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149 Astmh.Org 149 499 endosymbiont Wolbachia. Depletion of Wolbachia with the antibiotic doxycycline arrests development, fertility and viability and delivers potent ONCHOCERCA LUPI: AN EMERGING ZOONOSIS IN macrofilaricidal efficacy in clinical trials. In order to identify alternative NORTHERN EUROPE AND THE UNITED STATES? anti-Wolbachia drugs with a more rapid activity we have exploited the host nematodes immune regulation of Wolbachia populations through 1 2 Matthew Belenchia , David Adams autophagy to discover drugs with a wolbachiacidal mode of action. We 1AASU, Savannah, GA, United States, 2AASU/Medical College of Georgia, have screened libraries of 100 autophagy inducing drugs and compounds Savannah, GA, United States against B. malayi. Selected ‘hits’ are then screened against transgenic C. The geographical distribution of cases caused by Onchocerca lupi, a elegans and human embryonic kidney (HEK) cells expressing a fluorescent common parasitic worm in wolves, has widened during the last decade. autophagy marker, ATG8 to identify drugs, which are selectively more Moving from Southern European countries such as Greece and Turkey, an potent against nematode versus human autophagy activation. Hits ranked increasing number of cases have begun to emerge in Northern Europe and by relative nematode potency are progressed through the A·WOL drug the Americas. Common arthropod vectors for O. lupi include blackflies discovery and development screening pipeline to identify pre-clinical lead (e.g., Simulium yahense and midges (e.g., Culicoides spp.). O. lupi candidates and optimized combinations of anti-Wolbachia drugs to reduce commonly causes ocular problems such as conjunctivitis, photophobia, and treatment timeframes. excessive lacrimation. In recent years, however, clinicians have identified cases that have involved extra-ocular sites such as the spinal canal. As of 502 2014, more than sixty cases of O. lupi, ocular and otherwise, have been identified throughout Europe and the United States. This presentation PHARMACOKINETIC/PHARMACODYNAMIC MODELLING will examine not only the global epidemiology of Onchocerca lupi but OF ANTI- WOLBACHIA CHEMOTHERAPEUTIC AGENTS IN A potential surveillance measures for this emerging zoonosis. LYMPHATIC FILARIASIS MURINE INFECTION MODEL Raman Sharma, Ghaith Aljayoussi, Ana de Castro Guimaraes, 500 Jill Davies, Lousie Ford, Alison Shone, Joseph Turner, David OPTIMIZATION OF THE ACANTHACHEILONEMA VITEAE LIFE Waterhouse, Stephen A. Ward, Mark Taylor CYCLE FOR INTENSIVE PRODUCTION Liverpool School of Tropical Medicine, Liverpool, United Kingdom An estimated 120 million people are infected by lymphatic filariasis Steven Schaar1, Leah Mann1, Zachary Williams1, Katy Anderson2, 1 1 throughout the tropics leading to a profound public health and socio- Jordan Trentadue , Shelly Michalski economic burden in severely affected communities. Wolbachia is an 1University of Wisconsin Oshkosh, Oshkosh, WI, United States, 2Purdue essential endosymbiont of the filarial nematodes Wuchereria bancrofti, University, West Lafayette, IN, United States Brugia malayi the causative agents of lymphatic filariasis. Doxycycline is The filarial nematode Acanthacheilonema viteae is of great interest to currently the gold standard for the targeting of Wolbachia in lymphatic the filariasis community because it lacks the endosymbiotic Wolbachia filariasis chemotherapy. However, the current drug regimen is a 100-200 bacterium found in several filarial parasites of humans. The NIH/NIAID mg/day doxycycline dose given for 4 to 6 weeks to patients. The A·WOL Filariasis Research Reagent Resource Center (FR3) began distributing A. consortium plan to reduce the current treatment time to 7 days or less viteae to its users in 2011. The FR3 strain of A. viteae and of its tick vector to improve drug regimen adherence and to reduce drug resistance and Ornithodoros tartakowskyi were originally acquired from TRS Laboratories costs of treatment. To achieve a rapid 7-day or less kill rate of Wolbachia, and the life cycle is currently being optimized for intensive production to a number of drug combinations will be employed. These include different meet increasing demand. Variables targeted in this optimization include 1) tetracyclines (Doxycycline and minocycline) rifamycins (Rifampicin or the microfilaremia of gerbils used for infecting ticks, 2) the life stage and Rifapentine), Moxifloxacin as well as anti-helminthic drugs. The complexity sex of ticks used for infections, 3) the number of subcutaneous injections of multiple drug combinations necessitates a rational approach in the used to infect hamsters with third-stage infective larvae (L3), and 4) the identification and choice of the best treatments in in-vivo models and medium used for subcutaneous infection of gerbils with L3. By feeding translating the animal treatments in the lab into clinical trials on the field. only adult stage ticks on gerbils selected for high microfilaremias we We have done series of PK-PD models and simulations using parameter increased our yields from 13 L3/tick (n=497) to 126 L3/tick (n=107 ticks). and non-parameteric population PK-PD modelling software programs to Similarly, the average recovery of adult A. viteae from hamsters has risen further dissect and quantify the dynamics of anti-bacterial activity of these from 51 ± 45 (n=13) to 90 ± 48 (n=22), partially due to the finding that drugs in the treatment of Lymphatic Filariasis and Onchocerciasis. As an worms isolated in Hanks’ Balanced Salt Solution (HBSS) produce patent example here, we identified the PK parameters of doxycycline, minocycline infections whereas those isolated in RPMI 1640 do not. Methods to infect and rifampicin in in-vivo PK studies in the SCID Brugia malayi model and ticks with A. viteae via artificial membrane feeding and inoculation are used the PK data along with pharmacodynamic output to interpret the currently being developed, and a recent trial resulted in recovery of 208 PK-PD relationships in light of the effect of each drug upon Wolbachia L3s/tick inoculated by enema with microfilaremic gerbil blood (n=4 ticks). viability in parasites. The data display the power of PK-PD modelling in The increased recovery of A. viteae from host animals has allowed the FR3 quantifying the PK-PD relationships of different drugs whilst giving insight to meet higher demand from the filariasis research community for all life to predictions of drug dynamics and interaction with Wolbachia viability. stages of the worm. 501 ACTIVATING AUTOPHAGY AS A NOVEL ANTI-WOLBACHIA MODE OF ACTION FOR MACROFILARICIDAL DRUG DISCOVERY Amber Fanthome, Denis Voronin, Mark J. Taylor Liverpool School of Tropical Medicine, Liverpool, United Kingdom River blindness (Onchocerca volvulus) and elephantiasis (Wuchereria bancrofti and Brugia malayi) affect over 150 million people in more than 80 countries, with a further 1 billion at risk of infection. Each of these nematode parasites has evolved a mutualistic association with the bacterial astmh.org 150 503 developing alternative plans. It can also be used to simulate the impact of applying combination therapies, drugs with different pharmacokinetics DEVELOPMENT OF A SMALL ANIMAL MODEL parameters and different potencies and modes of action. As an example, OF ONCHOCERCIASIS FOR THE SCREENING OF we show a case study where the model predicts the treatment outcomes MACROFILARICIDAL DRUGS in endemic areas when using ivermectin alone or a combination of ivermectin and doxycycline. The modelling shows that the use of a 1 2 Alice Halliday , Haelly Mejane Metuge , Winston Patrick combination of ivermectin and doxycycline treatment vastly increases the 2 2 Chounna Ndongmo , Jonas Arnauld Kengne-Ouafo , Tayong chances of elimination success. The use of a macrofilaricidal drug such as 2 1 1 2 Dizzle Bita Kwenti , Ana Guimaraes , Hayley Tyrer , Peter Enyong , doxycycline shows a much higher potential in achieving overall elimination 1 2 1 Mark Taylor , Samuel Wanji , Joseph Turner than microfilaricidal drugs (such as ivermectin) alone which agrees with 1Liverpool School of Tropical Medicine, Liverpool, United Kingdom, previous computer models 2Research Foundation for Tropical Diseases and the Environment, Buea, Cameroon 505 Onchocerciasis affects >37 million people with approximately 800,000 suffering visual impairment (river blindness). Development LASER DIFFRACTION ASSAY FOR LOW RESOURCE of macrofilaricides for onchocerciasis is hampered by the lack of a QUANTIFICATION OF MICROFILARIAL BURDEN facile animal model. Our laboratories and others have demonstrated Philip J. Budge1, David W. Wright2 protracted survival of non-murine filariae in immune-compromised mice. 1Washington University School of Medicine, St. Louis, MO, United States, Therefore we tested whether cattle-sourced Onchocerca could persist 2Vanderbilt University, Nashville, TN, United States in Severe-Combined Immuno Deficient (SCID) mice. O. ochengi nodules were harvested from the skins of infected cattle while infective L3 were Efforts to eliminate lymphatic filariasis and to control onchocerciasis by produced from blackflies fed on cattle in Cameroon. Motile male O. mass drug administration (MDA) in West and Central Africa have been ochengi were isolated following culture of disrupted onchocercomas, prior hampered by the risk of severe adverse reactions among persons harboring to surgical implantation into the peritoneal cavity of SCID mice. Viable high-level loiasis infections (>8,000 microfilariae per mL of blood). A macrofilariae
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