Author(s): Rocha, Elerson Matos
Date: 2020
Origin: Oasisbr
Subject(s): Paratransg??nese; Anopheles darlingi; Bact??rias simbi??ticas; Mal??ria; Plasmodium vivax; CI??NCIAS BIOL??GICAS: BIOLOGIA GERAL; Bact??rias simbi??ticas; Anopheles darlingi; Paratransg??nese; Amaz??nia brasileira
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CAPES
Despite positive advances in the fight against malaria each year, this disease is still one of the most lethal among vector-borne parasitic diseases worldwide, with the highest death records in Africa, caused mainly by Plasmodium spp. and transmitted by Anopheles spp. Among the ways of controlling this disease, paratransgenesis stands out as a new and promising alternative that aims to inhibit the development of parasites within the vector through the action of genetically modified bacteria. This new approach is being successfully conducted against Plasmodium spp. in laboratory on the African continent. Due to the constant records of malaria cases in the Brazilian Amazon, this region it could be also the target of paratransgenic approaches, with the objective of blocking the development of Plasmodium vivax within the body of Anopheles darlingi, the main responsibles for this disease in this region. In this sense, the first step should be directed towards obtaining of symbionts bacteria of A. darlingi, which can be transmitted horizontally and vertically in mosquitoes and not being pathogenic to humans. Such characteristics are essential in paratransgenesis. In this sense, this work aimed to select cultivable bacterial species, symbionts of A. darlingi to control malaria by paratransgenesis. Bacterial isolations were performed on samples associated with the larval development environment, as well as on the body of A. darlingi, in order to identify recurrent species in the different samples. After sequencing the 16S rRNA gene of the isolated colonies, 179 species were identified in the samples from Coari and Manaus, which constitute 83 different species, 12 from Coari and 71 from Manaus. The predominant species in all samples, from Coari and Manaus, were: Acinetobacter nosocomialis; Enterobacter asburiae; Klebsiella pneumoniae; Serratia marcescens; Bacillus cereus; Pantoea dispersa; Elizabethkingia miricola; Klebsiella variicola; Stenotrophomonas pavanii and Pantoea agglomerans. After analyzing some criteria, fundamental for use in paratransgenesis, such as non-pathogenicity to humans, the bacterial strains S. marcescens-Adu40; E. asburiae-Adu24; P. dispersa-Adu38 and P. agglomerans-Ovo3, were selected to be transformed with a plasmid, containing a sequence of genes that expresses a green fluorescent protein, when exposed to UV light. Thus, the potential of two bacterial strains that went analyzed for horizontal and vertical transfer capacity in A. darlingi was revealed. Such strains were: S. marcescens-Adu40 and P. agglomerans-Ovo3. The results of these analyzes concluded that these two strains have the potential to be used to control malaria by paratransgenesis.
Apesar dos avan??os positivos na luta contra a mal??ria a cada ano, essa doen??a ainda ?? uma das mais letais entre as doen??as parasit??rias transmitidas por vetores em todo o mundo, com os maiores registros de mortes na ??frica, causados principalmente por Plasmodium spp. e transmitido por Anopheles spp. Dentre as formas de controle dessa doen??a, a paratransg??nese se destaca como uma alternativa nova e promissora que visa inibir o desenvolvimento de parasitas no vetor por meio da a????o de bact??rias geneticamente modificadas. Esta nova abordagem est?? sendo conduzida com sucesso contra o Plasmodium spp. em laborat??rio no continente africano. Devido aos constantes registros de casos de mal??ria na Amaz??nia brasileira, essa regi??o tamb??m poderia ser alvo de abordagens paratransg??nicas, com o objetivo de bloquear o desenvolvimento de Plasmodium vivax no corpo de Anopheles darlingi, os principais respons??veis por esta doen??a nesta regi??o. regi??o. Nesse sentido, o primeiro passo deve ser direcionado ?? obten????o de bact??rias simbiontes de A. darlingi, que podem ser transmitidas horizontal e verticalmente em mosquitos e n??o serem patog??nicas para os seres humanos. Tais caracter??sticas s??o essenciais na paratransg??nese. Nesse sentido, este trabalho teve como objetivo selecionar esp??cies bacterianas cultiv??veis, simbiontes de A. darlingi para controlar a mal??ria por paratransg??nese. Isolamentos bacterianos foram realizados em amostras associadas ao ambiente de desenvolvimento larval, bem como no corpo de A. darlingi, a fim de identificar esp??cies recorrentes nas diferentes amostras. Ap??s o sequenciamento do gene 16S rRNA das col??nias isoladas, foram identificadas 179 esp??cies nas amostras de Coari e Manaus, que constituem 83 esp??cies diferentes, 12 de Coari e 71 de Manaus. As esp??cies predominantes em todas as amostras, de Coari e Manaus, foram: Acinetobacter nosocomialis; Enterobacter asburiae; Klebsiella pneumoniae; Serratia marcescens; Bacillus cereus; Pantoea dispersa; Elizabethkingia miricola; Klebsiella variicola; Stenotrophomonas pavanii e Pantoea agglomerans. Ap??s an??lise de alguns crit??rios, fundamentais para o uso em paratransg??nese, como a n??o patogenicidade para humanos, as cepas bacterianas S. marcescens-Adu40; E. asburiae-Adu24; P. dispersa-Adu38 e P. agglomerans-Ovo3, foram selecionados para serem transformados com um plasm??deo, contendo uma sequ??ncia de genes que expressa uma prote??na verde fluorescente, quando exposta ?? luz UV. Assim, foi revelado o potencial de duas cepas bacterianas analisadas quanto ?? capacidade de transfer??ncia horizontal e vertical em A. darlingi. Tais cepas foram: S. marcescens-Adu40 e P. agglomerans-Ovo3. Os resultados dessas an??lises conclu??ram que essas duas linhagens t??m o potencial de serem usadas no controle da mal??ria por paratransg??nese.
