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Research article - Peer-reviewed, 2021

Culturable bacteria associated with Anopheles darlingi and their paratransgenesis potential

Rocha, Elerson Matos; Marinotti, Osvaldo; Serrao, Deidre Machado; Correa, Laura Viana; Katak, Ricardo de Melo; Juan, Campos-de-Oliveir; Muniz, Veranilce Alves; de Oliveira, Marta Rodrigues; Ferreira, Joaquim; Pessoa, Marcos Cezar Fernandes; Roque, Rosemary Aparecida; da Mota, Adolfo Jose; Onorati, Piero; Souza-Neto, Jayme A.; Terenius, Olle; Tadei, Wanderli Pedro

Abstract

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.

Keywords

Mosquito; Malaria; Microbiota; Vector-borne disease; Amazon forest

Published in

Malaria Journal
2021, volume: 20, number: 1, article number: 40

Authors' information

Rocha, Elerson Matos
Universidade Federal de Amazonas
Marinotti, Osvaldo
MTEKPrime
Serrao, Deidre Machado
Universidade do Estado do Amazonas
Correa, Laura Viana
Universidade do Estado do Amazonas
Katak, Ricardo de Melo
Univ Fed Amazonas PPGBIOTEC UFAM
Campos de Oliveira, Juan
Universidade Federal de Amazonas
Muniz, Veranilce Alves
Universidade do Estado do Amazonas
de Oliveira, Marta Rodrigues
No organisation
Ferreira, Joaquim
Institute Nacional de Pesquisas da Amazonia
Pessoa, Marcos Cezar Fernandes
Univ Fed Amazonas PPGBIOTEC UFAM
da Mota, Adolfo Jose
Univ Fed Amazonas PPGBIOTEC UFAM
Roque, Rosemary Aparecida
Institute Nacional de Pesquisas da Amazonia
Swedish University of Agricultural Sciences, Department of Ecology
Souza-Neto, Jayme A.
Universidade Estadual Paulista
Swedish University of Agricultural Sciences, Department of Ecology
Uppsala University
Tadei, Wanderli Pedro
Institute Nacional de Pesquisas da Amazonia

Sustainable Development Goals

SDG3 Good health and wellbeing

UKÄ Subject classification

Ecology
Microbiology

Publication Identifiers

DOI: https://doi.org/10.1186/s12936-020-03574-1

URI (permanent link to this page)

https://res.slu.se/id/publ/109965