Interactions between a fungal entomopathogen and malaria parasites within a mosquito vector.

BACKGROUND: Mosquitoes are becoming increasingly resistant to the chemical insecticides currently available for malaria vector control, spurring interest in alternative management tools. One promising technology is the use of fungal entomopathogens. Fungi have been shown to impact the potential for mosquitoes to transmit malaria by reducing mosquito longevity and altering behaviour associated with flight and host location. Additionally, fungi could impact the development of malaria parasites within the mosquito via competition for resources or effects on the mosquito immune system. This study evaluated whether co-infection or superinfection with the fungal entomopathogen Beauveria bassiana affected malaria infection progress in Anopheles stephensi mosquitoes. METHODS: The study used two parasite species to examine possible effects of fungal infection at different parasite development stages. First, the rodent malaria model Plasmodium yoelii was used to explore interactions at the oocyst stage. Plasmodium yoelii produces high oocyst densities in infected mosquitoes and thus was expected to maximize host immunological and resource demands. Second, fungal interactions with mature sporozoites were evaluated by infecting mosquitoes with the human malaria species Plasmodium falciparum, which is highly efficient at invading mosquito salivary glands. RESULTS: With P. yoelii, there was no evidence that fungal co-infection (on the same day as the blood meal) or superinfection (during a subsequent gonotrophic cycle after parasite infection) affected the proportion of mosquitoes with oocysts, the number of oocysts per infected mosquito or the number of sporozoites per oocyst. Similarly, for P. falciparum, there was no evidence that fungal infection affected sporozoite prevalence. Furthermore, there was no impact of infection with either malaria species on fungal virulence as measured by mosquito survival time. CONCLUSIONS: These results suggest that the impact of fungus on malaria control potential is limited to the well-established effects on mosquito survival and transmission behaviour. Direct or indirect interactions between fungus and malaria parasites within mosquitoes appear to have little additional influence.

Temperature alters Plasmodium blocking by Wolbachia.

Very recently, the Asian malaria vector (Anopheles stephensi) was stably transinfected with the wAlbB strain of Wolbachia, inducing refractoriness to the human malaria parasite Plasmodium falciparum. However, conditions in the field can differ substantially from those in the laboratory. We use the rodent malaria P. yoelii, and somatically transinfected An. stephensi as a model system to investigate whether the transmission blocking potential of wAlbB is likely to be robust across different thermal environments. wAlbB reduced malaria parasite prevalence and oocyst intensity at 28°C. At 24°C there was no effect on prevalence but a marked increase in oocyst intensity. At 20°C, wAlbB had no effect on prevalence or intensity. Additionally, we identified a novel effect of wAlbB that resulted in reduced sporozoite development across temperatures, counterbalancing the oocyst enhancement at 24°C. Our results demonstrate complex effects of temperature on the Wolbachia-malaria interaction, and suggest the impacts of transinfection might vary across diverse environments.

Mosquito ingestion of antibodies against mosquito midgut microbiota improves conversion of ookinetes to oocysts for Plasmodium falciparum, but not P. yoelii.

The mosquito midgut is a site of complex interactions between the mosquito, the malaria parasite and the resident bacterial flora. In laboratory experiments, we observed significant enhancement of Plasmodium falciparum oocyst production when Anopheles gambiae (Diptera: Culicidae) mosquitoes were membrane-fed on infected blood containing gametocytes from in vitro cultures mixed with sera from rabbits immunized with A. gambiae midguts. To identify specific mechanisms, we evaluated whether the immune sera was interfering with the usual limiting activity of gram-negative bacteria in An. gambiae midguts. Enhancement of P. falciparum infection rates occurred at some stage between the ookinete and oocyst stage and was associated with greater numbers of oocysts in mosquitoes fed on immune sera. The same immune sera did not affect the sporogonic development of P. yoelii, a rodent malaria parasite. Not only did antibodies in the immune sera recognize several types of midgut-derived gram-negative bacteria (Pseudomonas spp. and Cedecea spp.), but gentamicin provided in the sugar meal 3 days before an infectious P. falciparum blood meal mixed with immune sera eliminated the enhancing effect. These results suggest that gram-negative bacteria, which normally impair P. falciparum development between the ookinete and oocyst stage, were altered by specific anti-bacterial antibodies produced by immunizing rabbits with non-antibiotic-treated midgut lysates. Because of the differences in developmental kinetics between human and rodent malaria species, the anti-bacterial antibodies had no effect on P. yoelii because their ookinetes leave the midgut much earlier than P. falciparum and so are not influenced as strongly by resident midgut bacteria. While this study highlights the complex interactions occurring between the parasite, mosquito, and midgut microbiota, the ultimate goal is to determine the influence of midgut microbiota on Plasmodium development in anopheline midguts in malaria endemic settings.