Culturable bacteria associated with Anopheles darlingi and their paratransgenesis potential

Malaria Journal

Piero Onorati¹  Olle Terenius²  Rosemary Aparecida Roque³  Wanderli Pedro Tadei³  Joaquim Ferreira do Nascimento Neto³  Osvaldo Marinotti⁴  Elerson Matos Rocha⁵  Marcos Cézar Fernandes Pessoa⁵  Ricardo de MeloKatak⁵  Adolfo Jose da Mota⁵  Juan Campos de Oliveira⁵  Jayme A. Souza-Neto⁶  Marta Rodrigues de Oliveira⁷  Laura Viana Correa⁸  Deidre Machado Serrão⁸  Veranilce Alves Muniz⁸ 

[1] Department of Ecology, Swedish University of Agricultural Sciences (SLU), Box 7044, 750 07, Uppsala, Sweden;Department of Ecology, Swedish University of Agricultural Sciences (SLU), Box 7044, 750 07, Uppsala, Sweden;Department of Cell and Molecular Biology, Microbiology, Uppsala University, Box 596, 751 24, Uppsala, Sweden;Laboratório de Malária E Dengue, Instituto Nacional de Pesquisas da Amazônia, INPA, Manaus, Brazil;MTEKPrime, Aliso Viejo, CA, USA;Programa de Pós-Graduação em Biotecnologia, Universidade Federal do Amazonas - PPGBIOTEC / UFAM, Manaus, Brazil;School of Agricultural Sciences, Department of Bioprocesses and Biotechnology, Central Multi User Laboratory, São Paulo State University (UNESP), Botucatu, Brazil;Universidade Estadual do Amazonas - BIONORTE, UEA, Manaus, Brazil;Universidade Estadual do Amazonas - MBT, UEA, Manaus, Brazil;

关键词: Mosquito;    Malaria;    Microbiota;    Vector-borne disease;    Amazon forest;   

DOI  :  10.1186/s12936-020-03574-1

来源: Springer

【 摘 要 】

BackgroundMalaria 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.MethodsCulturable 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.ResultsA 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.ConclusionSerratia-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.

【 授权许可】

CC BY