Changing the way we control malaria in Africa Evaluation of long-lasting odor release formulations to increase the attractiveness of the passive Mosquito Contamination Device (MCD) for Anopheles mosquitoes Jennifer McCaw, Michael Cordel, Stella Kessy, Dr. Issa Lyimo, Dr. Ladlaus Mnyone, Dr. Andreas Rose

THE CHALLENGE: Mosquito Ecology is Changing ESOVE 17 October 2014 www.mcdproject.org THE CHALLENGE: Mosquito Ecology is Changing Urgent need for novel vector control tools that target outdoor biting mosquitoes Conventional control methods rely heavily on pyrethroids Pyrethroid Resistance Behavioral Adaptations Found in 64 countries Particular concern in sub-Saharan Africa Increase in outdoor biting Biting earlier in night Shift in species composition Residual transmition will be maintained by outdoor biting mosquitoes at dusk and dawn

THE MCD PROJECT Basic Info Goal Objectives ESOVE 17 October 2014 THE MCD PROJECT Basic Info Funded by EU Framework 7 Health Innovation Programme Length: 3 years, Started in 2012 5 consortium partners Goal Development of a mosquito contamination device (MCD) Effectively lure, infect and contaminate malaria mosquitoes in resource-poor settings Objectives Outdoor and Indoor mosquitoes in different life stages Use novel bioactives (PPF, Fungi) Develop long-lasting odor release formulations Low-tech devices using shared „software“ development of a mosquito contamination device (MCD) that can effectively lure, infect and contaminate malaria mosquitoes in tropical field settings and offer a novel way to combat mosquitoes in resource-poor settings www.mcdproject.org

All Prototypes utilize same „software“ ESOVE 17 October 2014 THE MCD PROTOTYPES Indoor Resting Device Outdoor Host-seeking Device Smart Patch 2 1 3 4 5 6 Oviposistioning Device Eave Screens Eave Tubes All Prototypes utilize same „software“ Objective: Evaluation of long-lasting odor release formulations to increase the attractiveness of the MCD Prototypes www.mcdproject.org

ESOVE 17 October 2014 RESULTS: Longevity of Natural Odors in Nylon Confirmed up to 34 Days Post Treatment BugDorm Bioassay Treatments: Negative Control: fresh nylon Positive Control: worn nylon with natural host odors Worn Sock: nylon sock worn 24hrs and exposed to env. conditions over time An. gambiae s.s. N=6, Mean values with standard deviation, p<0.05 n.s. * ** n.s . *** www.mcdproject.org

ESOVE 17 October 2014 RESULTS: Nylon Impregnated with Synthetic Blends Had a Repellent Effect in BugDorm Bioassay An. arabiensis Individual Compounds and Their Concentrations in the Tested Synthetic Blends Name Basic Blend* Mbita Blend** Ifakara Blend 1*** Ammonia aqueous 2.5% L-(+)-Lactic acid 85% Tetradecanoic acid 0.01% 0.00025% Propionic acid 0.1% Butanoic acid 1.0% Pentanoic acid 3-methyl butanoic acid 0.001% Heptanoic acid Octanoic acid 3-methyl-1-butanol 0.000001% Additional compound a b bc c c c N=10, p<0.05 * Smallegange, R. et al. (2009): The effect of Aliphatic Carboxylic Acids on Olfaction-Based Host-Seeking of the Malaria Mosquito Anopheles gambiae. J Chem Ecology. ** Mukabana, W. et al. (2012): A Novel Synthetic Odorant Blend for Trapping of Malaria and Other African Mosquito Species. J Chem Ecology. *** Okumu, F. et al. (2010): Develpment and Field Evaluation of a Synthetic Mosquito Lure That is More Attractive then Humans. PLoS ONE. www.mcdproject.org

ESOVE 17 October 2014 RESULTS: Nylon Impregnated with Dilutions of Mbita Blend and IB1 Blend Did Not Elicit Landing Responses Mbita Blend IB1 Blend An. arabiensis a An. arabiensis b a b b b bc b b b bc c bc b N=10, p<0.05 www.mcdproject.org

ESOVE 17 October 2014 RESULTS: Nylon Impregnated with Agar and Contaminated with Human Foot Bacteria Elicited Landing Responses Treatments: Negative Control: fresh nylon Positive Control: worn nylon with natural host odors Agar: nylon sock impregnated with agar solution Agar + Foot: nylon sock impregnated with agar solution and contaminated with human foot bacteria An. arabiensis a a a b N=10, p<0.05 www.mcdproject.org

ESOVE 17 October 2014 RESULTS: Polyethylene Dispensers filled with Agar Solutions Elicited Landing Responses Treatments: Negative Control: fresh nylon Positive Control: worn nylon with natural host odors Solvent: polyethylene dispenser filled with deionized water 1% Agar: polyethylene dispenser filled with 1% agar solution 3% Agar: polyethylene dispenser filled with 3% agar solution An. arabiensis a a a b c N=10, p<0.05 www.mcdproject.org

Room Tests with Outdoor Landing Device Prototype ESOVE 17 October 2014 Room Tests with Outdoor Landing Device Prototype Low-tech Cheap Outdoors Utilize existing cues Attractive landing surface Host-seeking mosquitoes Devices that can be placed outdoors, near or surrounding houses and cattle sheds, and target host-seeking mosquitoes at the moment they are in search of a bloodmeal. We envisage small, lowtech, portable and/or hanging devices that can be placed near house openings such as eaves or windows and thereby benefit from the attractive odours and in particular long-range cues emitted from the inhabitants. Outdoor devices will minimise human exposure and can thus utilise a broader range of (nastier) chemicals and/or bioactives. Assembly of the Outdoor Landing Device: 1.5l water bottle filled with water heated to 44°C and then placed in the fitted, black cotton bag and covered with a blue Safi Net bed net that was treated with 80% Bendiocarb. Afterwards it was hung at 2.1 meters at the center of the room. Explain Room Tests Physical Cues Heat Visual Cues Black color Olfactory Cues Synthetic Blends Killing Agent Bendiocarb www.mcdproject.org

nylon strips impregnated with 5 components ESOVE 17 October 2014 ROOM TEST RESULTS: Nylon Strips Impregnated with Mbita Blend Increased Attractiveness of Outdoor Landing Device Attractant: An. arabiensis Mbita Blend nylon strips impregnated with 5 components a a b c BC BC BC Worn Sock Mbita Blend N=5, Mean values with SD, p<0.05 www.mcdproject.org

polyethylene dispensers filled with 3% agar solution ESOVE 17 October 2014 ROOM TEST RESULTS: Agar Filled Polyethylene Dispensers Increased Attractiveness of Outdoor Landing Device Attractant: An. arabiensis a Agar Solution a polyethylene dispensers filled with 3% agar solution b c BC BC BC Worn Sock Agar Disp. N=7, Mean values with SD, p<0.05 www.mcdproject.org

ESOVE 17 October 2014 Summary www.mcdproject.org

Thank you for your attention! Contact: Dr. Bart GJ Knols In2Care BV info@in2care.org Dr. Andreas Rose Biogents AG info@biogents.com The research leading to these results has received funding from the European Community’s Seventh Framework Programme under grant agreement No. 306105 (Project MCD)