Sele????o de Bacillus spp. da Amaz??nia Brasileira portadores de genes Cry e/ou PhaC via s??ntese Polihidroxiacalnoatos (PHAs) para o controle de Aedes aegypti Linnaeus 1762

• Autor(es): Katak, Ricardo de Melo
• Data: 2017
• Origem: Oasisbr
• Assunto(s): Vetores de doen??as; Metab??litos; Prote??na CRY; Pilihidroxialcanoatos; Alternativa de controle; Prote??nas inseticidas; Biopolimeros; CI??NCIAS BIOL??GICAS
• Tipo de Documento: Dissertação de mestrado
• Idioma: Português

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CNPq - Conselho Nacional de Desenvolvimento Cient??fico e Tecnol??gico

Insect vectors of tropical diseases such as Ae. Aegypti, the main vector of dengue virus, chikungunya, zika and West Nile virus, are a major problem for public health. One of the major control measures is to control the vector. In this regard, we sought to investigate the biological control of Ae. aegypti; with the use of Bacillus spp .; Amazon isolated from different environments. From the different samples, there was obtained a total of 118 bacterial strains, and 41 strains identified by phenotypic characteristics such as bacilli. Of these, 38 were positive and gram 2 gram negative bacilli. The molecular identification of these strains allowed the identification of 29 strains were characterized in three different genres. The molecular characterization of patients bacilli of Cry4 genes, cry11 and PHAC. Cry4 genes were observed in BtAM41, 2R6.2.1LB lines with no toxicity. The Bio19LB lines, BSBioLB, Bio01LB and Bio011LB presented the cry11 genes, as the larvicidal activity were efficient in both phases of bioassays except Bio011 lineage that any result below 50% in the second stage of bioassays. The 2WISP2, K5NA lines, R8ISP2, R15ISP2, Bio16LB, BtAM125LB, BtAM49LB just presented the PhaC gene. Just BtAM49LB strain was effective in larvicidal activity. Considering the results of the first and second stage, the best results when there were interaction sterile lysed cell supernatant (SN), showing 100% in 72 h. In this sense, it is recommended to study the characterization of metabolites of these strains. The K4NA lines; 103PHAISP2, BtAM220NA; ALP2ISP2, BtAM74LB and PHA50ISP2 bear the cry11 and PHAC genes showed no larvicidal activity above 50% considering it was not observed correlation of genes associated with larvicidal activity. Only Bio19LB strain showed the cry11 gene and Cry4, with larvicidal activity above 50% in the supernatant of lysed cells with interaction with 70% mortality within 72 h. There was no correlation of the phaC gene and Cry isolated, but the best results were in the supernatant of lysed cells with consortium, possibitando the possibility of interaction of molecules with insecticidal activity. So before these results are needed more detailed studies to understand and elucidate the interaction of strains that showed higher larvicidal activity, it is concluded that the strains that showed larvicidal activity above 50% and the same carriers of cry11 genes, Cry4 and PHAC can be associated with other virulence and pathogenicity factors, becoming future potential lines for the control of Ae. Aegypti.

Insetos vetores de pat??genos tropicais como Ae. Aegypti, principal vetor do v??rus dengue, chikungunya, zika e v??rus do Nilo Ocidental, s??o um grande problema para sa??de p??blica. Uma das principais medidas de combate ?? o controle do vetor. Neste sentido, buscou-se investigar o controle biol??gico de Ae. aegypti; com uso de Bacillus spp.; isolados de diferentes ambientes Amaz??nicos. A partir de diferentes amostras, obteve-se o total de 118 linhagens bacterianas, sendo 41 linhagens identificadas por caracter??sticas fenot??picas como bacilos. Destes, 39 foram bacilos gram positivos e 2 gram negativos. A identifica????o molecular destas linhagens permitiu identificar 29 linhagens, sendo caracterizadas em tr??s g??neros distintos. Quanto a caracteriza????o molecular dos bacilos portadores dos genes Cry4, Cry11 e PhaC. Foram observados genes Cry4 nas linhagens BtAM41, 2R6.2.1LB, com aus??ncia de toxicidade. As linhagens Bio19LB, BSBioLB, Bio01LB e Bio011LB apresentaram os genes Cry11, quanto a atividade larvicida foram eficientes nas duas etapas dos bioensaios exceto a linhagem Bio011 que apresentou resultados abaixo de 50 % na segunda etapa dos bioensaios. As linhagens 2WISP2, K5NA, R8ISP2, R15ISP2, Bio16LB, BtAM125LB, BtAM49LB apresentaram apenas o gene PhaC. Apenas a linhagem BtAM49LB foi eficiente na atividade larvicida. Considerando os resultados dos bioensaios seletivos de extratos brutos de Bacillus spp., os melhores resultados foram quando houve a intera????o do sobrenadante est??ril com c??lulas lisadas (SN), apresentando 100 % em 72 h. Neste sentido, recomenda-se estudos da caracteriza????o dos metabolitos destas linhagens. As linhagens K4NA; 103PHAISP2, BtAM220NA; ALP2ISP2, BtAM74LB e PHA50ISP2 portadoras dos genes Cry11 e PhaC, n??o apresentaram atividade larvicida acima de 50 %, considerando que n??o foi observado correla????o dos genes associados com atividade larvicida. Somente a linhagem Bio19LB apresentou os genes Cry11 e Cry4, apresentando atividade larvicida acima de 50 % na intera????o de sobrenadante com c??lulas lisadas com 70 % de mortalidade em 72 h. N??o foi observado correla????o dos genes PhaC e Cry nos isolados, mas os melhores resultados foram no cons??rcio do sobrenadante + c??lulas lisadas, possibilitando a hip??tese de intera????o de mol??culas com atividade inseticida. Portanto, diante destes resultados s??o necess??rios mais estudos detalhados para compreender e elucidar a intera????o das linhagens que apresentaram maior atividade larvicida, conclui-se que as linhagens que apresentaram atividade larvicida acima de 50 % e as mesmas portadoras dos genes Cry11, Cry4 e PhaC podem estar associados a outros fatores de virul??ncia e patogenicidade, tornando-se futuramente linhagens potenciais para o controle de Ae. Aegypti no Estado do Amazonas.