are also being determined in this study. The activity of 10- alkylaminoartemisinin oxidant compounds were evaluated for their activity against in vitro cultured drug sensitive and drug resistant P. falciparum asexual parasites, using a SYBR Green-I fluorescence based proliferative assay and the parasite lactate-dehydrogenase metabolic assay. Following activity determination against asexual parasites, the oxidant compounds were tested against in vitro produced early and late stage gametocytes using a luciferase based viability assay. The dose- responses of these compounds against asexual P. falciparum drug resistant and non-resistant strains were determined in the low nanomolar ranges with negligible drug cross-resistance. Furthermore, activities in the nanomolar ranges against early and late gametocytes were also observed, indicating that these oxidant drugs are cross-active against both the asexual and gametocyte stages. Subsequent derivitization of lead drugs to improve compound solubility/stability also showed increased activity and selectivity for asexual and gametocyte stages. Therefore, the oxidative stress induced by these artemisinin derivatives against both asexual and gametocyte stages of P. falciparum parasites not only displays a high antiplasmodial potency but is also indicative of their potential transmission blocking activity.
Page 56 of 170 HETEROGENEITY OF PLASMODIUM FALCIPARUM MALARIA IN MATABELELAND SOUTH: ANALYSIS TRAVEL HISTORY REPORTED BY MALARIA CASES REPORTED IN ROUTINE SURVEILLANCE
Abstract Theme Regional malaria and crossborder issues affecting malaria
Authors and Affiliations Mberikunashe, J. (1); Maponga, A.B. (2); Chamberlain, H. (3); Alegana, V. (3); Tatem, A.J. (3), Pindolia, D. (4); Ahmed, J. (4); Woolheater, K. (4); Dolenz, C. (4); Le Menach, A. (4); Svisva, A. (4); Owen, J. (4); Madinga, M. (4); Gambinga, B. (4)
(1) Zimbabwe National Malaria Control Programme, Harare; (2) Ministry of Health and Child Care, Matabeleland South, Zimbabwe; (3) WorldPop, Geography and Environment, University of Southampton, UK; (4) Clinton Health Access Initiative, Boston, MA, USA
Abstract As malaria prevalence decreases, importation of malaria parasites poses a challenge to malaria programs due to its potential to instigate local transmission. Travel history collected from RDT- confirmed cases was used to estimate the potential for malaria importation into Matabeleland South province of Zimbabwe, which launched its malaria elimination agenda in 2013. Travel history records from 1,190 confirmed cases reported between September 2014 and June 2015 were analysed to determine the number of cases that travelled within Matabeleland South province, to other parts of Zimbabwe, and those that travelled outside Zimbabwe in the four weeks before diagnosis. Reported travel locations of cases were geo-located. Of the 1,190 cases, 185 (15.5%) reported to have travelled outside their home in the past 4 weeks, of which 99 (53.5%) travelled domestically and 86 (46.5%) travelled internationally. Of the seven districts in Matabeleland South province, Beitbridge (63%) and Gwanda (21%) reported the greatest proportions of cases that travelled. Of cases that travelled outside Matabeleland South, 44.3% were reported in low transmission risk areas (<1 case per 1,000 population) of the province and 20% were reported in high and medium transmission risk areas (>10 cases per 1,000 population). Environmental Health Technicians classified 80 (6.7%) of all 1,190 confirmed cases as imported (transmission occurred outside Matabeleland South), 11 (0.9%) as intraported (transmission occurred outside case’s home district but within Matabeleland South), and 744 (62.5%) as local (transmission occurred in home district of case). Locations reported in travel history data were often described by coarse spatial and temporal resolutions, which may have led to inaccurate case classification. Characteristics such as age, gender, and occupation will be explored to determine the demographic differences between travellers and non-travellers. Improvements in data collection, validation of travel history and its link to infection transmission (including clustering of local cases) are needed for a better understanding of parasite movement as a potential barrier to successful national and regional elimination of malaria.
Page 57 of 170 ASSESSING RISK FACTORS ASSOCIATED WITH PLASMODIUM FALCIPARUM EXPOSURE USING SEROLOGICAL DATA IN BUSHBUCKRIDGE MUNICIPALITY, MPUMALANGA PROVINCE, SOUTH AFRICA
Abstract Theme Epidemiology
Authors and Affiliations Biggs, J. (1); Cook, J. (1); Raman, J. (2); Agubuzo, E. (2, 3); Morris, N. (4); Seocharan, I. (4); Hlongwana, K. (5); Mabuza, A. (6); Coetzee, M. (2,3); Kleinschmidt, I. (1, 7).
(1) London School of Hygiene and Tropical Medicine, London, UK. (2) Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Johannesburg, South Africa. (3) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa (4) Medical Research Council, Durban, South Africa (5) University of Kwazulu/Natal, Durban, South Africa (6) Department of Health and Social Services, Mpumalanga Provincial Government, Nelspruit, South Africa. (7) School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa
Abstract Malaria transmission intensity in South Africa has reached the WHO recommended pre-elimination threshold. Monitoring the effectiveness of current control interventions is however proving challenging as commonly used tools such as entomological inoculation rate and parasite rate have reduced sensitivity in low transmission settings. In this study we evaluated the potential of assessing the prevalence of anti-bodies to malarial antigens using sero-prevalence data to identify potential risk factors associated with low transmission malaria endemic district of South Africa. Finger-prick filter paper blood spots and demographic data were collected from 2,897 individuals across 28 study clusters in Bushbuckridge, Mpumalanga Province, between April and June, 2015. Sera eluates from the filter paper blood spots were assayed for human anti-malaria antibodies against P. falciparum AMA-1 and MSP-119 antigens, using a quantitative ELISA. Of the individuals surveyed 21.3% were sero-positive for AMA-1, 16.3% for MSP-119,and 31.4% sero-positive for both antigens.The odds of being sero-positive increased with age (p<0.001) and if the individual had noformal secondary schooling (p=0.038). No association between sero-positivity and travel (p=0.309), out-door night- time activities (p=0.259), employment status (p=0.902) or vector control measures (p=0.815) was found in this study. Sero-positivity to AMA-1 was higher than sero-positivityto MSP-119. This correlates with a higher seroconversion rate for AMA-1 compared to MSP-1 demonstrating AMA-1, alone, is a more suitable serological marker to detect more malaria exposed individuals. Poor educational status was identified as a potential risk factor for malaria exposure, however due to reverse causality, more needs to be revealed to determine whether poor education causes malaria exposure or does malaria exposure result in attaining a poor level of education. Recent transmission was revealed due to a proportion of under 5 year olds being seropositive, therefore were exposed to malaria in the past 5 years highlighting transmission is ongoing. Data from this study supports the use of serology to monitor malaria transmission in low transmission South African settings.
Page 58 of 170 PREVALENCE OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PD) DEFICIENT PHENOTYPES AND GENOTYPES AT SELECT POINT OF CARE HEALTH SETTINGS IN VHEMBE DISTRICT, LIMPOPO PROVINCE, SOUTH AFRICA
Abstract Theme Epidemiology
Authors and Affiliations Awandu, S.S. (1); Raman, J. (1, 2, 3); Kruger, P. (4); Frean, J. (2, 3); Birkholtz, LM. (1)
(1) Malaria Parasite Molecular Laboratory, UP Institute for Sustainable Malaria Control University of Pretoria, Pretoria, South Africa; (2) Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Johannesburg, South Africa; (3) Wits Research Institute for Malaria Research, Faculty of Health Sciences, University of Witwatersrand, Johannesburg, South Africa; (4) Limpopo Malaria Control Program, Limpopo, South Africa
Abstract South Africa is targeting malaria elimination by the year 2018. The World Health Organization recently recommended the antimalarial, primaquine be added to standard malaria treatment in eliminating countries to further reduce transmission intensity by clearing gametocytes, the transmissible stage of Plasmodium species. Unfortunately, individuals with glucose-6-phosphate dehydrogenase deficiency (G6PDd) risk acute haemolytic anaemia if exposed to primaquine, particularly at high doses. We evaluated the prevalence of G6PDd in Vhembe District, Limpopo Province South Africa, using a commercial rapid diagnostic test (RDT) kit and compared its performance with genotypic analyses. A total of 248 individuals attending six primary health care facilities in Vhembe District, were G6PD phenotyped using CareStartTM G6PDd RDT kits and genotyped by polymerase chain reaction, restriction length fragment polymorphisms and DNA sequencing. Prevalence of the most common African G6PDd variants, namely G6PD A (A376G) and G6PD A- (G202A, A542T, G680T and T968C) was assessed. Prevalence of G6PDd (combination of both mild and severe deficiencies) in Vhembe District was 13% (33/248) according to the CareStart™ RDT kit. Although more males (18%:10/55) than females (12%: 23/193), were found to G6PDd by RDT, this difference was not statistically significant (P = 0.23). Genotypic analysis found a much lower G6PDd prevalence, with only 3% of the study participants carrying the A376G/G202A genotype (6/248; 2.42% [n=6] male hemizygous and 0.80% [n=2] female homozygous). The A542T, G680T or T968C variants were not detected in any of the analysed samples. Sensitivity and specificity of the commercial RDT compared to the genotypic analyses was 90% (95 % CI 85.54–94.03%) and 33% (95 % CI 18.57–49.13 %), respectively. The agreement between test results was poor with a Kappa value of 0.246 (95% CI 0.090-0.413). The low prevalence of individuals carrying the G6PDd A376G/G202A mutations in the study area corresponds to published data from nearby areas including Pedi and Tsonga populations. This suggests the deployment of single low dose primaquine possess a minimal risk to the Vhembe population. The poor specificity of the CareStart™ kit compared to the genotypic analyses needs further investigation.
Page 59 of 170 HETEROGENEITY OF PLASMODIUM FALCIPARUM TRANSMISSION IN NORTH-EASTERN SOUTH AFRICA: IMPLICATIONS FOR MALARIA ELIMINATION
Abstract Theme Epidemiology
Authors and Affiliations Biggs, J.(1); Raman, J.(2, 3, 4); Cook, J.(1); Morris, N.(5); Serocharan, I.(5); Hlongwana, K.(6); Agubuzo, E.(2, 3); Hall, T.(1); Drakeley, C.(1); Mabuza, A.(7); Kruger, P.(8); Zitha, A.(7); Machaba, E.(8); Zitha, M.(3); Coetzee, M.(2, 3); Kleinschmidt, I.(1, 3).
(1) Department of Infectious Disease Epidemiology, London School of Hygiene and Tropical Medicine, London, United Kingdom; (2) Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Johannesburg, South Africa; (3) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (4) Centre for Sustainable Malaria Control, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa; (5) Health GIS Centre, South African Medical Research Council, Durban, South Africa; (6) School of Nursing and Public Health, University of KwaZulu-Natal, Durban, South Africa; (7) Mpumalanga Provincial Malaria Control Programme, Nelspruit, South Africa ; (8) Limpopo Provincial Malaria Control Programme, Polokwane, South Africa
Abstract It is widely agreed that modifications to existing control interventions are needed if South Africa is to achieve malaria elimination. One modification currently being trialled in South Africa, is targeted indoor residual spraying (IRS) instead of annual generalised IRS. This sero-prevalence baseline investigation, which forms part of the trial, was undertaken to provide evidence into the suitability of targeted interventions by providing insights into potential risk factors, transmission dynamics, and spatial characteristics associated with malaria exposure in the Ba-Phalaborwa, Limpopo and Bushbuckridge, Mpumalanga. Finger-prick filter-paper blood spots (4 948) and 2710 completed questionnaires were collected from randomly selected households in selected wards across the study area during the baseline household survey between April and June 2015. IgG antibodies to both Plasmodium falciparum AMA-1 and MSP-119 blood stage antigens were eluted from the filter-paper blood spots and assayed by quantitative ELISA. Individuals were then categorised as either sero- negative or sero-positive to P. falciparum. The seroconversion rate (SCR) was calculated from a simple compartmental model. Serological data were then linked to household demographic, questionnaire and spatial data for further analysis. Across the study area, 25% of the sampled population were sero-positive to either antigen, with a SCR of 0.05 (95% CI: 0.032-0.077). After stratifying individuals by study site, SCRs of 0.172 (0.046-0.645) and 0.037 (0.019-0.074) were reported in Bushbuckridge and Ba-Phalaborwa respectively. Within both study sites geospatial analysis revealed statistically significant clusters (p value >0.05) of sero- positive households. Ba-Phalaborwa had higher sero-positive cluster relative risk values and more focal household averaged, age-adjusted, PfAMA-1 antibody responses compared to Bushbuckridge. Although transmission intensity and exposure to malaria across both study sites was low, there was significant transmission heterogeneity within and between study areas, characterised by the presence of malaria transmission hotspots. Ba-Phalaborwa exhibited a higher degree of focal clustering of seropositive households, implying a higher incidence of focal transmission compared to Bushbuckridge. The identified heterogeneous transmission intensity suggests targeted interventions maybe more appropriate in certain areas. However, more data generated from a more robust sample set is required before concrete recommendations targeted interventions can be made.
Page 60 of 170 INFECTION OF AFRICAN MALARIA VECTOR MOSQUITOES WITH PLASMODIUM FALCIPARUM MALARIA PARASITES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Bennett, A. (1, 2); Hunt, R. (1, 2); Coetzee, M. (1, 2); Coetzer, T. (1, 3); Engelbrecht, D. (1, 3); Shunmugam, S. (1, 3); Bezuidenhout, B. (1, 3); Nkosi, N. (1, 2); Koekemoer. L.L. (1, 2).
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of the Health Sciences, University of the Witwatersrand, Johannesburg; (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, NICD/NHLS, Johannesburg; (3) Wits Research Institute for Malaria, School of Pathology, Faculty of the Health Sciences, Department of Molecular Medicine and Haematology, University of the Witwatersrand and National Health Laboratory Service, Johannesburg
Abstract The malaria parasite life cycle in the adult female mosquito has received renewed attention in recent years with the development of transmission-blocking strategies. The aim of the project was to infect African malaria vectors in a laboratory setting with Plasmodium falciparum gametocytes, to facilitate research and collaboration with other project partners interested in transmission-blocking initiatives. Life-stages of the parasite in the vector, in particular ookinete formation in the midgut, with subsequent oocyst formation on the midgut wall, sporozoite formation and migration to the salivary glands of the mosquito, are important measurable variables in infection studies. Malaria vectors, Anopheles arabiensis and An. coluzzii were fed on P. falciparum infected blood under two experimental mosquito treatment solutions, i.e. 0.05% gentamicin sulfate (40mg/ml;) and 0.05% para-aminobenzoic acid (PABA) made up in 10% sucrose. Adult mosquitoes were fed on the experimental solutions from the time of emergence to the time of dissection. Dissection of mosquito midguts was performed on days 9-10 after infection to determine oocyst prevalence and intensity, and salivary glands were dissected after 14 days to determine sporozoite presence. Anopheles arabiensis exposed to gentamicin showed a higher percentage of specimens with oocysts, when compared to the PABA treatment and when compared to An. coluzzii. Dissected salivary glands were found to be infected with sporozoites in both species when treated with gentamicin. Positive sporozoite results confirmed that infection in these vectors can be achieved under laboratory conditions. The infection is dependent on parasite cultures that produce viable gametocytes, as well as an adequate number of mated 3-5 day old female mosquitoes. This is the first time that a South African laboratory has succeeded in infecting mosquitoes with P. falciparum and it paves the way to embark on new research initiatives concerning the testing of transmission-blocking compounds in the future.
Page 61 of 170 ANOPHELES ARABIENSIS HABITAT SUITABILITY MODEL SPATIALLY PREDICTS MALARIA HOTSPOTS IN ZIMBABWE
Abstract Theme Modeling towards elimination
Authors and Affiliations Gwitira, I. (1), Murwira, A. (1) Masocha, M. (1), Zengeya, F.Z. (1)
1) University of Zimbabwe, Department of Geography and Environmental Science
Abstract Malaria is a climate-sensitive protozoal disease caused by parasites of Plasmodium genus transmitted among humans through bites of female Anopheles mosquitoes. Despite recent progress in reducing malaria morbidity and mortality through intensification of malaria control programs, approximately half of the world remains at risk of malaria. Thus, developing and adopting the most effective ways to predict malaria occurrence, as a preamble to controlling the spread of malaria is critical as accurate mapping and prediction of malaria hotspots is an important step towards effective malaria control ultimately malaria elimination. We determined localised geographic clusters of malaria using the Getis Ord Gi* statistic based on malaria cases recorded at health facilities in Zimbabwe between1996 and 1999. We used Anopheles arabiensis habitat suitability models developed by Gwitira et al 2015. The Anopheles arabiensis habitat suitability was modelled using maximum entropy (Maxent) with altitude, isothermality, temperature seasonality, annual precipitation and precipitation of the wettest month being the significant predictors. We then used logistic regression to predict the probability of malaria hotspots as a function of Anopheles arabiensis habitat suitability. Our results show that Anopheles arabiensis habitat suitability positively and significantly (p<0.05) predicts malaria hotspots. Our results show that areas in the north, northeast, south and south eastern parts of Zimbabwe are characterised by statistically significant malaria hotspots. In fact, we observe that the probability of malaria hotspot occurrence is high in areas with high Anopheles arabiensis habitat suitability values. Based on the ROC values, we observe that habitat suitability of Anopheles arabiensis can successfully be used to predict malaria hotspots. Results of this study indicate that mosquito habitat suitability consistently and significantly predicts malaria hotspots. Specifically, results suggest that high Anopheles arabiensis habitat suitability results in high chances of malaria hotspot occurrence. This result is confirmed by studies that also found a significant association between malaria hotspots and environmental factors that are favourable for the malaria vectors elsewhere in Africa. Our results indicate a strong positive relationship between malaria hotspots and Anopheles arabiensis habitat suitability, which is expected in regions with relatively weak malaria control measures.
Page 62 of 170 IS ANOPHELES VANEEDENI A VECTOR OF MALARIA IN SOUTH AFRICA?
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Ashley Burke (1, 2), Maureen Coetzee (1, 2) & Basil Brooke (1, 2)
(1).Wits Research Institute for Malaria, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; (2) Centre for Opportunistic, Tropical & Hospital Infections, National Institute for Communicable Diseases, Johannesburg, South Africa
Abstract The major malaria vectors of South Africa are Anopheles arabiensis of the An, gambiae complex, and An. funestus of the An, funestus group. South Africa’s malaria affected areas include the low altitude border regions of Limpopo, Mpumamlanga and KwaZulu-Natal Provinces. Each of these provinces has developed well- coordinated malaria vector control operations which are based on the application of indoor residual insecticide spraying (IRS). Although IRS has proven efficacy, low-level residual malaria transmission continues and is likely caused by outdoor feeding and resting Anopheles vector mosquitoes that are unaffected by indoor applications of insecticide. Residual malaria transmission and burgeoning insecticide resistance in malaria vector populations within South Africa’s borders necessitate ongoing vector surveillance. Vector surveillance is needed to assess vector population composition (malaria vector incrimination) and abundance, and pertinent behavioural traits such as indoor vs outdoor resting and feeding. Vector surveillance was conducted for one year in two villages in Mpumalanga (Tonga - Block A and Vlakbult) which experience seasonal malaria transmission. Two passive methods of collection were used in order to compare their relative efficacies in terms of trapping adult Anopheles mosquitoes. These methods included the deployment of ceramic pots and modified buckets that were placed both inside and outside households and were cleared weekly. All anopheline mosquitoes from these traps were collected and preserved. These samples were later identified to species by morphology and PCR, and all females were screened for the presence of Plasmodium falciparum sporozoites by ELISA. Unfortunately, the sampling period (March 2015 – April 2016) coincided with the driest year South Africa has ever experienced on record. Vector population abundance and thus malaria transmission was significantly reduced, presenting a challenge for vector surveillance. Despite this, samples of Anopheles mosquitoes including potential vector species were still collected. The outdoor traps proved the most efficient passive collection method, while very few samples were collected indoors, which can be attributed to IRS operations in both villages. Anopheline species collected were An. merus, An. arabiensis, and An. quadriannulatus of the An. gambiae complex, and An. leesoni, An. rivulorum, and An. vaneedeni of the An. funestus group. The most abundant species collected was An. vaneedeni. All female samples tested negative for Plasmodium sporozoites, except for one An. vaneedeni female which tested positive through two ELISA assays, the second of which showed positive after boiling. This female mosquito was collected from an outdoor ceramic pot. Molecular work is currently underway to confirm this Plasmodium positive specimen using a nested PCR assay. This is the first time that An. vaneedeni has been tentatively implicated in malaria transmission. The outdoor resting trait of this species will likely have important implications for the efficacy of IRS based malaria vector control in South Africa.
Page 63 of 170 ANALYSIS OF THE ESTERASE ENZYMES ACTIVITY IN ANOPHELES FUNESTUS ADULTS
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Dahan-Moss, Y.L. (1, 2); Koekemoer, L. (1, 2)
(1) WITS Research Institute for Malaria, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, a division of the National Health Laboratory Service, Sandringham, Johannesburg, South Africa
Abstract Anopheles funestus is a major malaria vector in Africa and in order to apply effective control measures against this vector, it is necessary to understand the underlying physiological factors that play critical roles in metabolism, development, insecticide resistance, reproduction and fertility. Esterases are one class of enzymes involved in these biological pathways. The aim of our study was to analyse the esterases and their activity levels at different time points during the life span of adult An. funestus. Isoenzyme electrophoresis (IEE) was used to examine the esterase activity in the different ages and genders of the laboratory colonised and wild An. funestus adults. The esterase activity was semi quantified by densitometry analysis of the IEE gels. The esterases were further characterised according to their activity inhibition by organic phosphates, eserine sulphate and sulphydryl reagents. Nine esterases IEE profiles were shared between both the laboratory colonised and wild An. funestus adults. These esterases were further divided into acetylesterases, arylesterases, carboxylesterases and acetylcholinesterase. The activity level of certain specific esterases was primarily influenced by age and/or gender. The activity level of each specific esterases in the An. funestus adults was influenced by the age and/or gender of the An. funestus adult. The individual esterases were classified as either acetylcholinesterases, carboxylesterases, arylesterases or acetylesterases. This study provides an understanding of the esterases that are associated with the An. funestus adult, which could serve as valuable tools for effective control of this major African malaria vector.
Page 64 of 170 MODELLING THE SPATIO-TEMPORAL DYNAMICS OF MOSQUITOES AND ITS RESPONSE TO SIT CONTROL
Abstract Theme Modeling towards elimination
Authors and Affiliations Dufourd, C. (1); Anguelov, R. (1); Dumont, Y. (2)
(1) Department of Mathematics and Applied Mathematics, UP, Pretoria; and UP Institute for Sustainable Malaria Control (2) CIRAD, Umr AMAP, Montpellier, France
Abstract The aim of this presentation is to present some results on the distribution and abundance of a malaria-vector Anopheles mosquito population, taking into account environmental parameters such as wind and temperature, and its response to Sterile Insect Technique (SIT) control. To do so, we consider a compartmental approach to account for the interactions between groups of individuals within the population, and we model the dispersal of each compartment via an advection-diffusion- reaction process based on biological and behavioural assumptions. The resulting mathematical model is formulated by a system of partial differential equations which accounts for temporal variations of the population abundance and spatial heterogeneity. We then carry out numerical simulations for the dispersal of mosquitoes and simulate various SIT control scenarios. We show that environmental factors have a significant influence on the distribution of mosquitoes and the efficiency of the control method.
Page 65 of 170 THE EFFECTS OF AGROCHEMICALS ON THE LIFE HISTORY AND INSECTICIDE RESISTANCE PHENOTYPE OF THE MAJOR MALARIA VECTOR ANOPHELES ARABIENSIS (DIPTERA: CULICIDAE)
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Oliver, S.V. (1, 2); Moloi, K. (1, 2, 3) Brooke, B.D. (1, 2)
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases (3) School of Animal, Plants and Environmental Sciences, Faculty of Science, University of the Witwatersrand
Abstract Anopheles arabiensis is a major vector of malaria in southern Africa. This vector is associated with increased malaria incidence that is often associated with agricultural activities. Previous studies have associated increased larval access to food in agricultural breeding sites with increased adult mosquito fitness. In this study the effects of agricultural pollutants on the life history and insecticide resistance phenotype of two laboratory strains of An. arabiensis were examined. SENN, an unselected strain and SENN DDT, a selected strain displaying multiple resistance phenotypes, were exposed to various agricultural pollutants during the larval life stage. 0.05, 1 and 2% Nitrogen/Phosphate/Potassium (NPK) fertilizer and 0.02, 0.04 and 0.06% Urea, Superphosphate and Potassium Chloride and was used for fertilizer development and life history studies. 0.025, 0.05 and 0.1M glyphosphate was used for herbicide development study. Larvae were reared in 2% NPK and 0.1M glyphosphate for insecticide resistance studies, 3 day old, non bloodfed adults were used in standard WHO bioassays to assess changes in insecticide resistance phenotype. Glyphosphate inhibited the larval development of both strains in a concentration-dependent manner. Larval exposure to glyphosphate in SENN DDT augmented malathion and deltamethrin resistance. Nitrogen/Phosphate/Potassium (NPK) fertilizer did not significantly affect larval development in either strain. When examining the effects of single element fertilizers, potassium element fertilizer offered the greatest advantage to both strains in terms of larval development rate and size. Nitrogen element fertilizer resulted in the slowest development and the smallest adults. High concentrations of NPK fertilizer reduced adult male longevity in both strains. NPK fertilizer also augmented malathion resistance in adult male and female SENN DDT. The increased insecticide resistance phenotype appears to be mediated by increased detoxification and oxidative stress resistance enzyme activity. This study demonstrates that although agriculturally derived pollutants have variable effects on larval and adult life history characteristics, inorganic pollutants can increase the expression of pyrethroid and organophosphate resistance in adult mosquitoes. This study demonstrates that non-insecticidal pollutants can act a selection pressure that could drive the development of insecticide resistance. Therefore, pollutants have the potential to affect vector control efforts.
Page 66 of 170 MONITORING INSECTICIDE RESISTANCE IN THE AN. GAMBIAE COMPLEX FROM KWAZULU-NATAL, SOUTH AFRICA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Mofokeng, M. (1, 2); Munhenga, G. (1, 2); Koekemoer, L.L. (1, 2)
(1) Division of the National Health Laboratory Services, Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Private Bag X4, Johannesburg 2131 Sandringham, South Africa; (2) WITS Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa
Abstract Mosquitoes from the genus Anopheles are responsible for transmission of malaria, and in sub- Saharan Africa three of the four main vectors are species from the Anopheles gambiae complex, while the forth species are a member of the An. funestus group. Anopheles arabiensis is currently the main vector in countries such as South Africa, Sudan and Ethiopia. There have been many initiatives in controlling this vector which includes the use of insecticides. However, this is becoming problematic due to selection of resistance against insecticides being used. To maintain utility of insecticides and manage resistance there is need to continuously update information on the susceptibility status of vectors against insecticides being used against them. The aim of this study was to monitor An. arabiensis susceptibility to insecticides in a malaria endemic province in South Africa, KZN. Mosquitoes were sampled outdoors from Mamfene, northern KZN, South Africa (27°23’S, 32°12’E), using African clay-pots and trapping plastic buckets. Molecular techniques were used to identify collected specimens to species-specific level and determining Plasmodium infectivity. In addition, susceptibility to four classes of insecticides was analysed in pooled F1 progeny of An. arabiensis reared from wild-caught females using the standard WHO bioassay protocol. Species analysis showed that the majority of the samples collected were An. arabiensis contributing 96% of total specimens analysed. Of the 761specimens tested for Plasmodium falciparum infectivity none were positive. Insecticide susceptibility diagnostic tests showed resistance to DDT (77 % mean mortality) and suspected resistance to the pyrethroid deltamethrin, at a mean mortality of 95% and susceptible to both malathion (an organophosphate) and bendiocarb (a carbamate) with a mean mortality of 100%. These findings suggest that An. arabiensis population from KZN is resistant to DDT being used by the malaria control programme. Insecticide resistance management is important to prevent it impacting on the malaria control program.
Page 67 of 170 SELECTING MOST EFFICIENT TARGETS FOR SPATIALLY FOCUSED INTERVENTIONS: MALARIA INCIDENCE VERSUS POPULATION MOBILITY
Abstract Theme Modeling towards elimination
Authors and Affiliations Sallah, K. (1, 2); Giorgi, R. (1, 2); Gaudart, J. (1, 2)
(1) INSERM, UMR S912, « Sciences Economiques & Sociales de la Santé et Traitement de l’Information Médicale » (SESSTIM), F-13385, Marseille, France; (2) Aix-Marseille Université, SESSTIM UMR_S912, IRD, F-13385 Marseille, France.
Abstract Since the 2000s, malaria control programs have resulted in a significant decrease of incidences. Spatial heterogeneity of incidence and human mobility patterns are critical factors to handle towards elimination. Focusing interventions on geographical areas is actually a promising approach, but definition of the targeted areas is still needed. One approach is based on incidence estimations at local scale (needing spatial variability assessments). Another one is to target area where population mobility is important, with high potential of transmission dynamic (“hub” areas). Using simulation methods, this study aimed to compare mobility-based criteria and spatial incidence approaches in choosing geographical targets for any focused malaria intervention. Geographical fields were simulated 1000 times. Each field covered about one degree latitude and one degree longitude (110x110 km2) and was assigned 30 locations. Coordinates of locations were distributed according to uniform distributions. Population sizes, mobility rate in populations and malaria incidences followed Poisson distributions with respect to plausible ranges. Fields were classified according to WHO definition of malaria zones into control, pre-elimination and elimination stages (respectively > 5, between 1 and 5, and < 1 case per 1000/year). Local transmission was 1odelled according to a compartmental SIR (Susceptible, Infectious, and Recovered) design where mobility was 1odelled by using a radiation model. For each simulation, locations selected for intervention were defined on the basis of a mobility-based criteria and incidence criteria according to thresholds. Primary endpoint was relative decrease in cumulated malaria incidence within three year from the start of the intervention. Focused intervention efficacy was higher in pre-elimination/elimination phases 39%, 95% CI [36-41%], than in control phase 30%, 95% CI [23-35%], Student t-test p- value=0.03. Whatever the epidemiological context, focusing interventions on higher incidence locations yielded higher overall incidence decrease, than selecting targets on high mobility rate criteria, 37% versus 25%, Student t-test, p-value<0.001. In low incidence locations, better efficacy occurred targeting most mobile populations rather than highest incidences (23% versus 16%, p- value<0.001). Focused malaria interventions were more efficient in pre-elimination/elimination stages. To protect low incidence areas from malaria importation, interventions on mobile populations were more useful than hitting highest incidences.
Page 68 of 170 EVALUATING EFFECTS OF LANDSAT-DERIVED ENVIRONMENTAL COVARIATES FOR PREDICTING MALARIA DISTRIBUTION IN RURAL VILLAGES OF VHEMBE DISTRICT, SOUTH AFRICA
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Malahlela, O.E. (1, 2); Olwoch, J.M., (1, 2); Adjorlolo, C. (2)
(1) Department of Geography, Geoinformatics and Meteorology, University of Pretoria; and UP Institute for Sustainable Malaria Control (2) South African National Space Agency (SANSA), Earth Observation Directorate
Abstract Malaria in Africa is still a problem despite existing efforts to eradicate the disease. In South Africa, the Vhembe District Municipality (VDM) is continuously experiencing high malaria prevalence. Evaluating local environmental factors that affect malaria distribution is necessary for efficient disease management and control. Based on epidemiological data collected in 2005 and corresponding Landsat 5 data, we analysed the effect of Landsat-derived environmental covariates on malaria distribution in VDM area. A total of 9 remotely-sensed covariates were derived including the NDVI, SAVI, normalized difference water indices (NDWI1 and NDWI2), green index, yellowness index, water indices (a1 and a2) and pseudo-absences in the ratio of 1:2 (presence/absence) were drawn at buffer distances of 0.5km – 20km from known presence locations. A stepwise logistic regression model was applied to analyse spatial distribution of malaria. Results of logistic regression showed that Landsat-derived covariates are significant and could be used to model malaria distribution. A buffer distance of 10 km yielded the highest classification accuracy of 82% at a threshold of 0.9. This model was significant (ρ < 0.05) and yielded a deviance (D2) of 36%. The significantly positive relationship (ρ < 0.05) between soil-adjusted vegetation index (SAVI) and malaria distribution at all buffer distances suggests that malaria vectors (Anopheles arabiensis) prefer highly productive, greener vegetation. The significant negative relationship
between water index (a1) and malaria distribution in buffer distances of 0.5km, 10km and 20km suggest that malaria distribution increases with increase in absorption in vegetation moisture.
Landsat-derived SAVI and water sensitive index (a1) can be used to model malaria distribution in semi-arid rural areas with very close proximity (~20km). The study has shown that suitable habitats of malaria vector are generally found within a radius of 10km in semi-arid environments and this insight can be useful to aid efforts aimed at malaria eradication by 2018 in South Africa. The study has also demonstrated that the distribution of malaria vector (An. arabiensis) and its pathogen (Plasmodium falciparum) is in response to moisture content in vegetation which serves as a surrogate for rainfall.
Page 69 of 170 ANALYSING TRENDS AND FORECASTING MALARIA EPIDEMICS IN MADAGASCAR USING A SENTINEL SURVEILLANCE NETWORK: A WEB-BASED APPLICATION
Abstract Theme Epidemiology
Authors and Affiliations Florian Girond
Abstract In Madagascar, malaria incidence has decreased in recent decades mainly due to successful malaria control interventions. However, an upsurge of malaria outbreaks in recent years stressed the need for a Malaria Early Warning System (MEWS) adapted to Malagasy resources. A sentinel surveillance with high quality data and rapid reporting mechanisms is an alternative to relying solely on data collected through the country’s routine Health Management Information Systems. The development and implementation of a technology-based Early Warning System is a major step towards efficient early detection of malaria outbreaks and rapid responses. However the first stages of its development and implementation are marked by scientific and technical challenges mainly due to the lack of consensus on epidemic definition. Since 2007, malaria data from 34 sentinel sites throughout Madagascar are collected on a daily basis by Short Message Service (SMS). Concomitantly, site-specific satellite weather data (temperature, rainfall, Normalized Difference Vegetation Index) are routinely captured from the Internet, while malaria control interventions (Long Lasting Insecticidal Nets, Indoor Residual Spraying) are provided by Roll Back Malaria partners. Data are automatically analyzed to detect malaria trends and malaria outbreak alerts with automated feedback reports. We describe a system using various epidemic thresholds and a forecasting component with the support of new technologies to improve the performance of a sentinel-MEWS. Our system uses a percentile-based detection method and has already demonstrated reliability in detecting malaria outbreaks in the south-eastern part of Madagascar in 2014. Its Web-based surveillance system, with automated analysis and timely output, allowed real- time monitoring of malaria trends and control intervention with RBM partners. This work aims to maximize the usefulness of a sentinel surveillance system to predict and detect epidemics in limited- resource environments and to provide practical examples and suggestions for use in other systems or settings.
Page 70 of 170 PYRETHROID-IMPREGNATED WALL LININGS: THREE YEARS POST INSTALLATION DURABILITY, USER ACCEPTABILITY, PERCEIVED EFFECTIVENESS, AND LABORATORY CONFIRMED EFFICACY
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Ramothole, M. (1); Kruger, T. (1); de Jager, C. (1)
(1) University of Pretoria School of Health Systems and Public Health (SHSPH) and University of Pretoria Institute for Sustainable Malaria Control (UP ISMC), University of Pretoria, South Africa;
Abstract Vector control is one of the most important malaria control interventions that has a proven track record of effectively controlling transmission of malaria if done correctly. The WHO recommends Long-lasting Insecticide Treated Nets (LLINs) or Indoor Residual Spraying (IRS) or a combination of both be used to protect people at risk. Both these methods however, have their own respective disadvantages. IRS is costly and labour intensive while LLINs can increase discomfort in hot, humid environments by interrupting air flow. Insecticide impregnated indoor lining; a potentially safer and more sustainable vector control method combines certain benefits of IRS and LLINs. This study evaluated the long-term durability, perceived effectiveness and user acceptability of linings installed in Vhembe District, Limpopo Province. Linings were installed in 40 dwellings (20 huts and 20 houses) in Tshilivho village in October 2012. Following an initial six-month pilot study, linings were left in dwellings to determine long-term efficacy and user acceptance. Structured questionnaires were completed in October 2015, three years following installation, to establish user acceptance and lining durability. Lining efficacy was determined from installation to October 2015 according toWHO recommended bioassays to assess long-term efficacy. All participants responded positively towards the linings and said that they would purchase linings if they were available and affordable. Some changes or damage to the linings were noticed by 10/40 participants. Three linings were lost due to collapsed huts. Of the 37 participants that still had linings, 36 wanted to keep the lining for longer. Linings were reported as still effective against mosquitoes and other annoying insects by 33/37 participants. Participants who lost their linings noted lining effectiveness until they were lost. Efficacy was measured against the WHO efficacy criteria for IRS of >95% knockdown (KD) after test mosquitoes were exposed to lining samples for 30 minutes, and >80% mortality at 24 hours after initial exposure. KD and mortality for all analyses were above efficacy criteria. The linings are highly accepted among study participants and based on bioassays these linings could remain effective for at least three years providing protection against mosquitoes and other annoying insects.
Page 71 of 170 SUSTAINED RELEASE OF REPELLENT MIXTURES THROUGH ENGINEERING THE MOLECULAR INTERACTIONS
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Izadi, H. (1); Focke, W.W. (1); Maharaj, R. (1, 2)
(1) University of Pretoria Institute for Sustainable Malaria Control, Faculty of Health Sciences, University of Pretoria, South Africa; (2) Office of Malaria Research, Medical Research Council, Durban South Africa
Abstract Repellents are considered a key component to control malaria transmission by providing outdoor personal protection. To be effective, the repellent formulation should cost-effectively provide good repellence activity over extended times while being non-toxic and non-irritant to the user. Modulating the evaporation rate of repellents remains a challenge despite extensive efforts to implement controlled-release techniques. Engineering the molecular interactions between the components in repellent formulations is a novel, alternative approach, towards reducing the volatility of the repellent composition. Ethyl butylacetylaminopropionate, commercially known as IR3535, and nonanoic acid were selected as the model compounds. The evaporation of mixtures of different composition was studied by thermogravimetric methods in a convection oven set at 50°C. The mass loss as a function of time was recorded. The change in the composition of the liquid phase was determined using Fourier transform infrared spectroscopy. Repellency assays were performed with 3-5 day old female An. arabiensis that had been starved for 6 hours but previously fed on 10% sucrose solution. Repellent activity was assessed by topical application of the test substance to the human’s arm skin, and subsequent exposure of the treated area to unfed female mosquitoes. The oven tests indicated that a 75:25 molar ratio mixture of IR3535 with nonanoic acid formed a negative pseudo-azeotrope. This was confirmed by thermogravimetric analysis. This means that this composition evaporated at a significantly lower rate than the neat constituents and that other mixtures approaches this liquid composition over time as evaporation proceeded. Repellency tests showed that this pseudo-azeotrope mixture provided an improved and a longer lasting effect compared to DEET and IR3535. Suitable combinations of known repellents may form negative pseudo-azeotropes that feature a longer lasting, yet effective repellent performance. This was shown to be the case for IR3535 combined with nonanoic acid in a 75:25 molar ratio. It is recommended that other potential repellent combinations be studied in order to find cost effective solutions for the prevention of outdoor malaria transmission.
Page 72 of 170 MICROPOROUS POLYOLEFINS AS CONTROLLED RELEASE DEVICES FOR MOSQUITO REPELLENTS
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Sibanda, M.M. (1); Focke, W.W. (1); Braack, L. (1); Muatcho, J. (1) ; Smart, J. (1) and Androsch, R. (2)
(1) University of Pretoria Institute for Sustainable Malaria Control, Faculty of Health Sciences, University of Pretoria, South Africa; (2) Centre of Engineering Sciences, Martin Luther University Halle Wittenberg, Germany
Abstract Effective repellents such as N,N-diethyl-m-toluamide (DEET) may reduce the risk of malaria infection outdoors. However, the residual efficacy of topical applications lasts only a few hours. This problem may be solved by mosquito repellent ankle bracelets that store repellent active within a polymer matrix and slowly release it at effective levels over extended periods of time. Citronellal, DEET, ethyl- anthranilate, IR3535®, icaridin, citriodiol, and permethrin were evaluated for their relative volatility and oxidative thermal stability using thermo-gravimetric analysis and rancimat tests. The ability of the repellent actives to form micro-porous matrices, via spinodal decomposition, was explored. Repellents impregnated polymer strands were extruded via spinodal decomposition and hot glued together to form anklets and “footlets”. These devices were tested for repellent efficacy using foot- in-cage tests. Icaridin, DEET and IR3535® were found to be the least volatile and most thermally stable repellents considered. Scanning electron microscopy (SEM) indicated that all of the repellent actives, with the exception of citronellal and citriodiol, formed microporous polymer structures with EVA. Icaridin, DEET and IR3535® were successfully compounded into EVA (18% VA) to produce microporous strands. SEM studies showed that these strands had a microporous matrix structure. Fourier transform infrared spectroscopy indicated that the compounding process did not degrade these repellents. DEET impregnated “footlet” devices, which covered a greater area of the ankle and foot, showed markedly better protection efficacy with 68% protective efficacy after 55 days of manufacture compared to anklets. DEET, ethyl anthranilate, icaridin, IR3535® and permethrin can be incorporated into the polymer matrices via spinodal decomposition. Repellent efficacy tests indicate that high concentration of repellents impregnated into polymers offer long lasting protection. These polymers can be used to manufacture products such as sandals that can be worn in resource limited communities.
Page 73 of 170 PLANNING OF HOUSING IMPROVEMENT INTERVENTIONS FOR MALARIA ELIMINATION IN SWAZILAND
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Sibandze, M. (1); Dlamini, N. (1); Bhangu, K. (1); Dlamini M. (1); Soble, A. (2); Seethaler, T. (2); Dolenz, C. (2); Woolheater, K. (2); Pindolia D. (2); Kunene, S. (1)
(1) National Malaria Control Programme, Manzini, Swaziland; (2) Clinton Health Access Initiative, Mbabane, Swaziland
Abstract Through analysis of Swaziland’s individual-level surveillance data collected between 2010 and 2014, the National Malaria Control Programme (NMCP) identified poor housing quality as a potential risk factor for local malaria transmission. To support Swaziland’s malaria elimination goal, the NMCP plans to implement housing improvements as a long-lasting intervention to reduce human-vector contact and potential for local transmission. Geo-located local cases and poor quality houses (defined based on construction materials used for walls, roofs and windows) were mapped in Google Earth. Based on the presence of local cases and generally poor quality houses, an operationally feasible area within the Ka-Ngcamphalala locality was selected for an assessment of vector control intervention coverage, housing quality, structural dimensions, and acceptability of housing improvement measures. A household level, tablet-based questionnaire was developed and administered over two days in January 2016. Descriptive analysis was completed to assess housing characteristics, intervention acceptability and coverage of vector control interventions in the sampling area. 54 households with 120 sleeping structures were surveyed. Households had an average of 4 people sleeping inside each structure. 91% of heads of household (49 households) reported making no improvements (including re-plastering walls, replacement of doors and windows) to the sleeping structures in their households within the last 24 months. All windows and 99% of doors did not have screening, and 37% of structures had open spaces around eaves. The number of structural elements requiring screens and their dimensions varied significantly amongst and between modern and traditional structures. All respondents reported willingness to receive housing improvement interventions. The implementation of housing improvements will require custom improvements to meet the needs of each sleeping structure. Results from the field assessment illustrate that housing improvement interventions in areas of Swaziland with local malaria transmission and poor housing quality could be well accepted. For malaria programs considering housing improvements, systematic collection of structural data during implementation planning would provide detailed information into the types of improvements required to comprehensively mosquito-proof sleeping structures.
Page 74 of 170 ENVIRONMENTAL PROBLEMS WITH DDT AS USED IN MALARIA CONTROL
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Bouwman, H. (1), Bornman, R. (2)
(1) Research Unit: Environmental Sciences and Management, North-West University, Potchefstroom, South Africa; (2) University of Pretoria Institute for Sustainable Malaria Control (UP ISMC), School of Health, Systems and Public Health, University of Pretoria, Pretoria, South Africa
Abstract For decades DDT has been used at the forefront in the reduction and prevention of malaria transmission. This has been very successful in saving untold lives and reducing human suffering. However, the cost of the effects of DDT on the environment has been neglected, considered inconsequential, or even (seemingly) relegated as collateral consequences that must be borne. Here, we will list and discuss the results of the numerous studies conducted in South Africa on the environmental consequences and health problems related to DDT. We will show that DDT has been damaging the environment and animals in the ecosystems on which rural South Africans to a greater or lesser extend depend upon for their residence, sustenance and income. We will report on the very high concentrations of DDT found in wild bird eggs, fish, frogs and other animals, as well as soils and sediment. We will also report on the growing evidence of damage caused by DDT in these and other wildlife, and the possible consequences on the ecosystems. Finally, we will argue that with all the tools available now and in development, the pace of moving away from DDT to alternatives, be it chemical, physical, improvement of housing and economic conditions and combinations thereof, is too slow and should be speeded up to prevent further damage.
Page 75 of 170 THE POTENTIAL USE OF SERIOUS GAMES IN MALARIA CONTROL AND ELIMINATION PROGRAMMES
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Coleman, M. (1), South, A. (1), Hemingway, C. (1), Thomson, E. (1)
(1) Liverpool School of Tropical Medicine
Abstract Serious games have been used in public health for many years. They are an engaging method to facilitate learning and exploration of the complex nature of disease determinants and the most effective ways to control effects on human health. The question is, can serious games, as part of established training and communication be useful to malaria control programmes to address important translation gaps? Widespread and increased insecticide resistance threatens the continued efficacy of the main malaria control interventions. If this situation is not managed and strategies to mitigate the threat put into place it will have serious consequences. It is expected from malaria control programmes to monitor and manage resistance and to develop insecticide resistance management plans as part of their strategic priorities. The limited availability of these plans and the lack of following through from policy to practice is a major concern to the vector control community. To develop and operationalize plans a good understanding of insecticide resistance management is critical. We aim to describe the success stories of other serious game delivery projects and the potential use of a serious game under current development, ResistanceSim, to assist with the challenge of insecticide resistance management in operational settings. The game will allow players from multiple disciplines to manage a virtual vector control and entomological surveillance programme to better understand the advantage of implementing good practices over space and time. The process from initiation, consultation, game development, testing and future delivery will be explained.
Page 76 of 170 THE LARVICIDAL AND BRINE SHRIMP ACTIVITIES OF EUGENOL AND FIVE DERIVATIVES
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Shunmoogam-Gounden, N. (1, 2); van Zyl, R.L. (1, 2)
(1) Pharmacology Division, Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, South Africa; (2) Wits Research Institute for Malaria (WRIM), University of the Witwatersrand, South Africa
Abstract Eugenol, an essential oil constituent from the Ocimum genus of plants has been shown to exhibit biological activity against a variety of organisms, including bacteria, fungi and protozoa. Many mosquito species, including Anopheles, are vectors for pathogens and are controlled by chemical insecticides. Apart from being harmful to the environment and humans, many vectors have developed resistance to these chemicals, indicating a need for alternative vector management. To investigate five eugenol derivatives: o-eugenol, eugenyl acetate, methyl eugenol, methylisoeugenol and isoeugenol for larvicidal and toxicological activities. Anopheles arabiensis (KGB) mosquito larvae (3rd/4th instar) were treated with eugenol and its five derivatives, according to the World Health Organisation protocol using dichlorodiphenyltrichloroethane as a positive control. The brine shrimp lethality assay was used to determine the toxicological properties of the essential oils against Artemia franciscana nauplii, using potassium dichromate as the positive control. The mosquito larvae and the brine shrimp nauplii were exposed to the essential oils for 24 hours under optimal conditions. Probit analysis was used to calculate LC50 values. Methyl eugenol and
methylisoeugenol displayed the greatest larvicidal activity with LC50 values of 0.0064±0.0013% and 0.0062±0.0009%, respectively. Eugenyl acetate and isoeugenol showed the weakest larvicidal
activity compared to eugenol and the other three derivatives, with LC50 values greater than 0.01%. Isoeugenol was 5.8-fold more selective towards the brine shrimp nauplii (LC50=0.00178±0.0002%) than the mosquito larvae. Eugenol and o-eugenol displayed very similar results in both assays, and this is attributed to their close structural similarity. Interestingly, methylisoeugenol, which bears close structural similarity to isoeugenol, was the most effective of the eugenol derivatives against the Anopheles arabiensis mosquito larvae. The results of the brine shrimp lethality assay indicated that isoeugenol was the most toxic and eugenyl acetate was the least toxic eugenol derivative. Methylisoeugenol should be investigated further as a larvicide against Anopheles mosquitoes to aid in malaria control.
Page 77 of 170 FIELD EFFECTIVENESS OF MICROBIAL LARVICIDES ON MOSQUITO LARVAE IN MALARIA AREAS OF BOTSWANA AND ZIMBABWE
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Mpofu, M. (1); Becker, P. (1); De Jager, C. (1); Mudambo, K. (2)
(1) UP Institute for Sustainable Malaria Control, School of Health Systems and Public Health, Faculty of Health Sciences, University of Pretoria; (2) Roll Back Malaria, East and Southern Africa Secretariat, Gaborone, Botswana
Abstract The successful control of malaria vectors requires the control of both the larval and adult stages. The adult control methods through indoor residual spraying (IRS) and use of long lasting insecticide nets (LLINs) are widely used. Larval control methods are also being used by a number of national malaria control programs with limited understanding of its contribution. This experimental study was conducted to assess the field effectiveness of winter larviciding on the larval stages of the mosquito in Botswana and Zimbabwe. Two villages were selected in each of the countries, one as an intervention and the other as the control. Water bodies in the intervention villages were treated using the commercial product VectoBac® WG (Valent BioSciences Corporation, IL, USA) containing the active ingredient Bacillus thuringiensis var. israelensis (Bti), a WHO recommended bio-larvicide, applied at a rate of 300g per hectare. Treatment was conducted between June and October 2015 at two week intervals with larval sampling and counting being conducted before the next treatment. Random-effects Poisson regression was employed during data analysis to compare intervention with control sites with respect to larval counts. The average marginal effect of larviciding on the mosquito larvae taking interaction with time (period) into account, was -1.94 (95% CI:-2.42 to -1.46) with incidence rate ratio of 0.14, thus an 86% larval reduction attributable to the intervention for both countries combined. There was a 92% and 65% effect for Botswana and Zimbabwe respectively. The effect on the early larval and late stages was 77% (P<0.001) and 91% (P<0.001) respectively. Overall, intervention larval sampling points had five more larvae than the control at baseline and 26 less after 16 weeks. The effect on the different species also showed similar trends. Larval control using Bti showed a high effect on the population of the mosquito larvae. The higher reduction of the late stages is a predictor of the effect in adult mosquito emergence and adult mosquito population. Larviciding can be used to control mosquito population by supressing the larval stages thereby reducing adult emergence and malaria risk.
Page 78 of 170 A REVIEW OF THE STERILE INSECT TECHNIQUE FOR THE CONTROL OF MALARIA VECTORS IN SOUTH AFRICA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Munhenga, G. (1, 2); Brooke, B.D. (1, 2); Gilles, J.R.L. (3); Slabbert, K. (4); Kemp, A. (5); Dandalo, L. (1, 2); Wood, O.R. (1, 2); Lobb, L.N.R. (1, 2); Govender, D. (6, 7); Renke, M. (8); Koekemoer, L.L. (1, 2)
(1) Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Private Bag X4, Sandringham, Johannesburg, South Africa (2) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa (3) Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, Vienna, Austria (4) iThemba LABS (Laboratory for Accelerator Based Sciences), Somerset West, South Africa (5) Special Pathogens Unit, Center for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Sandringham, Johannesburg, South Africa (6) Scientific Services, South African National Parks, Private Bag X402, Skukuza, South Africa (7) Department of Paraclinical Sciences, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort , South Africa (8) Conservation Management, Kruger National Park, Private Bag X402, Skukuza, 1350
Abstract The backbone of malaria control in South Africa is vector control, mainly achieved through the insecticide based indoor residual spraying (IRS) technique. However, the effectiveness of IRS is under threat owing to the development of insecticide resistance in target malaria vector populations. Furthermore, as transmission intensity decreases, the use of large amounts of chemical insecticides is not economically, socially or environmentally acceptable. Additionally, IRS is less effective at controlling the behaviourally variable vector Anopheles arabiensis, which is responsible for low-level, seasonal malaria transmission in South Africa. Against this background there was a need to identify additional vector control interventions to supplement existing strategies. The use of the Sterile Insect Technique (SIT) was proposed. A multi-year plan for testing the feasibility of SIT for malaria control in South Africa was developed by a consortium representing several partners in 2010. Over the last six years prefeasibility studies have been carried out culminating in the identification of a pilot release site, and the development and characterisation of an An. arabiensis genetic sexing strain. Intensive baseline vector surveillance at the release site has yielded important information concerning the population dynamics of the target An. arabiensis population. The purpose of his presentation is to review the experience and knowledge gained in developing the SIT as a malaria vector control tool and to focus on the challenges and current status of the project. We also provide an insight into the pilot release phase of the project.
Page 79 of 170 ATTACKING THE MOSQUITO ON MULTIPLE FRONTS: INSIGHTS ON OPTIMAL COMBINATIONS OF VECTOR CONTROL INTERVENTIONS FOR MALARIA ELIMINATION FROM A MATHEMATICAL MODEL
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Kiware, S. (1, 2); Tatarsky, A. (2); Wu, S. (2, 3); Sánchez, C.H.M. (2, 4); Chitnis, N (5), Marshall, J. (3)
(1) Environmental Health and Ecological Sciences Thematic Group, Ifakara Health Institute, Dar es Salaam, Tanzania (2) The Malaria Elimination Initiative, Global Health Group, University of California, San Francisco, CA, United States (3) Divisions of Biostatistics and Epidemiology, School of Public Health, University of California, Berkeley, Berkeley, CA, United States (4) School of Medicine, Tecnol o gico de Monterrey, Mexico, Mexico (5) Department of Epidemiology and Public Health, Swiss Tropical and Public Health Institute, Basel, Switzerland
Abstract Despite great achievements by long-lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS), research demonstrates that these tools are insufficient to eliminate malaria transmission in many settings today. Protective coverage from these interventions is attenuated where mosquitoes can access blood resources from non-human hosts or from humans when they are outdoors. Fortunately, field experiments indicate that there are many promising vector control interventions that can be used to complement LLINs and/or IRS by targeting a wide range of biological and environmental mosquito resources. The majority of these experiments were performed to test a single vector control intervention in isolation; however, there is growing evidence and consensus that effective vector control will require a combination of interventions tailored to the target ecological and epidemiological settings. We propose a mathematical modeling framework designed to examine combination interventions prior to empirical field trials. The model framework incorporates all stages of the mosquito life cycle from egg, larva, pupa, adult, and, crucially, the female gonotrophic cycle whereby females blood feed and lay eggs.”). We describe how the framework may be used to evaluate the impact of combining existing and novel interventions in synergistic ways in areas where LLINs and/or IRS are widely used but where malaria transmission persists. We consider the following vector control interventions in addition to LLINs and IRS: larvaciding (conventional and aerial), attractive toxic sugar baits (ATSBs), insecticide spraying of male mating swarms, mosquito-proofed housing, spatial and topical mosquito repellents, systemic and topical insecticide-treatment of cattle, odor-baited traps and space spraying. We describe optimal combinations of these interventions needed to significantly reduce entomological inoculation rate (EIR), a widely accepted measure of malaria transmission, in a range of ecological and epidemiological settings. Preliminary model simulations recommend a combination of interventions using larvaciding, insecticide treatment for livestock, mosquito proofed housing, and baited traps to control and eliminate malaria in addition to scaling up LLINs and/or IRS in most transmission scenarios. The model framework developed may be used to simulate and optimize the impact of current and novel tools on malaria control and elimination.
Page 80 of 170 MATHEMATICAL MODELLING GUIDING ELIMINATION IN THE ASIA-PACIFIC REGION AND ITS POTENTIAL IN AFRICA
Abstract Theme Modeling towards elimination
Authors and Affiliations Silal, S.P. (1); White, L.J. (2, 3); Shretta, R. (4)
(1) Department of Statistical Sciences, University of Cape Town, Cape Town, South Africa; (2) Mahidol-Oxford Tropical Medicine Research Unit, Mahidol University, Bangkok, Thailand; (3) Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, Churchill Hospital, University of Oxford, Oxford, UK; (4) Malaria Elimination Initiative, Global Health Group, University of California, San Francisco, San Francisco, California, United States of America
Abstract Many countries in the Asia-Pacific region have made substantial progress in controlling malaria, reducing deaths from the disease by more than 25% since 2000. These gains have been achieved in part because of increased political and financial commitment allowing the scale-up of effective tools for preventing, diagnosing and treating malaria. Several countries are now working towards malaria elimination and a regional goal for a malaria-free Asia Pacific by 2030 has been recognized at the highest levels. Mathematical modelling has been used to represent the transmission dynamics of malaria for more than 100 years and the Malaria Eradication Research Agenda Consultative Group on Modelling has recognised the contribution mathematical modeling can make to the elimination of malaria globally. The study aims to use mathematical modelling to project rates of decline to elimination by 2030 and determine the costs for (and maintaining) elimination in the Asia Pacific region. Non-linear differential equations were used to develop a mathematical model to capture the dynamics of Plasmodium falciparum and Plasmodium vivax malaria for the 22 countries in the Asia- Pacific region. The models accounted for health system interventions, immunity, G6PD deficiency and mobility. Interventions such as scaling up existing measures and targeted transmission- interrupting measures were tested on the model. Continuing interventions at current levels will not achieve malaria elimination. Scaling up interventions to universal coverage will substantially reduce malaria in the region, but is insufficient to achieve malaria elimination. Targeted interventions that interrupt the transmission cycle may be used alongside current interventions to bring countries within reach of malaria elimination. A model such as this can be applied to Southern African region and the Elimination 8 region to assist in answering key questions in working towards the African Leaders Malaria Alliance goal of elimination by 2030.
Page 81 of 170 DYNAMIC EVALUATION OF POTENTIAL TRANSMISSION BLOCKING ANTIPLASMODIALS: RATE AND STAGE-SPECIFIC DESCRIPTIONS OF GAMETOCYTOCIDAL COMPOUNDS
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Reader, J. (1); Botha, M.E. (1); Birkholtz, L. (1).
(1) Malaria Parasite Molecular Laboratory, UP Institute for Sustainable Malaria Control, Department of Biochemistry, University of Pretoria, Private Bag x20, Hatfield, Pretoria 0028, South Africa
Abstract Despite global efforts, malaria is still highly prevalent. The current malaria elimination strategy focuses not only on the discovery of compounds that target the asexual stage of the parasite to treat malaria; but also on the description of compounds capable of blocking malaria transmission by killing the transmissible gametocyte stage of the parasite. Current gametocytocidal compounds are expected to target mature, stage IV/V gametocytes to fit the target candidate profile set for transmission blocking entities by the Medicines for Malaria venture. Differential sensitivities to compounds exist between asexual parasites and both early and late gametocytes, emphasizing the importance of verifying the stage-specificity of action of compounds against both early- and late- stage gametocytes. We report here the development of a simple and relatively quick gametocyte stage-specificity assay as well as a novel rate-of-action assay to interrogate hit gametocytocidal compounds, using a luciferase-based reporter line approach. The Luciferase Reporter Assay was established to enable accurate, reliable and quantifiable investigations of the stage-specific action of gametocytocidal compounds against each of the early and late gametocyte marker cell lines. The chemical susceptibility of a set of six promising compounds and four controls was investigated to determine the compound(s) kill rate over time when tested against all stages of the sexual parasite. Drug assays were set up on day 5 and 10 (representing >90% early stage I/II and >90% mature stage IV/V gametocytes, respectively), using 2-fold dilutions per compound for dose- response determination. Parasites were incubated under drug pressure for 24, 48 or 72 hours. The
IC50 over time of each compound on the early and late gametocytes was used to determine the stage specificity and the rate of action. This study resulted in the first description of the differential, temporal effect for different gametocytocidal compounds. The assays developed here will enable description of the rate and stage of activity of new gametocytocidal compounds, thereby defining the activity of transmission blocking leads.
Page 82 of 170 DIGITAL IMAGE ANALYSIS FOR DETERMINING MALARIA PARASITE DENSITY IN THICK BLOOD FILMS
Abstract Theme Case management
Authors and Affiliations Frean, J.A. (1, 2); van Deventer, J.M.G. (1); Poonsamy, B. (1)
(1) National Institute for Communicable Diseases, Johannesburg; (2) Wits Research Institute for Malaria, Faculty of Health Sciences, University of the Witwatersrand
Abstract Quantitation of malaria parasites in blood films is an important component of laboratory diagnosis of malaria. Inaccurate parasite density estimation may have adverse clinical and therapeutic implications for patients, and for endpoints of clinical trials of anti-malaria vaccines or drugs. Microscopy of Giemsa-stained thick films is the conventional method for parasite enumeration, as recommended by the World Health Organization. Accurate and reproducible parasite counts are difficult to achieve, because of inherent technical limitations and human inconsistency. Digital image analysis provides an opportunity to improve parasite density quantitation. Accurate manual parasite counts were done on 497 images of a range of thick blood films with varying density of malaria parasites, to establish a uniformly reliable standard against which to assess the digital technique. ImageJ, a Java-based open access image analysis program, was used. By utilising descriptive statistical parameters of parasite size frequency distributions, particle counting algorithms were semi-automatically adapted to variations in parasite size, shape, and staining characteristics, to produce optimum signal/noise ratios. A reliable counting process was developed that requires no operator decisions that might bias the outcome. Digital counts were highly correlated with manual counts for medium to high parasite densities, and slightly less well correlated with conventional counts. At low densities (fewer than 6 parasites per analysed image) signal/noise ratios were compromised and correlation between digital and manual counts was poor. Conventional counts were consistently lower than both digital and manual counts, suggesting that even experienced microscopists commonly underestimate parasite densities. Using open-access software and avoiding custom programming or any special operator intervention, accurate digital counts were obtained, particularly at high parasite densities that are difficult to count conventionally. The major constraint of the method is the time required to capture the necessary number of images to be analysed. The technique is potentially useful for laboratories that routinely perform malaria parasite enumeration. The requirements of good quality stained blood films, a digital microscope camera, and a personal computer with image analysis software, are reasonably easy to meet.
Page 83 of 170 USING A RISK BASED APPROACH TO DEVELOP A QUALITY MANUAL FOR PUBLIC HEALTH INSTITUTIONS UNDERTAKING INVESTIGATOR-LED ANTIMALARIAL DRUG CLINICAL RESEARCH IN SOUTH AFRICA
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Leisegang, C.K. (1); Raman, J (2); Barnes, K.I. (1); Allen, E.A. (1)
(1) UCT Collaborating Centre for Optimising Antimalarial Therapy; (2) National Institute for Communicable Diseases
Abstract Clinical research activities are core to informing the best practices for malaria elimination strategies. Unfortunately, the challenges facing antimalarial drug clinical researchers once the revised ICH E6 (R2) Good Clinical Practice guidelines are finalised, have yet to be addressed. In the revised guidelines, some aspects of quality management will shift from study sponsors to the investigational sites (e.g. clinics). This shift is likely to exacerbate existing challenges that investigators leading their own trials encounter as more of the sponsor's responsibilities now fall to them. Taking this into consideration, the UCT Collaborating Centre for Optimising Antimalarial Therapy (CCOAT) is developing a quality manual for academic and public health institutions undertaking investigator-led antimalarial drug clinical research studies in South Africa. This manual focuses on developing operational and clinic level procedures that will avoid unnecessary complexity whilst ensuring patient safety and data accuracy. In order to understand the elements essential to prevent or mitigate risk, a quick survey was designed to elicit input from key stakeholders, including malaria drug researchers, Malaria Control Programme personnel, clinic/hospital managers and staff, and other Department of Health personnel involved in such studies. Topics covered in the manual include: relevant standard operating procedures (SOPs), training processes/materials and quality metrics; the level of clinical/lab/operational oversight needed for each function; plans and processes to ensure a smooth corrective and preventive action (CAPA) process; and any additional tools, plans and or timelines to ensure adequate implementation. It is proposed, given the consultative nature of the process, and broad base of stakeholder involvement, the resultant quality manual will serve to mitigate risk and ensure quality in a wide range of antimalarial drug clinical research studies.
Page 84 of 170 BIOPROSPECTING TRADITIONALLY USED ANTIMALARIAL PLANTS BY MEANS OF NMR-BASED METABOLOMICS
Abstract Theme Phytomedicine and bioprospecting
Authors and Affiliations Bapela, M.J. (1); Meyer, J.J.M. (1); Heyman, H.M. (1)
(1) Department of Plant Science, UP Institute for Sustainable Malaria Control (UP ISMC), University of Pretoria, Pretoria, South Africa 0002
Abstract Despite the significant advances achieved in lessening the burden of malaria and other tropical diseases in recent years, malaria remains a major cause of mortality in endemic countries. This is especially the case in sub-Saharan Africa where 99% of the estimated global malaria deaths occurs on an annual basis. Emergence of resistant Plasmodium species and the lack of diversified chemotherapeutic agents provide the rationale for bioprospecting for novel antiplasmodial scaffolds. Crude extracts from twenty indigenous antimalarial plant species were screened for antimalarial activity and then subjected to 1H NMR-based metabolomic analysis. Ten plant extracts exhibited significant in vitro antiplasmodial activity (IC50 ≤ 5 µg/ml). The Principal Component Analysis (PCA) of the acquired 1H NMR spectra could not separate the analysed plant extracts according to the detected antiplasmodial bioactivity. Application of supervised Orthogonal Projections to Latent Structures–Discriminant Analysis (OPLS-DA) to the 1H NMR profiles resulted in a discrimination pattern that could be correlated to bioactivity. A contribution plot generated from the OPLS-DA scoring plot illustrated the classes of compounds responsible for the observed grouping. Given the preliminary in vitro results, Tabernaemontana elegans Stapf. (Apocynaceae) and Vangueria infausta Burch. subsp. infausta (Rubiaceae) were subjected to further phytochemical investigations. Two indole alkaloids, dregamine and tabernaemontanine possessing antiplasmodial activity were isolated from T. elegans. Two compounds were isolated from V. infausta subsp. infausta and identified as friedelin (IC50 = 3.01 µg/ml) and morindolide (IC50 = 18.5 µg/ml). While these compounds have been previously identified, this is the first account of their occurrence in the genus Vangueria and their antiplasmodial activity. Based on the results of the study, metabolomics can be used to globally identify classes of plant secondary metabolites that are responsible for antiplasmodial activity.
Page 85 of 170 NOVEL DRUG SCAFFOLDS FOR THE TREATMENT OF MULTI-DRUG RESISTANT PLASMODIUM FALCIPARUM
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Panayides, J-L. (1), van der Westhuyzen, C.W. (1), Mancama, D. (1)
(1) Pioneering Health Sciences, CSIR Biosciences, South Africa
Abstract Malaria is one of the most important infectious diseases in the world, with malaria due to Plasmodium falciparum the most deadly form [1]. In 2014, 97 countries and territories had ongoing malaria transmission, with an estimated 1.2 billion people considered at high risk of contracting malaria [2]. In 2013, there were an estimated 198 million cases of malaria worldwide, with approximately 584 000 deaths occurring, 90% of which were in Africa [2]. If left unaddressed, emerging parasite resistance to current antimalarial drugs could render the current treatment tools ineffective and trigger a global rise in malaria mortality. In recent years, parasite resistance to artemisinin has been detected in five countries: Cambodia, Lao People’s Democratic Republic, Myanmar, Thailand and Vietnam, with multi-drug resistance an increasing concern [2]. With the emergence of parasites that are resistant to the frontline antimalarial drugs, novel classes of compounds that can inhibit the spread of multi-drug resistant malaria are urgently needed. Our current research focuses on identifying novel chemical entities of structurally diverse classes: (i) producing reactive oxygen species, (ii) interfering in the parasite’s biological reduction-oxidation cascades and (iii) with transmission blocking ability. Novel synthetic compounds have been screened in vitro against both the asexual (chloroquine-susceptible, chloroquine-/pyremethamine- /artemisinin-resistant) and sexual (gametocyte) stages of the malaria causing parasite P. falciparum. A series of synthetic derivatives have been identified which represent extremely active “lead compounds” against malaria parasites, which are of interest for further investigation through drug discovery channels.
Financial Support: CSIR YREF, NRF PDP
References: [1] Snow, R.W., Trape, J.F., Marsh, K., Trends in Parasitology, 2001, 17, 593-597 [2] World Health Organization, WHO World Malaria Report, 2014, 1-2, 29
Page 86 of 170 THE MOLECULAR DETERMINANTS FOR TRAFFICKING OF AMA-1, A MALARIA INVASION PROTEIN AND VACCINE CANDIDATE
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Churchyard, A. (1, 2); Coetzer, T.L. (1, 2, 3)
(1) Wits Research Institute for Malaria (WRIM); (2) Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa; (3) National Health Laboratory Service, Johannesburg, South Africa
Abstract The malaria parasite, Plasmodium falciparum, relies on invasion proteins such as apical membrane antigen-1 (AMA-1) to enter human red blood cells, where the parasites proliferate and cause clinical malaria. AMA-1 is transported to apically located secretory organelles called micronemes before being translocated onto the parasite surface just prior to invasion to evade detection by the host immune system. This study aims to understand the trafficking of newly synthesized AMA-1 from the endoplasmic reticulum (ER) to the micronemes. Bioinformatic analysis revealed that the protein contains a signal peptide, a prodomain, an ectodomain, as well as transmembrane and cytoplasmic regions. Selected domains were amplified by PCR and overlap extension PCR to generate ama-1 mini-genes, which all contained the signal peptide that is required for transport through the ER. The mini genes were cloned into a pARL-mCherry plasmid encoding a red fluorescent tag and the mCherry-tagged constructs were used to transfect P. falciparum parasites by electroporation. Once transgenic parasites were established, co-localisation studies were performed to identify the location of each chimaeric protein relative to a fluorescently-tagged microneme marker. Unlike other invasion protein families, PfAMA-1 was shown to utilise the prodomain, instead of the cysteine-rich ectodomain, to target the micronemes, Because prodomains are not usually implicated in trafficking of invasion proteins, the possibility of a protein escorter was explored. The AMA-1 prodomain was expressed as a “prey” protein with a histidine tag and exposed to “bait” proteins in a P. falciparum phage display library. Isolated binding partners were sequenced and compared to the online Plasmodium database, which identified two interesting proteins. The first is a putative chaperone binding protein and the other is a putative P. falciparum formin2, a protein whose human orthologue is involved in intracellular vesicle transport. Studies are currently underway to express these binding partners as GST-tagged recombinant proteins to verify their interaction with AMA-1. Since humans don’t have apical organelles, these trafficking mechanisms are unique to P. falciparum and may be exploited as novel drug targets.
Page 87 of 170
POSTER LIST
Page 88 of 170 Poster # Poster Title Presenting Abs ID THE JOURNEY FROM DIAGNOSIS TO CASE INVESTIGATION FOR MALARIA ELIMINATION IN SWAZILAND: IS MC/01 Dlamini, N MC/072 REPORTING AND RESPONSE TIMELY? EPIDEMIOLOGY OF MALARIA IN MADAGASCAR: SPATIOTEMPORAL DISTRIBUTION OF UNCOMPLICATED AND MC/02 Ihantamalala, F MC/010 COMPLICATED MALARIA MC/03 WORKER ABSENCE DUE TO MALARIA IN A BANANA PLANTATION IN ZIMBABWE Lukwa, A MC/005 MC/04 PERFORMANCE OF TWO PFHRP2-BASED RDTS AS COMMUNITY MALARIA TESTS IN MUTASA DISTRICT ZIMBABWE Mudare, N MC/118 MC/05 A NEW METHOD FOR SAMPLING VOLATILE HUMAN SKIN CHEMICALS IN A MALARIA CONTEXT Roodt, A MC/023 ASSESSING THE PERFORMANCE OF RE-ACTIVE CASE DETECTION (RACD) AS AN ELIMINATION STRATEGY IN MC/06 Maponga, A MC/053 MATABELELAND SOUTH PROVINCE TOWARDS MALARIA ELIMINATION: AN EVALUATION OF THE SOUTH AFRICAN MALARIA SURVEILLANCE SYSTEM, MC/07 Baloyi, E MC/080 2015 EVALUATING THE UTILITY OF AMA-1 AND MSP-1 AS SEROLOGICAL MARKERS OF MALARIA EXPOSURE FOR MC/08 Chisenga, M MC/087 SURVEILLANCE IN THE LOW TRANSMISSION, PRE-ELIMINATION SETTING OF NAMIBIA THREE PARALLEL INFORMATION SYSTEMS FOR MALARIA ELIMINATION IN SWAZILAND, 2010-2015: ARE THE MC/09 Zulu, Z MC/073 NUMBERS THE SAME? A PROTOCOL TO EVALUATE THE EFFECTIVENESS AND FEASIBILITY OF REACTIVE TARGETED PARASITE ELIMINATION MC/10 Mkhonta, N MC/078 (TPE) TO REACTIVE CASE DETECTION (RACD) AS A COMMUNITY LEVEL INTERVENTION IN SWAZILAND MC/11 IMPACT OF INTENSE VECTOR SURVEILLANCE IN KWAZULU-NATAL Koekemoer, L MC/110 RAPID RESTORATION OF CHLOROQUINE SENSITIVITY IN PLASMODIUM FALCIPARUM MALARIA OF MUTASA MC/12 Zingoni, M MC/117 DISTRICT, ZIMBABWE USING DRIED PLASMODIUM FALCIPARUM-INFECTED BLOOD SAMPLES AS POSITIVE CONTROLS FOR MALARIA MC/13 Ming Sun, L MC/027 RAPID DIAGNOSTIC TESTS DECREASED PREVALENCE OF THE PLASMODIUM FALCIPARUM CHLOROQUINE RESISTANCE TRANSPORTER GENE MC/14 MUTATION K76T NINE YEARS AFTER CHLOROQUINE TREATMENT WITHDRAWAL IN CHIPATA AND KATETE Mwenda, M MC/056 DISTRICTS, EASTERN ZAMBIA. MC/15 MALARIA TRENDS SOUTH AFRICA, 2011-2015 Shandukani, M MC/046 PRELIMINARY STUDY ON THE PREVALENCE OF RESISTANCE ASSOCIATED POLYMORPHISM IN PLASMODIUM MC/16 Sijuade, O MC/038 FALCIPARUM FIELD ISOLATES PRESENTED IN GUATENG, SOUTH AFRICA MC/17 MOLECULAR DIAGNOSIS OF MALARIA IN THE ZAMBEZI REGION OF NAMIBIA MOVING TOWARDS ELIMINATION Tambo, M MC/095
Page 89 of 170 MC/18 SUMMARY OF FINDINGS MALARIA ELIMINATION OPERATIONAL CAPACITY ASSESSMENT SURVEY - ZIMBABWE 2015 Mberikunashe, J MC/055 DEVELOPMENT AND IMPLEMENTATION OF A MALARIA COMMODITY TRACKING AND ORDERING TOOL FOR MC/19 IMPROVED FORECASTING, PROCUREMENT AND SUPPLY MANAGEMENT OF MALARIA DIAGNOSTIC AND Mathobela, M MC/065 TREATMENT COMMODITIES DEVELOPMENT OF A PHARMACOVIGILANCE SAFETY MONITORING TOOL FOR THE ROLLOUT OF SINGLE LOW-DOSE MC/20 PRIMAQUINE AND ARTEMETHER-LUMEFANTRINE TO TREAT PLASMODIUM FALCIPARUM INFECTIONS IN Malambe, C MC/070 SWAZILAND: A PILOT STUDY MC/21 CASE MANAGEMENT OF MALARIA IN SWAZILAND, 2011 TO 2015: ON TRACK FOR ELIMINATION? Dlamini, S MC/100 COMMMUNITY ANTIMALARIAL USE PATTERNS AND RAPID TESTING FOR FAKE OR SUBSTANDARD DRUGS IN MC/22 Mharakurwa, S MC/116 MUTASA DISTRICT, ZIMBABWE MC/23 SIMPLIFYING MALARIA PARASITE COUNTS ON THIN BLOOD FILMS Poonsamy, B MC/044 MC/24 UNDERSTANDING CROSS-BORDER MALARIA: LESSONS LEARNED FROM EASTERN ZIMBABWE Kanyangarara, M MC/114 MC/25 OPERATIONAL LESSONS LEARNED FROM IRS PROGRAM IMPROVEMENTS IN NAMIBIA Angula, H MC/052 AN ASSESSMENT OF COMPLIANCE TO ENVIRONMENTAL SAFEGUARDS IN THE USE OF DDT IN CHONGWE IRS MC/26 Chilabi, A MC/003 PROGRAMME RURAL PART OF ZAMBIA EFFECT OF IRRADIATON ON FEMALE ANOPHELES ARABIENSIS: AN INVESTIGATIVE STUDY TOWARDS THE MC/27 Dandalo, L MC/012 DEVELOPMENT OF THE STERILE INSECT TECHNIQUE TO CONTROL MALARIA VECTORS IN SOUTH AFRICA EFFECT OF STABLE AND FLUCTUATING TEMPERATURES ON THE LIFE HISTORY TRAITS OF ANOPHELES ARABIENSIS MC/28 Davies, C MC/004 AND AN. QUADRIANNULATUS UNDER CONDITIONS OF INTER- AND INTRA-SPECIFIC COMPETITION MC/29 BIOLOGICAL CONSEQUENCES OF DDT EXPOSURE Delport, R MC/121 EXPOSURE TO DDT/DDE IN RELATION TO HBA1C IN PREGNANT WOMEN OF TSHILIDZINI HOSPITAL, LIMPOPO, MC/30 Ndwamato, N MC/093 SOUTH AFRICA THE MONITORING OF BASELINE STERILITY IN THE POPULATION OF ANOPHELES ARABIENSIS IN MAMFENE, KZN, MC/31 Lobb, L MC/037 SOUTH AFRICA THE LARVICIDAL EFFECTS OF BLACK PEPPER (PIPER NIGRUM LINN.) AND PIPERINE AGAINST INSECTICIDE RESISTANT MC/32 Samuel, M MC/007 AND SUSCEPTIBLE STRAINS OF ANOPHELES MALARIA VECTOR MOSQUITOES MC/33 ADDRESSING CURRENT CHOKEPOINTS IN ANOPHELES ARABIENIS MOSQUITO MASS REARING Wood, O MC/015 MC/34 FREQUENCY OF SEXUALLY MATURED MALES, IN A WILD ANOPHELES FUNESTUS SWARM IN BURKINA FASO Zawada, J MC/035 INVESTIGATING THE ANTIPLASMODIAL POTENTIAL OF (BIS)THIOUREA AND (BIS)UREA POLYAMINE ANALOGUES ON MC/35 Abrie, C MC/025 THE GAMETE FORM OF PLASMODIUM FALCIPARUM PARASITE
Page 90 of 170 AN EVALUATION OF THE GAMETOCYTOCIDAL PROPERTIES OF NATURAL PRODUCT COMPOUNDS AGAINST MC/36 Breedt, C MC/039 PLASMODIUM FALCIPARUM IMPLEMENTATION OF THE DIAGNOSIS QUALITY ASSURANCE PROGRAM FOR ACCURATE MALARIA DIAGNOSIS IN MC/37 Ntshalintshali, N MC/071 SWAZILAND MC/38 DEVELOPMENT OF SEROLOGIC ASSAYS TO ASSESS MALARIA EXPOSURE IN SWAZILAND Nhlabathi, N MC/074 DRUG TARGET DECONVOLUTION OF ANTIPLASMODIAL COMPOUNDS USING DRUG AFFINITY RESPONSIVE TARGET MC/39 Henn, D MC/030 STABILITY, FOR ASEXUAL AND SEXUAL STAGES OF PLASMODIUM FALCIPARUM PARASITES PHENOTYPIC PROFILING OF GROWTH PERTURBATIONS OF THE HUMAN MALARIA PARASITE PLASMODIUM MC/40 Joshua, A MC/031 FALCIPARUM EFFECT OF NOVEL 7-CHLOROQUINOLIN-4-YL PIPERAZINE-1-YL-ACETAMIDE DERIVATIVES IN THE MANAGEMENT OF MC/41 Kathrada, F MC/107 MALARIA MC/42 USING CRISPR-CAS9 TO MANIPULATE THE GENOME OF P. FALCIPARUM Liebenberg, D MC/008 EVALUATION OF THE PREVALENCE OF CYP 2D6 MUTATIONS IN VHEMBE DISTRICT, LIMPOPO PROVINCE, SOUTH MC/43 Makhanthisa, T MC/026 AFRICA MC/44 THE HIDDEN NON-BLOODSTREAM PARASITE RESERVOIR: A MALARIA CONTROL PROBLEM Markus, M MC/002 EPIGENETIC DRUGS TARGETING HISTONE POSTTRANSLATIONAL MODIFICATIONS DURING GAMETOCYTOGENESIS MC/45 Mmekwa, N MC/081 OF THE PLASMODIUM FALCIPARUM PARASITE MC/46 IN VITRO ACTIVITY OF NAPHTHYLISOQUINOLINE ALKALOIDS ON PLASMODIUM FALCIPARUM GAMETOCYTES Moyo, P MC/040 MC/47 MEMBRANE TRANSPORT IN THE SEXUAL, TRANSMISSIBLE STAGES OF PLASMODIUM FALCIPARUM PARASITES Naude, M MC/029 MC/48 APPLICATION OF NETWORK ANALYSIS ON P. FALCIPARUM TRANSCRIPTOMES Van Wyk, R MC/049 MC/49 CRISPR-CAS EVALUATION OF IMPORTANT CELL CYCLE REGULATORS Von Gruning, H MC/022 MC/50 INVESTIGATION OF DIFFERENTIAL DRUG RESPONSES IN DIFFERENT P. FALCIPARUM PARASITE LIFE CYCLE STAGES Weidemann, G MC/033 ASSESSING THE IN VITRO EFFICACY IN SILICO DESIGNED COMPOUNDS TARGETING THE MALARIAL QI SITE OF MC/51 Damadeu, L MC/062 CYTOCHROME BC1 IN VITRO ANTIMALARIAL ACTIVITY OF DIOSPYROS CHAMAETHAMNUS AND GUIBOURTIA COLEOSPERMA, FOUND IN MC/52 Du Preez, C MC/088 THE ZAMBEZI REGION OF NAMIBIA THE COPPER CHELATING, ANTIMALARIAL AND LARVICIDAL EFFECTS OF 8-HYDROXYQUINOLINE AND ITS MC/53 Jansen v Vuuren, N MC/109 DERIVATIVES IN SILICO DESIGNED SMALL-GATEKEEPER PROTEIN KINASE AND BROMODOMAIN SELECTIVE INHIBITORS EXHIBIT MC/54 Matlebjane, D MC/059 POTENTIAL ANTIPLASMODIAL ACTIVITY
Page 91 of 170 MC/55 SYNTHESIS AND MODIFICATION OF ANTIFOLATES Molatsane, T MC/094 THE ISOMERIC EFFECT OF ESSENTIAL OIL CONSTITUENTS ON ANOPHELES ARABIENSIS LARVAE AND ARTEMIA MC/56 Mustapha, O MC/111 FRANCISCANA NAUPLII MC/57 SYNTHESIS AND ANTIMALARIAL PROPERTIES OF SPIROINDOLONE ANALOGUES Naicker, A MC/092 MC/58 SYNTHESIS AND BIOLOGICAL EVALUATION OF ANTIMALARIAL ANTIFOLATES Seanego, D MC/060 MC/59 MULTITARGET ANTIMALARIAL POLYAMINE ANALOGUES: RESISTING RESISTANCE Verlinden, B MC/045 TOWARDS THE ELIMINATION OF MALARIA IN THE UNION OF COMOROS: GEOGRAPHICAL INSIGHT ON CONTROL MC/60 Attoumane, A MC/091 ACTIONS AND MAINTENANCE FACTORS TOWARDS MALARIA ELIMINATION: IMPLEMENTATION OF AN ALERT AND RESPONSE SYSTEM FOR MALARIA MC/61 Ngwenyama, B MC/079 SURVEILLANCE, SOUTH AFRICA, 2016 MC/62 MATHEMATICAL MODEL OF MALARIA TRANSMISSION IN CHILDREN UNDER FIVE YEARS IN AN ENDEMIC AREA Danquah, B MC/103 INVESTIGATING THE INFLUENCE OF METEOROLOGICAL DRIVERS ON THE UNUSUAL MALARIA TRANSMISSION MC/63 Seyama, E MC/077 DURING JULY – OCTOBER 2013 INTERACTIVE MALARIA EDUCATION INTERVENTION AND ITS EFFECT ON COMMUNITY PARTICIPANT KNOWLEDGE: MC/64 Cox, S MC/102 THE MALARIA AWARENESS PROGRAM (MAP) IN VHEMBE DISTRICT, LIMPOPO, SOUTH AFRICA
Page 92 of 170
POSTER ABSTRACTS
Page 93 of 170 THE JOURNEY FROM DIAGNOSIS TO CASE INVESTIGATION FOR MALARIA ELIMINATION IN SWAZILAND: IS REPORTING AND RESPONSE TIMELY?
Abstract Theme Epidemiology
Authors and Affiliations Dlamini, N. (1); Kunene, S. (1); Zulu, Z. (1); Geoffroy, E. (2); Ntshalintshali, N. (3); Soble, A. (3); Owiti, P. (4); Sikhondze, W. (5); Makadzange, K. (6); Zachariah, R. (7)
(1) National Malaria Control Programme (NMCP), Mbabane, Swaziland; (2) Global AIDS Interfaith Alliance, California, USA; (3) Clinton Health Access Initiative, Mbabane, Swaziland; (4) Academic Model Providing Access to Healthcare (AMPATH), Eldoret, Kenya; (5) National TB Control Program, Ministry of Health, Mbabane, Swaziland; (6) World Health Organization, Mbabane, Swaziland; (7) Médecins Sans Frontières, Luxembourg, Luxembourg
Abstract Swaziland is one of several countries in southern Africa targeting malaria elimination by 2020. In 2010, Swaziland introduced the Immediate Disease Notification System (IDNS) to enhance emergency preparedness and response by facilitating immediate reporting of notifiable diseases, including malaria. All health facilities are required to report confirmed malaria cases within 24 hours to the IDNS through a toll free number, 977. This triggers an alert for each new malaria case and initiates the NCMP’s active surveillance system to investigate each reported case at the household level within 48 hours. At a national level, we assessed reporting rate through IDNS, subsequent case investigation rate, within stipulated timelines. A cross-sectional study was conducted using routine national programme data from IDNS and the NMCP’s Malaria Surveillance Database System (MSDS) systems. Data from individual confirmed cases presenting at health facilities nationwide, cases reported through IDNS and investigated cases were analysed between July 2011 and June 2015. The analysis was performed using the case presentation date, case report date and case investigation date variables. There were 1,991 confirmed malaria cases during the 4 transmission seasons analysed. Of confirmed cases, 76% were reported through IDNS and 81% of these cases were reported within 24 hours. 88% of confirmed cases reported to IDNS were investigated by the NMCP and 33% were investigated within 48 hours of reporting. Poor reporting to IDNS was most pronounced in hospitals and private health facilities as 58% and 66% of confirmed cases were respectively reported. Reporting to IDNS is critical to activate the NMCP’s response, as only 63% of unreported cases were investigated and only 18% were investigated within 48 hours. The utilization of the IDNS for case reporting to trigger the active surveillance system in Swaziland must be improved for Swaziland to achieve malaria elimination. There is need to reduce delays in immediate reporting and case investigation response time to ensure that all confirmed malaria cases receives the appropriate and timely transmission-interrupting interventions
Page 94 of 170 EPIDEMIOLOGY OF MALARIA IN MADAGASCAR: SPATIOTEMPORAL DISTRIBUTION OF UNCOMPLICATED AND COMPLICATED MALARIA
Abstract Theme Epidemiology
Authors and Affliations Ihantamalala, H.F.A. (1)
1) Institut pasteur de Madagascar
Abstract Malaria is endemic in Madagascar and a leading cause of mortality and morbidity. Its geographical distribution is heterogeneous throughout the country, in relation to climatic, environmental and social factors. In this study, we analyze the spatiotemporal distribution of malaria caused by Plasmodium falciparum. The Service for Health and Demographic Statistics of the Ministry of Public Health provided monthly epidemiological data related to complicated and uncomplicated malaria cases from 2010 to 2014. We analyzed and integrated these data into a Geographic Information System to map malaria trends by year, by month and by age for each district. The incidence of malaria has increased since 2012 and remains high in some coastal districts. The highest peaks of reported cases are observed between January and April with especially high incidences along the Eastern coast. The percentage of uncomplicated malaria cases of children less than five years represent about 36% of cases for each year. Both complicated and uncomplicated malaria show similar patterns and trends. The quality of epidemiological data is discussed regarding the provision and access to health services. The connectivity between districts and the persistence of malaria on the coast could induce the emergence of malaria in central highlands following reintroduction by travelers. Thus, non-endemic areas are at risk of emergence with potential risk of complicated malaria form. Districts presenting significantly high incidences should be carefully monitored in order to reduce transmission.
Page 95 of 170 WORKER ABSENCE DUE TO MALARIA IN A BANANA PLANTATION IN ZIMBABWE
Abstract Theme Epidemiology
Authors and Affiliations Lukwa, N. (1); Mawoyo, R. (2); Lukwa, A.T. (3); Marange, (4); Nyadundu, S. (5); Zimba, M. (6)
(1) National Institute of Health Research, P. O. Box CY573, Causeway, Harare, Zimbabwe; (2) Mutare Provincial Hospital, P. O. Box 30, Mutare, Zimbabwe; (3) National Malaria Control Programme, P. O. Box CY1122, Causeway, Harare, Zimbabwe; (4) Burma Valley Rural Health Centre, P. A. Chitakatira, Mutare, Zimbabwe; (5) Provincial Medical Directorate, Manicaland province, Mutare, Zimbabwe; (6) University of Zimbabwe, Biological Sciences Department, P. O. Box MP167, Mount Pleasant, Harare, Zimbabwe
Abstract Absenteeism is one of the areas receiving attention because this results in lost man hours that have a direct impact on company productivity and performance. Malaria has caused several workers to be absent from work due to its nature that requires those affected to be given time off (during treatment). A study was carried out at a banana producing Matanuska farm in Burma Valley, Zimbabwe. The raw data was provided in retrospect by the Farm Manager. Presence of malaria was detected using the Rapid Diagnostic Test (RDT) kit. The following measures of absence were determined; incidence of absence (number of absentees divided by the total workforce), absence frequency (number of malaria spells), frequency rate (number of spells divided by the number of absentees), estimated duration of spells (number of days lost due to malaria), severity rate (number of days lost divided by number of spells), incapacity rate (number of days lost divided by the number of absentees), number of absent days (number of spells divided by the severity rate), number of scheduled working days (actual working days in 5 months multiplied by total number of employees), absenteeism rate (number of absent days divided by the number of scheduled days multiplied by 100). A total of 143 employees where followed up over a 5 month period. Malaria positivity was 21%, 31.5%, 44.8%, 35.7% and 12.6% in January, February, March, April and May 2014 respectively. One spell of absence [194 (86.6%)] was very common followed by 2 spells of absence [30 (13.4%)] for all employees. Duration of spells of absence due to malaria ranged from 1.5 days to 4.1 working days with general workers being the most affected. Incidence of absence was 143/155 (93.3%) with total of spells of absence over a 5 month period totalling 224. The frequency rate of absenteeism was 1.6 with severity rate of absence being 2.4. The incapacity rate was 3.7. Although the final absenteeism rate was 2.99% (below the 4% excessive rate), malaria contributed significantly to worker absenteeism, requiring employers to put measures that protect their workers
Page 96 of 170 PERFORMANCE OF TWO PFHRP2-BASED RDTS AS COMMUNITY MALARIA TESTS IN MUTASA DISTRICT ZIMBABWE
Abstract Theme Epidemiology
Authors and Affiliations: Mudare, N. (2); Makuwaza, A. (1); Matsena-Zingoni, Z (1); Mamini, E. (2); Musana, J. (2); Munyati, S (2); Gwanzura, L. (2, 4); Mutambu, S. (1); Mason, P. (2); Mharakurwa, S. (2, 3); Koboyashi, T. (5); Moss, W. (5)
(1) National Institute of Health Research, P.O. Box 573, Harare, Zimbabwe; (2) Biomedical Research Training Institute, Harare, Zimbabwe; (3) Africa University, Mutare; (4) University of Zimbabwe, Institute of Continuing Health research Education (ICHE); (5) Johns Hopkins School of Public Health, USA
Abstract Accurate identification of Plasmodium malaria infections in community surveys is essential to successful malaria control. Microscopy and rapid diagnostic tests (RDTs) are the main techniques used to diagnose malaria in field-based surveys. However, their sensitivity and specificity are proving a limitation as endemic countries scale up interventions for malaria elimination and sub microscopic reservoir detection becomes important. While microscopy is still considered the gold standard, RDTs have established popularity as they allow for rapid and inexpensive diagnosis. The diagnostic performance of Paracheck P.f. and (ICT) RDTs was compared against in a community malaria survey conducted in Mutasa District, Zimbabwe. The study was a cross-sectional design, based on representative households randomly selected using satellite imagery. Paracheck P.f®. and ICT RDT kits were used in parallel to screen for malaria in participants of all age groups from the selected households between October 2012 and December 2014. A total of 1062 participant’s samples were assayed for PCR, microscopy, Paracheck and ICT. Dried blood spots, thin and thick smears were simultaneously collected. The smears were Giemsa-stained and examined for malaria parasites by microscopy. Parasite was extracted from the DBS samples using the Chelex method and subjected to nested targeting the Plasmodium cytochrome b mitochondrial gene. The proportion of P. falciparum infections detected by PCR was 7.2%, microscopy was 1.9%, and both Paracheck- Pf® and ICT were 6.8%. There was no significant difference between the two RDT brands. The sensitivity (95% CI) of microscopy was 60.7% (40.6-78.5), with a specificity of 93.8%, (92.5-94.9). Microscopy had a positive predictive value of 14.5%, (8.7- 22.2). The sensitivity of both Paracheck- Pf® and ICT RDTs was 47.8% (37.1-58.6) and specificity of 95.3%, CI (94.1-96.3). There was no significant difference in malaria detection sensitivity and specificity between Paracheck- Pf and ICT RDT brands, so they can be used inter-changeably. Both RDTs showed lower sensitivity than microscopy, possibly owing to prevailing low parasitemia. However, further studies are needed to rule out the possibility of HRP2-deleted P. falciparum parasitemia.
Page 97 of 170 A NEW METHOD FOR SAMPLING VOLATILE HUMAN SKIN CHEMICALS IN A MALARIA CONTEXT
Abstract Theme Epidemiology
Authors and Affiliations Roodt, A. P. (1); Rohwer, E. R. (1); Naudé, Y. (1) (1) University of Pretoria, Department of Chemistry & UP Institute for Sustainable Malaria Control
Abstract
Volatile organic compounds (VOCs) emanating from the surface of human skin are of great interest to researchers in many fields. Applications include medical and forensic diagnostics, as well as studying the ecology of blood-feeding insect vectors of human disease. The latter application being of great interest in the ongoing search for new ways to efficiently control malaria in Africa. Research in these fields is currently still limited by the methodology used for sample collection and pre- concentration. There is a need for novel methods to collect and quantify trace volatile organic compounds originating from the surface of human skin. Passive sampling over long periods of time is an ideal approach yielding time weighted average concentrations of volatile analytes which is often of more value than acute sampling. Sampling over long periods allows for a more representative profile of the compounds emanating from the surface of human skin to be determined. This project proposes to investigate the use of silicone rubber (polydimethylsiloxane (PDMS)) tubes, in the forms of bracelets or anklets, as non-invasive sampling devices for the collection and concentration of volatile human skin emissions prior to comprehensive gas chromatographic time of flight mass spectrometric (GC x GC-TOFMS), analysis. The project aims to augment the current knowledge base on human skin emissions with the overall objective being collection and quantification of known olfactory cues origination from the surface of a subject’s skin that allow malaria vector mosquitos to identify their human blood-host.
Page 98 of 170 ASSESSING THE PERFORMANCE OF RE-ACTIVE CASE DETECTION (RACD) AS AN ELIMINATION STRATEGY IN MATABELELAND SOUTH PROVINCE
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Mberikunashe, J. (1); Dhliwayo, P. (1); Maponga, A.B. (2); Muchena, G. (2); Pindolia, D. (3); Ahmed, J. (3); Dolenz, C. (3); Le Menach, A. (3); Owen, J. (3); Madinga, M. (3); Gambinga, B. (3); Svisva, A. (3)
(1) Zimbabwe National Malaria Control Programme, Harare; (2) Ministry of Health and Child Care, Matabeleland South, Zimbabwe; (3) Clinton Health Access Initiative, Boston, MA, USA
Abstract Matabeleland South, a province in Zimbabwe that plans to eliminate malaria by 2017, adopted re- active case detection (rACD) as part of its active surveillance strategy in 2013. Surveillance guidelines currently require Environmental Health Practitioners (EHPs) to screen and test (with a Rapid Diagnostic Test (RDT)) household contacts of every investigated index case. Positive contacts (secondary cases) are referred to a health facility for treatment. Individual-level surveillance case data from September 2014 to August 2015 was extracted from the electronic database. Contact screening and contact positivity rate were estimated and compared over time and space to assess potential efficiency improvements to the rACD strategy. Of 1,045 investigated cases recorded in the electronic database, 453 (43.3%) had their contacts screened of which only 122 (26.9%) were investigated within 3 days of diagnosis of the index case. Of 1,405 contacts screened, 57 (4.1%) tested RDT positive. Of the 415 (29.5%) contacts screened within the recommended 3 day period, 5 were positive, representing a positivity rate of 1.2%. 47 (82.5%) of re-actively detected cases were found in Beitbridge district, the highest incidence district in the province (11.1 cases per 1000, compared to provincial average of 2.37 in the same time period). With a small sample size of 57 secondary cases, further stratification for example by demographic and socioeconomic group was not feasible. If rACD is to be used as a response strategy for malaria elimination, it needs to be effectively targeted to areas such as those with outbreaks, where rACD operations may be more feasible. Use of more sensitive tests may be explored to improve effectiveness of rACD. However, recent studies have questioned the usefulness of rACD altogether, suggesting alternative strategies such as Targeted Parasite Elimination (TPE) should be explored.
Page 99 of 170 TOWARDS MALARIA ELIMINATION: AN EVALUATION OF THE SOUTH AFRICAN MALARIA SURVEILLANCE SYSTEM, 2015
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Baloyi, R.E. (1, 2); Motsuku, L. (2); Shandukani, M.B. (3); Kruger, P. (4); Mabuza, A. (5); Raswiswi, E. (6); Moonasar, D. (3)
(1) National Institute for Communicable Diseases; (2) Field Epidemiology and Training Program; (3)National Department of Health; (4) Limpopo Provincial Department of Health; (5) Mpumalanga Provincial Department of Health; (6) KwaZulu-Natal Provincial Department of Health
Abstract There was an unexpected increase in malaria cases in the 2014/2015 season. As South Africa aims to eliminate malaria by 2018, the surveillance system needs to be evaluated and strengthened to prevent future occurrences of outbreaks and decrease malaria transmission to zero. The malaria surveillance system was evaluated to measure various attributes of surveillance system. A retrospective records review for the period April 2014 to September 2015 was conducted using the CDC guidelines for evaluating a public health surveillance system. Not all attributes outlined in the guidance document were used. Simplicity (not quantified, but assessed using the malaria literature), completeness (as measured by the percentage of missing data noted on the record), representativeness (as measured by the percentage of healthcare facilities that were included in the surveillance system), validity (as measured by the number of errors, inconsistency and discrepancies) and timeliness (as measured by median days from date of diagnosis to the case notified to the district, provincial and national level). We reviewed a total of 762 paper based records during the period of the study. The system is simple, diagnosed cases are notified to the district, the district activates case investigation, and the case is notified into the Malaria Information System, which feeds into the provincial and national levels. The data collection tool however differs in each province. Out of the 762 malaria case records reviewed, 87.9% (n=670) were complete and case- specific demographic variables were not missing. The surveillance system was representative, with public and private healthcare facilities in the endemic districts represented. The median timeliness was 5.1 days, ranging from <24 hours to 8 days. Data validity was at 24% (n=183) of errors were noted and 11% (n=84) of inconsistencies notes. The malaria surveillance system succeeded in capturing malaria cases from the healthcare facility level to the provincial and national level and for case investigation. However, a faster system that will be able to detect outbreaks in a timely manner should be developed.
Page 100 of 170 EVALUATING THE UTILITY OF AMA-1 AND MSP-1 AS SEROLOGICAL MARKERS OF MALARIA EXPOSURE FOR SURVEILLANCE IN THE LOW TRANSMISSION, PRE-ELIMINATION SETTING OF NAMIBIA
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Chisenga, M.P. (1); Medzihradsky, O. (2, 3); McCreesh, P. (2); Drakeley, C. (6); Tetteh, K. (6); Roberts, K. (2); Smith, J.(2); Gosling, R.(2); Greenhouse, B. (2); Mumbengegwi, D. R. (1) Hsiang, M. (2, 3, 4, 5)
(1) Multidisciplinary Research Center, University of Namibia, Windhoek, Namibia; (2) Malaria Elimination Initiative, Global Health Group, University of California, San Francisco (UCSF), USA; (3) Department of Medicine, University of California San Francisco, California; (4) Department of Pediatrics, UCSF Benioff Children's Hospital, UCSF; (5) University of Texas Southwestern Medical Center, Dallas, Texas; (6) Department of Immunology and Infection, London School of Hygiene and Tropical Medicine
Abstract Namibia aims to eliminate malaria by 2020. However, a recent unexpected outbreak in the North of the country threatens this goal. New diagnostic and surveillance methods are urgently needed to track transmission intensity and predict the geographic location of outbreaks. Malaria incidence measured through passive surveillance has limited utility due to challenges related to access to healthcare, health-seeking behaviour and case reporting. Prevelence surveys based on infection data capture a snapshot, showing only a handful of infections in a low transmission setting. Malaria serological studies are a potentially useful tool in these areas as they provide information on cumulative exposure over time and are simple and inexpensive to perform. It is, however, unclear whether standard serological markers Apical membrane antigen (AMA-1) and Merozoite surface protein (MSP-1), will be informative due to their persistence for many years. This longitudinal study aims to describe the serologic profiles of symptomatic and asymptomatic subjects in the generally low transmission area of western Zambezi region. Cases include PCR-confirmed malaria reported from six randomly selected health facilities between February and August 2015, as well as PCR- confirmed asymptomatic malaria cases identified through a cross-sectional survey from the same study area, each case with two age, gender and geography matched controls. Subjects would be followed up 6-12 months, and 12-18 months after initial entry to collect serum and dried blood spots via finger prick for PCR and for ELISA targeting AMA-1 and MSP-1. Raw optical densities will be converted to antibody titers based on standard curves from known positive controls. Data will be analysed to describe decay or persistence of the antibody titers in known cases compared to controls and stratified analyses will be performed by age group and endemicity of the geographic area. Samples from the first and second time point have been collected and are undergoing laboratory testing. The final data collection will occur from September to November 2016 and analysis will be completed thereafter. These results will be used to evaluate the utility of AMA-1 and MSP-1 for serological surveillance in the low transmission, albeit outbreak prone setting of Namibia.
Page 101 of 170 THREE PARALLEL INFORMATION SYSTEMS FOR MALARIA ELIMINATION IN SWAZILAND, 2010-2015: ARE THE NUMBERS THE SAME?
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Zulu, Z. (1); Kunene, S. (1); Mkhonta, N. (1); Owiti ,P (2); Sikhondze, W. (3); Mhlanga, M. (4); Simelane, Z. (5); Geoffroy, E. (6); Zachariah, R. (7)
(1) Ministry of Health, National Malaria Control Programme, Manzini, Swaziland; (2) Academic Model Providing Access to Healthcare (AMPATH) Eldoret,Kenya; (3) Ministry of Health, Mbabane, Swaziland; (4) Ministry of Health, Epidemic, Preparedness and Response Unit, Ezulwini, Swaziland; (5) Ministry of Health, Strategic Information Department, Mbabane, Swaziland; (6) Global AIDS Interfaith Alliance, California, USA; (7) Medecins Sans Frontiers’, Operational Centre Brussels, Luxembourg
Abstract In order to eliminate malaria, accurate and timely reporting and tracking of all confirmed malaria cases is crucial. Swaziland, a country in the pre-elimination stage has three parallel health information systems: the Health Management Information System (HMIS); the Immediate Disease Notification System (IDNS) and the Malaria Surveillance Database System (MSDS). We aimed to assess if there was any discrepancy between numbers of confirmed malaria cases reported in the HMIS and IDNS in relation to the MSDS (used as the bench mark). A cross-sectional study using country-wide program data (2010- 2015). The MSDS is a dedicated and comprehensive malaria database; the IDNS is meant to provide early warning and trigger case investigations to prevent onward malaria transmission and potential epidemics; the HMIS reports on all morbidity at health facility level. Discrepancies were stratified by health facility level and type. Consistent over reporting of 9% to 85% was noticed in the HMIS and principally at the primary health care level (clinic and/or health centre). In the IDNS, the discrepancy went from under reporting 12% of cases to over reporting 32% and this was also seen at the primary care level. At the hospital level, there was under reporting in both HMIS and IDNS. There is considerable discrepancy in numbers of confirmed malaria in the HMIS and IDNS in Swaziland. This may misrepresent malaria burden and importantly, delay case investigation of malaria cases predisposing the population to potential epidemics. There is an urgent need to improve data integrity in order to better guide and evaluate efforts towards malaria elimination.
Page 102 of 170 A PROTOCOL TO EVALUATE THE EFFECTIVENESS AND FEASIBILITY OF REACTIVE TARGETED PARASITE ELIMINATION (TPE) TO REACTIVE CASE DETECTION (RACD) AS A COMMUNITY LEVEL INTERVENTION IN SWAZILAND
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Dlamini, B. (1); Dufour, M. (2); Mkhonta, N. (3); Malambe, C. (3); Maphalala, G. (4); Ntshalintshali, N. (1); Helb, D. (5); Greenhouse, B. (6); Baltzell, K. (7); Sturrock, H. (2, 5); Dorsey, G. (5); Kandula, D. (1); Cohen, J.M. (1); LeMenach, A. (1); Gosling, R. (2, 5); Kunene, S. (3); Hsiang, M. (4, 8)
(1) Clinton Health Access Initiative; (2) Department of Epidemiology and Biostatistics, University of California, San Francisco (UCSF); (3) Swaziland National Malaria Control Program; (4) Malaria Elimination Initiative, Global Health Group, UCSF; (5) National clinical Laboratory Services; (6) Department of Medicine, UCSF; (7) Department of Family Health Care Nursing, UCSF; (8) Department of Pediatrics, University of Texas Southwestern Medical Center
Abstract Reactive case detection (RACD), which is the identification of infections in communities and/or households of cases identified by the passive surveillance system, is recommended for malaria elimination yet current available diagnostics have limited sensitivity resulting in low positivity field detection rates. Treatment without testing, known as targeted parasite elimination (TPE), may be a more effective and feasible approach for reducing and interrupting malaria transmission. As an operational research study embedded into the activities of Swaziland’s National Malaria Control Program (NMCP), a cluster-randomised control study was designed to evaluate reactive TPE in the low transmission setting of Swaziland. The primary aim of the study is to compare the impact of reactive TPE versus RACD on malaria incidence, with the hypothesis that TPE will result in lower cumulative malaria incidence. A total of 77 high risk localities or clusters were selected for the trial (cumulative population 124,737) and randomized to either TPE or RACD. Over 2 years (September 2015 to August 2017), individuals residing within a radius of 500 m from a passively-detected index case will be targeted for RACD and those within 200 m will be targeted for TPE within 1 week of index case presentation. Incidence will be measured using case reporting through Swaziland’s passive surveillance system with subsequent household follow-up to identify which intervention arm the case resides in. The study is powered to detect a difference in cumulative incidence of 1.1 cases per1000 population in the TPE arm versus 2.2 cases per1000 population in the RACD arm observed in 3 previous malaria seasons. The study’s primary outcome indicators are seroprevalence (in a cross sectional survey at the end of the study; 2017 and prevalence), proportion of imported incident cases, and transmission potential (utilizing genotypes). Secondary outcome measures of feasibility include: coverage, adherence, serious adverse events, acceptability, and cost-effectiveness. The findings of this study will inform malaria elimination strategies for Swaziland and other countries pursuing malaria elimination.
Page 103 of 170 IMPACT OF INTENSE VECTOR SURVEILLANCE IN KWAZULU-NATAL
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Dandalo, L. C. (1, 2); Brooke, B.D. (1, 2 ); Munhenga, G. (1, 2); Zikhali, J. (1) Lobb, L.N. (1,2); Ngxongo, S.P. (3); Raswiswi, E. (3); Wood, O.R. (1,2); Mofokeng, M. (1,2); Koekemoer, L.L. (1, 2)
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Center for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Sandringham, Johannesburg, South Africa; (3) Department of Health-KwaZulu-Natal, KwaZulu-Natal Provincial Government.
Abstract Anopheles arabiensis has been implicated as the main malaria vector in KwaZulu-Natal province. Elimination of this vector using insecticide residual house spraying (IRS) has been a challenge due to its variability in feeding and resting behaviour, whereby a proportion of the populations of this species tend to feed and rest outdoors. This has necessitated a search for additional vector control methods such as the Sterile Insect Technique (SIT). The aim of this study was to establish baseline and seasonal variation in density of an An. arabiensispopulation at Mamfene, KwaZulu-Natal (KZN). Surveillance activities were increased at three sentinel sites in Mamfene, KZN. Mosquitoes were collected between January 2014 and June 2016 using modified buckets, clay pots and window exit traps. All captured mosquitoes were morphologically identified and those belonging to the An. gambiae complex and An. funestus group were identified to species level using a Polymerase Chain Reaction (PCR) assay. All female mosquitoes from these taxa were tested for Plasmodium falciparum infectivity using the enzyme- linked immunosorbent assay (ELISA). Anopheles arabiensis was the most abundant species consisting of 78.8 % of all mosquito collections. The population density was highest during the rainy season and afterwards slowly declining, reaching its lowest in the winter months. Clay pots appeared to be the most productive collection method. An overall Plasmodium falciparum infection rate of 0.28% was detected in the An. arabiensis mosquitoes collected. This is the first record of An. arabiensis being infected with Plasmodium falciparum in South Africa. The study confirmed An. arabiensis is a vector of malaria in KwaZulu-Natal and that it occurs throughout the year at Mamfene. The novel collection method of clay pots appears to be a comparatively effective outdoor mosquito collection method although this needs further investigation. Intensive surveillance over a long period of time has provided valuable insight into the seasonal population dynamics of An. arabiensis in KZN.
Page 104 of 170 RAPID RESTORATION OF CHLOROQUINE SENSITIVITY IN PLASMODIUM FALCIPARUM MALARIA OF MUTASA DISTRICT, ZIMBABWE
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Matsena-Zingoni, Z. (1); Mudare, N. (2); Makuwaza, A. (1); Munyati, S. (2); Gwanzura, L. (2); Mutambu, S.L. (1); Mason, P. (2); Moss, W. (4); Kobayashi, T. (4); Mharakurwa, S. (3,4)
(1) National Institute of Health Research, Malaria Section, Harare, Zimbabwe (2) Biomedical Research and Training Institute, Avondale, Harare, Zimbabwe (3) Faculty of Health Sciences, Africa University, Old Mutare, Mutare, Zimbabwe (4) John Hopkins Bloomberg School of Public Health, Baltimore, USA
Abstract The emergence of Plasmodium falciparum drug resistance erodes the arsenal of effective antimalarials and poses one of the major threats for malaria control and elimination programmes. It is imperative to monitor parasite resistance profiles to available programme antimalarials, both old and new. It has been shown that P. falciparum resistance to chloroquine, the former safest and most effective antimalarial, may recede after years of drug withdrawal from use. The current study determined the levels of the chloroquine resistance-conferring PfCRT K76T mutation among falciparum malaria infections of Mutasa District, Zimbabwe in 2013, compared to 2003, when chloroquine monotherapy was suspended. Cross-sectional surveys were conducted in 2003 and 2013, on 408 and 373 study participants, respectively, from representative households of Mutasa District. Parasite DNA samples were collected using filter paper dry blood spots (DBS). DNA was extracted by the Chelex method and P. falciparum infections were genotyped at PfCRT amino acid codon 76 using nested PCR and restriction enzyme digestion. Of the participants screened in 2003 and 2013, 66 (17.7%) and 49 (12%), respectively were PCR-positive for P. falciparum and all were successfully genotyped at the key chloroquine resistance-conferring PfCRT codon 76. In 2003, there was a high prevalence (63.6%) of the K76T chloroquine resistance mutant, including 3% mixed mutated and wild type infections, leaving only 33% wild type infection. By contrast, 2013 malaria infections had only 3% K76T mutants, the rest (97%) being wild type. No K76T mutant was found among P. falciparum infections in the mosquito mid-gut phase. There has been a considerable return of chloroquine-sensitive P. falciparum malaria in Mutasa, less than ten years after suspension of chloroquine use. Resumption of indoor residual spraying, sporogonic selection in the local vector An. funestus and adoption ACTs since 2008 may be contributing to the rapid restoration of the wild type parasitaemia. Further monitoring is recommended as it may later be viable to consider incorporating chloroquine as a safe, effective and affordable ACT partner drug for the national malaria control and elimination programme.
Page 105 of 170 USING DRIED PLASMODIUM FALCIPARUM-INFECTED BLOOD SAMPLES AS POSITIVE CONTROLS FOR MALARIA RAPID DIAGNOSTIC TESTS
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Ming Sun, L.K. (1); Poonsamy, B. (1); Frean, J.A. (1, 2)
(1) Parasitology Reference Laboratory, National Institute for Communicable Diseases, National Health Laboratory Service; (2) Wits Research Institute for Malaria, Faculty of Health Sciences, University of the Witwatersrand
Abstract Quality control (QC) of any rapid diagnostic test (RDT) is important; for malaria RDTs no standardised QC product/procedure currently exists. Dried tube samples (DTS) have been successfully used for QC for other serological tests and have recently been tested for malaria. In this study, the suitability of dried Plasmodium falciparum-infected blood samples as QC samples for malaria RDTs in our laboratory was evaluated. Routine blood samples, confirmed by microscopy and RDT as Plasmodium falciparum-positive,were used. Four patient samples with varying parasitaemias (0.1% to 10.4%) were selected. Fifty microliters of the EDTA whole blood was aliquoted into at least 20 sterile plastic tubes, either deep conical or shallow round-bottom. Half of the tubes had 50 µl of sterile water added to lyse the blood; this generated eight batches. The aliquots were air dried inside a biosafety cabinet overnight, then sealed and stored at 4°C. The batches were tested at 1, 4, 12 and 56 weeks. For testing, two aliquots of each batch were rehydrated with 50 µl PBS/Tween and deionised water respectively.The DBS was allowed to reconstitute at room temperature for one hour, then mixed with a pipette and the P. falciparum RDT performed. The intensity of the band was recorded as intense, faint, or very faint. All samples tested yielded positive P. falciparum RDT results, even at 56 weeks of storage. There were some variations in band intensity at different time points but none were very faint. The shallow round-bottom tubes dried within 24 hours, whereas, the deep conical tubes took longer to dry. There was no difference in lysing or not lysing the blood before drying, or using deionised water instead of PBS/Tween. From our results, we suggest that for the preparation of the DTS use round-bottom tubes and for the reconstitution allow more than an hour. We find that the DBS is an acceptable method to use for the QC of P. falciparum RDTs. It is suited to low resource settings, as minimal equipment and reagents are needed, and the tubes are stable for up to 56 weeks.
Page 106 of 170 DECREASED PREVALENCE OF THE PLASMODIUM FALCIPARUM CHLOROQUINE RESISTANCE TRANSPORTER GENE MUTATION K76T NINE YEARS AFTER CHLOROQUINE TREATMENT WITHDRAWAL IN CHIPATA AND KATETE DISTRICTS, EASTERN ZAMBIA
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Mwenda C, M. (1); Mharakurwa, S. (2); Hamainza, B. (3); Moonga, H. (3); Kumar, N. (4); Chipeta, J. (5)
(1) University of Zambia, School of Medicine Department of Biomedical Sciences; (2) Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore MD 21205, USA; (3)Ministry of Health National Malaria control centre; (4) Department of Tropical Medicine and Director of Vector-Borne Infectious Diseases Research Centre, Tulane University School of Public Health and Tropical Medicine; (5) Department of Paediatrics and Child Health, School of Medicine, Malaria Research Unit (SMUTH- MRU), Lusaka, Zambia
Abstract Zambia, like many malaria endemic countries, withdrew chloroquine in 2003 as a first line treatment for uncomplicated Plasmodium falciparum malaria due to widespread chloroquine resistance and increased treatment failure. The artemisinin combination regimen artemether–lumefantrine, was adopted as the new first line treatment for uncomplicated malaria. In a cross section survey, conducted between May and June 2012 to investigate the prevalence of chloroquine resistance nine years after the removal of the drug, we assayed, by polymerase chain reaction, the prevalence of the K76T mutation in the Plasmodium falciparum chloroquine resistance transporter (Pfcrt) gene which is the most important mutation conferring chloroquine resistance in Plasmodium falciparum. Out of the 244 P. falciparum infected samples that gave valid PCR results, 84.8% had the chloroquine sensitive genotype, 14.3% expressed the chloroquine resistance genotype and 0.4% expressed both the sensitive and the resistant genotype. The removal of the drug pressure from the P. falciparum population seems to have resulted in the reduction of chloroquine resistant parasites over the past nine years.
Page 107 of 170 MALARIA TRENDS SOUTH AFRICA, 2011-2015
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Shandukani, M.B. (1); Misiani, E. (1); Ngwenyama, B. (1, 2); Raman, J. (2); Graffy, R. (3); Kruger; P. (4); Mabuza, A. (5); Raswiswi, E. (6); Moonasar, D. (1)
(1) National Department of Health (DOH); (2) National Institute for Communicable Diseases; (3) Clinton Health Access Initiative; (4) Limpopo Provincial Department of Health; (5) Mpumalanga Provincial Department of Health; (6) KwaZulu-Natal Provincial Department of Health
Abstract South Africa plans to eliminate malaria by achieving an incidence of zero cases by 2018. The feasibility of malaria elimination will be possible through monitoring of current interventions and assessing their impact on local transmission. The aim of this study was to describe seasonal malaria trends in South Africa, for 2011to 2015. This was a descriptive retrospective study, utilizing secondary seasonal (July to June) malaria data extracted from the Malaria Information System. STATA version 13 was used to analyse the data and assess for trends. Data from malaria endemic provinces of Mpumalanga, Limpopo and KwaZulu Natal. From July 2011 to June 2015 there were a total of 33 142 cases and 350 deaths reported. Local malaria cases were reported to contribute 44% of the overall cases between 2011-2015 seasonal years. Within the study period the year 2012/2013 showed the lowest number of local cases, with 1167 (21%) local cases reported from a total of 5587 cases. In years where there was a malaria spike, local cases as a percentage of the total number of cases increased. Total cases reported in 2011/12, 2012/13, 2013/14, and 2014/15 were 5546, 5587, 10599 and 11410 respectively. In these years the corresponding percentage of local cases was 36%, 21%, 42% and 61%. The programme needs to strengthen surveillance interventions, and work at the district level to prioritise identification and addressing the foci of transmission to halt malaria transmission. Malaria elimination requires adequate financial and human resources, which is threatened by a gradual reduction of malaria funding within the three endemic provinces.
Page 108 of 170 PRELIMINARY STUDY ON THE PREVALENCE OF RESISTANCE ASSOCIATED POLYMORPHISM IN PLASMODIUM FALCIPARUM FIELD ISOLATES PRESENTED IN GUATENG, SOUTH AFRICA
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Sijuade, A. (1); Botha, M. (1); Niemand, J. (1); Louw, A. (1); Birkholtz L. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control (UP ISMC) Pretoria, University of Pretoria, South Africa
Abstract The prevalence of resistant Plasmodium falciparum (Pf) parasites remains an obstacle to malaria control and elimination programmes. However, as part of malaria elimination efforts this study was designed to understand the polymorphism of resistance associated with genes in isolates of Pf infected patients who visited different hospitals in Gauteng Province. Blood samples spotted on filter paper from 17 patients with microscopic and polymerase chain reaction (PCR) confirmed P. falciparum mono-infections were collected between 2014-early 2015. Single nucleotide polymorphisms in the Pf resistance transporter (Pfcrt K76T), multi drug resistance (Pfmdr1 N86Y), dihydrofolate reductase (Pfdhfr N51I, C59R, S108N, I164L) and dihydropteroate synthetase (Pfdhps A436S, G437A and E540K) genes were assessed from chelex extracted DNA using mutation- specific nested PCR and RFLP based methods. The triple pure mutant (108N, 51I and 59R) was detected in 88% of the isolates, and 6% of the isolates were triple mixed. The DHPS double mutant Gly-437/Glu-540, both in mixed and pure form were found in 5% and 41% respectively. In addition, 41% and 12% were single mutation Gly-437 or Glu-540 and wild-type. Likewise, mixed infection in Pfmdr1 (N86 + Y86) was detected in 12% of the isolates and no Pfcrt 76T mutation was detected. The prevalence of quintuple mutant (Pfdhfr 108N, 51I and 59R and Pfdhps 437G and 540E), which is associated with high-level of sulphadoxine-pyrimethamine (SP) resistance in-vivo was identified in 41% of the isolates, while 35% carried quadruple mutation (Pfdhfr 108N, 51I and 59R and Pfdhps 437G) and 12% were triple mutation. There were no double mutants (437G and 540E) detected. The single use of sulphadoxine-pyrimethamine as seasonal IPT must be discouraged. The use of SP must be associated in combination with amodiaquine, artesunate or piperaquine for instance to prevent the increase of resistance to SP.
Page 109 of 170 MOLECULAR DIAGNOSIS OF MALARIA IN THE ZAMBEZI REGION OF NAMIBIA MOVING TOWARDS ELIMINATION
Abstract Theme Surveillance (vector, parasite, resistance)
Authors and Affiliations Tambo, M. (1); Dausab, L. (1); Mukosha, C. (1); McCreesh, P. (2); Roberts, K. (2); Murphy, M. (3); Greenhouse, B. (3); Cueto, C. (2); Smith, J. (2); Mumbengegwi, D.R. (1); Hsiang, M.S. (2, 4, 5); Gosling, R. (2)
(1) University of Namibia School, Multi-disciplinary Research Center, Windhoek, Namibia; (2) Malaria Elimination Initiative, Global Health Group, University of California, San Francisco, USA; (3) Department of Medicine, University of California, San Francisco, California; (4) Department of Pediatrics, UCSF Benioff Children's Hospital, University of California, San Francisco, California; (5) University of Texas Southwestern Medical Center, Dallas, Texas
Abstract Namibia, a low transmission setting for malaria, declared a goal to eliminate malaria within its borders by the year 2020. One challenge for low transmission settings is low-density asymptomatic infections which in other settings have been reported to make up 70 - 80% of the total number of malaria cases. This poses a challenge for active surveillance of asymptomatic infections as routinely used RDTs have difficulty in detecting malaria at low parasite density (<50 - 100 parasites per microlitre). When these cases go undetected, they can act as reservoirs that could perpetuate the spread of malaria. In this study, we performed a cross-sectional survey in the catchment areas of six randomly selected health facilities in the low transmission area of western Zambezi in Northern Namibia. We performed malaria testing using rapid diagnostic tests (RDTs) as well as highly sensitive molecular tools PCR and LAMP (loop mediated isothermal amplification). A total of 2017 individuals were enrolled, among which there were a total of 16 RDT positives (0.79%), 26 PCR positives (1.3%) and 43 LAMP positives (2.1%). PCR and LAMP detected about 1.6 fold and 2.7 fold more respectively the number of cases detected by RDTs. Using LAMP or PCR positivity as gold standard, the sensitivity, specificity, PPV and NPV values of RDTs were 7.4%/93.6%/1.5%/98.7% for PCR and, 2.3%/99.2%/6.3%/97.8% for LAMP. Although we did not have a definitive gold standard such as high volume qPCR, LAMP may be more sensitive than PCR for the detection of low density infections. A high proportion of infections in our study were missed by RDTs. In order to detect and treat asymptomatic infections which serve as reservoirs for persistent transmission, more sensitive molecular tools such as LAMP and PCR will be needed.
Page 110 of 170 SUMMARY OF FINDINGS MALARIA ELIMINATION OPERATIONAL CAPACITY ASSESSMENT SURVEY – ZIMBABWE 2015
Abstract Theme Case Management
Authors and Affiliations Mberikunashe, J. (1); Tangwena, A. (1); Dube, B. (1); Owen, J. (2); Madinga, M. (2); Gambinga, B. (2); Svisva, A. (2); Dolenz, C. (2); Pindolia, D. (2); Woolheater, K. (2); Ahmed, J. (2)
(1) National Malaria Control Program Zimbabwe; (2) Clinton Health Access Initiative, Boston, MA, USA
Abstract Zimbabwe is scaling up its sub-national malaria elimination campaign from the 7 districts in Matabeleland South province to an additional 13 low burden districts. The health system in the new districts must be reoriented for malaria elimination, with the focus on interrupting transmission and reaching zero local cases in these districts. A feasibility assessment was conducted in June 2015, highlighting infrastructure, personnel and training gaps to assess the operational readiness for the 13 new districts to successfully scale up interventions. The assessment was conducted in 18 low burden districts. The sampled districts were chosen based on their epidemiological pattern of low incidence over time. A mixed method data gathering approach was implemented, which included an entomological field survey, retrospective gathering and analysis of malaria indicator data from HMIS databases, and cross-sectional interviews with health facility-based staff, community-based health workers, and key district and province-level stakeholders. The survey was designed to gather key indicators aimed at assessing personnel, infrastructure and training related to malaria considered critical for an elimination campaign. Results from the survey were illustrative of both the strengths and weaknesses of the program and the ministry as a whole. Analysis of case management indicators revealed that 99% of suspected cases received a parasitological test in 2014, marking a major strength of the program. However, significant gaps in both infrastructure and personnel were highlighted. 44% of Environmental Health Practitioner (EHP) posts in the surveyed districts were vacant, illustrative of a significant gap given the importance of EHPs in conducting Indoor Residual Spraying and case-based surveillance activities. Furthermore an average of 10% of the facilities surveyed had access to IT equipment for data collection and reporting. Approximately, one functional motorbike was reported to serve four health facilities- ideally every facility should have a functional motorbike. Gaps in infrastructure and personnel pose a challenge for malaria elimination in Zimbabwe. Procurements have been planned to fill gaps in motorbikes and IT equipment, while personnel gaps remain a challenge.
Page 111 of 170 DEVELOPMENT AND IMPLEMENTATION OF A MALARIA COMMODITY TRACKING AND ORDERING TOOL FOR IMPROVED FORECASTING, PROCUREMENT AND SUPPLY MANAGEMENT OF MALARIA DIAGNOSTIC AND TREATMENT COMMODITIES
Abstract Theme Case Management
Authors and Affiliations Mathobela, M. (1); Chitundu, S. (1); Soble, A. (2); Vilakati, S. (1); Mthethwa, S. (1); Ntshalintshali, N. (2); Kunene, S. (1)
(1) National Malaria Control Programme, Manzini, Swaziland; (2) Clinton Health Access Initiative, Mbabane, Swaziland
Abstract To ensure that all health facilities providing malaria services are adequately stocked with malaria commodities the National Malaria Control Programme (NMCP) introduced a malaria commodity reporting and ordering tool, whereby all health facilities providing malaria services are required to utilize the tool to submit monthly malaria commodity consumption and order reports. These reports were intended to improve the forecasting, procurement and supply of malaria commodities and limit malaria commodity stock outs in health facilities. The NMCP developed the Monthly Malaria Stock Report and Order Form, comprised of: (1) monthly aggregated data on the number of uncomplicated and severe malaria cases, patients tested with RDT and patients receiving prophylaxis; (2) initial and final monthly stock totals for malaria diagnostic and treatment commodities, inclusive of quantities received or disbursed (3) days of zero stock for each malaria commodity and (4) average monthly consumption calculations used to determine commodity orders. Regional trainings were conducted in February 2015 for 95 health facilities. The NMCP subsequently trained all outstanding facilities (130 health facilities) through health facility visits between March and April 2015. Between March and December 2015, out of a total of 225 participating health facilities, only 31% of health facilities were consistently reporting commodity consumption and order reports. During this same period, 84% of reporting health facilities reported zero malaria commodity stock-outs. When aggregating these same reporting health facilities, there were only 5 occurrences where health facilities were understocked, 4 related to mefloquine, Swaziland’s primary malaria prophylaxis, and 1 related to quinine sulphate, a second-line malaria treatment. As Swaziland approaches malaria elimination, improved consumption tracking will be increasingly important to ensure all cases are parasitologically confirmed and effectively treated. Improved commodity tracking data will further allow the NMCP to determine the number of suspected malaria cases tested by an RDT, and calculate annual blood positivity rate, a WHO requirement for elimination certification. Swaziland’s preliminary implementation experience demonstrates that new data collection tools and reporting systems at the health facility level require regular training, supervision and feedback to ensure effective use and uptake.
Page 112 of 170 DEVELOPMENT OF A PHARMACOVIGILANCE SAFETY MONITORING TOOL FOR THE ROLLOUT OF SINGLE LOW- DOSE PRIMAQUINE AND ARTEMETHER-LUMEFANTRINE TO TREAT PLASMODIUM FALCIPARUM INFECTIONS IN SWAZILAND: A PILOT STUDY
Abstract Theme Case Management
Authors and Affiliations Malambe, C. (1); Poirot, E. (2, 3); Soble, A. (4); Ntshalintshali, N. (4); Mwandemele, A. (4); Dlamini, N. (1); Vilakati, S. (1); Pan, S. (4); Darteh, S. (6); Maphalala, G. (7); Brown, J. (2, 3); Hwang, J. (2, 8); Pace, C. (9); Stergachis, A. (10); Vittinghoff, E. (2); Kunene, S. (1); Gosling, R. (2, 3)
(1) National Malaria Control Programme, Manzini, Swaziland; (2) Global Health Group, University of California San Francisco, San Francisco, California, USA; (3) Department of Epidemiology and Biostatistics, University of California, San Francisco, California, USA; (4) Clinton Health Access Initiative, Mbabane, Swaziland; (5) University of Namibia, Windhoek, Namibia; (6) International Center for AIDS Care and Treatment Programs, Mbabane, Swaziland; (7) Swaziland Health Laboratory Services, Mbabane, Swaziland; (8) President’s Malaria Initiative, Malaria Branch, U.S. Centers for Disease Control and Prevention, Atlanta, GA, USA; (9) Liverpool School of Tropical Medicine, Liverpool, UK; (10) Departments of Pharmacy and Global Health, Schools of Pharmacy and Public Health, University of Washington, Seattle, USA
Abstract Countries remain reluctant to adopt the 2012 WHO policy on single low-dose (0.25 mg/kg) primaquine (SLD PQ) for Plasmodium falciparum transmission blocking due to concern over risk of drug-related hemolysis among glucose-6-phosphate dehydrogenase-deficient (G6PDd) people Additional context-specific evidence demonstrating that primaquine can be safely deployed is required. The use of active pharmacovigilance methods provides an opportunity to monitor rollout of SLD PQ and generate data that can add to the limited body of evidence on the safety of the drug. Swaziland NCMP developed and piloted the Primaquine Roll Out Monitoring Pharmacovigilance Tool (PROMPT) comprising of: a standardized form collecting a minimum set of data elements designed to support passive and active surveillance, a patient information card to enhance awareness of known adverse reactions of SLD PQ use and a database compiling recorded information. Active follow-up included repeat hemoglobin (Hb) testing and adverse event monitoring on day 7. A 13-month prospective pilot study in two hospital facilities was used to collect preliminary evidence on the feasibility and acceptability of PROMPT, and to collect drug safety data. Enrollment and adherence rates were used to measure feasibility, while trained nurses offered their perceptions of tool acceptability. Of the 102 patients enrolled and administered SLD PQ, 93 (91.2%) returned on or near day 7 for follow-up. Four (4.6%) patients had falls in Hb>25% from baseline, none of whom presented with signs or symptoms of anemia. No patient’s Hb fell to <7 g/dL. Eleven (11.8%) patients reported a total of 20 adverse events. Four non-serious adverse events were considered definitely, probably, or possibly related to SLD PQ. Three serious adverse events, including two deaths, were reported over the study period; none of which were causally related to SLD PQ. No patients required a blood transfusion. PROMPT was well received by nurses as a simple, pragmatic approach to active surveillance that facilitated the collection of SLD PQ safety data. As access to SLD PQ increases, improved pharmacovigilance to monitor and promote the drug safety will be needed. The successful application of PROMPT demonstrates its potential for rapidly generating locally acquired safety data and supporting pharmacovigilance in resource-limited settings.
Page 113 of 170 CASE MANAGEMENT OF MALARIA IN SWAZILAND, 2011 TO 2015: ON TRACK FOR ELIMINATION?
Abstract Theme Case Management
Authors and Affiliations Dlamini, S.V. (1)*; Kosgei, R.J. (2); Mkhonta, N. (3); Zulu, Z. (3); Makadzange, K. (4); Zhou, S. (5); Owiti, P. (6); Sikhondze, W. (7); Namboze, J. (8); Reid, A. (9); Kunene, S. (3)
(1) University of Swaziland, Faculty of Health Sciences; (2) Department of Obstetrics and Gynaecology, University of Nairobi; (3) National Malaria Control Programme (Swaziland); (4) World Health Organisation (Swaziland Country Office); (5) National Institute for Parasitic Diseases, Chinese Center for Disease Control and Prevention; (6) PATH, Nairobi (Kenya); (7) National Tuberculosis Control Programme (Swaziland); (8) World Health Organisation, African Regional Office and Inter-Country Support Team (IST); (9) Operational Research Unit, Operational Centre Brussels, Médecins Sans Frontières, Luxembourg
Abstract Prompt diagnosis using Rapid Diagnostic Tests (RDTs) and/or microscopy and treatment with artemisinin-based combination therapies (ACTs) of all malaria cases are considered key towards achievement of malaria elimination in any country. The Swaziland malaria treatment policy recommends artemether-lumefantrine (AL) for uncomplicated malaria and quinine or artesunate for severe malaria. RDTs and AL were deployed in 2010 and are provided by the National Malaria Control Programme to all health facilities. This paper aims to assess adherence to the diagnosis and treatment guidelines during management of malaria cases in all healthcare facilities in Swaziland between January 2011 and August 2015. A cross sectional descriptive study involving all healthcare facilities that diagnosed and managed malaria cases in Swaziland was conducted. Patient and clinical characteristics such as age, gender, method of diagnosis, and type of treatment were analysed. Quantitative data was sourced from the database of the National Malaria Control Programme. A total of 1 981 records of patients treated for severe and uncomplicated malaria were analysed. Fifty- six per cent these cases were uncomplicated and 14% were severe malaria. The type of malaria was not recorded for 30% of the cases. About 71% of the cases were confirmed by RDT alone, 3% by microscopy alone and 26% by both RDT and microscopy. Ninety-three per cent of the uncomplicated cases were treated with AL alone, 5% with quinine alone and 2% with AL and quinine. Eleven per cent of severe cases were treated with AL alone, 44% with quinine alone and 45% with AL and quinine. Clinics and health centres tended to prescribe AL alone more often in severe malaria, as compared to hospitals (13%; 12%; 4%; P=0.003), respectively. Evidence suggests RDTs or microscopy results are used at all facilities to inform treatment decisions. Poor recording of the type of malaria makes it difficult to adequately assess prescription of antimalarial drugs by healthcare providers. Malaria case management, malaria elimination, Artemisinin-based combination therapy, Rapid Diagnostic Tests, uncomplicated malaria, severe malaria.
Page 114 of 170 COMMMUNITY ANTIMALARIAL USE PATTERNS AND RAPID TESTING FOR FAKE OR SUBSTANDARD DRUGS IN MUTASA DISTRICT, ZIMBABWE
Abstract Theme Case Management
Authors and Affiliations Mharakurwa, S. (1, 2, 3); Mutambu, S. (4); Matsena, Z. (4); Mudare, N. (2); Makuwaza, A. (4); Munyati, S. (2); Gwanzura, L. (2); Mason, P. (2); Moss, W. (3); Cui, L. (5)
(1) Africa University, Old Mutare, Mutare, Zimbabwe; (2) Biomedical Research and Training Institute, Harare, Zimbabwe; (3) Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; (4) National Institute of Health Research, Ministry of Health and Child Care, Harare Zimbabwe; (5) Pennsylvania State University, Philadelphia, USA
Abstract Fake or substandard antimalarial drugs pose one of the important threats to malaria control and elimination programmes. A large share of global malaria deaths is attributable to the ineffective medicines and one third of antimalarial drugs in endemic countries, especially Asia and Africa are fake. There is an increasing market for fake or substandard antimalarials, flourishing with unregulated or informal drug use, which is also associated the concomitant emergence of artemisinine-tolerant malaria. Owing to anecdotal reports of informal drug vending in parts of Zimbabwe, the current study examined community antimalarial use patterns and tested drug samples from informal vendors for presence of fake/substandard regimens in Mutasa District. The objectives of current study were (i) to determine community antimalarial drug consumption practices and the magnitude, extent and aetiology of informal use; and (ii) to ascertain presence of fake/substandard antimalarials in Mutasa District. The study was based willing resident adults of either gender from Mutasa District. Data on community antimalarial consumption and informal drug use were collected by interview using a standard questionnaire. A novel rapid diagnostic test was used to detect fake/substandard antimalarials among drug samples purchased by the from community informal drug vendors. From a total of 168 respondents aged 16 – 76 years (mean [95% CI]: 39 [36.4 – 41.2]) 126 (75%) had previously had malaria and 88 (70%) were treated in the village. Most (78%) were treated by village health workers, 9% by school health masters, but 13% were treated by informal vendors that sold drugs from their homes. Top reasons for seeking treatment from informal vendors were accessibility any time (30 %), antimalarials always available (17 %), lower cost (13 %), proximity to home (13 %) and no need to be tested (13 %). Community appreciation of village heath workers and health facilities was high but both were generally considered too few and far between, compared to informal vendors. The study showed that most malaria cases are now treated at village level. Informal vending can be successfully tackled by matching up key comparative advantages fulfilled by the vendors, or empowering the vendors with accreditation and requisite training given to VHWs. So far no fakes/substandards were detected among all the antimalarial samples tested in the area. However, continued monitoring is recommended.
Page 115 of 170 SIMPLIFYING MALARIA PARASITE COUNTS ON THIN BLOOD FILMS
Abstract Theme Case Management
Authors and Affiliations Poonsamy, B. (1); Iyaloo, S. (1); van Deventer, R. (1); du Plessis, D. (1); Ming Sun, L. (1); Frean, J. (1, 2)
(1) Parasitology Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases (NICD) of the National Health Laboratory Service (NHLS), South Africa; (2) University of the Witwatersrand, Johannesburg, South Africa
Abstract The standard method for malaria diagnosis remains microscopy of stained blood films; this method has an advantage of providing an estimate of the patient’s parasitaemia level. Knowing the parasitaemia can assist with identifying cases of severe malaria and monitoring treatment efficacy. Parasite counts can be performed on thick or thin blood films. The former is more suited to lower parasite densities and the latter, higher ones. In South Africa, counts are generally performed on thin blood films, however, there is evidence that methods vary and results are often inaccurate. We simplified and evaluated the method of malaria parasite counting on thin blood films by using a red blood cell (RBC) multiplication factor (RMF) based on a standardised number of RBCs per high-power microscopic field. We randomly selected ten slides from Plasmodium falciparum-positive batches with varying parasite densities. Four experienced microscopists counted the slides using the existing Miller squares graticule and the new RMF methods, at different times. A digital image analysis-based method was used as the gold standard (GS), which was performed by an additional experienced microscopist. We compared the counts from the RMF and Miller squares methods against the GS method, and checked for inter-reader variability. The true counts (GS method) of the 10 slides ranged from 0.07% to 20.15%. We found a high concordance in counts among the readers when using the RMF or Miller squares methods (p<0.001 respectively). Results of both methods were not statistically different from the GS counts (p = 0.358 and 0.508, respectively). However, the RMF counts were slightly higher than the GS counts, while the Miller squares counts were slightly lower than the GS counts. The RMF method was also noted to be less time consuming, compared to the Miller squares method. In order for results to be comparable across laboratories and countries, it is important to have standardised methods. Furthermore, routine laboratories need to produce accurate results in the least amount of time possible. To achieve these objectives, we believe the speed and simplicity of the RMF method makes it the most suitable for routine use.
Page 116 of 170 UNDERSTANDING CROSS-BORDER MALARIA: LESSONS LEARNED FROM EASTERN ZIMBABWE
Abstract Theme Regional malaria and crossborder issues affecting elimination
Authors and Affiliations Kanyangarara, M. (1); Mamini, E. (2); Mharakurwa, S. (3, 4), Munyati, S (2); Gwanzura, L. (5); Kobayashi, T. (6); Shields, T. (4); Mutambu, S. (7); Mason, P.R. (2); Curriero, F. (6); Moss, W.J. (4,6) for the Southern Africa International Centers of Excellence for Malaria Research
(1) Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD; (2) Biomedical Research Training Institute, Harare, Zimbabwe; (3) The Malaria Institute at Macha, Choma, Zambia; (4) Department of Molecular Microbiology & Immunology, Johns Hopkins Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD; (5) Department of Medical Laboratory Sciences, University of Zimbabwe College of Health Sciences, Harare, Zimbabwe; (6) Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD; (7) National Institute of Health Research, Harare, Zimbabwe
Abstract Malaria control efforts are particularly difficult in areas along international borders of countries. Border areas have environmental, administrative and geographic characteristics that uniquely affect the epidemiology and control of malaria in those areas. Differences in malaria endemicity, the porosity of borders, frequent human population movement, and suitable climatic conditions for malaria transmission are major causes of the heavy transmission of malaria along international borders. The objective of this study was to understand the malaria situation in Mutasa District, Zimbabwe, which shares a border with Mozambique, and is characterized by resurgent malaria. Previous research indicated that malaria risk was significantly higher among individuals residing closer to the Zimbabwe-Mozambique border. Using cross-sectional household surveys conducted as part of Southern Africa International Centers of Excellence for Malaria Research (ICEMR) project, routine data from health-facility based surveillance and publicly available remotely sensed environmental data, this research attempts to elucidate the underlying mechanisms affecting the malaria situation along the Zimbabwe-Mozambique border. Firstly, a descriptive analysis of laboratory confirmed autochthonous and imported malaria cases using routine health facility data is presented. Secondly, household survey data is used to quantify the influence of population movement on malaria transmission after adjusting for environmental factors. Lastly, the utility and potential of innovative approaches such GPS data loggers is discussed in the context of providing more detailed information on population movement. Collectively, this research suggest the importance of a two-pronged approach that focuses on improving local malaria control while engaging in regional initiatives to ensure cross-border malaria is minimized. The high risk of malaria on the Mozambique-Zimbabwe border, brings to the fore the relevance and importance of cross- border initiatives to stakeholders in the region.
Page 117 of 170 OPERATIONAL LESSONS LEARNED FROM IRS PROGRAM IMPROVEMENTS IN NAMIBIA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations: Angula, H. (1); Didier, B. (2); Woolheater, K. (2); Pindolia, D. (2); Dolenz, C. (2); Seethaler, T. (2); Renn, S. (3); Pollard, D. (3); Uusiku, P. (1); Iitula, I. (1)
(1) National Vector-borne Diseases Control Programme, Directorate of Special Programmes, Ministry of Health and Social Services, Windhoek, Namibia; (2) Clinton Health Access Initiative, Boston, MA, USA; (3) Akros, Lusaka, Zambia
Abstract Indoor Residual Spraying (IRS) is a critical but resource-intensive component of malaria elimination programs. To improve IRS effectiveness and efficiency in 2015, Namibia’s National Vector-borne Diseases Control Programme (NVDCP) adopted two technological innovations based on Akros’ mSpray approach: remote enumeration to geo-locate homesteads pre-spraying and tablet-based data collection for real-time field operation monitoring. Enumeration of homesteads took place between June and July 2015. Operationally feasible target clusters of homesteads were then determined and prioritized based on predicted malaria incidence. During IRS implementation, spray team leaders used the tablet-based mSpray application to visualize and capture data, including the number of sprayable and unsprayable structures on the homestead, type of unsprayable structures, number of persons living there, reasons for not spraying, and GPS location. IRS coverage and the proportion of target reached were available to the program through an online dashboard. Following the spray season, a short feedback survey was administered to spray team leaders to assess acceptability of the new approach. 392,174 homesteads (approximately 1.2 million structures) were enumerated, 327,268 homesteads were targeted and 448,530 structures were sprayed. User feedback indicated that the electronic form was easy to use and interest was expressed in utilizing the platform for future IRS seasons however challenges with connectivity and GPS calibration were reported, which allowed only 61,655 of the 448,530 (13.7%) of total structures sprayed to be electronically reported. This method has the potential to provide a more robust denominator for IRS coverage, the principal metric the NVDCP relies on to evaluate the success of the IRS campaign however improvement in implementation and uptake of the new software is required. Namibia’s implementation of IRS cost-effectiveness innovations yields operational lessons for other countries implementing IRS.
Page 118 of 170 AN ASSESSMENT OF COMPLIANCE TO ENVIRONMENTAL SAFEGUARDS IN THE USE OF DDT IN CHONGWE IRS PROGRAMME RURAL PART OF ZAMBIA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Chilabi, A. (1)
(1) Ministry of Health - National Malaria Control Center
Abstract Indoor Residual Spraying (IRS) a vector control measure was introduced in Chongwe District of Zambia in 2005 to help reduce the incidences of malaria. This measure involves the use of chemicals for vector control. Among these chemicals, Dichloro-Diphenyl-Trichroethane (DDT) was used in the program. The use of DDT has over the years been promoted by the World Health Organisation (WHO). However, the chemical can only be used with some restrictions as outlined in WHO guidelines regarding IRS. This is because DDT is highly persistent, lipid soluble, and bio-accumulates through the food chain. These characteristics of DDT coupled with its toxicity make it a health risk. Therefore, to minimize the chances of being exposed to the health risks, deliberate policy and guidelines have been put in place. The production of training manuals which outline the procedures to follow to ensure safety when handling DDT is one of these policies. These training manuals are targeted for protecting spray operators, Community and the environment, therefore the guidelines contained therein are referred to as environmental safeguards. Adherence to these environmental safeguards is of great importance, thus the need to assess the level of compliance in the IRS programme running from 2005 to 2010 in Chongwe District. The overall objective of this study was to assess the level of compliance to environmental safeguards in the use of Dichloro-Diphenyl-Trichroethane (DDT) during the Indoor Residual Spraying (IRS) program in Chongwe District from 2005 to 2010. The study involved the use of both primary and secondary data. The source of the primary data was a cross section study consisting of 132 households. A questionnaire was the main research instrument used for primary data collection. This was done after the households were sampled using the multistage sampling technique. To gather the needed secondary data, several checklists were developed and used to collect data for each environmental safeguard. These data, primary and secondary, were entered in MS Excel which was also used for formatting tables and figures. The rigorous statistical analysis itself was performed in STATA version 12. The results of the study showed that, inventory of DDT received and used was 100%. This gives a good indication of the utilization of the chemical and accountability. However, waste management was below acceptable standards. This is evident from the non-compliance rate of 37% in terms of location and sitting of one evaporation tank. In relation to maintenance of spraying equipment and nozzle calibration, compliance levels to these parameters could not be assessed. This was due to lack of documentation pertaining to the two activities. Nevertheless, majority (97%) of the respondents were aware of IRS and acknowledged having heard of it. Likewise, most (90%), indicated mosquito control as the main objective of IRS. Further, 98% acknowledged their houses being sprayed before 2010. However, 83%, did not know the chemical used during this activity. In addition, 66% of the respondents did not observe the two hour rule after spraying. This is despite 93% acknowledging receipt of instructions not to enter their houses immediately after the spraying exercise. The study found that levels of compliance to environmental safeguards during the IRS programme in Chongwe District with regards to the use of DDT, were very low (36%). Despite having an effective stock inventory control and audit system, waste management of DDT residue was not as expected. Further, documentation to determine the levels of adherence with respect to the use recommended nozzles and maintenance of equipment was non-existent. Additionally, even though the contents of the information, education and communication (IEC) package were of high quality, there was no information specifically on DDT and its effects for the sprayer operators, community and supervisors. And therefore it led to little application of the knowledge obtained from these materials. This is on part of households who had their houses sprayed.
Page 119 of 170 EFFECT OF IRRADIATON ON FEMALE ANOPHELES ARABIENSIS: AN INVESTIGATIVE STUDY TOWARDS THE DEVELOPMENT OF THE STERILE INSECT TECHNIQUE TO CONTROL MALARIA VECTORS IN SOUTH AFRICA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Dandalo, L.C. (1, 2); Bennett, A. (1, 2); Engelbrecht, D. (1, 3); Kemp, A. (4); Munhenga, G. (1, 2); Coetzer, T. (1, 3) and Koekemoer, L.L. (1, 2).
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of the Health Sciences, University of the Witwatersrand, Johannesburg; (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, NICD/NHLS, Johannesburg; (3) Department of Molecular Medicine and Haematology, University of the Witwatersrand and National Health Laboratory Service, Johannesburg; (4) Special Viral Pathogens Laboratory, Center for Emerging and Zoonotic Diseases, National Institute for Communicable Diseases, Sandringham, Johannesburg, South Africa
Abstract Ideally a Sterile Insect Technique (SIT) programme requires the release of sterile males only. Various sexing strategies to separate males from females before SIT releases are available. However, on a large scale operation it is nearly impossible to guarantee 100% female elimination before field releases. Although in a mosquito SIT programme it is a prerequisite to release only males because females transmit diseases it is inevitable that a smaller percentage of females might be accidentally released together with the sterile males. Against this background the effect of irradiation on a local genetic sexing strain of An. arabiensis (GMK) which is ear-marked for SIT pilot releases was investigated to determine the safety and fate of accidentally released females. GMK female pupae were irradiated at 70Gy, an optimal dose which induces sterility in GMK males. Effect of irradiation on adult emergence, longevity, fecundity and fertility was then assessed. The blood feeding capability of the irradiated GMK females and subsequent P. falciparum infected blood were evaluated. Irradiation had negative effects on female adult emergence, fecundity, fertility and feeding capability. However, it did not have an effect on longevity. The major observations were that irradiated blood fed females failed to lay eggs and were more susceptible to P. falciparum infection, 13.2% ± 2.4980 tested positive for parasite oocysts compared to 6.8 % ± 8.7888 for the unirradiated cohorts. Irradiation had an effect on most of the physiological fitness parameters and reproductive capacity of irradiated females with the exception of adult survivorship. Irradiation seems to increase susceptibility to Plasmodium infection indicating that in a mosquito SIT programme it is an essential prerequisite to release only males.
Page 120 of 170 EFFECT OF STABLE AND FLUCTUATING TEMPERATURES ON THE LIFE HISTORY TRAITS OF ANOPHELES ARABIENSIS AND AN. QUADRIANNULATUS UNDER CONDITIONS OF INTER- AND INTRA-SPECIFIC COMPETITION.
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Davies, C. (1, 2); Coetzee, M. (1, 2); Lyons, C.L. (1, 2,*)
(1) Wits Research Institute for Malaria, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Sandringham, Johannesburg, South Africa; * Present address: Plant Protection Research (PPR), Agricultural Research Council, Vredenburg, Stellenbosch, South Africa
Abstract Constant and fluctuating temperatures influence important life-history parameters of malaria vectors which has implications for community organization and the malaria disease burden. The effects of environmental temperature on the hatch rate, survivorship and development rate of Anopheles arabiensis and An. quadriannulatus under different competitive scenarios are studied. The eggs and larvae of laboratory established colonies were reared under controlled conditions at one constant (25°C) and two fluctuating (20 - 30°C and 18 - 35°C) temperature treatments at a ratio of 1:0 or 1:1 (An. arabiensis: An. quadriannulatus). Monitoring of hatch rate, development rate and survival was done at three intervals, 6 to 8 hours apart depending on developmental stage. Parametric ANOVAs were used where assumptions of equal variances and normality were met, and a Welch ANOVA where equal variance was violated (α = 0.05). Temperature significantly influenced the measured life-history traits and importantly, this was evident when these species co-occurred. A constant temperature resulted in a higher hatch rate in single species, larval treatments (p < 0.05). The treatment 18 - 35°C generally reduced survivorship except for An. arabiensis in mixed, larval species treatments where it was similar to values reported for 25°C. Survivorship of both species at 20 - 30°C was not significantly impacted and the adult production was high across species treatments. The development rates at 25°C and 20 - 30°C were significantly different between species when reared alone and in mixed species from larvae and from eggs. The effect of temperature was more pronounced at 18 - 35°C with An. arabiensis developing faster under both competitive scenarios and An. quadriannulatus slower, notably when in the presence of its competitor (p < 0.05). The influence of temperature treatment on the development rate and survival from egg/ larvae to adult differed across species treatments. Fluctuating temperatures incorporating the extremes influence the key life-history parameters measured here with An. arabiensis outcompeting An. quadriannulatus under these conditions. The quantification of the response variables measured here improve our knowledge of the link between temperature and species interactions and provide valuable information for modelling of vector population dynamics.
Page 121 of 170 BIOLOGICAL CONSEQUENCES OF DDT EXPOSURE
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Delport, R. (1, 4); de Jager, C. (2, 3, 4); Aneck-Hahn, N.H. (2, 3, 4); Bornman, M.S. (3, 4)
(1) Department of Chemical Pathology, University of Pretoria, Pretoria, South Africa; (2) Department of Urology, University of Pretoria, Pretoria, South Africa; (3) School of Health Systems and Public Health, University of Pretoria; (4) University of Pretoria Institute for Sustainable Malaria Control
Abstract The objective of this study is to establish the degree of metabolic insult associated with DDT uptake. We have previously reported changes in Retinol-binding protein (RBP) and thyroid hormone concentrations, and impaired semen quality in male subjects (aged 18-40) residing in an endemic malaria area in Limpopo where DDT is used for indoor residual spraying (IRS). This study aims to establish the degree of metabolic insult associated with DDT uptake (duplication of objective). Samples from five of the visits of an on-going study on the health- and fertility-related effects of DDT exposure were analysed for evaluation of liver- and thyroid function, and hormonal status, using routine analysers. RBP concentrations were determined on the Immage TM nephelometer (Beckman-Coulter Ltd., Johannesburg, South Africa) using the Dade-Behring RBP Kit (Marburg, Germany). Due to analytical considerations subjects were categorized as having either normal (RBP ≥ 34 mg/L; n = 160) or relatively low (RBP < 34 mg/L; n = 162) concentrations. Plasma DDT isomers were determined with GC-MS. Manual semen analysis was performed according to World Health Organization (WHO) standards and Computer Assisted Sperm Analysis (CASA: Hamilton-Thorne Research, Beverly, MA; IVOS-version 12) was also employed. Between-group comparisons were performed employing non-parametric test statistics and logistic regression analyses were used for risk assessment. IRS was associated with significantly higher γ-Glutamyltransferase concentrations and significantly lower thyroid- and higher sex hormone concentrations. A higher percentage of sperm with structural abnormalities ‒ cytoplasmic droplets (CD) ‒ were observed in the low-RBP group, accompanied by poorer sperm motility. Crude risk analyses showed that DDT uptake, total testosterone (t-Test), Estradiol, RBP and Free T3 were significant risk factors for increased CD, while independent risk following correction for DDT uptake was established for t-Test, Free-T3 and RBP. Inclusion of DDT uptake, RBP and Free-T3 in the analyses showed a 2.3 fold increase in risk for CD with low Free-T3 (95% CI 1.01-5.16). Although the biochemical changes associated with IRS are subtle, poorer sperm quality is observed with DDT uptake. We postulate that DDT induces a decrease in RBP, which contributes to lower circulating Free-T3 and poorer sperm quality.
Page 122 of 170 EXPOSURE TO DDT/DDE IN RELATION TO HBA1C IN PREGNANT WOMEN OF TSHILIDZINI HOSPITAL, LIMPOPO, SOUTH AFRICA
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Ndwamato, N.N. (1)
1) University of Pretoria: School of Health Systems and Public Health, and University of Pretoria Institute for Sustainable Malaria Control
Abstract There has been growing concern regarding the use of dichlorodiphenyltrichloroethane (DDT) for Indoor Residual Spraying (IRS) to control malaria vectors. DDT is used for IRS in the Vhembe District of the Limpopo Province, South Africa. Exposure to DDT and its metabolites is associated with risk of diabetes in non-pregnant adult populations1,2. We postulated that there could be high number of undiagnosed diabetics and gestational diabetics due to many factors including exposure to DDT. This study was performed to evaluate the relationship between DDT and Hba1c in women at the time of delivery. Data from the Venda Health Examination of Mothers, Babies and their Environment (VHEMBE) were used for this study. Blood samples were collected for HbA1c and DDT/DDE measurements from women presenting at the Tshilidzini Hospital Labour Ward between August 2012 and December 2013. HbA1c was analysed using the Afinion™ AS100 Analyzer and p,p’ and o,p’ DDT/E were measured using gas chromatography-tandem mass spectrometry (GC- MS). A total of 751 mothers with mean age of 26.4 years (range: 18 to 47 years) were enrolled in the study and Hba1c was measured in 689 (92%). Using 6% as cut-off for normal Hba1c, the prevalence of diabetes was 9.6%, which is high for a population of young women. p,p’-DDT and -DDE were>LOD in ≥98% of the samples and the median p,p’-DDT and p,p’-DDE maternal concentrations were 55.3 (19.0-259.3) and 242.2 (91.8-878.7) ng/g-lipid, respectively. Using linear regression, p,p’- DDT, o,p’-DDT and o,p’-DDE concentrations were positively associated with HbA1c (p,p’-DDT: β=0.0021, 95% CI=-0.001, -0.005; o,p’-DDT: β=0.0266, 95% CI=-0.013, 0.067; o,p’-DDE: β=0.0289, 95% CI=-0.0368, 0.4263.96 respectively; p,p’-DDE: β=-0.0002, 95% CI=-5-0.002, 0.02;) and no association was statistically significant. There were no significant associations between HbA1C and DDT exposure in the mothers of the VHEMBE birth cohort study. Future studies of VHEMBE should follow the women for post-partum development of diabetes.
Page 123 of 170 THE MONITORING OF BASELINE STERILITY IN THE POPULATION OF ANOPHELES ARABIENSIS IN MAMFENE, KZN, SOUTH AFRICA.
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Lobb, L.N. (1, 2); Munhenga, G. (1, 2); Koekemoer, L.L. (1, 2)
(1) WITS Research Institute for Malaria, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Vector Control Reference Laboratory, Centre for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, a division of the National Health Laboratory Service, Sandringham, Johannesburg, South Africa
Abstract South Africa is investigating the possibility of the sterile insect technique (SIT) as a complementary tool for malaria vector control specifically targeting the control of Anopheles arabiensis. Any vector control strategy has systems to monitor and evaluate the effectiveness of the intervention. In an SIT programme one of the most reliable and powerful tools for evaluating the impact of sterile insect releases is measuring the level of induced sterility in the targeted population. Level of induced sterility can only be accurately assessed if pre-control sterility levels of the natural population are known. This study was carried out to gather baseline fertility levels of a natural An. arabiensis population targeted for SIT. Anopheline mosquitoes belonging to the An. gambiae complex were collected in Mamfene, KwaZulu-Natal, South Africa using two types of outdoor traps Wild collected females were individually placed into oviposition vials and induced to lay eggs. Soon after oviposition females were identified to species level using PCR and only eggs from specimens positively identified as An. arabiensis were retained and used to determine fecundity and fertility. Mean number of eggs laid per female and percentage hatch were calculated monthly. A total of 167 egg-laying females were collected from February 2015 to January 2016, excluding May to August, and used to determine fertility and fecundity. The percentage hatch rate each month ranged between 9% (four females; average of 90.8 eggs per female) in September and 84% (13 females; average of 97.5 eggs per female) in March. In October 2015 the highest average of 140.3 eggs per female was recorded from three egg-laying females. The highest number of egg-laying females collected was 69, in January 2016, giving an average 115.8 eggs per female with a 48% hatch rate. Fertility in the wild population of Anopheles arabiensis varied between months and seasons during this period. Baseline sterility data for the population is necessary for quantifying the efficacy of sterile male release and must be considered for implementation. Additional data need to be gathered to determine the impact of weather conditions on fertility and fecundity.
Page 124 of 170 THE LARVICIDAL EFFECTS OF BLACK PEPPER (PIPER NIGRUM LINN.) AND PIPERINE AGAINST INSECTICIDE RESISTANT AND SUSCEPTIBLE STRAINS OF ANOPHELES MALARIA VECTOR MOSQUITOES
Authors and Affiliations Samuel, M. (1, 2), Oliver, S.V. (1, 2), Coetzee, M. (1, 2) & Brooke, B.D. (1, 2)
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Centre for Opportunistic, Tropical & Hospital Infections, National Institute for Communicable Diseases, Johannesburg, South Africa
Abstract Insecticide resistance carries the potential to undermine the efficacy of insecticide-based malaria vector control strategies. Therefore, there is an urgent need for new, effective insecticidal compounds. Black pepper (dried fruit from the vine,Piper nigrum), commonly used as a food additive and spice, and its principle alkaloid piperine, have previously been shown to have larvicidal properties. The aim of this study was to investigate the larvicidal effects of ground black pepper and piperine against third and fourth instar Anopheles larvae drawn from several laboratory-reared insecticide resistant and susceptible strains of Anopheles arabiensis, An. coluzzii, An. gambiae, An. quadriannulatus (all of which belong to the An. gambiae species complex) and An. funestus. Larvae were fed with mixtures of standard larval food and either ground black pepper or piperine in different proportions. Mortality was recorded 24hrs after black pepper and 48hrs after piperine mixtures were applied to the larval containers. Black pepper and piperine mixtures caused high mortality in the An. gambiae complex strains, with black pepper proving significantly more toxic than piperine. The An. funestus strains were substantially less sensitive to black pepper and piperine. Insecticide resistant and susceptible strains by species proved equally susceptible to black pepper and piperine. The difference in mortality between An. gambiae complex strains and An. funestus strains may reflect a marked difference in the feeding habits of these species, or, a difference in food metabolism possibly associated with the different breeding habitats used for rearing the species. Black pepper consistently induced high levels of mortality in Anopheles larvae compared to piperine and as such, it is concluded that black pepper may potentially be used as a larvicide for the control of certain malaria vector species.
Page 125 of 170 ADDRESSING CURRENT CHOKEPOINTS IN ANOPHELES ARABIENIS MOSQUITO MASS REARING
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Wood, O.R. (1, 2); Munhenga, G. (1, 2); Koekemoer, L.L. (1, 2); Brooke, B.D. (1, 2)
(1) Vector Control Reference Laboratory, National Institute for Communicable Diseases; (2) Wits Research Institute for Malaria (WRIM), University of the Witwatersrand
Abstract The Sterile Insect Technique (SIT) has proved to be an effective supportive tool in the control of various insect pest species including disease vectors. With the rise of insecticide resistance, or for insect species where complete insecticidal control is difficult, this role is becoming increasingly more important. While other insect species have been controlled effectively using SIT for decades, its use for the control of malaria vectors is still in the early stages of system development. It is therefore to be expected that the first generation of equipment and techniques developed to date will still need to be further developed, refined, or even radically overhauled in the quest to effectively and efficiently produce large numbers of competitive sterile male mosquitoes for release. Through preliminary, small-scale mass-rearing mock-up operations using existing technologies, workflow and operation have been evaluated and various production chokepoints identified that need to be addressed. From this experience, new equipment, as well as modifications to the current equipment have been designed, and are currently in the prototyping and evaluation stage. The improvements in equipment design, operation and work-flow should greatly reduce the number of man-hours and staff required for mass rearing, as well as significantly reduce the cost of equipment manufacture. The success of these designs and alterations should reduce the ultimate chokepoint of overall project cost, rendering SIT more accessible.
Page 126 of 170 FREQUENCY OF SEXUALLY MATURED MALES, IN A WILD ANOPHELES FUNESTUS SWARM IN BURKINA FASO
Abstract Theme Vector biology, control and IVM
Authors and Affiliations Zawada, J.W. (1, 2); Dahan-Moss, Y. (1, 2); Dabire, R (3), Koekemoer, L.L. (1, 2)
(1) Wits Research Institute for Malaria, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) Center for Opportunistic, Tropical and Hospital Infections, National Institute for Communicable Diseases, Sandringham, Johannesburg, South Africa; (3) Institut de Recherche en Science de la Santé, Direction Régionale de Babo-Dioulasso, Burkina Faso
Abstract The World Malaria Report states that approximately 438 000 deaths occurred in 2015 where 90% of these occurred in Africa. The main malaria vector, Anopheles funestus as many other anophelines, mate through swarming. Newly emerged male mosquitoes need to undergo an important physical change prior mating. This involves the rotation of the male genitalia between 135° and 180°, also referred to as inversion. The male genitalia consist of the abdominal segments 8 through 10, which include claspers that are tipped with claws. These claspers need to rotate180° to enable the male to grasp the female during copulation. Claspers that have not rotated are referred to as “T0” while
“T4” refers to claspers where the rotation has been completed (between 135° and 180°).There is a limited amount of understanding and knowledge of the An. funestus species when it comes to the adult male’s sexual characteristics and physiology. Limited published work on the fundamental biology of the male An. funestus is available. The aim of this study was to discover the frequency of sexually matured (rotated genitalia) males in a wild Anopheles funestus swarm in Burkina Faso. Collections were made in Bobo-Dioulasso during dusk and dawn periods to maximize the collection of An. funestus specimens from 3 swarming groups. Anopheles funestus group samples were collected from swarms and rotation of genitalia analyzed. All the male mosquitoes were morphologically identified and members of the An. funestus group were identified to species level using a standard multiplex PCR. All the samples were molecularly identified as An. funestus through a multiplex PCR. The frequency of sexually mature and sexually immature males was compared. Anopheles funestus is one of the main malaria vectors on the African continent, yet there is a very limited understanding and knowledge on the adult male’s sexual characteristics and physiology and this study has provided some insight into their biology. These results suggest that An. funestus goes through a stage of sexual maturation and a full transcriptome analysis can be performed to try and isolate the transcripts that are associated in this maturation process.
Page 127 of 170 INVESTIGATING THE ANTIPLASMODIAL POTENTIAL OF (BIS)THIOUREA AND (BIS)UREA POLYAMINE ANALOGUES ON THE GAMETE FORM OF PLASMODIUM FALCIPARUM PARASITE
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Abrie, C. (1); Verlinden, B.K. (1); Worster, P.M. (2); Birkholtz, L. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa; (2) Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, Charleston, SC 29425, USA
Abstract Malaria is a vector borne disease responsible for approximately half a million deaths of which an estimated 90% of deaths occur in Africa. The evolution and development of drug resistant strains of the Plasmodium falciparum parasite to current antimalarial drugs, is responsible for the difficulty of controlling malaria. The main focus of leading antimalarial agents has been against the asexual form and to a lesser extent sexual forms of the malaria parasite. Global malaria eradication is still the main goal therefore interventions to interrupt P. falciparum parasite transmission by targeting novel pathways in sexual gametocytes in humans and its subsequent gamete stages in the mosquito midgut could potentially contribute to eventual eradication of malaria. Polyamines are essential in cell differentiation and division, macromolecule synthesis and gene regulation in P. falciparum parasites. Polyamine analogues have similar structure to natural polyamines without substituting polyamine functions. Polyamine analogues have shown to inhibit both the asexual and sexual forms of P. falciparum parasites, however the activity of these compounds has not been investigated on gamete stages. The aim of this study was to further explore the antiplasmodial activity of (bis)thiourea and (bis)urea polyamine analogues against both male and female gamete forms of the P. falciparum parasite. The transmission-blocking activity of these polyamine analogues towards the gamete form of P. falciparum parasites was determined through quantitative PCR analysis using sex- specific gamete markers Pfs25, Pfs28, Pfs45/48 and Alpha tubulin II. As the male gamete increases its nuclear content to 8n through three consecutive mitotic divisions during gametogenesis we could further validate the transmission-blocking activity of the leading analogues against the male gametes through drug dose response assays measuring DNA content using the fluorescent dye SYBR Green I. Here we discuss the quantification of sex-specific gamete markers after polyamine analogue drug treatment. These polyamine analogues were validated in vitro for growth inhibitory activity against the male gamete forms of the P. falciparum parasite using a fluorescence-based assay. Here the antiplasmodial activity of the leading (bis)thiourea and (bis)urea polyamine analogues against the gamete form of P. falciparum parasites is assessed with regards to well established controls.
Page 128 of 170 AN EVALUATION OF THE GAMETOCYTOCIDAL PROPERTIES OF NATURAL PRODUCT COMPOUNDS AGAINST PLASMODIUM FALCIPARUM
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Breedt, C.H. (1); Moyo, P. (1); Birkholtz, L.M. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa.
Abstract Malaria remains a threat to over three billion people worldwide, with an estimated 214 million new cases and 438 000 deaths annually. Front-line treatments for malaria consist of fixed-dose artemisinin-based combination therapies (ACTs). However, due to the spread of resistant strains of Plasmodium falciparum parasites new strategies are needed to control the spread of the disease. ACTs have been shown in clinical studies not to block transmission or relapse, necessitating for follow-up treatment with primaquine. However, primaquine has shown to cause haemolytic anaemia in patients suffering from glucose-6-phosphate dehydrogenase deficiency. Therefore, greater strides need to be taken towards antimalarial drug research and discovery, particularly towards compounds that inhibit the transmission of the malaria parasite. Transmission from the human host to the mosquito vector depends on the differentiation of asexual parasites into the sexual gametocyte stages. Natural product compounds have historically been shown to be suitable stepping-stones towards identifying and developing new antimalarial drugs, through providing new chemical scaffolds from which drugs with improved activity can be developed. The most valued of these natural product compounds that have served as scaffolds include quinine and artemisinin, which have become the mainstay drug treatments for the clinical symptomatic asexual blood stages. The potential of natural product compounds to inhibit gametocyte development and therefore block transmission has, however, not been studied as extensively. This study therefore reports on the gametocytocidal in vitro activity of eight natural product compounds of plant and/or microbial origin. Compounds were selected for the study based on their activity against identified druggable gametocyte targets. The Malaria SYBR Green I-based Fluorescent assay was performed to confirm the activities against the asexual stages, and the Luciferase-based Cell Line assay was used to determine the activities against early and late stage gametocytes. The activity of the selected natural product compounds against asexual forms of malaria parasites were confirmed in the nanomolar range. Data on their efficacy on early and late stage gametocytes will be presented. Most of these compounds have encouragingly been shown to have activity against asexual parasite stages making them potential multistage active compounds for malaria control.
Page 129 of 170 IMPLEMENTATION OF THE DIAGNOSIS QUALITY ASSURANCE PROGRAM FOR ACCURATE MALARIA DIAGNOSIS IN SWAZILAND
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Chitundu, S (1); Soble, A (2); Darteh, S (3); Novotny, J (2); Ntshalintshali, N (2); Kunene, S (1)
(1) National Malaria Control Programme, Manzini, Swaziland; (2) Clinton Health Access Initiative, Mbabane, Swaziland; (3) International Center for AIDS Care and Treatment Programs, Mbabane, Swaziland
Abstract In 2009, the Swaziland Ministry of Health introduced a definitive diagnosis policy for malaria, whereby all suspected cases of malaria are required to be confirmed by parasitological diagnosis, either rapid diagnostic test (RDT) and/or microscopy, in support of the country’s national malaria elimination strategy; RDTs were rolled out in 2010 to all public and private health facilities. A comprehensive quality assurance program was established to monitor and ensure the efficacy of malaria diagnostics employed in Swaziland. Between November 2009 and January 2010, health care workers were trained on the use of RDTs and quality assurance procedures during cadre-specific trainings. The quality assurance program was implemented in Swaziland’s Lubombo region, where health facilities would collect an RDT, microscopy slide and dried blood spot (DBS) using diagnostic bundles for all suspected cases. Sample collection for confirmed cases was expanded nationally in 2013. Collected samples were labeled, stored individually and to be transported to the central level on a weekly basis. Samples were sorted by each health facility and 100% of positive results and 10% of negative results were analyzed, with the RDT and DBS being sent for PCR analysis and slides being read by expert microscopy readers for cross-checking Implementation experience of the program demonstrated that quality assurance of RDTs was challenging due to user error, inconsistent sample collection and infrequent use of microscopy for secondary confirmation due to capacity, including poor slide taking, and logistical constraints, and lack of actionable feedback provided to health facilities. The coverage and use of RDTs for definitive diagnosis in Swaziland’s passive surveillance system is high and critical for its elimination target, but the implementation of the quality assurance program to support accurate and reliable RDT results requires strengthening.
Page 130 of 170 DEVELOPMENT OF SEROLOGIC ASSAYS TO ASSESS MALARIA EXPOSURE IN SWAZILAND
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Nhlabathi, N. (1, 4); Helb, D. (2, 5); Ntshalintshali, N. (3); Mkhonta, N. (1); Shabalala, C. (1,4); Soble, A. (3); Maphalala, G. (4); Gosling, R. (5); Greenhouse, B. (5); Hsiang, M. (5, 6); Kunene, S. (1)
(1) National Malaria Control Programme, Manzini, Swaziland; (2) University of California, San Francisco; (3) Clinton Health Access Initiative, Mbabane, Swaziland; (4) Swaziland Laboratory Health Services, Mbabane, Swaziland; (5) University of California, San Francisco; (6) Department of Pediatrics, University of Texas Southwestern Medical Center
Abstract Antibody responses can provide information on prior exposure to malaria, potentially identifying high-risk groups and foci of transmission. However, in areas of declining transmission it is important to distinguish recent from more distant exposure. Swaziland NMCP measured antibody responses to Pf antigens apical membrane antigen 1 (AMA1), merozoite surface protein 1 (MSP1), circumsporozoite protein (CSP), and schizont extract (SE) via ELISA in dried blood spot samples from 208 symptomatic cases and 40 asymptomatic cases diagnosed by RDT, 38 uninfected individuals who reported malaria diagnosis in the past year (recent cases), and 560 uninfected controls who reported no history of malaria in the past year. Samples came from passive and active surveillance activities conducted in Swaziland from October 2012 to June 2014. Responses to CSP and SE provided poor discrimination between cases and controls. Reponses to AMA1 provided some discrimination for asymptomatic and recent cases in children, especially those 5 years and younger. At specificity of 99%, sensitivity for symptomatic cases was 33% in 1-5 year olds and 29% in 5-15 year olds. For asymptomatic cases, sensitivity was 78% in 1-5 year olds and 67% in 5-15 year olds. For recent cases sensitivity was 100% in 1-5 year olds and 83% in 5-15 year olds. Poor discrimination was found for all comparisons in adults. Results were better for MSP1.Sensitivity for symptomatic cases was 71% in 1- 5 year olds and 74% in 5-15 year olds. For asymptomatic cases, sensitivity was 100% in 1-5 year olds and 67% in 5-15 year olds. While serology has potential to provide more sensitive stratification of Pf exposure risk than parasite prevalence in low transmission areas such as Swaziland, antibody responses to commonly used antigens were not adequate in adults and only marginally adequate in children over the age of 5. For serology to support strategic programmatic planning and intervention targeting, additional work is required to identify alternative anti-Pf antibodies, such as those with reliable boosting but relatively short half-lives, to increase diagnostic precision.
Page 131 of 170 DRUG TARGET DECONVOLUTION OF ANTIPLASMODIAL COMPOUNDS USING DRUG AFFINITY RESPONSIVE TARGET STABILITY, FOR ASEXUAL AND SEXUAL STAGES OF PLASMODIUM FALCIPARUM PARASITES.
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Henn, D. (1); Verlinden, B. K. (1); Birkhotz, L.M. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, Centre for Sustainable Malaria Control, University of Pretoria, South Africa.
Abstract Malaria is a parasitic disease caused by the Plasmodium parasite, which, if untreated can result in the death of an infected individual. Due to the disease severity and the continuous emerging resistance of the parasite to current antimalarials, the list of novel drugs with known biochemical targets needs to be expanded. The mode of action of compounds that show antiplasmodial activity in vitro, first needs to be uncovered to further develop potential drugs. Drug affinity responsive target stability (DARTS) is an established technique for target deconvolution. DARTS has been successfully used to determine the protein target(s) of several promising antiplasmodial compounds in the asexual and late-stage gametocyte forms of the P. falciparum parasite. To perform DARTS the half maximal inhibitory concentrations (IC50s) of the compounds must first be determined. The IC50s were determined using a fluorescent based and a luciferase reporter assay respectively. DARTS was performed on both the asexual stages and late-stage gametocytes of P. falciparum by treating the parasite lysate with the selected antiplasmodial compounds, before addition of pronase. The binding of the antiplasmodial compound to its target, protects the target from digestion by pronase. The protein targets of the compounds were visualized on SDS-PAGE gel and identified using mass spectrometry. Here we discuss the antiplasmodial activity and the protein target(s) of promising single target compounds, such as MMV 390048, an aminopyridine derivative, and pleiotropic compounds such as Compound 30, a polyamine analogue on the asexual stage and late-stage gametocytes of P. falciparum parasites. DARTS can be used to determine the protein target(s) of antiplasmodial compounds for the asexual stage and late-stage gametocytes of P. falciparum parasites.
Page 132 of 170 PHENOTYPIC PROFILING OF GROWTH PERTURBATIONS OF THE HUMAN MALARIA PARASITE PLASMODIUM FALCIPARUM
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Joshua, A. (1), van Biljon, R. (1), Birkholtz, L.M. (1) and Niemand, J (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa
Abstract Currently the World Health Organization recommends Artemisinin -based combination therapies for the treatment of uncomplicated Plasmodium falciparum infections. However, the emergence of multi-drug resistant strains of Plasmodium species has highlighted the need for the development of antimalarials with novel modes of action (MOA). While high-throughput whole cell based assays can identify compounds with antiplasmodial activities, the MOA of these compounds are often unknown. Transcriptional profiling, proteomics, genomics and metabolomics have all been used to determine the MOA of compounds discovered to have antiplasmodial activity. These techniques fall short however, in that they provide a static picture of the parasite’s response, disregarding the physiological response of the organism to environmental perturbations. The aim of this research was to determine the baseline phenotypic profiles of P. falciparum parasites upon exposure to well- characterized antiplasmodial compounds. Following confirmation of inhibition of parasite proliferation using an in vitro fluorescence based assay, phenotype microarrays were used to quantitatively assess the biochemical responses of the parasite to chemical perturbations. Data generated from this study will allow for the classification of compounds with similar MOA by comparing their phenotypic profiles thereby increasing our understanding of the biological response of the parasite and aid in the development of novel antiplasmodial agents required for malaria elimination.
Page 133 of 170 EFFECT OF NOVEL 7-CHLOROQUINOLIN-4-YL PIPERAZINE-1-YL-ACETAMIDE DERIVATIVES IN THE MANAGEMENT OF MALARIA
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Kathrada, F. (1); Van Zyl, R.L. (1); Coetzee, M. (2); Oliver, S. (2); Hayat, F. (3); Inam, A. (3); Azam, A. (3)
(1) Pharmacology Division, Department of Pharmacy & Pharmacology; WITS Research Institute for Malaria (WRIM), Faculty of Health Science, University Of Witwatersrand, Johannesburg, South Africa; (2) WITS Research Institute for Malaria (WRIM), Faculty of Health Science University of Witwatersrand; Vector Control Reference Laboratory, National Health Laboratory Services, Sandringham, South Africa; (3) Department of Chemistry, Jamia Millia Islamia, New Delhi, India
Abstract Antimalarial drugs and vector control methods have played a crucial role in the elimination of malaria. With the decreasing efficacy of the current therapies, the development of resistant strains of Plasmodium and Anopheles. As such the development of novel compounds is vital. This study is aimed at investigating the effect of novel 7-chloroquinolin-4-yl piperazine-1-yl-acetamide derivatives in managing malaria. Twenty seven 7-chloroquinolin-4-yl piperazine-1-yl-acetamide derivatives were synthesized and characterised by NMR/IR. The parasite lactate dehydrogenase (pLDH) assay was used to assess the inhibitory properties of the derivatives on the chloroquine-sensitive NF54 strain of P.falciparum in comparison to quinine. The haemolytic potential of the compounds (50 µM) was evaluated against a 1% (v/v) haematocrit of fresh red blood cells after 48 hrs incubation at 37˚C. The percentage haemolysis was compared to 0.2% Triton X100 and quinine. The toxicity of the derivatives and positive controls were further evaluated using the MTT cell viability assay with human epithelial kidney (HEK-293) cells. The cell viability was evaluated in comparison to controls. The brine shrimp lethality test was used to screen derivatives for toxicity against Artemia fransiscana nauplii for 24 hrs compared to potassium dichromate and DDT. The larvicidal activity of the derivatives was determined against the fourth instar larvae of Anopheles arabiensis (KGB) for 24 hrs, in comparison to DDT. All twenty seven compounds inhibited malaria growth (IC50 range: 1.29 –
53.98 µM), with compound CQPA-25 and CQPA-26 (IC50: 1.72 µM and 1.29µM) the most active
compared to quinine (IC50: 0.18 µM). The inhibitory effects appear to be directed against the intra- erythrocytic parasite rather than the red blood cell host as no haemolysis was observed (< 1.5% lysis). The twenty seven derivatives showed a good safety profile, with low toxicity to human epithelial kidney cells (% inhibition average: ~45%) and minimal lethality to the brine shrimp (< 5%). The twenty seven derivatives displayed no larvicidal activity compared to DDT (100% dead) at 0.50 µM. Initial results indicate that these novel 7-chloroquinolin-4-yl piperazine-1-yl-acetamide derivatives are ineffective against the Anopheles larvae but show potential against the intra- erythrocytic stages of parasite growth, warranting further combination studies.
Page 134 of 170 USING CRISPR-CAS9 TO MANIPULATE THE GENOME OF P. FALCIPARUM
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Liebenberg, D. (1); Coetzer, T.L. (1, 2)
(1) Wits Research Institute for Malaria (WRIM), Department of Molecular Medicine and Haematology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa; (2) National Health Laboratory Service, Johannesburg, South Africa
Abstract The discovery of clustered regularly-interspaced short palindromic repeats (CRISPR) and the development of CRISPR-Cas9 techniques have had a revolutionary effect on genome editing. CRISPR-Cas9 technology has recently been shown to be effective in Plasmodium and in this study it is used to investigate the in vivo role of P. falciparum inhibitor of apoptosis (IAP) protein (PfIAP; PF3D7_0519600). IAP proteins are important in mammalian programmed cell death (PCD), inhibiting pro-apoptotic caspases. PCD is hypothesised to be used by P. falciparum to control its population, although the network of proteins involved in this process needs to be elucidated. The aim of this study is to modify a CRISPR-Cas9 vector to 1) knock-out the pfiap gene to determine if it is essential for parasite survival and 2) tag the PfIAP protein to localise it under normal and heat-shock conditions, which induce PCD. To delete the pfiap gene the 5’ and 3’ untranslated regions (UTRs) were amplified by PCR and sequentially cloned into the pL6-GOI vector, either side of the human dihydrofolate reductase (hDHFR) positive selection cassette. A protospacer adjacent motif (PAM) site was identified in the gene and the upstream nucleotides were synthesised and inserted into the modified vector using Gibson Assembly, to create the targeting custom single guide RNA (sgRNA) for the knock-out experiment. PfIAP-GFP tagging involved PCR amplifying the 3’ end of the pfiap gene, a GFP sequence and the 3’ UTR of the gene; assembling the fragments using Gibson Assembly and ligating the construct into the pL6-GOI vector. Another PAM site towards the 3’ end of the gene was identified and a new sgRNA was created and inserted into the vector. P. falciparum parasites were transfected with the two vector constructs and following cleavage of the genomic DNA by the Cas9 nuclease at the sites specified by the sgRNA, homologous recombination will result in the replacement of the pfiap gene with the hdhfr cassette or the insertion of the GFP tag at the 3’ end of the gene. These studies will enhance our knowledge of parasite biology and may provide a new drug target if the pfiap gene is essential.
Page 135 of 170 EVALUATION OF THE PREVALENCE OF CYP 2D6 MUTATIONS IN VHEMBE DISTRICT, LIMPOPO PROVINCE, SOUTH AFRICA
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Makhanthisa, T.I. (1); Awandu, S.S. (1); Niemand, J. (1); Birkholtz, L.M. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa
Abstract South Africa is aiming for malaria elimination by 2018. Primaquine is an 8-aminoquinoline gametocytocidal drug that is capable of clearing all circulating late stage gametocytes. The World Health Organization recommends a single low dose administration of primaquine in addition to artemisinin combination therapy in countries on the brink of malaria elimination. However, the efficacy of primaquine could be affected by the metabolism of primaquine by CYP 2D6. The CYP 2D6 is the most polymorphic drug metabolizing enzyme and its polymorphisms have been linked with reduced capacity to metabolize drugs among individuals with different poor metaboliser phenotypes. The aim of this study is to determine the distribution of cytochrome P450 2D6 (CYP 2D6) mutations in Vhembe District, Limpopo Province, South Africa. Blood samples from Vhembe District were genotyped for CYP 2D6 variants (CYP 2D6*3, CYP 2D6*4, CYP 2D6*5 and CYP 2D6*17) by polymerase chain reaction and restriction length polymorphisms. The CYP 2D6 distribution was measured and results between different study sites will be presented. The genotyping results predict a low prevalence of CYP 2D6 mutations in Vhembe district, Limpopo Province. The findings of this study thus contribute to the much needed evidence to inform the malaria elimination agenda by the National Malaria control programme.
Page 136 of 170 THE HIDDEN NON-BLOODSTREAM PARASITE RESERVOIR: A MALARIA CONTROL PROBLEM
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Markus, M.B. (1)
(1) University of the Witwatersrand
Abstract Since the 1980s, it has conventionally been stated that hypnozoites (a term coined by me 40 years ago) are found in the life-cycles of Plasmodium vivax and P. ovale but not in those of P. falciparum, P. knowlesi or P. malariae; and that hypnozoites are therefore the source of relapse-type recurrent malaria in P. vivax and P. ovale infections only. In the light of particular research findings during the past 20 years, but especially of studies reported in some recent publications, this is in retrospect almost certainly an inadvertently simplistic evaluation of the overall situation. Hypnozoites might also occur in the life-cycles of species of Plasmodium that are traditionally thought not to have them. That matter aside, there is indirect evidence that some latent non-hypnozoite parasites probably have extra-hepatic, non-bloodstream human cells as their habitat. There is also evidence that these stages are not necessarily always killed by drugs. How, then, will this “additional” hidden reservoir of infection in human populations be dealt with? It might not be possible to do much about it. These two subjects (concerning hypnozoites and extra-vascular, non-hypnozoite quiescent parasites, respectively) need to be addressed. I recently advocated cutting-edge work concerning them, involving the use of primatized mice; and in 2016, published in Trends in Parasitology the biological and genetic rationale (which is not immediately obvious) associated with these suggestions.
Page 137 of 170 EPIGENETIC DRUGS TARGETING HISTONE POSTTRANSLATIONAL MODIFICATIONS DURING GAMETOCYTOGENESIS OF THE PLASMODIUM FALCIPARUM PARASITE
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Mmekwa, N.E. (1); Coetzee, N (1); Birkholtz, L. (1)
(1) Department of Biochemistry, UP Institute for Sustainable Malaria control, University of Pretoria, Private Bag X20, Hatfield, 0028, South Africa
Abstract The Plasmodium falciparum parasite strongly depends on epigenetic processes for development during gametocytogenesis in the human host. Malaria parasite transmission from human host to mosquito vector is mediated by gametocytes, the sexual stage of the parasite. Due to the lack of effective transmission blocking drugs, novel drugs with new modes of action and activity against the sexual stages of the parasite are needed. The epigenetic gene regulatory processes are therefore seen as an attractive target for novel drugs. Epi-drugs target various epigenetic processes such as DNA methylation and histone posttranslational modifications. In this study we investigated the potential of a collection of epi-drugs, from the cancer field, to inhibit gametocytogenesis during P. falciparum parasite development. Epigenetic drugs from the Cayman’s Chemical Epigenetic Screening Library were tested for activity against transgenic P. falciparum NF54 strains, expressing the pfs16-GFP-Luc and mal8p1.16-GFP-Luc luciferase transgenes, during early gametocyte and late gametocyte development, respectively. Measurement of gametocytocidal activity was performed, using a luciferase reporter assay, while the asexual blood stages were tested for activity using SYBR
Green l-based fluorescence assay. Previously screened compounds that showed IC50 values below 0.1 µM were selected as lead compounds. The lead compounds from the gametocyte were then further evaluated using Western blot. Analysis of the Western blot showed that, compounds did have an effect on the histone posttranslational modifications of P. falciparum gametocytes. This data will help to further expand our understanding of drug targets in histone posttranslational modifications of P. falciparum parasite gametocyte.Identification of such novel compounds with the ability to block transmission will help design therapies, to ultimately eradicate malaria.
Page 138 of 170 IN VITRO ACTIVITY OF NAPHTHYLISOQUINOLINE ALKALOIDS ON PLASMODIUM FALCIPARUM GAMETOCYTES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Moyo, P. (1); Eloff, J.N. (2); Bringmann, G. (3); Niemand, J. (1); Maharaj, V.J. (4); Louw, A.I. (1); Birkholtz, L. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, Faculty of Natural and Agricultural Sciences, University of Pretoria, Private Bag x20, Pretoria 0028, South Africa (2) Phytomedicine Programme, Department of Paraclinical Sciences, Faculty of Veterinary Science, University of Pretoria, Private Bag x04, Onderstepoort 0110, Pretoria, South Africa (3) Institut fur Organische Chemie, Universitat Wurzburg Am Hubland, 97074 Wurzburg, Germany (4) Department of Chemistry, Centre for Sustainable Malaria Control, Faculty of Natural and Agricultural Sciences, University of Pretoria, Private Bag x20, Pretoria 0028, South Africa
Abstract The multiple stages of the protozoan Plasmodium in both the human and mosquito hosts present a unique and complex challenge in the management of malaria. For the elimination of the disease to be achieved, there is need for the development of single multi stage active drugs. However, historically emphasis has been skewed towards the search for compounds active against the symptomatic asexual parasite stages. The Medicines for Malaria Venture has outlined the properties of an ideal malarial drug candidate. In addition to its curative abilities, it states that these also need to be active against the liver and the non-proliferative sexual or transmittable gametocyte stages. No such compound is currently available commercially among the antimalarial arsenal. Naphthylisoquinoline alkaloids are a unique class of natural product compounds that occur abundantly in the Dioncophyllaceae and Ancistrocladaceae plant families. This class of compounds has been shown to be potent against the liver and intra-erythrocytic asexual stages of the Plasmodium parasite; active on both chloroquine sensitive and resistant strains, in vitro and in vivo. However, no investigations have been performed to examine its gametocytocidal properties. In the present study, an in vitro phenotypic screen was undertaken on five naphthylisoquinoline alkaloids for activity against immature and mature stage P. falciparum parasite gametocytes. In vitro activity of five selected compounds on P. falciparum NF54 parasite immature and mature gametocyte stages was interrogated using the parasite lactate dehydrogenase assay (pLDH) (72 h – with drugs + 72 h – without drug incubation). Dual point screens at 1 and 5 µM were carried out
for both stages followed by full IC50 determination for the most active compounds. Our study showed that naphthylisoquinoline alkaloids have dual activity against both immature and mature gametocyte developmental stages. This activity was significantly more pronounced on immature stages in comparison to the mature stages. Out of five compounds screened, Dioncophylline C, Jozimine A2 and Ancistrocladanium A, showed good activity (<1 µM IC50) against immature stages with IC50 values in the nanomolar range. However,
only Jozimine A2, gave over 50% and 70% inhibition of mature stage gametocytes in vitro viability at 1 and 5 µM, respectively. The findings from our investigation not only show the potential transmission blocking ability of the naphthylisoquinoline alkaloid chemical compound class but extend to its potential development into the much sought after pan-reactive antimalarial drug.
Page 139 of 170 MEMBRANE TRANSPORT IN THE SEXUAL, TRANSMISSIBLE STAGES OF PLASMODIUM FALCIPARUM PARASITES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Naude, M. (1); van Biljon, R. (1); Birkholtz, L.M. (1); Niemand, J. (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa
Abstract Malaria elimination requires that the sexual, gametocyte stage of the Plasmodium parasite is targeted, as this is the form that is transmitted from the human host to the mosquito vector, leading to the spread of the disease. While single-dose primaquine has been suggested as a gametocytocidal therapy, haemolytic side effects may occur with G6PD deficient patients. By understanding the biology of the gametocyte, which includes the metabolism and transport of metabolites, new transmission blocking compounds can be developed with novel biological targets. The asexual intra- erythrocytic P. falciparum parasite has adapted to obtain essential metabolites required for parasite survival from the extracellular environment through the use of both endogenous RBC and parasite- derived membrane transporters, channels and uptake processes. Limited information is however available about these uptake processes in the gametocyte stage of P. falciparum. A variety of in silico analyses were used where transmembrane domains were identified. The presence of the various transcripts during gametocytogenesis was validated by DNA microarray analysis of stage I-V gametocytes. We identified proteins with transmembrane domains in the gametocyte stage, which was classified as ion channels, permeases, ABC transporters, electrochemical potential-driven transporters, pumps and MDR transporters. The various transcripts during gametocytogenesis were validated by DNA microarray analysis. The identification of metabolite uptake mechanisms within P. falciparum gametocytes may ultimately identify new targets for transmission blocking strategies.
Page 140 of 170 APPLICATION OF NETWORK ANALYSIS ON P. FALCIPARUM TRANSCRIPTOMES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Van Wyk, R.D.J (1); Van Biljon, R.A (1); Niemand, J (1); Birkholtz, L (1)
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa
Abstract In the era of “Big data”, integration of large data sets in order to extract usable information has become a useful research tool. However, this large amount of information does present with a number of challenges. For example, there are over 4900 data sets currently available for Plasmodium falciparum on Gene Expression Omnibus (GEO) housing several transcriptomes. Extracting useful information from this data becomes an almost Sisyphean task. One method of simplifying large data sets would be the use of interaction networks such as; gene interaction networks, gene regulation networks, protein-protein interaction (PPI) networks and protein-drug interaction networks. These interaction networks all query different aspects of the parasite biology and give a systems view of individual genes and their relationship with other genes given certain conditions. In 2005 one of the first interaction networks were established for P. falciparum from an experimentally determined PPI network. Since then network analysis have expanded to include more PPI data, predicted gene interactions, homology-based interaction, co-expression analysis and co-occurrence data, adding to the complexity of the networks. The application of big data in network analysis now becomes the new challenge. We have constructed gene regulation networks for both asexual and gametocyte stages of the parasite from transcriptomic data. These networks were done using Gene Regulatory Network Inference Using Time Series (GRENITS) in R. Drug response networks were constructed from the STRING database and drug treated transcriptomes. The GRENITS analysis yielded 19 potential gene regulators for gametocytogenesis, while the drug perturbed networks identified key genes that may pertain to the drugs mode-of-action. The networks produced by this study illustrated that the use of a systems biology approach provide valuable insights into gene interactions for P. falciparum. The drug perturbed transcriptome networks become part of a larger project where the mode-of-action for several drugs and potential drug targets may be elucidated.
Page 141 of 170 CRISPR-CAS EVALUATION OF IMPORTANT CELL CYCLE REGULATORS
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations von Grüning, H. (1); van Biljon, R. (1); Niemand, J. (1); Birkholtz, L. (1).
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa.
Abstract In the light of the current growing antimalarial resistance against first-line drugs, functional genomics tools can be used to interrogate features of parasite biology that could be marked in future intervention strategies. One such feature is the cell cycle the P. falciparum parasite, and the key regulators governing cell and life cycle progression. Considering that the parasite’s life cycle involves two hosts and multiple life cycle stages, gene expression is tightly regulated in Plasmodium parasites. Genes encoding for transcription factors and other kinases have previously been implicated as major role players in the unusual cell cycle governing both asexual and sexual development of the parasite, which leads to a vast expansion of parasite numbers during a malaria infection. The majority of these transcription factor encoding genes are putative, with no functional roles as of yet described. The aim of the study was to functionally describe putative cell cycle regulator genes of the P. falciparum parasite through the powerful new genome editing tool, CRISPR-Cas. Prior to transfection, in silico design of guides targeted towards the genes of interest, and the template DNA used for repair of the Cas9 induced double stranded break, is achieved through bioinformatical tools including Benchling and PlasmoDB. Guides were optimised for efficiency and specificity. The oligo guides/repair templates are cloned into Cas9- and eGFP carrying vectors followed by drug selection. Integration of guides and repair templates into vectors are confirmed through PCR and partial DNA sequencing. Ten guide sequences and repair templates that were designed in silico show high on- and off-target scores. The successful integration into vectors as confirmed by PCR and partial DNA sequencing. The constructs’ suitability for future transfection was evaluated and confirmed. In the study, the highly versatile genome editing CRISPR-Cas tool was applied to elucidate the functional roles of putative transcription factors and other cell cycle regulators, their roles in the cell cycle regulation of both asexual and sexual forms of P. falciparum.
Page 142 of 170 INVESTIGATION OF DIFFERENTIAL DRUG RESPONSES IN DIFFERENT P. FALCIPARUM PARASITE LIFE CYCLE STAGES
Abstract Theme Parasite biology, control, transmission blocking and gametocyte studies
Authors and Affiliations Weidemann, G.J. (1); Van Biljon, R.A. (1); Niemand, J. (1); Birkholtz, L. (1)
(1) Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa.
Abstract The most lethal form of malaria in humans is caused by the P. falciparum parasite. The parasite’s life cycle in the human host is divided into an asexual and sexual developmental cycle that differs in morphological appearance and biological function. The asexual parasites proliferate rapidly and rupture the erythrocytes, causing the clinical symptoms of the disease. Approximately one-tenth of the asexual parasite population will commit to sexual reproduction and progress through five stages of development to mature into female and male gametocytes that can transmit to Anopheles mosquitoes. New intervention strategies therefore require novel chemotypes that clear all persistent pathogenic asexual blood-stage forms of P. falciparum as well as targeting the long-lived transmissible forms of the parasite (mature-stage V gametocytes). It is presently unclear where in this developmental cycle antimalarial efforts should be focused. Currently, high throughput screening programmes that test possible antimalarial compounds screen against asexual parasites and both early and late stage gametocytes. From these programmes, it has been observed that early and late gametocytes have differential susceptibility to a variety of compounds. Two such compounds, Compound A and Compound B were previously investigated by DNA microarray to investigate the global transcriptomic responses following treatment with these compounds and were further investigated here. The antiplasmodial activity of two compounds, Compound A and Compound B were determined against asexual stage P. falciparum parasites using a fluorescence based proliferation assay and against sexual stage P. falciparum parasites using a luciferase reporter assay. Using quantitative real-time PCR (qPCR), unique transcriptional responses previously identified with DNA microarray were validated for the sexual stage parasites and these responses were additionally probed for asexual stage parasites. Compounds A and B were found to act with differential potency against asexual, early and late stage gametocytes. Likewise, differential transcriptional responses by different life cycle stages following exposure with compounds A and B were determined using qPCR, validated the previously observed DNA microarray analyses. This study contributes biological information on the difference in drug responses of the proliferating, pathogenic asexual stages and the inactive, transmissible gametocyte stages of the P. falciparum parasite.
Page 143 of 170 ASSESSING THE IN VITRO EFFICACY IN SILICO DESIGNED COMPOUNDS TARGETING THE MALARIAL QI SITE OF CYTOCHROME BC1
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Laura, K. (1); Cromarty, D. (1); Birkholtz, L. (2); Stander, A. (3)
(1) Department of Pharmacology; (2) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control; (3) Department of Physiology, University of Pretoria, South Africa
Abstract Malaria is a global parasitic disease caused by parasites of the genus Plasmodium. Widespread emergence of clinical resistance in marketed antimalarial drugs, demands an urgent need to synthesizncompounds. The mitochondrial electron transport chain (mtETC) is recently being explored for the design of new antimalarial therapy. Interestingly, the mtETC is a valuable target in antimalarial development because it differs significantly from that in mammals. The inhibition of enzymes involved in the mtETC stops the flow of electrons hence, no metabolic processes can subsequently occur in P. falciparum parasites affected in this manner. In silicodrug design is the synthesis of small molecule compounds, which would bind to the protein target and modulate protein function. Using in silico design methods, 6 new compounds were sythesized. These compounds are aimed to target the Qi site of cytochrome bc1 on the mtETC. In this study, we aimed at testing these 6 newly synthesized in silico designed compounds on P. falciparum parasites in vitro.A fluorescent-based assay using SYBR Green I dye will be used to assess Plasmodium parasite proliferation and to determine the IC50, speed and stage specificity of the compounds. To determine the transmission blocking potential of the compounds, the luciferase based assay will be used to assess the viabilty of the gametocytes after treatment with the compounds. Cytotoxicity of the compounds will be assessed on the HEpG2 hepatocarcinoma cells, using the lactate dehydrogenase assay. Primary screen data of the compounds on P. falciparum parasites in vitro will be presented, with full dose-response determination of compounds passing primary filters at >50-70% activity at 1 and 5 µM evaluated. Compounds inhibiting some of the complexes of the mtETC have been shown to have potential as antimalarial compounds. However, many aspects have to be investigated to assess the suitability of these compounds in terms of efficiency and efficacy.
Page 144 of 170 IN VITRO ANTIMALARIAL ACTIVITY OF DIOSPYROS CHAMAETHAMNUS AND GUIBOURTIA COLEOSPERMA, FOUND IN THE ZAMBEZI REGION OF NAMIBIA
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Du Preez, C.I. (1); Mumbengegwi, D.R. (1)
(1) Multidisciplinary Research Centre, University of Namibia
Abstract As a result of the implementation of multiple interventions, malaria cases are on the decline in Namibia. This is in line with the national goal of having a malaria free status by 2020. However, because of reported cases of decreased sensitivity of parasites to current treatments, and the fact that some communities in rural areas cannot afford or access allopathic medicine or prefer to use herbal remedies; the malaria elimination process might be implicated. Two plants, Diospyros chamaethamnus and Guibourtia coleosperma are used in an ethnomedicinal setting in Namibia to treat symptoms of malaria. It is pertinent to show that these ethnomedicines are effective and safe for long term use, for their integration into mainstream malaria case management to aid in eliminating malaria within the country’s borders. The main aim of this study was to valorize their use as antimalarials in an in vitro model. Crude plant extracts were prepared using aqueous and organic (MeOH:DCM 1:1 v/v) solvents, and were screened for known classes of antiplasmodial compounds using TLC. GC-MS was also used. Radical scavenging abilities of the plant extracts were ascertained by the DPPH method. Growth inhibition of Plasmodium falciparum D10 was determined using parasitaemia. Similarly, in vitro cytotoxic effects of the plant extracts were conducted using a fibroblast cell line (W138). The crude root extracts contained alkaloids, anthraquinones, flavonoids, steroids and terpenoids. Secondary metabolites with antiplasmodial, antibacterial and antioxidant properties identified using GC-MS were 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one, phloroglucinol, stigmasterol, glycerol 2-hexadecanoate, α-amyrin, 9,12-octadecdenoic acid (Z,Z)-, hexadecanoic acid, oleic acid, lanosterol, spiculesporic acid, squalene, campesterol and vitamin E. D. chamaethamnus organic (DCOE) and aqueous extracts (DCAE) exhibited the highest antioxidant activities (IC50=7.63 and 10.74µg/mL), whilst the organic extract of G. coleosperma (GCOE) exerted antioxidant activities of 22.03µg/mL, and 36.05 µg/mL for the aqueous extract (GCAE), respectively. Moderate in vitro antiplasmodial activity (IC50<50 µg/ml) was observed for the two plants, GCOE (28.17µg/mL) and GCAE (31.61µg/mL), and DCOE (19.51µg/mL) and DCAE (18.30µg/mL). All plant extracts showed no cytotoxicity with IC50 values above 100µg/mL, except for DCOE (CC50=29.73). The results of this study corroborate the uses of these plants. Further studies will provide in vivo and pharmacological data to advice on dosage regimens.
Page 145 of 170 THE COPPER CHELATING, ANTIMALARIAL AND LARVICIDAL EFFECTS OF 8-HYDROXYQUINOLINE AND ITS DERIVATIVES
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Jansen van Vuuren, N. (1); van Zyl, R.L. (1)
(1) Pharmacology Division, Department of Pharmacy and Pharmacology; Malaria Research Institute for Malaria (WRIM), Faculty of Health Sciences, University of the Witwatersrand, South Africa.
Abstract Resistance of Plasmodium falciparum and its vector Anopholes to antimalarials and insecticides requires the development of novel compounds and combinations. Inhibiting the malaria vector in its larvae stage decreases the spread of the Plasmodium parasite. Derivatives of 8-hydroxyquinoline and the compound itself have demonstrated metal chelating, insecticidal, antibacterial, antifungal and antiparasitic activity. These derivatives may prove favourable in the control of malaria. The aim of this investigation was to determine the antimalarial, larvicidal and toxicological properties of 8- hydroxyquinoline and its derivatives. The copper chelating, in vitro antimalarial, haemolytic, whole organism toxicity and larvicidal activity of 8-hydroxyquinoline and 18 of its derivatives were determined. The copper ion homeostasis properties of the derivatives were determined using the copper chelation assay. The in vitro antimalarial activity was tested against the 3D7 parasite strain using the Plasmodium lactate dehydrogenase assay and the larvicidal activity of the derivatives was determined against Anopheles arabiensis (KGB) larvae. The lethality effect of the derivatives was determined on Artemia franciscana nauphlii. Eleven derivatives demonstrated notable copper (I) chelation activity, with 5,7-diiodo-8-OH, 5-HSO3-7-iodo-8-OH, 5,7-dichloro-8-OH, 5-chloro-7-iodo-8- OH and N-butyl-2,2-imino-di-(8-OH) comparable to the tetrathiomolybdate control. The derivatives preferentially chelated copper (I) rather than copper (II). 2-Amino-8-OH, 5,7-diiodo-8-OH, 5,7- dimethyl-8-OH and 5,7-dichloro-8-OH inhibited the intra-erythrocytic parasite (% Parasite inhibited: 100.0±6.1; 93.9±6.2; 87.1±1.3; 84.9±0.7 and 82.5±0.2%, respectively) similarly to the chloroquine control (86.7±6.8%). The derivatives induced less haemolysis compared to chloroquine and had a low lethal effect against the Artemia franciscana nauplii with the exception of three derivatives (5,7- dimethyl-8-OH, N-butyl-2,2-imino-di-(8-OH) and 2-benzyl-8-OH). Derivatives that demonstrated larvicidal activity included 5,7-dichloro-2-methyl-8-OH, 2-benzyl-8-OH and 5-chloro-7-bromo-8-OH (% Mortality: 100.0±0.0, 100.0±0.0 and 88.0±17.7%, respectively) at 200 µM compared to DDT (100.0±0.0%). Compounds with a functional group at position 2, 5 and 7 on the hydroxyquinoline ring and those with a large functional group demonstrated a high affinity to chelate copper, inhibit the malaria parasite and possessed larvicidal activity. The copper chelating, antimalarial and larvicidal properties of 8-hydroxyquinoline derivatives indicate their potential for use in the control of malaria.
Page 146 of 170 IN SILICO DESIGNED SMALL-GATEKEEPER PROTEIN KINASE AND BROMODOMAIN SELECTIVE INHIBITORS EXHIBIT POTENTIAL ANTIPLASMODIAL ACTIVITY
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Matlebjane, D.M.A. (1); Birkholtz, L. (2); Stander, B.A. (3); Cromarty, A.D (1)
(1) Department of Pharmacology, Faculty of Health Sciences, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa; (2) Department of Biochemistry, Faculty of Natural and Agricultural Sciences, Centre for Sustainable Malaria Control, University of Pretoria, South Africa; (3) Department of Physiology, Faculty of Health Sciences, Centre for Sustainable Malaria Control, University of Pretoria, South Africa
Abstract The malaria parasite is developing resistance to current antimalarial drugs. To combat the emerging resistance, artemisinin combination therapy is recommended as the first line antimalarial treatment by the World Health Organisation. There is an urgent need for the development of new antimalarial drugs with novel mechanisms of action that can treat malaria and reduce transmission, to replace drugs lost to resistance. Protein kinases are involved in regulation of gene expression. Protein kinase inhibitors (PKIs) have been used successfully in anticancer therapy and studies suggest that PKIs can be used as targets for malaria chemotherapeutics. Furthermore, bromodomain proteins, which contain a hydrophobic acetyl-lysine binding site, mediate the recruitment of proteins into macromolecular complexes. These are promising targets as they represent attractive binding pockets for small active molecules. Recently five potential antimalarial compounds were synthesised, following in silico structure-based drug design to target P. falciparum small-gatekeeper protein kinases and bromodomain proteins. The aim of this study is to determine the antiplasmodial activity of these compounds. The antiplasmodial activity of the compounds was tested in vitro against the P. falciparum 3D7 (drug-sensitive), K1 and W2 (drug-resistant), strains using a standardised parasite culture methodology. A fluorescence-based assay using SYBR Green 1 dye was used. Fluorescence was measured using Glomax-Multi+ Detection System at 520 nm. The IC50 of each compound was obtained for the different blood stages for the different parasite strains. The compounds were also tested at the early and late stage gametocytes, at 1 and 5 µM. Compounds
W11, W12, and W14 all had IC50 values less than 1 µM when tested against the asexual 3D7 strain. These compounds have a resistance index of less than 3, indicating low cross-resistance amongst the resistant strains compared to chloroquine. Early and late stage gametocytes were not sensitive to the compounds at either 1 or 5 µM. The present study demonstrates that in silico drug design is a valuable tool to identify antiplasmodial lead compounds in a cost-effective manner. Although three of the five compounds show activity against the asexual stages of the parasite none demonstrated transmission blocking activity and requires further optimisation.
Page 147 of 170 SYNTHESIS AND MODIFICATION OF ANTIFOLATES
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Molatsane, T.; Rousseau, A.
Abstract The synthesis of novel antifolates initially designed against Plasmodium falciparum dihydrofolate reductase (PfDHFR) began with dihydrotriazine analogues based on the structure of cycloguanil which was succeeded by pyrimidine analogues all of which had hit compounds which showed comparable potency when compared to cycloguanil. Due to time constraints very few pyrimidine analogues were made, as such, we want to synthesize a series of analogues of the potent flexible pyrimidine, prepared in our laboratories, in order to gain an understanding of the structure activity relationships of this class of compounds. In particular, we will vary the length of the flexible linker, and the nature and position of substituents on the aromatic ring. Biological data obtained from the first series will be used to determine which compounds will be prepared in the subsequent series of compounds. The compounds will be prepared using a multicomponent reaction to prepare the core pyrimidine. Subsequent functional group interconversion to produce an aldehyde will enable us to carry out our key step, reductive amination, to afford the desired flexible pyrimidines. As only a limited number of compounds were prepared in this series, we now wish to expand the collection of compounds prepared and explore the structure-activity relationships of this class of compounds. In this synthesis we intend on using substituted anilines, benzylamines and phenethylamines which are commercially available in the first series of compounds. The synthesis of complex precursors that are not commercially available would be considered for subsequent series of compounds.
Page 148 of 170 THE ISOMERIC EFFECT OF ESSENTIAL OIL CONSTITUENTS ON ANOPHELES ARABIENSIS LARVAE AND ARTEMIA FRANCISCANA NAUPLII
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Mustapha, O. (1); Van Zyl, R.L. (1); Coetzee, M. (2); Oliver, S. (2)
(1) Pharmacology Division, Department of Pharmacy & Pharmacology; WITS Research Institute for Malaria (WRIM), Faculty of Health Science University of the Witwatersrand, Johannesburg; (2) WITS Research Institute for Malaria (WRIM), Faculty of Health Science University of Witwatersrand; Vector Control Reference Laboratory, National Health Laboratory Services, Sandringham
Abstract Due to the increase in resistance to antimalarial and larvicidal agents, new compounds are required to ensure adequate management of malaria. Traditional plants are commonly used in South Africa, where the essential oils and their constituents (EOCs) have promising antimalarial activity against the intra-erythrocytic parasite and vector. As such the aim of this study was to investigate the larvicidal and toxicological properties of isomeric EOCs. The EOCs isomers were commercially purchased. An. arabiensis larvae (KGB strain) in the 3rd/4th instar stage and Artemia franciscana nauplii were incubated under optimal conditions with 0.01% EOC or test control. After 24 hrs, the percentage mortality was recorded in comparison to an untreated control and positive control, namely, DDT and potassium dichromate for the larvae and nauplii, respectively. If found to be active (>60% inhibition), the 50% lethality concentration (LC50 values) were determined using probit analysis. Cis-nerolidol was the most active of the evaluated EOCs, and twice as active as trans-nerolidol; but cis-nerolidol was the most toxic towards the nauphlii. (R)-(+)-citronella and (S)-(-)-citronellal were both active against Anopheles and both were non-toxic to the Artemia nauphii at 0.01%. The negative pinene isomers ((-)-α-pinene, (-)-β-pinene)) were more active against the larvae than the positive isomers ((+)-α-pinene, (+)-β-pinene)); whilst only (-)-β-pinene and (+)-β- pinene displayed toxicity to the nauplii. The isomeric nature of the EOCs conferred varying larvicidal activity and toxicological profiles. The promising inhibitory larvicidal activity and preliminary safety profile, warrants further investigations.
Page 149 of 170 SYNTHESIS AND ANTIMALARIAL PROPERTIES OF SPIROINDOLONE ANALOGUES
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Naicker, A. (1), de Koning, C.B. (1) Rousseau, A.L. (1)
(1) Molecular Sciences Institute, School of Chemistry, University of the Witwatersrand, Private Bag 3, P.O. WITS, 2050, South Africa
Abstract Widespread drug resistance has extended to most classes of antimalarial drugs resulting in the need for the development of novel drugs. The focus of the discovery process is on new medicines that are structurally different from existing drugs and that act by novel mechanisms. Cipargamin, a spiroindolone, is a promising synthetic antimalarial candidate has been shown to exert transmission- blocking activity1. In addition, it has an IC50 of 1nM against P. falciparum strains, including those that are drug resistant1. Using standard medicinal chemistry rationale, the structure- activity relationship of a series of analogue compounds bearing reduced functionality, has been explored. One strategy included the synthesis of analogues with modifications that increased the free rotation of the compound by removing rigid bonds to allow for flexibility within the active site. Another strategy included the synthesis of analogues with a reduced number of aromatic rings in order to assess the effect of reduced hydrophobic interactions on biological activity. The synthesis of these analogues will be described.
Page 150 of 170 SYNTHESIS AND BIOLOGICAL EVALUATION OF ANTIMALARIAL ANTIFOLATES
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Seanego, T.D. (1); de Koning, C.B. (1); Rousseau, A.L. (1); Zyl, R.L. (2)
(1) Molecular Sciences Institute, School of Chemistry, University of the Witwatersrand, Private Bag 3, PO WITS, 2050, South Africa, [email protected]; (2) Pharmacology Division, Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown 2193, South Africa
Abstract Increasing resistance of P. falciparum to chloroquine and the dihydrofolate reductase (DHFR) inhibitors pyrimethamine and cycloguanil has shown the need for the development of new antimalarial drugs. Studies have shown that resistance to pyrimethamine is caused by its rigid structure, resulting in steric clashes with mutant amino acid residues within the PfDHFR active site1. In the search of new antimalarial antifolates, a series of novel flexible pyrimethamine analogues were prepared and screened for antimalarial activity in an in vitro P. falciparum screen on the Gambian FCR-3 strain (chloroquine and cycloguanil resistant strain) with dihydroartemisinin, methotrexate and quinine as controls. Biological results of a small set of flexible pyrimethamine analogues prepared will be described.
Page 151 of 170 MULTITARGET ANTIMALARIAL POLYAMINE ANALOGUES: RESISTING RESISTANCE
Abstract Theme Pharmacology, drug discovery and development
Authors and Affiliations Verlinden, B.
(1) Malaria Parasite Molecular Laboratory, Department of Biochemistry, UP Institute for Sustainable Malaria Control, University of Pretoria, South Africa; (2) Department of Pharmaceutical and Biomedical Sciences, Medical University of South Carolina, Charleston, South Carolina 29425, USA
Abstract Currently, any widely used antimalarial drug has a limited therapeutic life time. The central focus of this research is to drastically increase the therapeutic life time of an antimalarial drug. There are currently nine promising antimalarial candidates with molecularly defined targets in phase I and II clinical trials. A major concern is that all of these candidates are directed at single targets within the malaria parasite. Treating patients with a combination of antimalarial drugs that are directed at single targets will inevitably lead to more drug resistance. Antimalarial drugs such as artemisinin and chloroquine have demonstrated lengthened therapeutic usability due their pleiotropic nature. This study investigated a pleiotropic polyamine scaffold that inhibits asexual parasites in the low nM- range with selectivity indexes >7000-fold. The multitarget nature of the polyamine scaffold was investigated on P. falciparum 3D7 asexual parasites using flow cytometric analysis and immunofluorescent imaging of nuclear division. The epigenetic status of treated parasites with regards to histone post-translational modification was studied with western blot analysis, and protein targets were determined with DARTS (drug affinity responsive target stability). The polyamine scaffold produced numerous effects in various aspects of the malaria parasite’s life cycle. The lead inhibitor investigated targeted (1) parasite DNA synthesis, (2) schizont microtubule structures, (3) the polyamine related enzyme, S-Adenosylmethionine synthetase and (4) the parasite epigenetic enzyme, LSD-1. Furthermore, this scaffold demonstrated transmission blocking activity, targeting late stage P. falciparum gametocytes. Microarray analysis indicates that this multitarget scaffold is not directed at established antimalarial targets with >80% of the differential transcripts unique to those of polyamine treatment with no strong correlations to 20 well-established antimalarial compounds, suggesting novel target/s. When asexual parasites are exposed to this multitarget scaffold no recrudescence or viable resistant mutants are generated, suggesting that these promising multitarget inhibitors may serve as “resistance-resistant” antimalarial candidates.
Page 152 of 170 TOWARDS THE ELIMINATION OF MALARIA IN THE UNION OF COMOROS: GEOGRAPHICAL INSIGHT ON CONTROL ACTIONS AND MAINTENANCE FACTORS
Abstract Theme Modeling towards elimination
Authors and Affiliations Attoumane, A. (1*); Bacar, A. (2); Silai, R. (2); Cardinale, E. (3); Pennober, G. (1); Herbreteau, V. (1*)
(1) UMR ESPACE-DEV (IRD, UM, UR, UAG), Saint-Pierre, La Réunion, France; (2) Programme National de Lutte Contre le Paludisme, Ministère de la santé publique, Union des Comores; (3) CIRAD, UMR 15 CMAEE, Sainte Clotilde, La Réunion, France;
Abstract Malaria has long been endemic in the Union of Comoros. Since 2007, the Comorian government has been conducting a national strategy for eradicating malaria by 2016. Between 2010 and 2014, the number of malaria cases has radically fallen by more than 90%. This study aims at describing the spatial patterns of the epidemiology of malaria and at identifying risk factors through a geographical approach. Monthly cases, as reported by the “Programme National de lutte Contre le paludisme” (PNLP) from 2010 to 2014, were geo-referenced in the three islands (i.e. Grande Comore, Moheli and Anjouan) at two different scales: by sanitary district and by health center. In Grande Comore, malaria cases confirmed by microscopic exam during the active control from 2012 to 2014 were geo- localized at the village scale. Data on environmental factors, including meteorology and land use (analyzed by remote sensing of SPOT 5 satellite images) but also information on malaria control actions were integrated into a spatial database. We realized statistical and spatial analyses to show the relationships between this dataset and malaria information. The mapping of malaria incidence reveals spatial and temporal variations between and within islands. In the 7 districts of Anjouan and in 2 of the 3 districts of Moheli, the incidence is less than 1 case per 100,000 inhabitants in 2014. In contrast, the incidence remains higher in every health district of Grande Comore with more than 4 cases per 100,000 inhabitants in 2014. This is consistent with the reduction of prevalence of Plasmodium falciparum that decreased from 24% in 2012 to 2 in 2014 in Grande Comore. Also, Long before the two other islands, Moheli has benefited from a campaign of indoor insecticide spraying and a population mass treatment by Artequick and Primaquine between 2007 and 2009. In addition, we highlight spatial relationships between malaria incidences and environmental factors in Grande Comore. This study clarifies the impact of control actions on malaria epidemiology and the environmental factors that contributed to its maintenance.
Page 153 of 170 TOWARDS MALARIA ELIMINATION: IMPLEMENTATION OF AN ALERT AND RESPONSE SYSTEM FOR MALARIA SURVEILLANCE, SOUTH AFRICA, 2016
Abstract Theme Modeling towards elimination
Authors and Affiliations Ngwenyama, B. (1, 2); Baloyi, R.E. (2, 3); Shandukani, M.B. (1); Graffy, R. (4); Kruger, P. (5); Mabuza, A. (6); Raswiswi, E. (7); Moonasar,D. (1)
(1) National Department of Health (DOH); (2) National Institute for Communicable Diseases; (3) Field Epidemiology and Training Program; (4)Clinton Health Access Initiative; (5) Limpopo Provincial Department of Health; (6) Mpumalanga Provincial Department of Health; (7) KwaZulu-Natal Provincial Department of Health
Abstract South Africa’s surveillance system has historically been decentralized, with each province independently operating its own information systems. The soloed databases limits the ability of stakeholders to make timorous decisions based upon complete and accurate data. In 2015, the National and Provincial Departments took a first step toward resolving this by developing MalariaConnect, an electronic tool for real-time case notification from healthcare facilities. MalariaConnect has been deployed in 297 healthcare facilities across all five malaria endemic districts (Ehlanzeni, Mopani, Umkhanyakude, Uthungulu, and Vhembe). MalariaConnect has reduced the time to report a case from the healthcare facility to district, provincial, and national level staff. The next step will be holistic surveillance system strengthening, which will include: development of a single centralized National Malaria Information System (NMIS) into which stakeholders at all levels have access. Integration of MalariaConnect into the National Malaria Information System and development of mobile tools to inform, expedite, and target program response. Operational effectiveness and impact of South Africa’s surveillance system upgrade will be measured by assessment of the following indicators: stability (down time of the NMIS for data entry and data reporting), accessibility (users at all levels are able to access the system for data entry and reporting), reporting functionality (the NMIS is able to generate all required reports), analytic functionality ( the NMIS is able to facilitate analysis across data sets) and operational effectiveness( the NMIS upgrade decreases the time spent capturing, cleaning, analysing and reporting upon data for the program holistically) and program response (the NMIS upgrade improve the ability for the program to deliver upon its response targets). Surveillance system strengthening will improve the overall response and management for the Malaria program.
Page 154 of 170 MATHEMATICAL MODEL OF MALARIA TRANSMISSION IN CHILDREN UNDER FIVE YEARS IN AN ENDEMIC AREA
Abstract Theme Modeling towards elimination
Authors and Affiliations Danquah, B. (1); Chirove, F. (1); Banasiak, J. (2)
(1) School of Mathematics, Statistics and Computer Science, University of KwaZulu Natal; (2) Department of Mathematics and Applied Mathematics, University of Pretoria
Abstract The population of children aged under five years are the most vulnerable to the malaria infection and mortality. In the endemic area, they make up more than 90% of the disease death. However, few models have concentrated on this population. A deterministic model that considers the transmission of malaria infection in variable population of mosquitoes and children aged under five with recovery due to effective treatment is formulated and analyzed to investigate the impact of effective treatment and disease mortality on the dynamics of the disease. Using the next generation matrix, the reproduction number is derived. The disease free and endemic equilibria are found. The stability of the equilibria are investigated. Sensitivity analysis is performed. The results demonstrate that the disease free equilibrium is globally asymptotically stable when the reproduction number is less than one, the density dependent death rate of mosquitoes and the disease induced mortality in children are zero. The model reveals the absence of backward bifurcation, when the disease induced mortality is zero due to effective treatment. The reproduction number is most sensitive to the contact rate between mosquitoes and the children, the probability of transmission of infection from an infectious mosquito to a susceptible child and vice versa. Thus, to eliminate malaria in the long term, the combination of control interventions that will keep the disease reproduction number less than one must be used. First reducing contact rate between the two populations, early diagnosis and effective treatment that block transmission of infection at the early stages, death due to infection and development of gametocytes must be used. Last but not the least, vaccine that give total recovery and immunity is highly recommended.
Page 155 of 170 INVESTIGATING THE INFLUENCE OF METEOROLOGICAL DRIVERS ON THE UNUSUAL MALARIA TRANSMISSION BETWEEN JULY AND OCTOBER 2013 IN SWAZILAND
Abstract Theme Modeling towards elimination
Authors and Affiliations Dlamini, M. (1); Dlamini, B. (2); Kunene, S. (1); Seyama, E. (3); Chuang, T. (4, 5)
(1) National Malaria Control Programme, Swaziland Ministry of Health, Swaziland; (2) Clinton Health Access Initiative, Swaziland; (3) Swaziland Meteorology Services, Swaziland; (4) Department of Molecular Parasitology and Tropical Medicine, College of Medicine, Taipei Medical University, Taiwan; (5) Center for International Tropical Medicine, College of Medicine, Taipei Medical University, Taiwan
Abstract Weather patterns have a profound effect on mosquito life cycle, longevity (survival) and also on the development of malaria parasites. Understanding weather patterns can offer suitable indicators for malaria early warning before outbreaks and also contribute to the reversal of malaria incidence. Malaria outbreaks in a locality in Swaziland in the months of August and September 2013 warranted the investigation of weather anomalies that could be associated to the sudden increase in malaria cases. Daily meteorological data derived from Mhlume and Big Bend weather stations between January 1991 and December 2012 was used to investigate associations between weather patterns and rising incidence of malaria. Geo-statistical methods were used to analyze climate data that included rainfall, temperature ranges, and relative humidity to determine departures and uncommon patterns including deviations from the long term mean patterns. A ten day decadal analysis of climatic parameters revealed departures from the long term mean for the time periods attributed to the occurrence of malaria cases. High minimum relative humidity (> 60%) experienced during most decads of 2013 contributed to the longevity of the mosquito life span resulting in increased vectoral capacity. The year 2013 experienced rainfall in almost all decads including the winter months. The effects and impacts of climate change can be deduced from this investigation as indicated by unusual occurrences of higher than normal minimum relative humidity and rainfall in most decads of 2013. Evidence of change in weather and the occurrence of malaria cases were observed. Climate change is likely to increase malaria cases as observed from malaria cases recorded in the winter season of 2013. The patterns observed in this analysis can form a basis for early warning for malaria in Swaziland.
Page 156 of 170 INTERACTIVE MALARIA EDUCATION INTERVENTION AND ITS EFFECT ON COMMUNITY PARTICIPANT KNOWLEDGE: THE MALARIA AWARENESS PROGRAM (MAP) IN VHEMBE DISTRICT, LIMPOPO, SOUTH AFRICA
Abstract Theme Education & health promotion
Authors and Affiliations Cox, S. (1, 2); Guidera, K. (1, 3); Simon, M. (1)
(1) One Sun Health; (2) Johns Hopkins Bloomberg School of Public Health; (3) Harvard Business School
Abstract Malaria is preventable and treatable, yet remains the most prevalent parasitic endemic disease in Africa. Although South Africa exceeded the 2015 Millennium Development Goals for malaria control, Limpopo Province recorded the lowest reduction in malaria cases compared to other endemic provinces within the country between 2000 and 2010. This paper analyzes prospective observational data from the Malaria Awareness Program (MAP), a multi-week interactive education program led by local home-based care (HBC) workers to improve knowledge of malaria in the Mutale Municipality, Vhembe District, Limpopo, South Africa. MAP education sessions were conducted in 19 villages by HBC workers and representatives from the Department of Health’s Malaria Control Division. The effectiveness of MAP was measured through pre/post-participation surveys assessing general malaria knowledge in four categories: transmission, symptoms, prevention, and treatment. All analyses were performed using Stata 13. Baseline characteristics of participants were compared through chi-squared tests using a threshold p=0.05 as a cut-off for significance. From 2012 – 2015, 1063 individuals participated in MAP. Data was collected and analyzed from 723 participants. Over 41% of participants previously experienced malaria themselves, whereas 77.8% reported that someone in their household had experienced malaria. Surveys demonstrated a 11.2% and 12.9% increase in the number of participants who cited correct methods for transmission and prevention of malaria, respectively. After participating in 2-4 educational workshops, the adjusted odds of correct knowledge score versus the combined partially correct and incorrect categories was 3.6 (95% CI: 2.4 – 5.5, p-value <0.001) times greater for transmission and 3.2 (95% CI: 2.3 – 4.4, p-value <0.001) times greater for prevention, with all other variables in the model held constant. Participants perceived MAP as rewarding and expressed interest in future programming. Findings from this study suggest that MAP is an effective approach to increasing understanding of malaria, however, it is essential to tailor interventions and health promotion strategies based on results presented. Future research should assess behavioral changes toward malaria prevention and treatment as a result of an intervention, and examine changes in overall incidence in the region.
Page 157 of 170
LIST OF CONFERENCE PARTICIPANTS
Page 158 of 170 Abs ID Abstract title Presenting Type Plenary SETTING THE RESEARCH AGENDA AROUND MALARIA ELIMINATION Blumberg, L Oral Plenary TOWARDS MALARIA ELIMINATION IN THE MOSASWA (MOZAMBIQUE, SOUTH AFRICA AND SWAZILAND) REGION Moonasar, P Oral MALARIA CASE MANAGEMENT IN THE CONTEXT OF MALARIA ELIMINATION: THE ROLE OF THE POPULATION AT Plenary Ukpe, S Oral RISK RESIDUAL MALARIA: CHANGED MIND-SETS TO QUELL THE SEEDS OF FAILURE Plenary RESIDUAL MALARIA: CHANGED MIND-SETS TO QUELL THE SEEDS OF FAILURE Braack, L Oral SEEING THE WOOD FOR THE TREES: MALARIA PARASITE SYSTEMS BIOLOGY AND HOW IT AIDS ANTIMALARIAL Plenary Birkholtz, L Oral DRUG DISCOVERY Plenary MUTANT MALARIA PARASITES: AN OVERVIEW ON THEIR GENETICS, ORIGINS AND SPREAD Raman, J Oral Plenary PERSPECTIVES ON TRANSLATING RESEARCH INTO MALARIA VECTOR CONTROL Brooke, B Oral Plenary SPATIO-TEMPORAL DYNAMIC OF ASYMPTOMATIC MALARIA IN A SAHELIAN ENVIRONMENT Gaudart, J Oral Plenary NEW APPROACHES TOWARDS MALARIA VECTOR CONTROL Focke, W Oral IMPACT OF THE LUBOMBO SPATIAL DEVELOPMENT INITIATIVE AFTER TWELVE YEARS IN SOUTHERN Plenary Maharaj, R Oral MOZAMBIQUE Plenary ANTIMALARIAL DRUG DISCOVERY AT H3D Chibale, K Oral Plenary CURRENT STATE OF DEVELOPMENT OF PANTOTHENAMIDES AS ANTI-PLASMODIAL AGENTS Strauss, E Oral MC/001 A REVIEW OF THE STERILE INSECT TECHNIQUE FOR THE CONTROL OF MALARIA VECTORS IN SOUTH AFRICA Munhenga, G Oral MC/002 THE HIDDEN NON-BLOODSTREAM PARASITE RESERVOIR: A MALARIA CONTROL PROBLEM Markus, M Poster AN ASSESSMENT OF COMPLIANCE TO ENVIRONMENTAL SAFEGUARDS IN THE USE OF DDT IN CHONGWE IRS MC/003 Chilabi, A Poster PROGRAMME RURAL PART OF ZAMBIA EFFECT OF STABLE AND FLUCTUATING TEMPERATURES ON THE LIFE HISTORY TRAITS OF ANOPHELES ARABIENSIS MC/004 Davies, C Poster AND AN. QUADRIANNULATUS UNDER CONDITIONS OF INTER- AND INTRA-SPECIFIC COMPETITION MC/005 WORKER ABSENCE DUE TO MALARIA IN A BANANA PLANTATION IN ZIMBABWE Lukwa, A Poster THE LARVICIDAL EFFECTS OF BLACK PEPPER (PIPER NIGRUM LINN.) AND PIPERINE AGAINST INSECTICIDE MC/007 Samuel, M Poster RESISTANT AND SUSCEPTIBLE STRAINS OF ANOPHELES MALARIA VECTOR MOSQUITOES MC/008 USING CRISPR-CAS9 TO MANIPULATE THE GENOME OF P. FALCIPARUM Liebenberg, D Poster FACILITATING FACTORS AND BARRIERS TO MALARIA RESEARCH UTILIZATION FOR POLICY DEVELOPMENT IN MC/009 Mwendera, C Oral MALAWI
Page 159 of 170 EPIDEMIOLOGY OF MALARIA IN MADAGASCAR: SPATIOTEMPORAL DISTRIBUTION OF UNCOMPLICATED AND MC/010 Ihantamalala, F Poster COMPLICATED MALARIA MC/011 COMPARING CANINE BABESIA ROSSI WITH HUMAN FALCIPARUM MALARIA Leisewitz, A Oral EFFECT OF IRRADIATON ON FEMALE ANOPHELES ARABIENSIS: AN INVESTIGATIVE STUDY TOWARDS THE MC/012 Dandalo, L Poster DEVELOPMENT OF THE STERILE INSECT TECHNIQUE TO CONTROL MALARIA VECTORS IN SOUTH AFRICA QUANTITATIVE PROFILING OF THE PLASMODIUM FALCIPARUM PARASITE’S HISTONE POSTTRANSLATIONAL MC/013 Coetzee, N Oral MODIFICATION LANDSCAPE DURING SEXUAL DIFFERENTIATION SELECTING MOST EFFICIENT TARGETS FOR SPATIALLY FOCUSED INTERVENTIONS: MALARIA INCIDENCE VERSUS MC/014 Kankoe, S Oral POPULATION MOBILITY MC/015 ADDRESSING CURRENT CHOKEPOINTS IN ANOPHELES ARABIENIS MOSQUITO MASS REARING Wood, O Poster CROSS-REACTIVITY OF OXIDANT DRUGS AGAINST ASEXUAL AND GAMETOCYTE STAGES IN PLASMODIUM MC/016 Coertzen, D Oral FALCIPARUM PARASITES ASSESSING RISK FACTORS ASSOCIATED WITH PLASMODIUM FALCIPARUM EXPOSURE USING SEROLOGICAL DATA MC/017 Agubuzo, E Oral IN BUSHBUCKRIDGE MUNICIPALITY, MPUMALANGA PROVINCE, SOUTH AFRICA INFECTION OF AFRICAN MALARIA VECTOR MOSQUITOES WITH PLASMODIUM FALCIPARUM MALARIA MC/018 Bennett, A Oral PARASITES MC/020 MICROPOROUS POLYOLEFINS AS CONTROLLED RELEASE DEVICES FOR MOSQUITO REPELLENTS Sibanda, M Oral MC/021 EFFECT OF MICROBIAL LARVICIDES ON MOSQUITO LARVAE IN MALARIA AREAS OF BOTSWANA AND ZIMBABWE Mpofu, M Oral MC/022 CRISPR-CAS EVALUATION OF IMPORTANT CELL CYCLE REGULATORS Von Gruning, H Poster MC/023 A NEW METHOD FOR SAMPLING VOLATILE HUMAN SKIN CHEMICALS IN A MALARIA CONTEXT Roodt, A Poster MC/024 IS ANOPHELES VANEEDENI A VECTOR OF MALARIA IN SOUTH AFRICA? Burke, A Oral INVESTIGATING THE ANTIPLASMODIAL POTENTIAL OF (BIS)THIOUREA AND (BIS)UREA POLYAMINE ANALOGUES MC/025 Abrie, C Poster ON THE GAMETE FORM OF PLASMODIUM FALCIPARUM PARASITE EVALUATION OF THE PREVALENCE OF CYP 2D6 MUTATIONS IN VHEMBE DISTRICT, LIMPOPO PROVINCE, SOUTH MC/026 Makhanthisa, T Poster AFRICA USING DRIED PLASMODIUM FALCIPARUM-INFECTED BLOOD SAMPLES AS POSITIVE CONTROLS FOR MALARIA MC/027 Ming Sun, L Poster RAPID DIAGNOSTIC TESTS MC/029 MEMBRANE TRANSPORT IN THE SEXUAL, TRANSMISSIBLE STAGES OF PLASMODIUM FALCIPARUM PARASITES Naude, M Poster DRUG TARGET DECONVOLUTION OF ANTIPLASMODIAL COMPOUNDS USING DRUG AFFINITY RESPONSIVE MC/030 Henn, D Poster TARGET STABILITY, FOR ASEXUAL AND SEXUAL STAGES OF PLASMODIUM FALCIPARUM PARASITES
Page 160 of 170 PHENOTYPIC PROFILING OF GROWTH PERTURBATIONS OF THE HUMAN MALARIA PARASITE PLASMODIUM MC/031 Joshua, A Poster FALCIPARUM MC/032 MONITORING INSECTICIDE RESISTANCE IN THE AN. GAMBIAE COMPLEX FROM KWAZULU-NATAL, SOUTH AFRICA Mofokeng Oral MC/033 INVESTIGATION OF DIFFERENTIAL DRUG RESPONSES IN DIFFERENT P. FALCIPARUM PARASITE LIFE CYCLE STAGES Weidemann, G Poster MC/034 SYNTHESIS AND BIOLOGICAL EVALUATION OF HETEROAROMATIC INHIBITORS OF PLASMODIUM FALCIPARUM Rousseau, A Oral MC/035 FREQUENCY OF SEXUALLY MATURED MALES, IN A WILD ANOPHELES FUNESTUS SWARM IN BURKINA FASO Zawada, J Poster THE MONITORING OF BASELINE STERILITY IN THE POPULATION OF ANOPHELES ARABIENSIS IN MAMFENE, KZN, MC/037 Lobb, L Poster SOUTH AFRICA PRELIMINARY STUDY ON THE PREVALENCE OF RESISTANCE ASSOCIATED POLYMORPHISM IN PLASMODIUM MC/038 Sijuade, O Poster FALCIPARUM FIELD ISOLATES PRESENTED IN GUATENG, SOUTH AFRICA AN EVALUATION OF THE GAMETOCYTOCIDAL PROPERTIES OF NATURAL PRODUCT COMPOUNDS AGAINST MC/039 Breedt, C Poster PLASMODIUM FALCIPARUM MC/040 IN VITRO ACTIVITY OF NAPHTHYLISOQUINOLINE ALKALOIDS ON PLASMODIUM FALCIPARUM GAMETOCYTES Moyo, P Poster MC/041 ANALYSIS OF THE ESTERASE ENZYMES ACTIVITY IN ANOPHELES FUNESTUS ADULTS Koekemoer, L Oral THE MOLECULAR DETERMINANTS FOR TRAFFICKING OF AMA-1, A MALARIA INVASION PROTEIN AND VACCINE MC/042 Churchyard, A Oral CANDIDATE THE EFFECTS OF AGRICULTURAL POLLUTANTS ON THE LIFE HISTORY AND INSECTICIDE RESISTANCE PHENOTYPE MC/043 Oliver, S Oral OF THE MAJOR MALARIA VECTOR ANOPHELES ARABIENSIS (DIPTERA: CULICIDAE) MC/044 SIMPLIFYING MALARIA PARASITE COUNTS ON THIN BLOOD FILMS Poonsamy, B Poster MC/045 MULTITARGET ANTIMALARIAL POLYAMINE ANALOGUES: RESISTING RESISTANCE Verlinden, B Poster MC/046 MALARIA TRENDS SOUTH AFRICA, 2011-2015 Shandukani, M Poster MC/047 ANOPHELES ARABIENSIS HABITAT SUITABILITY MODEL SPATIALLY PREDICTS MALARIA HOTSPOTS IN ZIMBABWE Gwitira, I Oral A COMPARATIVE, TRANSCRIPTOMIC ANALYSIS OF THE EFFECTS OF A LEAD CLINICAL CANDIDATE ON THE MC/048 Connacher, J Oral ASEXUAL AND GAMETOCYTE STAGES OF PLASMODIUM FALCIPARUM PARASITES MC/049 APPLICATION OF NETWORK ANALYSIS ON P. FALCIPARUM TRANSCRIPTOMES Van Wyk, R Poster PLASMODIUM FALCIPARUM GAMETOCYTES SHOW STAGE-SPECIFIC, CHEMOTYPE SPECIFIC TRANSCRIPTIONAL MC/050 Van Biljon, R Oral RESPONSE TO DRUG PERTURBATION ANALYSING TRENDS AND FORECASTING MALARIA EPIDEMICS IN MADAGASCAR USING A SENTINEL MC/051 Girond, F Oral SURVEILLANCE NETWORK: A WEB-BASED APPLICATION MC/052 OPERATIONAL LESSONS LEARNED FROM IRS PROGRAM IMPROVEMENTS IN NAMIBIA Angula, H Poster
Page 161 of 170 ASSESSING THE PERFORMANCE OF RE-ACTIVE CASE DETECTION (RACD) AS AN ELIMINATION STRATEGY IN MC/053 Maponga, A Poster MATABELELAND SOUTH PROVINCE HETEROGENEITY OF PLASMODIUM FALCIPARUM MALARIA IN MATABELELAND SOUTH: ANALYSIS TRAVEL MC/054 Maponga, A Oral HISTORY REPORTED BY MALARIA CASES REPORTED IN ROUTINE SURVEILLANCE SUMMARY OF FINDINGS MALARIA ELIMINATION OPERATIONAL CAPACITY ASSESSMENT SURVEY - ZIMBABWE MC/055 Mberikunashe, J Poster 2015 DECREASED PREVALENCE OF THE PLASMODIUM FALCIPARUM CHLOROQUINE RESISTANCE TRANSPORTER GENE MC/056 MUTATION K76T NINE YEARS AFTER CHLOROQUINE TREATMENT WITHDRAWAL IN CHIPATA AND KATETE Mwenda, M Poster DISTRICTS, EASTERN ZAMBIA. MATHEMATICAL MODELLING GUIDING ELIMINATION IN THE ASIA-PACIFIC REGION AND ITS POTENTIAL IN MC/057 Silal, S Oral AFRICA HETEROGENEITY OF PLASMODIUM FALCIPARUM TRANSMISSION IN NORTH-EASTERN SOUTH AFRICA: MC/058 Biggs, J Oral IMPLICATIONS FOR MALARIA ELIMINATION IN SILICO DESIGNED SMALL-GATEKEEPER PROTEIN KINASE AND BROMODOMAIN SELECTIVE INHIBITORS EXHIBIT MC/059 Matlebjane, D Poster POTENTIAL ANTIPLASMODIAL ACTIVITY MC/060 SYNTHESIS AND BIOLOGICAL EVALUATION OF ANTIMALARIAL ANTIFOLATES Seanego, D Poster MC/061 DIGITAL IMAGE ANALYSIS FOR DETERMINING MALARIA PARASITE DENSITY IN THICK BLOOD FILMS Frean, J Oral ASSESSING THE IN VITRO EFFICACY IN SILICO DESIGNED COMPOUNDS TARGETING THE MALARIAL QI SITE OF MC/062 Damadeu, L Poster CYTOCHROME BC1 PREVALENCE OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PD) DEFICIENT PHENOTYPES AND GENOTYPES MC/063 Awandu, S Oral AT SELECT POINT OF CARE HEALTH SETTINGS IN VHEMBE DISTRICT, LIMPOPO PROVINCE, SOUTH AFRICA MC/064 PLANNING OF HOUSING IMPROVEMENT INTERVENTIONS FOR MALARIA ELIMINATION IN SWAZILAND Sibandze, M Oral DEVELOPMENT AND IMPLEMENTATION OF A MALARIA COMMODITY TRACKING AND ORDERING TOOL FOR MC/065 IMPROVED FORECASTING, PROCUREMENT AND SUPPLY MANAGEMENT OF MALARIA DIAGNOSTIC AND Mathobela, M Poster TREATMENT COMMODITIES USING A RISK BASED APPROACH TO DEVELOP A QUALITY MANUAL FOR PUBLIC HEALTH INSTITUTIONS MC/066 Leisegang, C Oral UNDERTAKING INVESTIGATOR-LED ANTIMALARIAL DRUG CLINICAL RESEARCH IN SOUTH AFRICA MC/068 TARGETING FOCI OF TRANSMISSION TOWARDS ELIMINATION OF MALARIA IN SOUTH AFRICA Morris, N Oral TRAVEL PATTERNS AND DEMOGRAPHIC CHARACTERISTICS OF MALARIA CASES, THE CASE STUDY OF MC/069 Pindolia, D Oral SWAZILAND'S ACTIVE SURVEILLANCE SYSTEM, 2010-2014
Page 162 of 170 DEVELOPMENT OF A PHARMACOVIGILANCE SAFETY MONITORING TOOL FOR THE ROLLOUT OF SINGLE LOW- MC/070 DOSE PRIMAQUINE AND ARTEMETHER-LUMEFANTRINE TO TREAT PLASMODIUM FALCIPARUM INFECTIONS IN Malambe, C Poster SWAZILAND: A PILOT STUDY IMPLEMENTATION OF THE DIAGNOSIS QUALITY ASSURANCE PROGRAM FOR ACCURATE MALARIA DIAGNOSIS IN MC/071 Ntshalintshali, N Poster SWAZILAND THE JOURNEY FROM DIAGNOSIS TO CASE INVESTIGATION FOR MALARIA ELIMINATION IN SWAZILAND: IS MC/072 Dlamini, N Poster REPORTING AND RESPONSE TIMELY? THREE PARALLEL INFORMATION SYSTEMS FOR MALARIA ELIMINATION IN SWAZILAND, 2010-2015: ARE THE MC/073 Zulu, Z Poster NUMBERS THE SAME? MC/074 DEVELOPMENT OF SEROLOGIC ASSAYS TO ASSESS MALARIA EXPOSURE IN SWAZILAND Nhlabathi, N Poster OPERATIONAL FACTORS ASSOCIATED WITH INCREASED DETECTION OF INFECTIONS FOR REACTIVE CASE MC/075 Bhangu, K Oral DETECTION (RACD) IN SWAZILAND. MC/076 ENVIRONMENTAL PROBLEMS WITH DDT AS USED IN MALARIA CONTROL Bouwman, H Oral INVESTIGATING THE INFLUENCE OF METEOROLOGICAL DRIVERS ON THE UNUSUAL MALARIA TRANSMISSION MC/077 Seyama, E Poster DURING JULY – OCTOBER 2013 A PROTOCOL TO EVALUATE THE EFFECTIVENESS AND FEASIBILITY OF REACTIVE TARGETED PARASITE MC/078 ELIMINATION (TPE) TO REACTIVE CASE DETECTION (RACD) AS A COMMUNITY LEVEL INTERVENTION IN Mkhonta, N Poster SWAZILAND TOWARDS MALARIA ELIMINATION: IMPLEMENTATION OF AN ALERT AND RESPONSE SYSTEM FOR MALARIA MC/079 Ngwenyama, B Poster SURVEILLANCE, SOUTH AFRICA, 2016 TOWARDS MALARIA ELIMINATION: AN EVALUATION OF THE SOUTH AFRICAN MALARIA SURVEILLANCE SYSTEM, MC/080 Baloyi, E Poster 2015 EPIGENETIC DRUGS TARGETING HISTONE POSTTRANSLATIONAL MODIFICATIONS DURING GAMETOCYTOGENESIS MC/081 Mmekwa, N Poster OF THE PLASMODIUM FALCIPARUM PARASITE EVALUATING EFFECTS OF LANDSAT-DERIVED ENVIRONMENTAL COVARIATES FOR PREDICTING MALARIA MC/082 Malahlela, O Oral DISTRIBUTION IN RURAL VILLAGES OF VHEMBE DISTRICT, SOUTH AFRICA THE CONTRIBUTION OF A SUSTANABLE VOLUNTEER BASE ON CROSS-BORDER MALARIA ELIMINATION: AN MC/083 Njovu, C Oral IMPACT ASESSMENT A STUDY TO IDENTIFY HIGH MALARIA RISK GROUPS AND THE ASSOCIATED RISK FACTORS USING A MC/084 Haindongo, E Oral PROGRAMMATIC TOOL: MALARIA ELIMINATION RISK FACTOR ASSESSMENT TOOL (MERFAT)
Page 163 of 170 MC/085 THE POTENTIAL USE OF SERIOUS GAMES IN MALARIA CONTROL AND ELIMINATION PROGRAMMES Coleman, M Oral EVALUATING THE UTILITY OF AMA-1 AND MSP-1 AS SEROLOGICAL MARKERS OF MALARIA EXPOSURE FOR MC/087 Chisenga, M Poster SURVEILLANCE IN THE LOW TRANSMISSION, PRE-ELIMINATION SETTING OF NAMIBIA IN VITRO ANTIMALARIAL ACTIVITY OF DIOSPYROS CHAMAETHAMNUS AND GUIBOURTIA COLEOSPERMA, FOUND MC/088 Du Preez, C Poster IN THE ZAMBEZI REGION OF NAMIBIA ATTACKING THE MOSQUITO ON MULTIPLE FRONTS: INSIGHTS ON OPTIMAL COMBINATIONS OF VECTOR MC/089 Kiware, S Oral CONTROL INTERVENTIONS FOR MALARIA ELIMINATION FROM A MATHEMATICAL MODEL STARVING MALARIA: TARGETING NUTRIENT UPTAKE IN THE SEXUAL, TRANSMISSIBLE STAGES OF PLASMODIUM MC/090 Niemand, J Oral FALCIPARUM PARASITES TOWARDS THE ELIMINATION OF MALARIA IN THE UNION OF COMOROS: GEOGRAPHICAL INSIGHT ON CONTROL MC/091 Attoumane, A Poster ACTIONS AND MAINTENANCE FACTORS MC/092 SYNTHESIS AND ANTIMALARIAL PROPERTIES OF SPIROINDOLONE ANALOGUES Naicker, A Poster EXPOSURE TO DDT/DDE IN RELATION TO HBA1C IN PREGNANT WOMEN OF TSHILIDZINI HOSPITAL, LIMPOPO, MC/093 Ndwamato, N Poster SOUTH AFRICA MC/094 SYNTHESIS AND MODIFICATION OF ANTIFOLATES Molatsane, T Poster MC/095 MOLECULAR DIAGNOSIS OF MALARIA IN THE ZAMBEZI REGION OF NAMIBIA MOVING TOWARDS ELIMINATION Tambo, M Poster DYNAMIC EVALUATION OF POTENTIAL TRANSMISSION BLOCKING ANTIPLASMODIALS: RATE AND STAGE- MC/096 Reader, J Oral SPECIFIC DESCRIPTIONS OF GAMETOCYTOCIDAL COMPOUNDS IMPORTED CASES, SPORADIC OUTBREAKSAND RESURGENCE ARE THE HIGHEST THREATS TO MALARIA MC/098 Mudambo, K Oral ELIMINATION IN THE E8 REGION MC/100 CASE MANAGEMENT OF MALARIA IN SWAZILAND, 2011 TO 2015: ON TRACK FOR ELIMINATION? Dlamini, S Poster MC/101 FOCUS ON MALARIA IN EDUCATION IN SOUTH AFRICA Petersen, J Oral INTERACTIVE MALARIA EDUCATION INTERVENTION AND ITS EFFECT ON COMMUNITY PARTICIPANT MC/102 Cox, S Poster KNOWLEDGE: THE MALARIA AWARENESS PROGRAM (MAP) IN VHEMBE DISTRICT, LIMPOPO, SOUTH AFRICA MC/103 MATHEMATICAL MODEL OF MALARIA TRANSMISSION IN CHILDREN UNDER FIVE YEARS IN AN ENDEMIC AREA Danquah, B Poster MC/105 SUSTAINED RELEASE OF REPELLENT MIXTURES THROUGH ENGINEERING THE MOLECULAR INTERACTIONS Izadi, H Oral MC/106 NOVEL DRUG SCAFFOLDS FOR THE TREATMENT OF MULTI-DRUG RESISTANT PLASMODIUM FALCIPARUM Panayides, J-L Oral EFFECT OF NOVEL 7-CHLOROQUINOLIN-4-YL PIPERAZINE-1-YL-ACETAMIDE DERIVATIVES IN THE MANAGEMENT MC/107 Kathrada, F Poster OF MALARIA MC/108 MODELLING THE SPATIO-TEMPORAL DYNAMICS OF MOSQUITOES AND ITS RESPONSE TO SIT CONTROL Dufourd, C Oral
Page 164 of 170 THE COPPER CHELATING, ANTIMALARIAL AND LARVICIDAL EFFECTS OF 8-HYDROXYQUINOLINE AND ITS MC/109 Jansen v Vuuren, N Poster DERIVATIVES MC/110 IMPACT OF INTENSE VECTOR SURVEILLANCE IN KWAZULU-NATAL Koekemoer, L Poster THE ISOMERIC EFFECT OF ESSENTIAL OIL CONSTITUENTS ON ANOPHELES ARABIENSIS LARVAE AND ARTEMIA MC/111 Mustapha, O Poster FRANCISCANA NAUPLII THE IMPACT OF MALARIA PROGRAM INTERVENTIONS AND CLIMATE CHANGE ON MORTALITY AND MORBIDITY MC/112 Motsuku, L Oral RATES BETWEEN 2009 AND 2016 IN LIMPOPO PROVINCE: PRELIMINARY FINDINGS MC/113 MALARIA DEATHS: MEETING THE 2015 NEAR ZERO TARGET IN SOUTH AFRICA Misiani, E Oral MC/114 UNDERSTANDING CROSS-BORDER MALARIA: LESSONS LEARNED FROM EASTERN ZIMBABWE Kanyangarara, M Poster MC/115 THE LARVICIDAL AND BRINE SHRIMP ACTIVITIES OF EUGENOL AND FIVE DERIVATIVES Shunmoogam-Gounden, N Oral COMMMUNITY ANTIMALARIAL USE PATTERNS AND RAPID TESTING FOR FAKE OR SUBSTANDARD DRUGS IN MC/116 Mharakurwa, S Poster MUTASA DISTRICT, ZIMBABWE RAPID RESTORATION OF CHLOROQUINE SENSITIVITY IN PLASMODIUM FALCIPARUM MALARIA OF MUTASA MC/117 Zingoni, M Poster DISTRICT, ZIMBABWE MC/118 PERFORMANCE OF TWO PFHRP2-BASED RDTS AS COMMUNITY MALARIA TESTS IN MUTASA DISTRICT ZIMBABWE Mudare, N Poster COULD A SONG ABOUT MALARIA PREVENTION HELP PROTECT CHILDREN UNDER 5 IN VENDA RURAL MC/119 Anderson, C Oral COMMUNITIES? PYRETHROID-IMPREGNATED WALL LININGS: THREE YEARS POST INSTALLATION DURABILITY, USER MC/120 Ramothole , M Oral ACCEPTABILITY, PERCEIVED EFFECTIVENESS, AND LABORATORY CONFIRMED EFFICACY MC/121 BIOLOGICAL CONSEQUENCES OF DDT EXPOSURE Delport, R Poster MC/122 BIOPROSPECTING TRADITIONALLY USED ANTIMALARIAL PLANTS BY MEANS OF NMR-BASED METABOLOMICS Bapela, J Oral
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CONFERENCE SPONSORS
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CONFERENCE RELATED FUNCTIONS
Page 168 of 170 2nd Malaria Conference Welcoming Cocktail
Venue: Normaalsaal, Groenkloof Campus Date: 31 July 2016
Programme
Programme director Prof Tiaan de Jager Function Host & Director: UP ISMC
17:35 - 17:45 Prof Norman Duncan Welcoming of delegates to UP UP Vice-Principal Academic
17:45 - 17:55 Prof Jeffrey Mphahlele The MRC & malaria Vice President SA MRC: Research (Pretoria)
17:55 - 18:00 Dr Taneshka Kruger Introduction to the live entertainment Senior Project Coordinator: UP ISMC
18:00 - 18:45 2nd and 3rd year UP Drama students A Malaria Awareness production
Cocktail Networking
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University of Pretoria • Universiteit van Pretoria • Yunibesithi ya Pretoria Private Bag/Privaatsak/Mokotla wa Poso X20 Hatfield 0028 South Africa • Suid-Afrika • Afrika-Borwa Tel: +27 (0)12 420 4111 • Fax/Faks/Fekse: +27 (0)12 420 4555 Venue: Protea Manor Hotel, Hatfield 2nd Malaria Conference ~ 1050 Burnett Street, Corner of Burnett and Gala Dinner Festival Streets, Hatfield ~ Date: 1 August 2016, 17:30 for 18:00
Programme
Programme director Prof Tiaan de Jager Function Host & Director: UP ISMC
18:05 - 18:15 Prof Rajendra Maharaj Welcoming of delegates Director: MRC Office of Malaria Researchice President SA
18:15 - 18:30 Dr Richard Kamwi Introducing Malaria Elimination 8 Ambassador for Malaria Elimination 8 (E8) Countries
Sit down dinner Sponsored by Elimination 8 and UP ISMC
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University of Pretoria • Universiteit van Pretoria • Yunibesithi ya Pretoria Private Bag/Privaatsak/Mokotla wa Poso X20 Hatfield 0028 South Africa • Suid-Afrika • Afrika-Borwa Tel: +27 (0)12 420 4111 • Fax/Faks/Fekse: +27 (0)12 420 4555