Culturable bacteria associated with Anopheles darlingi and their paratransgenesis potential.

BACKGROUND: Malaria remains a major public health problem in South America, mostly in the Amazon region. Among newly proposed ways of controlling malaria transmission to humans, paratransgenesis is a promising alternative. Paratransgenesis aims to inhibit the development of parasites within the vector through the action of genetically modified bacteria. The first step towards successful paratransgenesis in the Amazon is the identification of Anopheles darlingi symbiotic bacteria, which are transmitted vertically among mosquitoes, and are not pathogenic to humans. METHODS: Culturable bacteria associated with An. darlingi and their breeding sites were isolated by conventional microbiological techniques. Isolated strains were transformed with a GFP expressing plasmid, pSPT-1-GFP, and reintroduced in mosquitoes by feeding. Their survival and persistence in the next generation was assessed by the isolation of fluorescent bacteria from eggs, larvae, pupae and adult homogenates. RESULTS: A total of 179 bacterial strains were isolated from samples from two locations, Coari and Manaus. The predominant genera identified in this study were Acinetobacter, Enterobacter, Klebsiella, Serratia, Bacillus, Elizabethkingia, Stenotrophomonas and Pantoea. Two isolated strains, Serratia-Adu40 and Pantoea-Ovo3, were successfully transformed with the pSPT-1-GFP plasmid and expressed GFP. The fluorescent bacteria fed to adult females were transferred to their eggs, which persisted in larvae and throughout metamorphosis, and were detected in adult mosquitoes of the next generation. CONCLUSION: Serratia-Adu40 and Pantoea-Ovo3 are promising candidates for paratransgenesis in An. darlingi. Further research is needed to determine if these bacteria are vertically transferred in nature.

Analysis of the transcription of genes encoding heat shock proteins (hsp) in Aedes aegypti Linnaeus, 1762 (Diptera: Culicidae), maintained under climatic conditions provided by the IPCC (Intergovernmental Panel On Climate Change) for the year 2100.

Human actions intensify the greenhouse effect, aggravating climate changes in the Amazon and elsewhere in the world. The Intergovernmental Panel on Climate Change (IPCC) foresees a global increase of up to 4.5 °C and 850 ppm CO2 (above current levels) by 2100. This will impact the biology of the Aedes aegypti mosquito, vector of Dengue, Zika, urban Yellow Fever and Chikungunya. Heat shock proteins are associated with adaptations to anthropic environments and the interaction of some viruses with the vector. The transcription of the hsp26, hsp83 and hsc70 genes of an A. aegypti population, maintained for more than forty-eight generations, in the Current, Intermediate and Extreme climatic scenario predicted by the IPCC was evaluated with qPCR. In females, highest levels of hsp26, hsp83 and hsc70 expression occurred in the Intermediate scenario, while in males, levels were high only for hsp26 gene in Current and Extreme scenarios. Expression of hsp83 and hsc70 genes in males was low under all climatic scenarios, while in the Extreme scenario females had lower expression than in the Current scenario. The data suggest compensatory or adaptive processes acting on heat shock proteins, which can lead to changes in the mosquito's biology, altering vectorial competence.