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dc.contributor.authorOliveira, Juan Campos de-
dc.date.available2020-03-12-
dc.date.available2020-03-12T14:29:27Z-
dc.date.issued2018-08-29-
dc.identifier.urihttp://repositorioinstitucional.uea.edu.br//handle/riuea/2249-
dc.description.abstractThe Aedes aegypti mosquito is an important vector in the transmission of dengue virus, urban yellow fever, zika and chikungunya. The main tool used to control this vector is conventional chemical insecticides. However, continuous use causes many negative factors, such as environmental contamination and selection of resistant populations. In this sense, the biological control by microorganisms is a biotechnological alternative in the control of vectors, mainly the bioinseticidas formulated with bacteria of the genus Bacillus. Considering the diversity of Bacillus, members of this genus are capable of synthesizing bioinsecticidal biomolecules against different orders of insects. In addition, they are able to synthesize biopolymers, antibiotics and enzymes such as chitinase expressed by the chi gene. These enzymes have the ability to hydrolyze β-1,4 glycosidic bonds between the chitin polymer units. After ingestion by the larvae of the insect, it causes degradation of the peritrophic membrane that separates the epithelium from the middle intestine of the intestinal lumen, whose main component is chitin. This factor is a favorable requirement for its use in the control programs of insect vectors and agricultural pests. In this study, 124 bacterial strains isolated from soil, water, plant and insect from different Amazonian environments were used. All strains were identified by phenotypic characteristics as belonging to the bacilli group, being one hundred and fourteen (91.4%) Gram positive and ten (8.6%) Gram negative. In the molecular identification by amplification and sequencing of the 16S (rDNA) gene, 33 lines were identified, 27 belonging to the genus Bacillus, 4 to the genus Serratia, 1 to the genus Paenibacillus and 1 to the genus Achromobacter. In the molecular detection of the chi gene, six lineages (4.5%), BTAM27LB, 103PHAISP2, BTAM138LB, BTAM18NA, K2NA and R11ISP2 showed positive amplification, of which two were identified, BTAM 27- Bacillus sp. and R11ISP2-Bacillus amyloliquefaciens. In the selective bioassays of 124 lineages, 21 showed activity in Aedes aegypti larvae. The bioassays with the metabolic extracts produced by these 21 lineages cultivated in NA, LB and ISP2 medium demonstrated that 7 presented pathogenicity in the lyophilized culture and only 3 in the filtered supernatant. In another step, metabolic extracts produced by 9 strains cultured in chitin medium were evaluated, of which only 1 presented mortality in both the supernatant and lyophilized culture, presenting values corresponding to 100%. Thus, the selected strains present potential for the development of microbial bioinsecticides.pt_BR
dc.languageporpt_BR
dc.publisherUniversidade do Estado do Amazonaspt_BR
dc.rightsAcesso Abertopt_BR
dc.rightsAtribuição-NãoComercial-SemDerivados 3.0 Brasil*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/br/*
dc.subjectBioprospecçãopt_BR
dc.subjectMicrobiota amazônicapt_BR
dc.subjectEnzimas hidrolíticaspt_BR
dc.subjectInsetos vetorespt_BR
dc.titleSeleção de bacillus spp. da Amazonia brasileira portadores do gene Chi (quitinase) para o controle biológico de Aedes (stegomyia) aegypti(Linnaeus, 1762).pt_BR
dc.typeDissertaçãopt_BR
dc.date.accessioned2020-03-12T14:29:27Z-
dc.contributor.advisor1Tadei, Wanderli Pedro-
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/6806722604010480pt_BR
dc.contributor.referee1Tadei, Wanderli Pedro-
dc.contributor.referee1Latteshttp://lattes.cnpq.br/6806722604010480pt_BR
dc.contributor.referee2Fonseca, Maria Dolores Pinheiro-
dc.contributor.referee2Latteshttp://lattes.cnpq.br/2306799349043803pt_BR
dc.contributor.referee3Procópio, Rudi Emerson de Lima-
dc.contributor.referee3Latteshttp://lattes.cnpq.br/2478199435796976pt_BR
dc.creator.Latteshttp://lattes.cnpq.br/2020048717853869pt_BR
dc.description.resumoO mosquito Aedes aegypti é um importante vetor na transmissão do vírus dengue, febre amarela urbana, zika e chikungunya. A principal ferramenta utilizada no controle deste vetor são os inseticidas químicos convencionais, porém, o uso contínuo provoca fatores negativos destacando-se a contaminação do meio ambiente e a seleção de populações resistentes. Neste sentido, o controle biológico por microrganismos é uma alternativa biotecnológica no controle de vetores, principalmente os bioinseticidas formulados à base de bactérias do gênero Bacillus. Considerando a diversidade de Bacillus, estes apresentam capacidade de síntese de biomoléculas bioinseticidas contra diferentes ordens de insetos. Além disso, sintetizam biopolimeros, antibióticos e enzimas como, por exemplo, as quitinases expressas pelo gene Chi. Estas enzimas hidrolisam as ligações glicosídicas β-1,4 entre as unidades constituintes da quitina. Após a ingestão das quitinases pelas larvas do inseto, provoca a degradação da membrana peritrófica que separa o epitélio do intestino médio do lúmen intestinal, cujo principal componente é a quitina. Este fator é um requisito favorável para a sua utilização nos programas de controle de insetos vetores e pragas agrícolas. Neste estudo, foram utilizadas 124 linhagens bacterianas isoladas do solo, água, planta e inseto de diferentes ambientes amazônicos. Todas foram identificadas pelas características fenotípicas como pertencentes ao grupo dos bacilos, sendo, cento e quatorze (91,4%) Gram positivos e dez (8,6%) Gram negativos. Na identificação molecular pela amplificação e sequenciamento do gene 16S (rDNA), 33 linhagens foram identificadas, 27 pertencendo ao gênero Bacillus, 4 ao gênero Serratia, 1 ao gênero Paenibacillus e 1 ao gênero Achromobacter. Na detecção molecular do gene Chi, seis (4,5%) linhagens, BTAM27LB, 103PHAISP2, BTAM138LB, BTAM18NA, K2NA e R11ISP2, apresentaram amplificação positiva para este gene, dentre estas, duas foram identificadas, BTAM 27- Bacillus sp. e R11ISP2- Bacillus amyloliquefaciens. No bioensaio seletivo de 124 linhagens, 21 apresentaram atividade larvicida em Aedes aegypti. Os bioensaios com os extratos metabólicos produzidos por estas 21 linhagens ativas, cultivadas em meio NA, LB e ISP2, demonstraram que sete apresentaram patogenicidade nas células lisadas e apenas 3 no sobrenadante filtrado. Em outra etapa, foi avaliado os extratos metabólicos produzidos por 9 linhagens ativas cultivadas em meio contendo quitina, das quais apenas 1 apresentou mortalidade tanto no sobrenadante quanto em células liofilizadas, apresentando valores correspondentes a 100%. Sendo assim as linhagens selecionadas apresentaram potencial para o desenvolvimento de bioinseticidas microbianos.pt_BR
dc.publisher.countryBrasilpt_BR
dc.publisher.programPROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA E RECURSOS NATURAIS DA AMAZÔNIApt_BR
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