PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/3954 |
Resumo: | Recently, a decline of pollinators, especially the bees Apis mellifera L. (Hymenoptera: Apidae), has affected beekeeping and consequently agricultural harvests in some countries, although the damage to ecosystems has not been effectively accounted. Among the causes listed for this unexplained phenomenon, pathogens are among the possible factors responsible. There are several pathogens that attack the bees A. mellifera around the world, as bacteria Paenibacillus larvae (White) and the fungi Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir, Nosema apis Zander and Nosema ceranae Fries et al. Their distributions in some parts of the world, such as Brazil, are not exactly known, not only because of the difficulty of collecting samples in such a large country, and also because of the time and cost of diagnoses involved. At this point, it is important to standardize techniques for rapid diagnosis and to facilitate the safe conduct of epidemiological surveys, and therefore assist in controlling the spread of these microorganisms. The aim of this work was standardize a multiplex PCR for simultaneous detection of the spores of A. apis, N. ceranae and P. larvae present in honey and use it in the analysis of honey samples from some regions. The multiplex PCR was standardized using specific primers and DNA was extracted from honey samples positive for each of the pathogens. The recommendations of existing national legislation were used for the preparation of the solutions of honey to be submitted to the technique developed. The standard technique in this study was effective for diagnosing three pathogens to A. mellifera in honey, A. apis, N. ceranae and P. larvae. The detection threshold of monospecific PCR was of 10 CFU/mL of honey, and of 10 and 100 spores/mL of honey for A. apis and N. ceranae, respectively. The detection sensitivity of multiplex PCR was of 10 CFU/mL of honey for P. larvae, and of 100 spores/mL of honey for A. apis and for N. ceranae. Did not match any of those pathogens in 120 honey samples analyzed with standardized multiplex PCR. Thus this method was suitable for simultaneous detection of pathogens to A. mellifera in honey, but can probably be used in other bee products with minor modifications. |
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Puker, Andersonhttp://lattes.cnpq.br/7920341302303692Paula, Sérgio Oliveira dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767540P4Santana, Weyder Cristianohttp://lattes.cnpq.br/8704112718246122Message, Dejairhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783671P8Moreira, Maria Aparecida Scatamburlohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J62015-03-26T13:30:42Z2013-05-132015-03-26T13:30:42Z2011-02-22PUKER, Anderson. Multiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honey. 2011. 86 f. Dissertação (Mestrado em Ciência entomológica; Tecnologia entomológica) - Universidade Federal de Viçosa, Viçosa, 2011.http://locus.ufv.br/handle/123456789/3954Recently, a decline of pollinators, especially the bees Apis mellifera L. (Hymenoptera: Apidae), has affected beekeeping and consequently agricultural harvests in some countries, although the damage to ecosystems has not been effectively accounted. Among the causes listed for this unexplained phenomenon, pathogens are among the possible factors responsible. There are several pathogens that attack the bees A. mellifera around the world, as bacteria Paenibacillus larvae (White) and the fungi Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir, Nosema apis Zander and Nosema ceranae Fries et al. Their distributions in some parts of the world, such as Brazil, are not exactly known, not only because of the difficulty of collecting samples in such a large country, and also because of the time and cost of diagnoses involved. At this point, it is important to standardize techniques for rapid diagnosis and to facilitate the safe conduct of epidemiological surveys, and therefore assist in controlling the spread of these microorganisms. The aim of this work was standardize a multiplex PCR for simultaneous detection of the spores of A. apis, N. ceranae and P. larvae present in honey and use it in the analysis of honey samples from some regions. The multiplex PCR was standardized using specific primers and DNA was extracted from honey samples positive for each of the pathogens. The recommendations of existing national legislation were used for the preparation of the solutions of honey to be submitted to the technique developed. The standard technique in this study was effective for diagnosing three pathogens to A. mellifera in honey, A. apis, N. ceranae and P. larvae. The detection threshold of monospecific PCR was of 10 CFU/mL of honey, and of 10 and 100 spores/mL of honey for A. apis and N. ceranae, respectively. The detection sensitivity of multiplex PCR was of 10 CFU/mL of honey for P. larvae, and of 100 spores/mL of honey for A. apis and for N. ceranae. Did not match any of those pathogens in 120 honey samples analyzed with standardized multiplex PCR. Thus this method was suitable for simultaneous detection of pathogens to A. mellifera in honey, but can probably be used in other bee products with minor modifications.Recentemente, o declínio global dos polinizadores, especialmente das abelhas Apis mellifera L. (Hymenoptera: Apidae), tem acometido a atividade apícola e agrícola de alguns países causando prejuízos econômicos e ambientais, notadamente para os ecossistemas, ainda não efetivamente contabilizados. Dentre as causas elencadas para esse inexplicável fenômeno os patógenos estão entre os principais possíveis responsáveis. Vários são os patógenos que acometem as abelhas A. mellifera pelo mundo, entre eles a bactéria Paenibacillus larvae (White), e os fungos Ascosphaera apis (Maassen ex Claussen) Olive e Spiltoir, Nosema apis Zander e Nosema ceranae Fries et al. Suas distribuições em algumas partes do mundo, como o Brasil, são pouco conhecidas, não apenas pela dificuldade de coleta de amostras em um país com tamanha dimensão, mas também em virtude da morosidade e custo dos diagnósticos envolvidos. Diante disso, torna-se importante a padronização de técnicas para o diagnóstico rápido e seguro que facilite a realização de levantamentos epidemiológicos e que, consequentemente, auxilie no controle da disseminação desses micro-organismos. O objetivo desse trabalho foi padronizar uma técnica de PCR multiplex para detecção simultânea da presença de A. apis, N. ceranae e P. larvae em mel, bem como empregá-la na análise de amostras de mel provenientes de algumas regiões brasileiras. A PCR multiplex foi padronizada com primers específicos e DNA dos micro-organismos obtidos de amostras de mel positivas para cada um dos patógenos. Utilizou-se as recomendações da legislação nacional vigente para o preparo das soluções de mel a serem submetidas à técnica desenvolvida. A técnica padronizada neste estudo foi eficiente para diagnosticar simultaneamente três patógenos de A. mellifera em mel: A. apis, N. ceranae e P. larvae. O limiar de detecção da PCR monoespecífica foi 10 UFC/mL de mel para P. larvae e de 10 e 100 esporos/mL de mel para A. apis e N. ceranae, respectivamente. A sensibilidade de detecção da PCR multiplex foi de 10 UFC/mL de mel para P. larvae e 100 esporos/mL de mel para A. apis e N. ceranae. Não foram encontrados nenhum dos referidos patógenos nas 120 amostras de mel que foram analisadas com a PCR multiplex padronizada. A PCR multiplex foi adequada para detecção simultânea de patógenos de A. mellifera em mel, mas, provavelmente, poderá ser utilizada em outros produtos apícolas com pequenas modificações.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaMestrado em EntomologiaUFVBRCiência entomológica; Tecnologia entomológicaMultiplex-PCRSanidade apícolaApis melliferaAbelhas africanizadasMultiplex-PCRHealth beekeepingApis melliferaAfricanized beesCNPQ::CIENCIAS BIOLOGICAS::PARASITOLOGIA::ENTOMOLOGIA E MALACOLOGIA DE PARASITOS E VETORESPCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em melPCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em melMultiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honeyMultiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honeyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf1210474https://locus.ufv.br//bitstream/123456789/3954/1/texto%20completo.pdf54a2d68e22373c9be043b1a04f343b11MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain111584https://locus.ufv.br//bitstream/123456789/3954/2/texto%20completo.pdf.txtea46f75853e856eed8f2968beb79aaefMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3531https://locus.ufv.br//bitstream/123456789/3954/3/texto%20completo.pdf.jpg8230fc66f265ca99f85f6502c6964a23MD53123456789/39542016-04-09 23:11:31.147oai:locus.ufv.br:123456789/3954Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-10T02:11:31LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
dc.title.alternative.eng.fl_str_mv |
Multiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honey Multiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honey |
title |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
spellingShingle |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel Puker, Anderson Multiplex-PCR Sanidade apícola Apis mellifera Abelhas africanizadas Multiplex-PCR Health beekeeping Apis mellifera Africanized bees CNPQ::CIENCIAS BIOLOGICAS::PARASITOLOGIA::ENTOMOLOGIA E MALACOLOGIA DE PARASITOS E VETORES |
title_short |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
title_full |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
title_fullStr |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
title_full_unstemmed |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
title_sort |
PCR multiplex para detecção de patógenos de Apis mellifera L. (Hymenoptera, Apidae) em mel |
author |
Puker, Anderson |
author_facet |
Puker, Anderson |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/7920341302303692 |
dc.contributor.author.fl_str_mv |
Puker, Anderson |
dc.contributor.advisor-co1.fl_str_mv |
Paula, Sérgio Oliveira de |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767540P4 |
dc.contributor.advisor-co2.fl_str_mv |
Santana, Weyder Cristiano |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://lattes.cnpq.br/8704112718246122 |
dc.contributor.advisor1.fl_str_mv |
Message, Dejair |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783671P8 |
dc.contributor.referee1.fl_str_mv |
Moreira, Maria Aparecida Scatamburlo |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J6 |
contributor_str_mv |
Paula, Sérgio Oliveira de Santana, Weyder Cristiano Message, Dejair Moreira, Maria Aparecida Scatamburlo |
dc.subject.por.fl_str_mv |
Multiplex-PCR Sanidade apícola Apis mellifera Abelhas africanizadas |
topic |
Multiplex-PCR Sanidade apícola Apis mellifera Abelhas africanizadas Multiplex-PCR Health beekeeping Apis mellifera Africanized bees CNPQ::CIENCIAS BIOLOGICAS::PARASITOLOGIA::ENTOMOLOGIA E MALACOLOGIA DE PARASITOS E VETORES |
dc.subject.eng.fl_str_mv |
Multiplex-PCR Health beekeeping Apis mellifera Africanized bees |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::PARASITOLOGIA::ENTOMOLOGIA E MALACOLOGIA DE PARASITOS E VETORES |
description |
Recently, a decline of pollinators, especially the bees Apis mellifera L. (Hymenoptera: Apidae), has affected beekeeping and consequently agricultural harvests in some countries, although the damage to ecosystems has not been effectively accounted. Among the causes listed for this unexplained phenomenon, pathogens are among the possible factors responsible. There are several pathogens that attack the bees A. mellifera around the world, as bacteria Paenibacillus larvae (White) and the fungi Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir, Nosema apis Zander and Nosema ceranae Fries et al. Their distributions in some parts of the world, such as Brazil, are not exactly known, not only because of the difficulty of collecting samples in such a large country, and also because of the time and cost of diagnoses involved. At this point, it is important to standardize techniques for rapid diagnosis and to facilitate the safe conduct of epidemiological surveys, and therefore assist in controlling the spread of these microorganisms. The aim of this work was standardize a multiplex PCR for simultaneous detection of the spores of A. apis, N. ceranae and P. larvae present in honey and use it in the analysis of honey samples from some regions. The multiplex PCR was standardized using specific primers and DNA was extracted from honey samples positive for each of the pathogens. The recommendations of existing national legislation were used for the preparation of the solutions of honey to be submitted to the technique developed. The standard technique in this study was effective for diagnosing three pathogens to A. mellifera in honey, A. apis, N. ceranae and P. larvae. The detection threshold of monospecific PCR was of 10 CFU/mL of honey, and of 10 and 100 spores/mL of honey for A. apis and N. ceranae, respectively. The detection sensitivity of multiplex PCR was of 10 CFU/mL of honey for P. larvae, and of 100 spores/mL of honey for A. apis and for N. ceranae. Did not match any of those pathogens in 120 honey samples analyzed with standardized multiplex PCR. Thus this method was suitable for simultaneous detection of pathogens to A. mellifera in honey, but can probably be used in other bee products with minor modifications. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011-02-22 |
dc.date.available.fl_str_mv |
2013-05-13 2015-03-26T13:30:42Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:30:42Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
PUKER, Anderson. Multiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honey. 2011. 86 f. Dissertação (Mestrado em Ciência entomológica; Tecnologia entomológica) - Universidade Federal de Viçosa, Viçosa, 2011. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/3954 |
identifier_str_mv |
PUKER, Anderson. Multiplex PCR for detection of pathogens of Apis mellifera L. (Hymenoptera, Apidae) in honey. 2011. 86 f. Dissertação (Mestrado em Ciência entomológica; Tecnologia entomológica) - Universidade Federal de Viçosa, Viçosa, 2011. |
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Universidade Federal de Viçosa |
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Mestrado em Entomologia |
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UFV |
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BR |
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Ciência entomológica; Tecnologia entomológica |
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Universidade Federal de Viçosa |
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