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Isolation of an acetochlor detoxifying bacterium and cloning of an associated gene.Martin, Darren Patrick. 05 July 2013 (has links)
A Pseudomonas strain, AI08, which was capable of detoxifying the herbicide acetochlor (2-
chloro-N-ethoxymethyl-6'-ethylacet-o-toluide) was isolated from soils. The microbe was
isolated using a combination of batch culture enrichment techniques, phenotypic agar plate
based assays and a qualitative bioassay for detecting acetochlor detoxification. With the aid
of a bioassay developed specifically for the quantification of acetochlor concentrations, it was
determined that over a 21 day period Al 08 was capable of detoxifying 20 % of the acetochlor
present in a medium containing no other organic carbon and 53 % of the herbicide in a
medium containing glucose and yeast extract at concentrations of 0.02 g.l-l and 0.005 g.l-l
respectively. A fragment of A108 DNA was cloned in Escherichia coli which produced
recombinant cells with both elevated acetochlor resistance and the ability to detoxify 15 %
of the acetochlor present in a minimal nutrient medium (containing 0.02 g.l-l glucose and
0.005 g.l-l yeast extract) over a 21 day period. Partial sequencing of the cloned A108 DNA
revealed that it encoded an amino acid sequence with significant homology with the
dihydrolipoyltransacetylase component of the pyruvate dehydrogenase complexes of
Azotobacter vinlandii, E. coli and Alcaligenes eutrophus. Theories are proposed as to the
possible biochemical mechanisms whereby expression of the dihydrolipoyltransacetylase gene
of Al 08 in recombinant E. coli cells may function in the detoxification of acetochlor. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1995.
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Estudo da resposta antioxidativa de linhagens bacterianas na presença do herbicida Acetochlor / Study of the response antioxidative of bacterial strains in the presence of herbicide AcetochlorCarvalho, Giselle de 21 January 2008 (has links)
Atualmente existe uma grande preocupação com relação aos efeitos a curto e longo prazo que muitos herbicidas podem ter sobre a saúde pública e o meio ambiente. Estes podem causar sérios danos ao metabolismo celular, levando geralmente a inibição do crescimento em microrganismos. Em exposição a herbicidas, esses microrganismos aumentam significativamente a geração de EAOs (Espécies Ativas de Oxigênio), causando estado de estresse oxidativo. O objetivo desse trabalho foi estudar a relação entre o efeito do herbicida acetochlor e a resposta do sistema antioxidante. Três linhagens de Klebsiella oxytoca foram submetidas ao crescimento em meio nutritivo com este herbicida nas concentrações de 0, 62 e 620 mM. O perfil protéico foi avaliado em SDS-PAGE, o qual revelou diferenças de intensidade das bandas, bem como a presença e ausência das mesmas em meio com herbicida. O herbicida causou estresse oxidativo nas três linhagens, o qual foi avaliado através da peroxidação de lipídeos, observado principalmente na concentração de 620 mM. As atividades da Catalase (CAT) e Superóxido Dismutase (SOD) foram altas para as três linhagens na concentração de 62 mM seguida de queda em 620 mM. Para a atividade da Glutationa Redutase (GR) foi observado decréscimo gradativo da atividade com o respectivo aumento da concentração de herbicida para as três linhagens. Porém para a atividade da Glutationa-S-transferase (GST), as três linhagens apresentaram comportamento diferencial para os mesmos tratamentos. O estudo da resposta antioxidativa de microrganismos na presença de xenobióticos são importantes para a compreensão de mecanismos de tolerância, biodegradação bem como para aplicação na biorremediação de ambientes contaminados. / Currently there is a great concern with respect to short and long term effects that many herbicides may cause on public health and the environment. They may seriously damage the cellular metabolism, generally leading to inhibition of the microbial growth. With the exposure to herbicides, these microorganisms significantly increase the generation of EAOs (Active Oxygen Species), generating a state of oxidative stress. The aim of this work was to study the effect of acetochlor herbicide in the response of antioxidant system. Three strains of the bacteria Klebsiella oxytoca were grown in a nutritional media containing the herbicide in concentrations of 0, 62 and 620 mM. The protein profile was evaluated in SDS-PAGE, which showed differences in the intensity of the bands, as well as the presence and absence in the media with herbicide. The oxidative stress was evaluated through the lipid peroxidation, and herbicide caused stress in all the three strains, mainly at the concentration of 620 mM. The activities of Catalase (CAT) and Superoxide Dismutase (SOD) were higher for the three strains in the concentration of 62 mM followed by a decrease at the concentration of 620 mM. For Glutathione Reductase (GR) it was observed a gradual reduction in its activity with the increase of herbicide concentration for all the three strains. However, in the case of Glutathione-S-transferase (GST) activity, the three strains showed differential behavior for the same treatments. The study of the antioxidative response of microorganisms in the presence of xenobiotics is important for the understanding of tolerance mechanisms, biodegradation as well as for application in the bioremediation of contaminated environments.
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Estudo da resposta antioxidativa de linhagens bacterianas na presença do herbicida Acetochlor / Study of the response antioxidative of bacterial strains in the presence of herbicide AcetochlorGiselle de Carvalho 21 January 2008 (has links)
Atualmente existe uma grande preocupação com relação aos efeitos a curto e longo prazo que muitos herbicidas podem ter sobre a saúde pública e o meio ambiente. Estes podem causar sérios danos ao metabolismo celular, levando geralmente a inibição do crescimento em microrganismos. Em exposição a herbicidas, esses microrganismos aumentam significativamente a geração de EAOs (Espécies Ativas de Oxigênio), causando estado de estresse oxidativo. O objetivo desse trabalho foi estudar a relação entre o efeito do herbicida acetochlor e a resposta do sistema antioxidante. Três linhagens de Klebsiella oxytoca foram submetidas ao crescimento em meio nutritivo com este herbicida nas concentrações de 0, 62 e 620 mM. O perfil protéico foi avaliado em SDS-PAGE, o qual revelou diferenças de intensidade das bandas, bem como a presença e ausência das mesmas em meio com herbicida. O herbicida causou estresse oxidativo nas três linhagens, o qual foi avaliado através da peroxidação de lipídeos, observado principalmente na concentração de 620 mM. As atividades da Catalase (CAT) e Superóxido Dismutase (SOD) foram altas para as três linhagens na concentração de 62 mM seguida de queda em 620 mM. Para a atividade da Glutationa Redutase (GR) foi observado decréscimo gradativo da atividade com o respectivo aumento da concentração de herbicida para as três linhagens. Porém para a atividade da Glutationa-S-transferase (GST), as três linhagens apresentaram comportamento diferencial para os mesmos tratamentos. O estudo da resposta antioxidativa de microrganismos na presença de xenobióticos são importantes para a compreensão de mecanismos de tolerância, biodegradação bem como para aplicação na biorremediação de ambientes contaminados. / Currently there is a great concern with respect to short and long term effects that many herbicides may cause on public health and the environment. They may seriously damage the cellular metabolism, generally leading to inhibition of the microbial growth. With the exposure to herbicides, these microorganisms significantly increase the generation of EAOs (Active Oxygen Species), generating a state of oxidative stress. The aim of this work was to study the effect of acetochlor herbicide in the response of antioxidant system. Three strains of the bacteria Klebsiella oxytoca were grown in a nutritional media containing the herbicide in concentrations of 0, 62 and 620 mM. The protein profile was evaluated in SDS-PAGE, which showed differences in the intensity of the bands, as well as the presence and absence in the media with herbicide. The oxidative stress was evaluated through the lipid peroxidation, and herbicide caused stress in all the three strains, mainly at the concentration of 620 mM. The activities of Catalase (CAT) and Superoxide Dismutase (SOD) were higher for the three strains in the concentration of 62 mM followed by a decrease at the concentration of 620 mM. For Glutathione Reductase (GR) it was observed a gradual reduction in its activity with the increase of herbicide concentration for all the three strains. However, in the case of Glutathione-S-transferase (GST) activity, the three strains showed differential behavior for the same treatments. The study of the antioxidative response of microorganisms in the presence of xenobiotics is important for the understanding of tolerance mechanisms, biodegradation as well as for application in the bioremediation of contaminated environments.
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Studium průběhu degradace xenobiotik a biologicky aktivních látek s využitím oxidu titaničitého / Study of xenobiotics and biologically active compounds degradation on titanium dioxidePíšťková, Veronika January 2012 (has links)
Heterogenous photocatalysis using titanium dioxide seems to be a promising method for disposal xenobiotics from the environment. The aim of this diploma thesis is the study of degradation of selected xenobiotics and biologically active substrances applying this method. Theoretical part of diploma thesis deals with the principals of heterogenous photocatalysis by means of a semiconductor TiO2 and the examples of its possible application are mentioned too. The compounds which could be appropriate for a study of degradation were selected from the group of pharmaceuticals and pesticides. The properties of target substances and their environmental impact were described. Furthermore, a bibliographic search focused on the possibilities of their analytical determination was conducted. The experimental part of the thesis describes the experiments with photocatalyst in a form of powder as well as with immobilized photocatalyst in thin layer on a carrier. Identification and quantification of analytes was realized by high performance liquid chromatography with mass spectrometic detection.
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Using Contaminant Photoreactivity as a Holistic Indicator to Monitor Changes in Wetland Water CharacteristicsLanglois, Maureen Connell 21 October 2011 (has links)
No description available.
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