Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini)
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/13486 |
Resumo: | The mercury (Hg) is a non-essential trace metal, that is highly toxic to living beings, is a major contaminant of water bodies. Despite the genotoxic importance of Hg, there are no studies on its genotoxicity in native neotropical bees, using effective biomarkers. Therefore, a study of the genotoxicity of Hg in Bombus atratus bees was carried out through the comet assay. This assay determines the extent of DNA damage to individual cell nuclei by differential migration of DNA fragments in microelectrophoresis. Despite the effectiveness and sensitivity of the technique, there are no records in the literature about the applicability of this assay to Bombus neotropical bees, which have many endangered species. B. atratus workers were collected from Sorocaba, SP, (23º34'53.1''S 47º31'29.5'') during the months of February to March 2019. After capture, the workers were individually placed in special boxes for the toxicological test and kept in an incubator at 26 ºC, 70% humidity, in the dark, for 48 hours, according to OECD recommendations. Eleven workers were used for the group exposed to Hg at a concentration of 0.2 µg L -1, concentration considered safe by CONAMA (nº 357, 2005), and the same number for the control group. Both groups were maintained with a 70% sucrose solution. In addition to the exposed group, 0.2 µg L -1 Hg solution was offered, and to the control group, uncontaminated water. After 48 hours, the tissues that compose the Hepato-Nephrocytic System (HNS), dorsal vessel, hemocytes, fat body/pericardial cells, and Malpighi tubules (MT), were collected and processed for the comet assay. The results showed that Hg promoted genotoxicity in these animals. Regarding the control group, the HNS of workers exposed to Hg was sensitive to contamination due to the abundant presence of different levels of comets, which indicate the fragmentation of genetic material. While MT, in turn, were less responsive to the trace metal, presenting fewer damaged nuclei. Statistical analysis compared between level 0 and total damage parameters showed higher genotoxicity promoted by Hg in HNS of the exposed group, compared to the control group. While genotoxicity in MT was similar between control and exposed groups. Therefore, it can be concluded that HNS is more responsive to Hg exposure than MT. It was concluded that the concentration of Hg considered safe by CONAMA compromised the cells that compose the HNS, impairing the general physiology of the studied species, even if the TM were not affected. |
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Ceschi-Bertoli, LetíciaAbdalla, Fábio Camargohttp://lattes.cnpq.br/7770803221041927http://lattes.cnpq.br/6218507280198988840c0f61-23a5-4340-9c77-f9ea13ac06b82020-11-26T11:59:07Z2020-11-26T11:59:07Z2020-03-03CESCHI-BERTOLI, Letícia. Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini). 2020. Dissertação (Mestrado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/13486.https://repositorio.ufscar.br/handle/ufscar/13486The mercury (Hg) is a non-essential trace metal, that is highly toxic to living beings, is a major contaminant of water bodies. Despite the genotoxic importance of Hg, there are no studies on its genotoxicity in native neotropical bees, using effective biomarkers. Therefore, a study of the genotoxicity of Hg in Bombus atratus bees was carried out through the comet assay. This assay determines the extent of DNA damage to individual cell nuclei by differential migration of DNA fragments in microelectrophoresis. Despite the effectiveness and sensitivity of the technique, there are no records in the literature about the applicability of this assay to Bombus neotropical bees, which have many endangered species. B. atratus workers were collected from Sorocaba, SP, (23º34'53.1''S 47º31'29.5'') during the months of February to March 2019. After capture, the workers were individually placed in special boxes for the toxicological test and kept in an incubator at 26 ºC, 70% humidity, in the dark, for 48 hours, according to OECD recommendations. Eleven workers were used for the group exposed to Hg at a concentration of 0.2 µg L -1, concentration considered safe by CONAMA (nº 357, 2005), and the same number for the control group. Both groups were maintained with a 70% sucrose solution. In addition to the exposed group, 0.2 µg L -1 Hg solution was offered, and to the control group, uncontaminated water. After 48 hours, the tissues that compose the Hepato-Nephrocytic System (HNS), dorsal vessel, hemocytes, fat body/pericardial cells, and Malpighi tubules (MT), were collected and processed for the comet assay. The results showed that Hg promoted genotoxicity in these animals. Regarding the control group, the HNS of workers exposed to Hg was sensitive to contamination due to the abundant presence of different levels of comets, which indicate the fragmentation of genetic material. While MT, in turn, were less responsive to the trace metal, presenting fewer damaged nuclei. Statistical analysis compared between level 0 and total damage parameters showed higher genotoxicity promoted by Hg in HNS of the exposed group, compared to the control group. While genotoxicity in MT was similar between control and exposed groups. Therefore, it can be concluded that HNS is more responsive to Hg exposure than MT. It was concluded that the concentration of Hg considered safe by CONAMA compromised the cells that compose the HNS, impairing the general physiology of the studied species, even if the TM were not affected.O mercúrio (Hg) é um metal traço não essencial, altamente tóxico aos seres vivos e um dos principais contaminantes dos corpos d’água. Apesar da importância genotóxica do Hg, não há estudos sobre sua genotoxicidade em abelhas nativas neotropicais, utilizando biomarcadores de efeito. Pelo exposto, realizou-se um estudo da genotoxicidade do Hg em abelhas da espécie Bombus atratus por meio do ensaio cometa. Este ensaio determina a extensão do dano ocorrido no DNA de núcleos celulares individualmente, através da migração diferencial de fragmentos do DNA em microeletroforese. Apesar da efetividade e sensibilidade da técnica, não há registros na literatura da aplicabilidade desse ensaio em abelhas neotropicais do gênero Bombus, o qual apresenta muitas espécies ameaçadas de extinção. Operárias de B. atratus foram coletadas no munícipio de Sorocaba, SP, (23º34'53.1''S 47º31'29.5'') durante os meses de fevereiro a março de 2019. Após a coleta, as operárias foram colocadas, individualmente, em caixas especiais para o ensaio toxicológico e mantidas em incubadora a 26 oC, 70% de umidade, no escuro, por 48 horas, conforme recomendações da OECD. Foram utilizadas 11 operárias para o grupo exposto ao Hg à concentração de 0,2 µg L -1, concentração de Hg considerada segura pelo CONAMA (nº 357, 2005), e mesmo número para o grupo controle. Ambos os grupos foram mantidos com uma solução de sacarose a 70%, adicionalmente ao grupo exposto ofertou-se solução de 0,2 µg L -1 de Hg e ao grupo controle, água não contaminada. Após 48h, os tecidos que compõe o Sistema Hepatonefrocítico (SHN), vaso dorsal, hemócitos, corpo gorduroso e células pericárdicas e os túbulos de Malpighi (TM) foram coletados e processados para a realização do Ensaio Cometa. Os resultados mostraram que o Hg promoveu genotoxicidade nestes animais. Com relação ao grupo controle, o SHN das operárias expostas ao Hg mostrou-se sensível à contaminação pela presença abundante de diferentes níveis de cometas, os quais indicam a fragmentação do material genético, enquanto os TM, por sua vez, foram menos responsivos ao metal traço, apresentando menor quantidade de núcleos danificados. A análise estatística comparada entre os parâmetros nível 0 e dano total mostrou maior genotoxicidade promovida pelo Hg no SHN do grupo exposto com relação ao grupo controle. Por outro lado, a genotoxicidade nos TM foi semelhante entre os grupos controle e exposto. Portanto, conclui-se que o SHN é mais responsivo à exposição ao Hg do que os TM. Também se concluiu que a concentração de Hg considerada segura pelo CONAMA comprometeu as células que compõe o SHN, prejudicando a fisiologia geral da espécie estudada, mesmo que os TM não tenham sido afetados.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)porUniversidade Federal de São CarlosCâmpus SorocabaPrograma de Pós-Graduação em Biotecnologia e Monitoramento Ambiental - PPGBMA-SoUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessDano DNAAbelhasMetalGenotoxicidadeDNA damageBeesMetalGenotoxicityCIENCIAS BIOLOGICAS::BIOLOGIA GERALPadronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini)Standardisation and application of the comet assay to analyze the genotoxicity of mercury in bees of the Bombus atratus species (Hymenoptera, Bombini)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis600600c4f0703e-797c-491e-889d-533cc39b3edbreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdfDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdfDissertação de mestradoapplication/pdf2905864https://repositorio.ufscar.br/bitstream/ufscar/13486/1/Disserta%c3%a7%c3%a3o%20de%20mestrado%20-%20Let%c3%adcia%20Ceschi%20Bertoli%202020.pdfeb543fd27f263b0331c3a31f053efdfeMD51Carta Comprovante - Fabio Abdalla.docCarta Comprovante - Fabio Abdalla.docCarta comprovante do orientadorapplication/msword284160https://repositorio.ufscar.br/bitstream/ufscar/13486/3/Carta%20Comprovante%20-%20Fabio%20Abdalla.doc9a2259d5a823ed5386bb3328235c2452MD53CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufscar.br/bitstream/ufscar/13486/4/license_rdfe39d27027a6cc9cb039ad269a5db8e34MD54TEXTDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdf.txtDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdf.txtExtracted texttext/plain209099https://repositorio.ufscar.br/bitstream/ufscar/13486/5/Disserta%c3%a7%c3%a3o%20de%20mestrado%20-%20Let%c3%adcia%20Ceschi%20Bertoli%202020.pdf.txt21071f6c543ede6467ae192ed09ee396MD55Carta Comprovante - Fabio Abdalla.doc.txtCarta Comprovante - Fabio Abdalla.doc.txtExtracted texttext/plain1429https://repositorio.ufscar.br/bitstream/ufscar/13486/7/Carta%20Comprovante%20-%20Fabio%20Abdalla.doc.txt3705f8570089a6c181904b13997b8d79MD57THUMBNAILDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdf.jpgDissertação de mestrado - Letícia Ceschi Bertoli 2020.pdf.jpgIM Thumbnailimage/jpeg7285https://repositorio.ufscar.br/bitstream/ufscar/13486/6/Disserta%c3%a7%c3%a3o%20de%20mestrado%20-%20Let%c3%adcia%20Ceschi%20Bertoli%202020.pdf.jpg83280c8ca74180c261ef66db1b0ba6b7MD56ufscar/134862023-09-18 18:32:05.003oai:repositorio.ufscar.br:ufscar/13486Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:32:05Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
dc.title.alternative.por.fl_str_mv |
Standardisation and application of the comet assay to analyze the genotoxicity of mercury in bees of the Bombus atratus species (Hymenoptera, Bombini) |
title |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
spellingShingle |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) Ceschi-Bertoli, Letícia Dano DNA Abelhas Metal Genotoxicidade DNA damage Bees Metal Genotoxicity CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
title_full |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
title_fullStr |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
title_full_unstemmed |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
title_sort |
Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini) |
author |
Ceschi-Bertoli, Letícia |
author_facet |
Ceschi-Bertoli, Letícia |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/6218507280198988 |
dc.contributor.author.fl_str_mv |
Ceschi-Bertoli, Letícia |
dc.contributor.advisor1.fl_str_mv |
Abdalla, Fábio Camargo |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7770803221041927 |
dc.contributor.authorID.fl_str_mv |
840c0f61-23a5-4340-9c77-f9ea13ac06b8 |
contributor_str_mv |
Abdalla, Fábio Camargo |
dc.subject.por.fl_str_mv |
Dano DNA Abelhas Metal Genotoxicidade DNA damage Bees Metal Genotoxicity |
topic |
Dano DNA Abelhas Metal Genotoxicidade DNA damage Bees Metal Genotoxicity CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
The mercury (Hg) is a non-essential trace metal, that is highly toxic to living beings, is a major contaminant of water bodies. Despite the genotoxic importance of Hg, there are no studies on its genotoxicity in native neotropical bees, using effective biomarkers. Therefore, a study of the genotoxicity of Hg in Bombus atratus bees was carried out through the comet assay. This assay determines the extent of DNA damage to individual cell nuclei by differential migration of DNA fragments in microelectrophoresis. Despite the effectiveness and sensitivity of the technique, there are no records in the literature about the applicability of this assay to Bombus neotropical bees, which have many endangered species. B. atratus workers were collected from Sorocaba, SP, (23º34'53.1''S 47º31'29.5'') during the months of February to March 2019. After capture, the workers were individually placed in special boxes for the toxicological test and kept in an incubator at 26 ºC, 70% humidity, in the dark, for 48 hours, according to OECD recommendations. Eleven workers were used for the group exposed to Hg at a concentration of 0.2 µg L -1, concentration considered safe by CONAMA (nº 357, 2005), and the same number for the control group. Both groups were maintained with a 70% sucrose solution. In addition to the exposed group, 0.2 µg L -1 Hg solution was offered, and to the control group, uncontaminated water. After 48 hours, the tissues that compose the Hepato-Nephrocytic System (HNS), dorsal vessel, hemocytes, fat body/pericardial cells, and Malpighi tubules (MT), were collected and processed for the comet assay. The results showed that Hg promoted genotoxicity in these animals. Regarding the control group, the HNS of workers exposed to Hg was sensitive to contamination due to the abundant presence of different levels of comets, which indicate the fragmentation of genetic material. While MT, in turn, were less responsive to the trace metal, presenting fewer damaged nuclei. Statistical analysis compared between level 0 and total damage parameters showed higher genotoxicity promoted by Hg in HNS of the exposed group, compared to the control group. While genotoxicity in MT was similar between control and exposed groups. Therefore, it can be concluded that HNS is more responsive to Hg exposure than MT. It was concluded that the concentration of Hg considered safe by CONAMA compromised the cells that compose the HNS, impairing the general physiology of the studied species, even if the TM were not affected. |
publishDate |
2020 |
dc.date.accessioned.fl_str_mv |
2020-11-26T11:59:07Z |
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2020-11-26T11:59:07Z |
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2020-03-03 |
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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 |
CESCHI-BERTOLI, Letícia. Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini). 2020. Dissertação (Mestrado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/13486. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/13486 |
identifier_str_mv |
CESCHI-BERTOLI, Letícia. Padronização e aplicação do ensaio cometa para análise de genotoxicidade do mercúrio em abelhas da espécie Bombus atratus (Hymenoptera, Bombini). 2020. Dissertação (Mestrado em Biotecnologia e Monitoramento Ambiental) – Universidade Federal de São Carlos, Sorocaba, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/13486. |
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https://repositorio.ufscar.br/handle/ufscar/13486 |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
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openAccess |
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Universidade Federal de São Carlos Câmpus Sorocaba |
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Programa de Pós-Graduação em Biotecnologia e Monitoramento Ambiental - PPGBMA-So |
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Universidade Federal de São Carlos Câmpus Sorocaba |
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