BackgroundThe use of gravid mosquitoes as vehicles to auto-disseminate larvicides was recently demonstrated for the transfer of pyriproxyfen (PPF) by container-breeding Aedes mosquitoes and presents an appealing idea to explore for other disease vectors. The success of this approach depends on the female’s behaviour, the time of exposure and the amount of PPF that can be carried by an individual. We explore the effect of PPF exposure at seven time points around blood feeding on individual Anopheles gambiae sensu stricto and Culex quinquefasciatus fecundity and ability to transfer in laboratory assays.MethodMosquitoes were exposed to 2.6 mg PPF per m2 at 48, 24 and 0.5 hours before and after a blood meal and on the day of egg-laying. The proportion of exposed females (N = 80-100) laying eggs, the number of eggs laid and hatched was studied. Transfer of PPF to oviposition cups was assessed by introducing 10 late instar insectary-reared An. gambiae s.s. larvae into all the cups and monitored for adult emergence inhibition.ResultsExposure to PPF between 24 hours before and after a blood meal had significant sterilizing effects: females of both species were 6 times less likely (Odds ratio (OR) 0.16, 95% confidence interval (CI) 0.10-0.26) to lay eggs than unexposed females. Of the few eggs laid, the odds of an egg hatching was reduced 17 times (OR 0.06, 95% CI 0.04-0.08) in Anopheles but only 1.2 times (OR 0.82, 95% CI 0.73-0.93) in Culex. Adult emergence inhibition from larvae introduced in the oviposition cups was observed only from cups in which eggs were laid. When females were exposed to PPF close to egg laying they transferred enough PPF to reduce emergence by 65-71% (95% CI 62-74%).ConclusionPPF exposure within a day before and after blood feeding affects egg-development in An. gambiae s.s. and Cx. quinquefasciatus and presents a promising opportunity for integrated control of vectors and nuisance mosquitoes. However, sterilized females are unlikely to visit an oviposition site and therefore do not transfer lethal concentrations of PPF to aquatic habitats. This suggests that for successful auto-dissemination the optimum contamination time is close to oviposition.

BackgroundRecently research has shown that larviciding can be an effective tool for integrated malaria vector control. Nevertheless, the uptake of this intervention has been hampered by the need to re-apply larvicides frequently. There is a need to explore persistent, environmentally friendly larvicides for malaria vector control to reduce intervention efforts and costs by reducing the frequency of application. In this study, the efficacy of a 0.5% pyriproxyfen granule (Surmilarv®0.5G, Sumitomo Chemicals) was assessed for the control of Anopheles gambiae sensu stricto and Anopheles arabiensis, the major malaria vectors in sub-Saharan Africa.MethodsDose–response and standardized field tests were implemented following standard procedures of the World Health Organization’s Pesticide Evaluation Scheme to determine: (i) the susceptibility of vectors to this formulation; (ii) the residual activity and appropriate retreatment schedule for field application; and, (iii) sub-lethal impacts on the number and viability of eggs laid by adults after exposure to Sumilarv®0.5G during larval development.ResultsAnopheles gambiae s.s. and An. arabiensis were highly susceptible to Sumilarv®0.5G. Estimated emergence inhibition (EI) values were very low and similar for both species. The minimum dosage that completely inhibited adult emergence was between 0.01-0.03 parts per million (ppm) active ingredient (ai). Compared to the untreated control, an application of 0.018 ppm ai prevented 85% (95% confidence interval (CI) 82%-88%) of adult emergence over six weeks under standardized field conditions. A fivefold increase in dosage of 0.09 ppm ai prevented 97% (95% CI 94%-98%) emergence. Significant sub-lethal effects were observed in the standardized field tests. Female An. gambiae s.s. that were exposed to 0.018 ppm ai as larvae laid 47% less eggs, and females exposed to 0.09 ppm ai laid 74% less eggs than females that were unexposed to the treatment. Furthermore, 77% of eggs laid by females exposed to 0.018 ppm ai failed to hatch, whilst 98% of eggs laid by females exposed to 0.09 ppm ai did not hatch.ConclusionAnopheles gambiae s.s. and An. arabiensis are highly susceptible to Sumilarv®0.5G at very low dosages. The persistence of this granule formulation in treated habitats under standardized field conditions and its sub-lethal impact, reducing the number of viable eggs from adults emerging from treated ponds, enhances its potential as malaria vector control tool. These unique properties warrant further field testing to determine its suitability for inclusion in malaria vector control programmes.

BackgroundPersistent monomolecular surface films could benefit larval source management for malaria control by reducing programme costs and managing insecticide resistance. This study evaluated the efficacy of the silicone-based surface film, Aquatain® Mosquito Formulation (AMF), for the control of the Afrotropical malaria vectors, Anopheles gambiae sensu stricto and Anopheles arabiensis in laboratory dose–response assays and standardized field tests.MethodsTests were carried out following guidelines made by the World Health Organization Pesticide Evaluation Scheme (WHOPES). Sub-lethal effects of AMF were evaluated by measuring egg-laying and hatching of eggs laid by female An. gambiae s.s. that emerged from habitats treated with a dose that resulted in 50% larval mortality in laboratory tests.ResultsBoth vector species were highly susceptible to AMF. The estimated lethal doses to cause complete larval mortality in dose–response tests in the laboratory were 1.23 (95% confidence interval (CI) 0.99-1.59) ml/m2 for An. gambiae s.s. and 1.35 (95% CI 1.09-1.75) ml/m2 for An. arabiensis. Standardized field tests showed that a single dose of AMF at 1 ml/m2 inhibited emergence by 85% (95% CI 82-88%) for six weeks. Females exposed as larvae to a sub-lethal dose of AMF were 2.2 times less likely (Odds ratio (OR) 0.45, 95% CI 0.26-0.78) to lay eggs compared to those from untreated ponds. However, exposure to sub-lethal doses neither affected the number of eggs laid by females nor the proportion hatching.ConclusionAMF provided high levels of larval control for a minimum of six weeks, with sub-lethal doses reducing the ability of female mosquitoes to lay eggs. The application of AMF provides a promising novel strategy for larval control interventions against malaria vectors in Africa. Further field studies in different eco-epidemiological settings are justified to determine the persistence of AMF film for mosquito vector control and its potential for inclusion in integrated vector management programmes.

Background Larviciding is an effective supplementary tool for malaria vector control, but the identification and accessibility of aquatic habitats impedes application. Dissemination of the insect growth regulator, pyriproxyfen (PPF), by gravid Anopheles might constitute a novel application strategy. This study aimed to explore the feasibility of using an attractive bait-station to contaminate gravid Anopheles gambiae sensu stricto with PPF and subsequently transfer PPF to larval habitats. Methods A bait-station was developed comprising of an artificial pond containing water treated with 20 ppm cedrol, an oviposition attractant, and a netting-cover treated with PPF. Three identical semi-field cages were used to assess the potential of gravid Anopheles to transfer PPF from the bait-station to ponds. Gravid females were released in two semi-field cages, one with PPF on its bait-station (test) and one without PPF (control). No mosquitoes were released in the third cage with a PPF-treated station (control). Transfer of PPF to open ponds was assessed by monitoring emergence of late instar insectary-reared larvae introduced into the ponds. The amount of PPF carried by a mosquito and transferred to water was quantified using liquid chromatography-mass spectrometry. Results In the controls, 86% (95% CI 81–89%) of larvae introduced into open ponds developed into adults, indicating that wind did not distribute PPF in absence of mosquitoes. Emergence inhibition was observed in the test cage but was dependent on the distance between pond and bait-station. Only 25% (95% CI 22–29%) of larvae emerged as adults from ponds 4 m from the bait-station, but 92% (95% CI 89–94%) emerged from ponds 10 m away. Each mosquito was contaminated on average with 112 μg (95% CI 93–123 μg) PPF resulting in the transfer of 230 ng/L (95% CI 180–290 ng/L) PPF to 100 ml volumes of water. Conclusions The bait-stations successfully attracted gravid females which were subsequently dusted with effective levels of PPF. However, in this study design, attraction and dissemination was limited to short distances. To make this approach feasible for malaria vector control, stronger attractants that lure gravid females from longer distances, in landscapes with many water bodies, and better PPF delivery systems are needed.

Background Larval source management is an effective supplementary tool for malaria vector control although it is not used widely in sub-Saharan Africa. This study explored whether an attract-and-kill strategy could contaminate gravid Anopheles gambiae sensu stricto with the insect growth regulator, pyriproxyfen, at a bait-station, for dissemination to larval habitats. Methods A bait-station comprising an artificial pond, containing water was treated with 20 ppm cedrol, an oviposition attractant, was covered with pyriproxfen-treated netting. Three identical semi-field cages were used to assess the potential of gravid Anopheles gambiae sensu stricto to transfer pyriproxyfen from the bait-station to three open ponds. Gravid females were released in the test and one of the control cages that had no pyriproxyfen on its bait-station. No mosquitoes were released in the third cage with a pyriproxyfen-treated station. Transfer of pyriproxyfen to open ponds was assessed by monitoring emergence of late instar insectary-reared An. gambiae sensu stricto larvae introduced into the open ponds. Liquid chromatography-mass spectrometry was used to quantify the amount of pyriproxyfen carried by a mosquito and the amount transferred to water. Results 86% (95% CI 81-89%) of larvae introduced into the open ponds in the two control cages developed into adults. Transfer of pyriproxyfen to the test cage depended on the distance of the pond from the bait-station. While only 25% (95% CI 22-29%) adult emergence was observed in larvae introduced into ponds 4.4 m from the bait-station, the emergence rates increased to 92% (95% CI 89-94%) in larvae introduced in ponds 10.3 m away. Each mosquito was contaminated with 112 µg (95% CI 93-123 µg) pyriproxyfen, whilst 230 ng/L (95% CI 180-290 ng/L) was transferred by a single female to 100 ml of water. Conclusions Pyriproxyfen was auto-disseminated by gravid females from attractive bait-stations, but mainly to aquatic habitats near the bait station. To make this approach feasible for malaria vector control, stronger attractants and better pyriproxyfen delivery systems are needed.

Background Larval source management is an effective supplementary tool for malaria vector control although it is not used widely in sub-Saharan Africa. This study explored whether an attract-and-kill strategy could contaminate gravid Anopheles gambiae sensu stricto with the insect growth regulator, pyriproxyfen, at a bait-station, for dissemination to larval habitats. Methods A bait-station comprising an artificial pond, containing water was treated with 20 ppm cedrol, an oviposition attractant, was covered with pyriproxfen-treated netting. Three identical semi-field cages were used to assess the potential of gravid Anopheles gambiae sensu stricto to transfer pyriproxyfen from the bait-station to three open ponds. Gravid females were released in the test and one of the control cages that had no pyriproxyfen on its bait-station. No mosquitoes were released in the third cage with a pyriproxyfen-treated station. Transfer of pyriproxyfen to open ponds was assessed by monitoring emergence of late instar insectary-reared An. gambiae sensu stricto larvae introduced into the open ponds. Liquid chromatography-mass spectrometry was used to quantify the amount of pyriproxyfen carried by a mosquito and the amount transferred to water. Results 86% (95% CI 81-89%) of larvae introduced into the open ponds in the two control cages developed into adults. Transfer of pyriproxyfen to the test cage depended on the distance of the pond from the bait-station. While only 25% (95% CI 22-29%) adult emergence was observed in larvae introduced into ponds 4.4 m from the bait-station, the emergence rates increased to 92% (95% CI 89-94%) in larvae introduced in ponds 10.3 m away. Each mosquito was contaminated with 112 µg (95% CI 93-123 µg) pyriproxyfen, whilst 230 ng/L (95% CI 180-290 ng/L) was transferred by a single female to 100 ml of water. Conclusions Pyriproxyfen was auto-disseminated by gravid females from attractive bait-stations, but mainly to aquatic habitats near the bait station. To make this approach feasible for malaria vector control, stronger attractants and better pyriproxyfen delivery systems are needed.

Background Larval source management is an effective supplementary tool for malaria vector control although it is not used widely in sub-Saharan Africa. This study explored whether an attract-and-kill strategy could contaminate gravid Anopheles gambiae sensu stricto with the insect growth regulator, pyriproxyfen, at a bait-station, for dissemination to larval habitats. Methods A bait-station comprising an artificial pond, containing water was treated with 20 ppm cedrol, an oviposition attractant, was covered with pyriproxfen-treated netting. Three identical semi-field cages were used to assess the potential of gravid Anopheles gambiae sensu stricto to transfer pyriproxyfen from the bait-station to three open ponds. Gravid females were released in the test and one of the control cages that had no pyriproxyfen on its bait-station. No mosquitoes were released in the third cage with a pyriproxyfen-treated station. Transfer of pyriproxyfen to open ponds was assessed by monitoring emergence of late instar insectary-reared An. gambiae sensu stricto larvae introduced into the open ponds. Liquid chromatography-mass spectrometry was used to quantify the amount of pyriproxyfen carried by a mosquito and the amount transferred to water. Results 86% (95% CI 81-89%) of larvae introduced into the open ponds in the two control cages developed into adults. Transfer of pyriproxyfen to the test cage depended on the distance of the pond from the bait-station. While only 25% (95% CI 22-29%) adult emergence was observed in larvae introduced into ponds 4.4 m from the bait-station, the emergence rates increased to 92% (95% CI 89-94%) in larvae introduced in ponds 10.3 m away. Each mosquito was contaminated with 112 µg (95% CI 93-123 µg) pyriproxyfen, whilst 230 ng/L (95% CI 180-290 ng/L) was transferred by a single female to 100 ml of water. Conclusions Pyriproxyfen was auto-disseminated by gravid females from attractive bait-stations, but mainly to aquatic habitats near the bait station. To make this approach feasible for malaria vector control, stronger attractants and better pyriproxyfen delivery systems are needed.