Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório do Centro Universitário Braz Cubas |
Texto Completo: | https://repositorio.cruzeirodosul.edu.br/handle/123456789/2606 |
Resumo: | In order to minimize the significant environmental impact consequent to the indiscriminate use of chemicals on crops, biopesticides have been proposed increasingly for pest control and integrated management. Despite the proven Bacillus thuringiensis efficiency in these products, there are still limitations to its use due to the cost of production and lack of knowledge regarding the correct utilization of the formulations. This study aimed at developing a biopesticide product based on Bacillus thuringiensis subsp. kurstaki HD-1, which attends the quality criteria and is viable industrially. Initially the influence of the culture medium composition on the growth and sporulation was evaluated. The modification in the nitrogen source, adding peptone to the yeast exteract present in the original medium, did not present significant effect. However the modification in the carbon source, from glucose to sucrose, resulted in significantly higher sporulation (4.83.108 UFC/mL) in agitated flasks. For the cultivation in 5 L bottles comparing glucose and sucrose, there was no difference between the maximum concentrations of spores obtained in both media, between 2.107 and 3.107 UFC/mL after 34 h, however, no consumption of sucrose by the microorganism was verified. In a bench-scale bioreactor (7.5 L) in batch mode using glucose as carbon source, the sporulation peak occured in 30 h (4.83.1010 UFC/mL). In fed-batch mode with glucose, using the same medium, the maximum value of sporulation was obtained after 72 h of cultivation (4.44.1012 UFC/mL). The presence of proteins with estimated molecular weights of 70 kDa and 130 kDa, charactertistic of the toxins of interest, was observed in electrophoresis gel, visually in higher concentration in the samples of the fedbatch process. A formulation was proposed for the concentrated broth of Bacillus thuringiensis subsp. kurstaki HD-1 produced in batch and fed-batch modes. The formulations were tested by bioassay with the lepidoptera Spodoptera frugiperda in order to verify the lethal concentration (LC50). The LC50 of the batch formulation was 635.121 spores/mL and the one of the fed-batch formulation was 10.994 spores/mL. |
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Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1Inseticida - biológicoEndotoxinasCinética de crescimentoCNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERALIn order to minimize the significant environmental impact consequent to the indiscriminate use of chemicals on crops, biopesticides have been proposed increasingly for pest control and integrated management. Despite the proven Bacillus thuringiensis efficiency in these products, there are still limitations to its use due to the cost of production and lack of knowledge regarding the correct utilization of the formulations. This study aimed at developing a biopesticide product based on Bacillus thuringiensis subsp. kurstaki HD-1, which attends the quality criteria and is viable industrially. Initially the influence of the culture medium composition on the growth and sporulation was evaluated. The modification in the nitrogen source, adding peptone to the yeast exteract present in the original medium, did not present significant effect. However the modification in the carbon source, from glucose to sucrose, resulted in significantly higher sporulation (4.83.108 UFC/mL) in agitated flasks. For the cultivation in 5 L bottles comparing glucose and sucrose, there was no difference between the maximum concentrations of spores obtained in both media, between 2.107 and 3.107 UFC/mL after 34 h, however, no consumption of sucrose by the microorganism was verified. In a bench-scale bioreactor (7.5 L) in batch mode using glucose as carbon source, the sporulation peak occured in 30 h (4.83.1010 UFC/mL). In fed-batch mode with glucose, using the same medium, the maximum value of sporulation was obtained after 72 h of cultivation (4.44.1012 UFC/mL). The presence of proteins with estimated molecular weights of 70 kDa and 130 kDa, charactertistic of the toxins of interest, was observed in electrophoresis gel, visually in higher concentration in the samples of the fedbatch process. A formulation was proposed for the concentrated broth of Bacillus thuringiensis subsp. kurstaki HD-1 produced in batch and fed-batch modes. The formulations were tested by bioassay with the lepidoptera Spodoptera frugiperda in order to verify the lethal concentration (LC50). The LC50 of the batch formulation was 635.121 spores/mL and the one of the fed-batch formulation was 10.994 spores/mL.Visando minimizar o impacto ambiental importante consequente ao uso indiscriminado de produtos químicos nas lavouras, cada vez mais bioinseticidas têm sido propostos para controle e manejo integrado de pragas. Apesar da comprovada eficiência de Bacillus thuringiensis nesses produtos, ainda existem limitações ao seu uso devido ao custo de produção e pouco conhecimento referente à utilização correta das formulações. O presente trabalho teve como objetivo desenvolver um produto bioinseticida à base de Bacillus thuringiensis subsp. kurstaki HD-1, que atenda aos critérios de qualidade e que seja viável industrialmente. Foi avaliada inicialmente a influência da composição do meio de cultivo no crescimento e esporulação. A modificação da fonte de nitrogênio, acrescentando-se peptona ao extrato de levedura do meio original, não apresentou efeito significativo. Já a modificação da fonte de carbono de glicose para sacarose resultou em esporulação significativamente maior (4,83.108 UFC/mL) em frascos agitados. Para o cultivo em garrafões de 5 L comparando glicose e sacarose, não houve diferença entre as concentrações máximas de esporos obtidas nos dois meios, entre 2.107 e 3.107 UFC/mL após 34h, no entanto, verificouse que não houve consumo da sacarose pelo micro-organismo. Em biorreator de bancada (7,5 L) em batelada utilizando-se glicose como fonte de carbono, o pico de esporulação ocorreu em 30 h (4,83.1010 UFC/mL). Já em batelada alimentada com glicose, utilizando-se o mesmo meio, obteve-se valor máximo de esporulação após 72 h de cultivo (4,44.1012 UFC/mL). Foi observada em gel de eletroforese a presença de proteínas com pesos moleculares estimados em 70 kDa e 130 kDa, característicos das toxinas de interesse, visualmente em maior concentração nas amostras da batelada alimentada. Uma formulação foi proposta para o caldo concentrado de Bacillus thuringiensis subsp. kurstaki HD-1 produzido em batelada e batelada alimentada. As formulações foram testadas por meio de bioensaios com a lepidóptera Spodoptera frugiperda para verificação da concentração letal (CL50). A CL50 da formulação produzida em batelada foi 635.121 esporos/mL e da formulação de batelada alimentada foi 10.994 esporos/mL.Universidade PositivoBrasilPós-GraduaçãoPrograma de Pós-Graduação em Biotecnologia IndustrialUPKarp, Susan Gracehttp://lattes.cnpq.br/8158732509142272Barga, Marcelo CalideMartins, Luciane Müller2021-08-17T19:56:11Z20162021-08-17T19:56:11Z2016info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://repositorio.cruzeirodosul.edu.br/handle/123456789/2606porinfo:eu-repo/semantics/openAccessreponame:Repositório do Centro Universitário Braz Cubasinstname:Centro Universitário Braz Cubas (CUB)instacron:CUB2021-08-23T17:32:33Zoai:repositorio.cruzeirodosul.edu.br:123456789/2606Repositório InstitucionalPUBhttps://repositorio.brazcubas.edu.br/oai/requestbibli@brazcubas.edu.bropendoar:2021-08-23T17:32:33Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)false |
dc.title.none.fl_str_mv |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
title |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
spellingShingle |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 Martins, Luciane Müller Inseticida - biológico Endotoxinas Cinética de crescimento CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
title_full |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
title_fullStr |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
title_full_unstemmed |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
title_sort |
Desenvolvimento de um produto bioinseticida à base de bacillus thuringiensis subsp. kurstaki HD-1 |
author |
Martins, Luciane Müller |
author_facet |
Martins, Luciane Müller |
author_role |
author |
dc.contributor.none.fl_str_mv |
Karp, Susan Grace http://lattes.cnpq.br/8158732509142272 Barga, Marcelo Calide |
dc.contributor.author.fl_str_mv |
Martins, Luciane Müller |
dc.subject.por.fl_str_mv |
Inseticida - biológico Endotoxinas Cinética de crescimento CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
topic |
Inseticida - biológico Endotoxinas Cinética de crescimento CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
In order to minimize the significant environmental impact consequent to the indiscriminate use of chemicals on crops, biopesticides have been proposed increasingly for pest control and integrated management. Despite the proven Bacillus thuringiensis efficiency in these products, there are still limitations to its use due to the cost of production and lack of knowledge regarding the correct utilization of the formulations. This study aimed at developing a biopesticide product based on Bacillus thuringiensis subsp. kurstaki HD-1, which attends the quality criteria and is viable industrially. Initially the influence of the culture medium composition on the growth and sporulation was evaluated. The modification in the nitrogen source, adding peptone to the yeast exteract present in the original medium, did not present significant effect. However the modification in the carbon source, from glucose to sucrose, resulted in significantly higher sporulation (4.83.108 UFC/mL) in agitated flasks. For the cultivation in 5 L bottles comparing glucose and sucrose, there was no difference between the maximum concentrations of spores obtained in both media, between 2.107 and 3.107 UFC/mL after 34 h, however, no consumption of sucrose by the microorganism was verified. In a bench-scale bioreactor (7.5 L) in batch mode using glucose as carbon source, the sporulation peak occured in 30 h (4.83.1010 UFC/mL). In fed-batch mode with glucose, using the same medium, the maximum value of sporulation was obtained after 72 h of cultivation (4.44.1012 UFC/mL). The presence of proteins with estimated molecular weights of 70 kDa and 130 kDa, charactertistic of the toxins of interest, was observed in electrophoresis gel, visually in higher concentration in the samples of the fedbatch process. A formulation was proposed for the concentrated broth of Bacillus thuringiensis subsp. kurstaki HD-1 produced in batch and fed-batch modes. The formulations were tested by bioassay with the lepidoptera Spodoptera frugiperda in order to verify the lethal concentration (LC50). The LC50 of the batch formulation was 635.121 spores/mL and the one of the fed-batch formulation was 10.994 spores/mL. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016 2016 2021-08-17T19:56:11Z 2021-08-17T19:56:11Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.cruzeirodosul.edu.br/handle/123456789/2606 |
url |
https://repositorio.cruzeirodosul.edu.br/handle/123456789/2606 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Positivo Brasil Pós-Graduação Programa de Pós-Graduação em Biotecnologia Industrial UP |
publisher.none.fl_str_mv |
Universidade Positivo Brasil Pós-Graduação Programa de Pós-Graduação em Biotecnologia Industrial UP |
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reponame:Repositório do Centro Universitário Braz Cubas instname:Centro Universitário Braz Cubas (CUB) instacron:CUB |
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Centro Universitário Braz Cubas (CUB) |
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CUB |
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CUB |
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Repositório do Centro Universitário Braz Cubas |
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Repositório do Centro Universitário Braz Cubas |
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Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB) |
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bibli@brazcubas.edu.br |
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1798311347817021440 |