Table 2_Laboratory An. gambiae s.l. mosquito colonies show sustained high transmission of Microsporidia sp. MB and a small decrease in egg viability.docx

IntroductionMicrosporidia sp. MB, a microsporidian symbiont found naturally in Anopheles mosquitoes, has potential as a novel malaria control tool since it can inhibit Plasmodium development and transmission. The most feasible MB-based Plasmodium control strategy would involve dissemination through live mosquito releases, or release of spores infective to mosquito larvae. To implement either strategy, establishment of stable mosquito colonies carrying MB at a high frequency is likely to be essential.
MethodsThe progeny of field caught An. gambiae s.l from Burkina Faso were isolated for individual egg laying and tested for MB. The progeny of the MB positive females were pooled and this process was repeated for multiple generations. The relative density of MB in different life stages and tissues of the An. coluzzii host was examined using a novel duplex qPCR assay. We also examined the impact of MB on egg production through individualization for egg laying and counting of eggs. Finally, we examined laid eggs for presence of MB spores.
ResultsThree An. coluzzii colonies and one An. gambiae s.l hybrid colony were established with high prevalence and density of MB and were maintained for more than two years with minimal intervention. MB prevalence and density was highest in eggs and adult females and lowest in L4 larvae; in adults density was highest in the gonads. Additionally, MB density increased in ovary samples following blood feeding which was likely due to the activation of sporogony.
DiscussionThe production of spores is the reason why MB-carrying females lay more white non-hatching eggs and show a small reduction in the median number of eggs laid. Establishment of several stable MB carrying An. gambiae s.l colonies and understanding the impact of spores on egg production are significant steps forward in developing MB as a malaria control tool.