The mosquito gut represents an ecosystem that accommodates a complex, intimately associated microbiome. It is increasingly clear that the gut microbiome influences a wide variety of host traits, such as fitness and immunity. Understanding the microbial community structure and its dynamics across mosquito life is a prerequisite for comprehending the symbiotic relationship between the mosquito and its gut microbial residents. Here we characterized gut bacterial communities across larvae, pupae and adults of Anopheles gambiae reared in semi-natural habitats in Kenya by pyrosequencing bacterial 16S rRNA fragments. Immatures and adults showed distinctive gut community structures. Photosynthetic Cyanobacteria were predominant in the larval and pupal guts while Proteobacteria and Bacteroidetes dominated the adult guts, with core taxa of Enterobacteriaceae and Flavobacteriaceae. At the adult stage, diet regime (sugar meal and blood meal) significantly affects the microbial structure. Intriguingly, blood meals drastically reduced the community diversity and favored enteric bacteria. Comparative genomic analysis revealed that the enriched enteric bacteria possess large genetic redox capacity of coping with oxidative and nitrosative stresses that are associated with the catabolism of blood meal, suggesting a beneficial role in maintaining gut redox homeostasis. Interestingly, gut community structure was similar in the adult stage between the field and laboratory mosquitoes, indicating that mosquito gut is a selective eco-environment for its microbiome. This comprehensive gut metatgenomic profile suggests a concerted symbiotic genetic association between gut inhabitants and host.

BackgroundMalaria vector intervention and control programs require reliable and accurate information about vector abundance and their seasonal distribution. The availability of reliable information on the spatial and temporal productivity of larval vector habitats can improve targeting of larval control interventions and our understanding of local malaria transmission and epidemics. The main objective of this study was to evaluate two methods of estimating larval habitat productivity in the western Kenyan highlands, the aerial sampler and the emergence trap.MethodsThe study was conducted during the dry and rainy seasons in 2008, 2009 and 2010. Aerial samplers and emergence traps were set up for sixty days in each season in three habitat types: drainage ditches, natural swamps, and abandoned goldmines. Aerial samplers and emergence traps were set up in eleven places in each habitat type. The success of each in estimating habitat productivity was assessed according to method, habitat type, and season. The effect of other factors including algae cover, grass cover, habitat depth and width, and habitat water volume on species productivity was analysed using stepwise logistic regressionResultsHabitat productivity estimates obtained by the two sampling methods differed significantly for all species except for An. implexus. For for An. gambiae s.l. and An. funestus, aerial samplers performed better, 21.5 and 14.6 folds, than emergence trap respectively, while the emergence trap was shown to be more efficient for culicine species. Seasonality had a significant influence on the productivity of all species monitored. Dry season was most productive season. Overall, drainage ditches had significantly higher productivity in all seasons compared to other habitat types. Algae cover, debris, chlorophyll-a, and habitat depth and size had significant influence with respect to species.ConclusionThese findings suggest that the aerial sampler is the better of the two methods for estimating the productivity of An. gambiae s.l. and An. funestus in the western Kenya highlands and possibly other malaria endemic parts of Africa. This method has proven to be a useful tool for monitoring malaria vector populations and for control program design, and provides useful means for determining the most suitable sites for targeted interventions.

BackgroundOutdoor malaria transmission is becoming an increasingly important problem in malaria control in Africa. Larval control is a promising intervention as it can target both indoor and outdoor biting mosquitoes. However, the currently available biolarvicide formulations have a short effective duration, and consequently larval control incurs a high operational expense due to the requirement for frequent re-treatment of larval habitats. Formulations of biolarvicides with long-lasting effects is highly desired. A recently developed FourStar® slow-release briquet formulation of Bacillus thuringiensis israelensis and Bacillus sphaericus was evaluated to test its efficacy on malaria vectors.MethodsThe study evaluated FourStar™ briquets 180-days formulation under semi-natural and natural conditions to test their efficacy in reducing the mosquito population in western Kenya. The semi-natural habitats used the formulation dissolved in rainwater with appropriate concentrations, and second-instar larvae of Anopheles gambiae were introduced and the number of surviving larvae and pupae produced was recorded daily as the outcome. The briquets formulation was then tested in natural habitats for efficacy on pupal productivity reduction in highland and lowland sites in western Kenya. The formulation was finally tested for efficacy in reducing adult mosquito populations in randomized clusters in western Kenya highland.ResultsIn semi-natural conditions, the FourStar™ briquets 180-days formulation completely inhibited mosquito pupal production in the first 3 months, and then reduced pupal productivity by 87–98% (P < 0.001) 4–6 months after application. In natural habitats, during the first 2 months no pupae were detected from any of the treated habitats in highland sites, and Anopheles spp. pupal density was reduced by 60–90% in the next 3–5 months (P < 0.001). In the lowland site, pupal productivity reduction was 100% in the first 3 months, and 75–90% in the next 4–5 months (P < 0.001). The randomized cluster trial found that the application of the briquets formulation reduced mean densities of indoor-biting mosquitoes by 76–82% (P < 0.001) and by 67–75% (P < 0.001) for outdoor-biting mosquitoes.ConclusionThis study demonstrated that long-lasting biological larviciding was effective in reducing pupal productivity of larval habitats, and reducing indoor and outdoor resting mosquitoes. The study suggests that long-lasting microbial larviciding may be a promising complementary malaria vector control tool and warrants further large-scale evaluation.

BackgroundThe current status of insecticide resistance in mosquitoes and the effects of insecticides on non-target insect species have raised the need for alternative control methods for malaria vectors. Predation has been suggested as one of the important regulation mechanisms for malaria vectors in long-lasting aquatic habitats, but the predation efficiency of the potential predators is largely unknown in the highlands of western Kenya. In the current study, we examined the predation efficiency of five predators on Anopheles gambiae s.s larvae in 24 hour and semi- field evaluations.MethodsPredators were collected from natural habitats and starved for 12 hours prior to starting experiments. Preliminary experiments were conducted to ascertain the larval stage most predated by each predator species. When each larval instar was subjected to predation, third instar larvae were predated at the highest rate. Third instar larvae of An. gambiae were introduced into artificial habitats with and without refugia at various larval densities. The numbers of surviving larvae were counted after 24 hours in 24. In semi-field experiments, the larvae were counted daily until they were all either consumed or had developed to the pupal stage. Polymerase chain reaction was used to confirm the presence of An. gambiae DNA in predator guts.ResultsExperiments found that habitat type (P < 0.0001) and predator species (P < 0.0001) had a significant impact on the predation rate in the 24 hour evaluations. In semi-field experiments, predator species (P < 0.0001) and habitat type (P < 0.0001) were significant factors in both the daily survival and the overall developmental time of larvae. Pupation rates took significantly longer in habitats with refugia. An. gambiae DNA was found in at least three out of ten midguts for all predator species. Gambusia affins was the most efficient, being three times more efficient than tadpoles.ConclusionThese experiments provide insight into the efficiency of specific natural predators against mosquito larvae. These naturally occurring predators may be useful in biocontrol strategies for aquatic stage An. gambiae mosquitoes. Further investigations should be done in complex natural habitats for these predators.

BackgroundThe effective measures for the control of malaria and filariasis vectors can be achieved by targeting immature stages of anopheline and culicine mosquitoes in productive habitat. To design this strategy, the mechanisms (like biotic interactions with conspecifc and heterospecific larvae) regulating mosquito aquatic stages survivorship, development time and the size of emerging adults should be understood. This study explored the effect of co-habitation between An. gambiae s.s. and Cx. quinquefasciatus on different life history traits of both species under different densities and constant food supply in the habitats of the same size under semi-natural conditions.MethodsExperiments were set up with three combinations; Cx. quinquefasciatus alone (single species treatment), An. gambiae s.s. alone (single species treatment); and An. gambiae s.s. with Cx. quiquefasciatus (co-habitation treatment) in different densities in semi field situation.ResultsThe effect of co-habitation of An. gambiae s.s. and Cx. quinquefasciatus was found to principally affect three parameters. The wing-lengths (a proxy measure of body size) of An. gambiae s.s. in co-habitation treatments were significantly shorter in both females and males than in An. gambiae s.s single species treatments. In Cx. quinquefasciatus, no significant differences in wing-length were observed between the single species and co-habitation treatments. Daily survival rates were not significantly different between co-habitation and single species treatments for both An. gambiae s.s. and Cx. quinquefasciatus. Developmental time was found to be significantly different with single species treatments developing better than co-habitation treatments. Sex ratio was found to be significantly different from the proportion of 0.5 among single and co-habitation treatments species at different densities. Single species treatments had more males than females emerging while in co-habitation treatments more females emerged than males. In this study, there was no significant competitive survival advantage in co-habitation.ConclusionThese results suggest that co-habitation of An. gambiae s.s. and Cx. quinquefasciatus in semi-natural conditions affect mostly An. gambiae s.s. body size. Hence, more has to be understood on the effects of co-habitation of An. gambiae s.s. and Cx. quinquefasciatus in a natural ecology and its possible consequences in malaria and filariasis epidemiology.