Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa

Detalhes bibliográficos
Autor(a) principal: Aragão, Francielen Barroso
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/9936
Resumo: The increase in food demand due to world population growth, results in increased use of synthetic substances in order to increase food productivity. However, the growing advance of monoculture stands out for significant volume of waste dispersed in environment, such as pesticides. These substances cause environmental impacts, because when leached can affect groundwater and surface water, damaging micro and macro local biota. Thus, the search for elucidation of hereditary or lethal effects of toxic substances in living organisms, various evaluation methods have been developed in order to understand the properties and mechanisms of action thereof. Therefore, the study aimed to evaluate the effect of tebuconazole fungicides (TBZ) and difenoconazole (DFZ) through different analysis tools in the plant model Lactuca sativa. Lactuca sativa the seeds were germinated in distilled water to achieve 1-2 mm, after reaching the minimum size were treated with different concentration of fungicides: TBZ (25; 50; 100; 200 e 400 g/mL) and DFZ (15,62; 31,25; 62,5; 125 e 250 g/mL). Trifluralin (0,84 g/L) and MMS (4x10-4 mol L-1 ) was used as a positive control (C +) and distilled water as a negative control (C-). Analysis of cytogenotoxic potential was performed through cell cycle analysis (mitotic index, chromosomal aberrations (CA) and nuclear aberration (NA), quantification of DNA damage by comet test and Tunnel test, as well as evaluation of oxidative stress through antioxidant enzymes quantification as: CAT, APX, SOD and POD. The means were submitted to ANOVA and compared by the Kruskal Wallis and Tukey tests (p <0.05). The different dilutions of TBZ and DFZ showed reduction in root growth compared to negative control. Compared with mitotic index of C-, the concentrations of DFZ showed no statistical difference, whereas the concentrations of TBZ showed a significant reduction in the concentration C1, and a significant increase in the other concentrations. However, both DFZ and TBZ showed a significant increase in CA in the highest concentrations. Both fungicides induced formation of CA as cmetaphases, bridges, and sticky chromosomes. The higher concentrations of the main principles increased the NA percentage. It was observed increased the DNA fragmentation evaluated by the comet and Tunel test in fungicide TBZ, and only for the TUNEL in the fungicide DFZ. Oxidative stress analysis showed that only the APX enzyme presented a statistical difference of C- for TBZ, whereas in DFZ only the SOD enzyme was statistically different from C-. Concerning to the tests carried out, it was observed that fungicides TBZ and DFZ were cytotoxic by reducing root growth, genotoxic by increasing the occurrence of NA and CA as well as increasing DNA fragmentation, and altered the enzymatic activity.
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spelling Matsumoto, Silvia TamieAragão, Francielen BarrosoCorte, Viviana BorgesAndrade-Vieira, Larissa Fonseca2018-08-02T00:16:05Z2018-08-012018-08-02T00:16:05Z2017-02-22The increase in food demand due to world population growth, results in increased use of synthetic substances in order to increase food productivity. However, the growing advance of monoculture stands out for significant volume of waste dispersed in environment, such as pesticides. These substances cause environmental impacts, because when leached can affect groundwater and surface water, damaging micro and macro local biota. Thus, the search for elucidation of hereditary or lethal effects of toxic substances in living organisms, various evaluation methods have been developed in order to understand the properties and mechanisms of action thereof. Therefore, the study aimed to evaluate the effect of tebuconazole fungicides (TBZ) and difenoconazole (DFZ) through different analysis tools in the plant model Lactuca sativa. Lactuca sativa the seeds were germinated in distilled water to achieve 1-2 mm, after reaching the minimum size were treated with different concentration of fungicides: TBZ (25; 50; 100; 200 e 400 g/mL) and DFZ (15,62; 31,25; 62,5; 125 e 250 g/mL). Trifluralin (0,84 g/L) and MMS (4x10-4 mol L-1 ) was used as a positive control (C +) and distilled water as a negative control (C-). Analysis of cytogenotoxic potential was performed through cell cycle analysis (mitotic index, chromosomal aberrations (CA) and nuclear aberration (NA), quantification of DNA damage by comet test and Tunnel test, as well as evaluation of oxidative stress through antioxidant enzymes quantification as: CAT, APX, SOD and POD. The means were submitted to ANOVA and compared by the Kruskal Wallis and Tukey tests (p <0.05). The different dilutions of TBZ and DFZ showed reduction in root growth compared to negative control. Compared with mitotic index of C-, the concentrations of DFZ showed no statistical difference, whereas the concentrations of TBZ showed a significant reduction in the concentration C1, and a significant increase in the other concentrations. However, both DFZ and TBZ showed a significant increase in CA in the highest concentrations. Both fungicides induced formation of CA as cmetaphases, bridges, and sticky chromosomes. The higher concentrations of the main principles increased the NA percentage. It was observed increased the DNA fragmentation evaluated by the comet and Tunel test in fungicide TBZ, and only for the TUNEL in the fungicide DFZ. Oxidative stress analysis showed that only the APX enzyme presented a statistical difference of C- for TBZ, whereas in DFZ only the SOD enzyme was statistically different from C-. Concerning to the tests carried out, it was observed that fungicides TBZ and DFZ were cytotoxic by reducing root growth, genotoxic by increasing the occurrence of NA and CA as well as increasing DNA fragmentation, and altered the enzymatic activity.O aumento na demanda de alimentos, devido ao crescimento da população mundial, acarreta no uso intensificado de substâncias sintéticas com a finalidade de ampliar a produtividade de alimentos. Entretanto, o crescente avanço de monoculturas destaca-se pelo significativo volume de resíduos dispersos no ambiente, como os agrotóxicos. Estas substâncias ocasionam impactos ambientais, pois quando lixiviadas podem afetar lençóis freáticos e águas superficiais, prejudicando, e consequentemente, a micro e macro biota local. Assim, na busca pela elucidação dos efeitos hereditários ou letais dessas substâncias tóxicas nos organismos vivos, diversos métodos de avaliação tem sido desenvolvidos com o intuito de compreender as propriedades e mecanismos de ação dos mesmos. Logo, o trabalho objetivou avaliar o efeito dos fungicidas tebuconazol (TBZ) e difenoconazol (DFZ) por meio de diferentes ferramentas de análise no modelo vegetal Lactuca sativa. As sementes de Lactuca sativa foram germinadas em água destilada até atingirem 1 a 2 mm, após alcançarem o tamanho mínimo foram tratadas com diferentes diluições dos fungicidas: Tebuconazol (12,5; 25; 50; 100 e 200 µL) e Difenoconazol (6,25; 12,5; 25; 50 e 100µL) em 100 ml de água destilada. A trifluralina foi utilizada como controle positivo (C+) e água destilada como controle negativo (C-). Foram realizadas análises de crescimento radicular, avaliação do potencial citogenotóxico por meio da análise de ciclo celular (índice mitótico, alterações cromossômicas (AC) e nucleares (AN)), quantificação dos danos ao DNA por meio do teste do cometa e teste do Túnel, bem como avaliação do estresse oxidativo por meio da quantificação das enzimas antioxidantes CAT, APX, SOD e POD. As médias foram submetidas a ANOVA e comparadas pelos testes de Kruskal Wallis e Tukey (p<0.05). As diferentes diluições do TBZ e DFZ apresentaram redução no crescimento radicular, com inibição de 36,87% e 83,04% para a maior concentração testada (C5) em relação ao controle negativo. Comparando-se com o índice mitótico do C-, as concentrações avaliadas de DFZ não apresentaram diferença estatística, enquanto as concentrações de TBZ apresentaram redução significativa na menor concentração, e aumento significativo nas demais concentrações. Todavia, tanto DFZ quanto TBZ apresentaram aumento significativo nas AC nas maiores concentrações. Ambos fungicidas induziram a formação de AC como cromossomos perdidos e quebrados, c-metáfases, pontes, c-metáfase poliplóides e cromossomos pegajosos. As maiores concentrações dos dois princípios aumentaram a porcentagem de AN. Observou-se que os tratamentos dos fungicidas aumentaram a fragmentação de DNA avaliados pelo teste do cometa e TUNEL no fungicida TBZ, onde todas as concentrações foram diferentes do C-. Enquanto no fungicida DFZ pelo teste TUNEL, houve aumento na quantidade de danos leves e diminuição de células sem danos, em relação ao C-. A análise do estresse oxidativo demonstrou que apenas a enzima APX apresentou uma diferença estatística do C- para o TBZ, enquanto no DFZ apenas a enzima SOD foi diferente estatisticamente do C-. Diante dos testes realizados observou-se que os fungicidas TBZ e DFZ foram citotóxicos por diminuir o crescimento radicular, genotóxicos por aumentar a ocorrência de AN e AC bem como elevar a fragmentação de DNA, e alteraram a atividade enzimática.TextARAGÃO, Francielen Barroso. Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa. 2017. 93 f. Dissertação (Mestrado em Biologia Vegetal) - Universidade Federal do Espírito Santo, Centro de Ciências Humanas e Naturais, Vitória, 2017.http://repositorio.ufes.br/handle/10/9936porUniversidade Federal do Espírito SantoMestrado em Biologia VegetalPrograma de Pós-Graduação em Biologia VegetalUFESBRCitogenotoxicityPlant comet assayFungicidesOxidative stressRoot growthCitogenotoxicidadeCometaEstresse oxidativoCrescimento radicularTUNELFungicidasMutagêneseEnzimasAlfaceAgronomia57Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESORIGINALPPGBV_FRANCIELEN ARAGAO_VERSÃO BANCA.pdfapplication/pdf1424885http://repositorio.ufes.br/bitstreams/97363a42-67fc-4dcd-9eb7-a3bab59d3837/download7d951bfbe1cd246be5a1e3e57f4992b2MD5110/99362024-07-01 16:12:27.755oai:repositorio.ufes.br:10/9936http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-07-01T16:12:27Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false
dc.title.none.fl_str_mv Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
title Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
spellingShingle Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
Aragão, Francielen Barroso
Citogenotoxicity
Plant comet assay
Fungicides
Oxidative stress
Root growth
Citogenotoxicidade
Cometa
Estresse oxidativo
Crescimento radicular
TUNEL
Agronomia
Fungicidas
Mutagênese
Enzimas
Alface
57
title_short Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
title_full Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
title_fullStr Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
title_full_unstemmed Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
title_sort Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa
author Aragão, Francielen Barroso
author_facet Aragão, Francielen Barroso
author_role author
dc.contributor.advisor1.fl_str_mv Matsumoto, Silvia Tamie
dc.contributor.author.fl_str_mv Aragão, Francielen Barroso
dc.contributor.referee1.fl_str_mv Corte, Viviana Borges
dc.contributor.referee2.fl_str_mv Andrade-Vieira, Larissa Fonseca
contributor_str_mv Matsumoto, Silvia Tamie
Corte, Viviana Borges
Andrade-Vieira, Larissa Fonseca
dc.subject.eng.fl_str_mv Citogenotoxicity
Plant comet assay
Fungicides
Oxidative stress
Root growth
topic Citogenotoxicity
Plant comet assay
Fungicides
Oxidative stress
Root growth
Citogenotoxicidade
Cometa
Estresse oxidativo
Crescimento radicular
TUNEL
Agronomia
Fungicidas
Mutagênese
Enzimas
Alface
57
dc.subject.por.fl_str_mv Citogenotoxicidade
Cometa
Estresse oxidativo
Crescimento radicular
TUNEL
dc.subject.cnpq.fl_str_mv Agronomia
dc.subject.br-rjbn.none.fl_str_mv Fungicidas
Mutagênese
Enzimas
Alface
dc.subject.udc.none.fl_str_mv 57
description The increase in food demand due to world population growth, results in increased use of synthetic substances in order to increase food productivity. However, the growing advance of monoculture stands out for significant volume of waste dispersed in environment, such as pesticides. These substances cause environmental impacts, because when leached can affect groundwater and surface water, damaging micro and macro local biota. Thus, the search for elucidation of hereditary or lethal effects of toxic substances in living organisms, various evaluation methods have been developed in order to understand the properties and mechanisms of action thereof. Therefore, the study aimed to evaluate the effect of tebuconazole fungicides (TBZ) and difenoconazole (DFZ) through different analysis tools in the plant model Lactuca sativa. Lactuca sativa the seeds were germinated in distilled water to achieve 1-2 mm, after reaching the minimum size were treated with different concentration of fungicides: TBZ (25; 50; 100; 200 e 400 g/mL) and DFZ (15,62; 31,25; 62,5; 125 e 250 g/mL). Trifluralin (0,84 g/L) and MMS (4x10-4 mol L-1 ) was used as a positive control (C +) and distilled water as a negative control (C-). Analysis of cytogenotoxic potential was performed through cell cycle analysis (mitotic index, chromosomal aberrations (CA) and nuclear aberration (NA), quantification of DNA damage by comet test and Tunnel test, as well as evaluation of oxidative stress through antioxidant enzymes quantification as: CAT, APX, SOD and POD. The means were submitted to ANOVA and compared by the Kruskal Wallis and Tukey tests (p <0.05). The different dilutions of TBZ and DFZ showed reduction in root growth compared to negative control. Compared with mitotic index of C-, the concentrations of DFZ showed no statistical difference, whereas the concentrations of TBZ showed a significant reduction in the concentration C1, and a significant increase in the other concentrations. However, both DFZ and TBZ showed a significant increase in CA in the highest concentrations. Both fungicides induced formation of CA as cmetaphases, bridges, and sticky chromosomes. The higher concentrations of the main principles increased the NA percentage. It was observed increased the DNA fragmentation evaluated by the comet and Tunel test in fungicide TBZ, and only for the TUNEL in the fungicide DFZ. Oxidative stress analysis showed that only the APX enzyme presented a statistical difference of C- for TBZ, whereas in DFZ only the SOD enzyme was statistically different from C-. Concerning to the tests carried out, it was observed that fungicides TBZ and DFZ were cytotoxic by reducing root growth, genotoxic by increasing the occurrence of NA and CA as well as increasing DNA fragmentation, and altered the enzymatic activity.
publishDate 2017
dc.date.issued.fl_str_mv 2017-02-22
dc.date.accessioned.fl_str_mv 2018-08-02T00:16:05Z
dc.date.available.fl_str_mv 2018-08-01
2018-08-02T00:16:05Z
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dc.identifier.citation.fl_str_mv ARAGÃO, Francielen Barroso. Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa. 2017. 93 f. Dissertação (Mestrado em Biologia Vegetal) - Universidade Federal do Espírito Santo, Centro de Ciências Humanas e Naturais, Vitória, 2017.
dc.identifier.uri.fl_str_mv http://repositorio.ufes.br/handle/10/9936
identifier_str_mv ARAGÃO, Francielen Barroso. Prospecção da toxicidade e atividade enzimática de fungicidas por meio de bioensaios com Lactuca sativa. 2017. 93 f. Dissertação (Mestrado em Biologia Vegetal) - Universidade Federal do Espírito Santo, Centro de Ciências Humanas e Naturais, Vitória, 2017.
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Mestrado em Biologia Vegetal
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Mestrado em Biologia Vegetal
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