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Mechanisms and DNA specificity in site-specific recombination of integron cassettes /Johansson, Carolina, January 2007 (has links)
Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 4 uppsatser.
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Caracterização de cepas de Pseudomonas spp isoladas de efluente hospitalar não tratado : resistência a beta-lactâmicos e presença de integrons / Characterization of Pseudomonas spp strains isolated from untreated hospital effluents: beta-lactams resistance and presence of integronsSpindler, Aline January 2009 (has links)
As bactérias contendo este elemento de mobilidade genética podem contribuir para a disseminação de determinantes de resistência, bem como servir de reservatório em potencial para estes genes. / Pseudomonas genus, widely distributed in the environment, is often found associated to determinants of beta-lactams resistance and presence of integrons. The present study was conducted to investigate the resistance profile of Pseudomonas spp strains isolated from untreated hospital effluents, likewise beta-lactamase genes and integrons. Untreated hospital effluents were collected from four hospitals in Porto Alegre, RS. Non-fermenter bacteria were isolated; the identification was confirmed by 16S rRNA PCR. Susceptibility testing was determined by the disk-diffusion method using 11 different betalactams antimicrobials. The beta-lactamase genes blaIMP, blaVIM, blaSPM-1, blaOXA-23-like, blaOXA-24-like, blaOXA-51-like and blaGES-like genes and the integrons genes intI1, intI2 and intI3 were determined by PCR using specific primers. One hundred and twenty-eight isolates were recovered; the most common species was P. pseudoalcaligenes. The resistance level found was considered high, 62 (50%) isolates were multi-resistants. No isolate carrying the beta-lactamase genes tested was found among the strains. Of 68 isolates considered positives for integrons, 52 were identified as carrying the class 1 integron gene, intI1. No isolate carrying class 1 or class 2 integrons was found among the strains. Untreated hospital effluents could be a source of environmental contamination due to discharge of antimicrobial resistant bacteria, in the case of the present study, also integron positives. Bacteria carrying this genetic mobile element can contribute for the dissemination of resistance determinants and also act like a potential reservoir for many types of resistance genes.
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Caracterização de cepas de Pseudomonas spp isoladas de efluente hospitalar não tratado : resistência a beta-lactâmicos e presença de integrons / Characterization of Pseudomonas spp strains isolated from untreated hospital effluents: beta-lactams resistance and presence of integronsSpindler, Aline January 2009 (has links)
As bactérias contendo este elemento de mobilidade genética podem contribuir para a disseminação de determinantes de resistência, bem como servir de reservatório em potencial para estes genes. / Pseudomonas genus, widely distributed in the environment, is often found associated to determinants of beta-lactams resistance and presence of integrons. The present study was conducted to investigate the resistance profile of Pseudomonas spp strains isolated from untreated hospital effluents, likewise beta-lactamase genes and integrons. Untreated hospital effluents were collected from four hospitals in Porto Alegre, RS. Non-fermenter bacteria were isolated; the identification was confirmed by 16S rRNA PCR. Susceptibility testing was determined by the disk-diffusion method using 11 different betalactams antimicrobials. The beta-lactamase genes blaIMP, blaVIM, blaSPM-1, blaOXA-23-like, blaOXA-24-like, blaOXA-51-like and blaGES-like genes and the integrons genes intI1, intI2 and intI3 were determined by PCR using specific primers. One hundred and twenty-eight isolates were recovered; the most common species was P. pseudoalcaligenes. The resistance level found was considered high, 62 (50%) isolates were multi-resistants. No isolate carrying the beta-lactamase genes tested was found among the strains. Of 68 isolates considered positives for integrons, 52 were identified as carrying the class 1 integron gene, intI1. No isolate carrying class 1 or class 2 integrons was found among the strains. Untreated hospital effluents could be a source of environmental contamination due to discharge of antimicrobial resistant bacteria, in the case of the present study, also integron positives. Bacteria carrying this genetic mobile element can contribute for the dissemination of resistance determinants and also act like a potential reservoir for many types of resistance genes.
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Caracterização de cepas de Pseudomonas spp isoladas de efluente hospitalar não tratado : resistência a beta-lactâmicos e presença de integrons / Characterization of Pseudomonas spp strains isolated from untreated hospital effluents: beta-lactams resistance and presence of integronsSpindler, Aline January 2009 (has links)
As bactérias contendo este elemento de mobilidade genética podem contribuir para a disseminação de determinantes de resistência, bem como servir de reservatório em potencial para estes genes. / Pseudomonas genus, widely distributed in the environment, is often found associated to determinants of beta-lactams resistance and presence of integrons. The present study was conducted to investigate the resistance profile of Pseudomonas spp strains isolated from untreated hospital effluents, likewise beta-lactamase genes and integrons. Untreated hospital effluents were collected from four hospitals in Porto Alegre, RS. Non-fermenter bacteria were isolated; the identification was confirmed by 16S rRNA PCR. Susceptibility testing was determined by the disk-diffusion method using 11 different betalactams antimicrobials. The beta-lactamase genes blaIMP, blaVIM, blaSPM-1, blaOXA-23-like, blaOXA-24-like, blaOXA-51-like and blaGES-like genes and the integrons genes intI1, intI2 and intI3 were determined by PCR using specific primers. One hundred and twenty-eight isolates were recovered; the most common species was P. pseudoalcaligenes. The resistance level found was considered high, 62 (50%) isolates were multi-resistants. No isolate carrying the beta-lactamase genes tested was found among the strains. Of 68 isolates considered positives for integrons, 52 were identified as carrying the class 1 integron gene, intI1. No isolate carrying class 1 or class 2 integrons was found among the strains. Untreated hospital effluents could be a source of environmental contamination due to discharge of antimicrobial resistant bacteria, in the case of the present study, also integron positives. Bacteria carrying this genetic mobile element can contribute for the dissemination of resistance determinants and also act like a potential reservoir for many types of resistance genes.
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Intégrons de classe 3 : aspects mécanistiques et épidémiologiques / Class 3 integrons : machanistical and epidemiological aspectsSimo Tchuinte, Pierrette Landrie 24 March 2016 (has links)
Les intégrons sont des supports génétiques bactériens de capture, d’expression et de dissémination de gènes de résistance aux antibiotiques sous forme de cassettes. Ils sont majoritairement décrits chez les bactéries à Gram négatif chez qui ils confèrent généralement un phénotype de multirésistance. Les intégrons de résistance (IR) jouent un rôle majeur dans l’acquisition de la résistance dans le monde bactérien. Il existe 3 principales classes d’IR ; les IR de classe 1, les IR de classe 2 et les IR de classe 3 (IR3). Contrairement aux 2 premières classes, les IR3 représentent la classe d’intégrons de résistance la moins étudiée. Très peu de travaux s’intéressent à leur étude et on dénombre actuellement moins de 10 IR3 entièrement caractérisés. Les objectifs de ce travail de thèse étaient (i) d’effectuer une étude épidémiologique des IR3 en France et au Cameroun et (ii) d’étudier les modalités d’expression de l’intégrase et des cassettes de ces intégrons. Nos travaux ont permis d’isoler puis de décrire 3 nouveaux IR3 présents au sein de bactéries environnementales appartenant aux genres Aeromonas, Acinetobacter et Citrobacter. Les cassettes de ces IR3 codent des résistances aux bétalactamines, aminosides et ammoniums quaternaires. De plus, nous avons caractérisé des IR3 dans 3 souches de Delftia spp. (2 D.acidovorans et 1 D. tsuruhatensis) isolées en Afrique ; les cassettes de ces intégrons ne codent pas de résistance aux antibiotiques. L’axe plus fondamental de ce travail de thèse a permis de montrer que le PintI3(1) est le promoteur impliqué dans l’expression du gène intI3. De plus, nous avons montré que les variants du promoteur Pc, ainsi que les variants du promoteur PintI3(1) sont fonctionnels et de force différente. Il ressort de nos travaux que l’environnement constituerait un réservoir d’intégrons de classe 3 et que ces supports génétiques pourraient jouer un rôle important dans la dissémination de la résistance au sein de cet écosystème. / Integrons are bacterial genetic elements able to capture and express genes embedded within gene cassettes. They are widely described among Gram-negative bacteria and generally confer a multidrug resistance phenotype. Resistance integrons (RI) play an important role in the acquisition of antibiotic resistance. There are 3 main classes of RI. Class 3 RI has been poorly studied class with less than ten fully IR3 characterized. Objectives of this thesis were (i) to conduct an epidemiological study of class 3 RI in France and Cameroon and (ii) to better understand the modes of expression of the integrase and cassettes of IR3. We described 3 new class 3 RI isolated from environmental bacteria belonging to genus Aeromonas, Acinetobacter and Citrobacter. Gene cassettes encoded resistance to betalactams, aminoglycosides and quaternary ammonium compounds. We also described IR3 from three Delftia strains (2 D.acidovorans and 1 D.tsuruhatensis) in Africa containing cassettes that do not encode antibiotic resistance. The fundamental part of the work showed that the PintI3(1) promoter is involved in the expression of the intI3 gene. Furthermore, we demonstrated that variants of the Pc promoter and variants of the PintI3(1) promoter are functional with different strengths. These results showed that the environment may constitute a reservoir of class 3 integrons and that these genetic elements could play an important role in the spread of the resistance in this ecosystem.
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Wind Cave: Direct Access to a Deep Subsurface Aquifer Reveals a Diverse Microbial Community and Unusual Manganese MetabolismHershey, Olivia Suzanne 30 November 2021 (has links)
No description available.
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Análise molecular de mecanismos determinantes de resistência a antibióticos em Pseudomonas aeruginosa e Acinetobacter ssp. / Molecular evaluation of the mechanisms that determine antimicrobial resistance in Pseudomonas aeruginosa and Acinetobacter spp.Clímaco, Eduardo Carneiro 19 August 2011 (has links)
P. aeruginosa e espécies de Acinetobacter são causas comuns de diversas infecções em pacientes hospitalizados, principalmente nos internados em centros de tratamento intensivo. Além disso, esses microrganismos se destacam por apresentarem resistência, intrínseca e adquirida, a várias classes de antibióticos, conferindo à bactéria fenótipos de multirresistência e panresistência. O objetivo deste estudo foi avaliar a participação de integrons (elementos genéticos que carreiam genes de resistência), de genes codificadores de metalo--lactamases, da perda de porinas (canais protéicos da membrana externa), e da atividade de efluxo aumentada, como determinantes do fenótipo de multirresistência e panresistência. Foram estudadas 147 P. aeruginosa e 57 Acinetobacter spp. isolados de pacientes hospitalizados no Hospital Universitário da Universidade Federal de Juiz de Fora, no período de 2003 a 2006. O perfil de sensibilidade destes isolados foi determinado por disco de difusão e utilizado para classificá-las como multirresistentes (MDR) e não multirresistentes (n-MDR). A variabilidade clonal dos isolados foi investigada por PFGE. Os isolados pertencentes aos grupos MDR e n-MDR foram investigados quanto a presença de integrons de classe 1, 2 e 3, por PCR e análise de RFLP. Os cassetes gênicos contidos nestes integrons, assim como genes codificadores de carbapenemases (ex. IMP, VIM e SPM), foram detectados por PCR e identificados por seqüenciamento. Avaliação da expressão gênica de bombas de efluxo (mexB, mexY, mexD e adeB) e de porina (OprD) foi conduzida por real-time RT-PCR. Os dados apresentados para os isolados do grupo MDR foram comparados àqueles do grupo n-MDR e a associação entre os determinantes de resistência e o fenótipo MDR foi calculada estatisticamente. Fenótipo de multiresistência foi observado em 42,2% e 84,2% das P. aeruginosa e Acinetobacter spp. estudadas. Nenhum isolado bacteriano apresentou fenótipo panresistente. Em 65 (44,2%) dos isolados de P. aeruginosa, foram detectados integrons de classe 1. Esses elementos apresentaram relação estatisticamente significativa com fenótipos MDR em P. aeruginosa. Entretanto, a maioria desses integrons não carreava nenhum cassete gênico (43/65) ou continham apenas cassetes gênicos de resistência a aminoglicosídeos (19/65). Entre os isolados de Acinetobacter spp., 11 (17,5%) apresentaram integrons de classe 1 e 30 (47,6%) integrons de classe 2. Apenas os últimos foram estatisticamente associados com fenótipos MDR. A pesquisa de metalo--lactamase (MBL) revelou a produção de enzimas SPM em 24 isolados de P. aeruginosa. Os estudos de expresão gênica demonstraram que, entre os sistemas de efluxo mais relatados para P. aeruginosa, MexXY-OprM foi o que mostrou maior diferença entre o nível de expressão dos grupos MDR e n-MDR, sugerindo que este sistema de efluxo desempenha importante papel no fenótipo MDR. Diminuição, em média de 66,4%, da produçãode OprD também foi um padrão encontrado nos isolados MDRem relação aos n-MDR. Dois grupos clonais de P. aeruginosa e dois de Acinetobacter spp. foram predominantes e tiveram relação com presença de integrons, produção de SPM-1 e com fenótipo MDR. Portanto, esse fenótipo pode ser consequência de acúmulo de determinantes de resistência em clones específicos. / The non-fermenting pathogenic bacteria Pseudomonas aeruginosa and Acinetobacter spp. are important causes of nosocomial infections. Theses species are often associated with a multidrug resistance (MDR) phenotype, due to intrinsic and acquired resistance genes. Some determinants of resistance, such as integrons, carbapenemases, overexpression of efflux systems and porins loss may be associated with the MDR phenotype. The aim of this study was to evaluate the association of non-MDR and MDR phenotypes in P. aeruginosa and Acinetobacter spp. to the presence of integrons and carbapenemases encoding genes, the overexpression of mexY, mexB, mexD and adeB genes and loss of the outer membrane protein, OprD. These resistance determinants were evaluated in 147 P. aeruginosa and 57 Acinetobacter spp., isolated from in-patients of University Hospital of UFJF. Isolates with different PFGE and non-susceptibility profiles were grouped according to MDR or non-MDR phenotypes. PCR and real-time RT-PCR were used to investigate the presence of class 1, 2 and 3 integrons and carbapenemase encoding genes and the expression of mexY, mexB, mexD and adeB efflux pumps and OprD porin, respectively. Class 1 integrons were one of the most common genetic elements present in MDR P. aeruginosa (44,2%), but the phenotype could not be attributed to these elements, since they showed empty (43/65) or only aminoglycoside gene cassettes (19/65). Class 2 integrons were the most common genetic elements in MDR Acinetobacter spp., and this association was statistically significant. SPM encoding gene was the only carbapenemase gene found in P. aeruginosa and, predominantly, in the PFGE cluster A. Expression of MexXY-OprM determined by real-time RT-PCR was the highest variable between MDR and non-MDR P. aeruginosa isolates (almost 10-fold). Reduction of 66.4% in OprD expression was observed in MDR P. aeruginosa, in comparison with non-MDR ones. It is concluded that the most important genetic determinants in the MDR phenotype of P. aeruginosa were SPM-1 production, followed by MexXY-OprM over expression and diminished production of OprD, while class 2 integrons was the most important genetic determinant of MDR phenotype in A. baumannii.
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Análise molecular de mecanismos determinantes de resistência a antibióticos em Pseudomonas aeruginosa e Acinetobacter ssp. / Molecular evaluation of the mechanisms that determine antimicrobial resistance in Pseudomonas aeruginosa and Acinetobacter spp.Eduardo Carneiro Clímaco 19 August 2011 (has links)
P. aeruginosa e espécies de Acinetobacter são causas comuns de diversas infecções em pacientes hospitalizados, principalmente nos internados em centros de tratamento intensivo. Além disso, esses microrganismos se destacam por apresentarem resistência, intrínseca e adquirida, a várias classes de antibióticos, conferindo à bactéria fenótipos de multirresistência e panresistência. O objetivo deste estudo foi avaliar a participação de integrons (elementos genéticos que carreiam genes de resistência), de genes codificadores de metalo--lactamases, da perda de porinas (canais protéicos da membrana externa), e da atividade de efluxo aumentada, como determinantes do fenótipo de multirresistência e panresistência. Foram estudadas 147 P. aeruginosa e 57 Acinetobacter spp. isolados de pacientes hospitalizados no Hospital Universitário da Universidade Federal de Juiz de Fora, no período de 2003 a 2006. O perfil de sensibilidade destes isolados foi determinado por disco de difusão e utilizado para classificá-las como multirresistentes (MDR) e não multirresistentes (n-MDR). A variabilidade clonal dos isolados foi investigada por PFGE. Os isolados pertencentes aos grupos MDR e n-MDR foram investigados quanto a presença de integrons de classe 1, 2 e 3, por PCR e análise de RFLP. Os cassetes gênicos contidos nestes integrons, assim como genes codificadores de carbapenemases (ex. IMP, VIM e SPM), foram detectados por PCR e identificados por seqüenciamento. Avaliação da expressão gênica de bombas de efluxo (mexB, mexY, mexD e adeB) e de porina (OprD) foi conduzida por real-time RT-PCR. Os dados apresentados para os isolados do grupo MDR foram comparados àqueles do grupo n-MDR e a associação entre os determinantes de resistência e o fenótipo MDR foi calculada estatisticamente. Fenótipo de multiresistência foi observado em 42,2% e 84,2% das P. aeruginosa e Acinetobacter spp. estudadas. Nenhum isolado bacteriano apresentou fenótipo panresistente. Em 65 (44,2%) dos isolados de P. aeruginosa, foram detectados integrons de classe 1. Esses elementos apresentaram relação estatisticamente significativa com fenótipos MDR em P. aeruginosa. Entretanto, a maioria desses integrons não carreava nenhum cassete gênico (43/65) ou continham apenas cassetes gênicos de resistência a aminoglicosídeos (19/65). Entre os isolados de Acinetobacter spp., 11 (17,5%) apresentaram integrons de classe 1 e 30 (47,6%) integrons de classe 2. Apenas os últimos foram estatisticamente associados com fenótipos MDR. A pesquisa de metalo--lactamase (MBL) revelou a produção de enzimas SPM em 24 isolados de P. aeruginosa. Os estudos de expresão gênica demonstraram que, entre os sistemas de efluxo mais relatados para P. aeruginosa, MexXY-OprM foi o que mostrou maior diferença entre o nível de expressão dos grupos MDR e n-MDR, sugerindo que este sistema de efluxo desempenha importante papel no fenótipo MDR. Diminuição, em média de 66,4%, da produçãode OprD também foi um padrão encontrado nos isolados MDRem relação aos n-MDR. Dois grupos clonais de P. aeruginosa e dois de Acinetobacter spp. foram predominantes e tiveram relação com presença de integrons, produção de SPM-1 e com fenótipo MDR. Portanto, esse fenótipo pode ser consequência de acúmulo de determinantes de resistência em clones específicos. / The non-fermenting pathogenic bacteria Pseudomonas aeruginosa and Acinetobacter spp. are important causes of nosocomial infections. Theses species are often associated with a multidrug resistance (MDR) phenotype, due to intrinsic and acquired resistance genes. Some determinants of resistance, such as integrons, carbapenemases, overexpression of efflux systems and porins loss may be associated with the MDR phenotype. The aim of this study was to evaluate the association of non-MDR and MDR phenotypes in P. aeruginosa and Acinetobacter spp. to the presence of integrons and carbapenemases encoding genes, the overexpression of mexY, mexB, mexD and adeB genes and loss of the outer membrane protein, OprD. These resistance determinants were evaluated in 147 P. aeruginosa and 57 Acinetobacter spp., isolated from in-patients of University Hospital of UFJF. Isolates with different PFGE and non-susceptibility profiles were grouped according to MDR or non-MDR phenotypes. PCR and real-time RT-PCR were used to investigate the presence of class 1, 2 and 3 integrons and carbapenemase encoding genes and the expression of mexY, mexB, mexD and adeB efflux pumps and OprD porin, respectively. Class 1 integrons were one of the most common genetic elements present in MDR P. aeruginosa (44,2%), but the phenotype could not be attributed to these elements, since they showed empty (43/65) or only aminoglycoside gene cassettes (19/65). Class 2 integrons were the most common genetic elements in MDR Acinetobacter spp., and this association was statistically significant. SPM encoding gene was the only carbapenemase gene found in P. aeruginosa and, predominantly, in the PFGE cluster A. Expression of MexXY-OprM determined by real-time RT-PCR was the highest variable between MDR and non-MDR P. aeruginosa isolates (almost 10-fold). Reduction of 66.4% in OprD expression was observed in MDR P. aeruginosa, in comparison with non-MDR ones. It is concluded that the most important genetic determinants in the MDR phenotype of P. aeruginosa were SPM-1 production, followed by MexXY-OprM over expression and diminished production of OprD, while class 2 integrons was the most important genetic determinant of MDR phenotype in A. baumannii.
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Implication des intégrons dans l’adaptation des communautés bactériennes / Integron implication into bacterial community adaptationAbella, Justine,Marie 09 December 2015 (has links)
Les intégrons sont des éléments génétiques bactériens. Découverts récemment dans le contexte clinique, ils sont présents dans le génome d’un certain nombre de bactéries provenant d’environnements très variés. Ils sont composés d’un gène codant une intégrase et d’une succession de cassettes de gènes. L’activité de l’intégrase permet l’acquisition, la perte ou le réarrangement des cassettes. Par ailleurs un promoteur permet l’expression des cadres de lecture contenus dans les cassettes de gène. Ainsi les intégrons sont à la fois des réservoirs de gènes et des systèmes d'expression de ces gènes. Dans le contexte clinique, ils sont connus pour être impliqués dans l'adaptation des bactéries pathogènes. Ils sont en effet capables d'acquérir et de diffuser des gènes conférant un avantage sélectif face à la pression exercée par l’usage des antibiotiques et des biocides, et par ailleurs d’être mobilisés afin d’être transférés horizontalement. Quelques études ont porté sur les intégrons en dehors des environnements cliniques. Elles ont permis de caractériser de nombreuses cassettes de gènes, sans toutefois en atteindre toute la diversité, à partir de bactéries ou de communautés bactériennes issues d’environnements soumis à différents niveaux de contaminations. Cependant, la diversité de l'intégrase a été peu étudiée, car le plus souvent les études se sont limitées aux séquences d’intégrons cliniques. Ainsi, les intégrons environnementaux sont encore mal connus et mal caractérisés. Les objectifs de ma thèse étaient de caractériser la diversité des intégrons environnementaux, avec un focus particulier sur les intégrases, à partir d’environnements soumis à des contaminations chimiques variables, dans le but d’évaluer le rôle possible des intégrons dans l'adaptation des bactéries face à des perturbations environnementales. Au cours de ces travaux, environ 800 séquences d’intégrases différentes, pour la plupart encore inconnues, ont été obtenues à partir de différents sédiments d’eau douce et côtiers. Des études in situ et en microcosmes, d’environnements d’eau douce ou de milieux côtiers, et avec différents types et niveaux de polluants, ont permis de mettre en évidence un impact des contaminations du milieu sur la diversité des intégrons, sur l’intégrase comme sur les cassettes de gène, de manière indépendante à la structure de la communauté bactérienne. Enfin, lors de cette thèse a été réalisée la caractérisation d’un intégron potentiellement adaptatif face à une pollution pétrolière, porteur de la séquence intIOPS mise en évidence et nommée par Lionel Huang lors de sa thèse. Finalement, les résultats obtenus lors de cette thèse ont apporté de nouveaux éléments qui viennent soutenir notre hypothèse principale que les intégrons environnementaux seraient impliqués dans l’adaptation des communautés bactériennes en réponse à la présence de contaminants dans les milieux non cliniques. / Integrons are bacterial genetic elements. Recently discovered in the clinical context, they are present in the genome of a number of bacteria from a variety of environments. They are composed of a gene encoding an integrase and a succession of gene cassettes. The activity of integrase allows the acquisition, loss or rearrangement of cassettes. Furthermore, a promoter allows expression reading frames contained in the gene cassettes. Thus, integrons are both reservoirs of genes and these gene expression systems. In the clinical context, they are known to be involved in the adaptation of pathogenic bacteria. They are able to acquire and disseminate genes conferring a selective advantage over the pressure exerted by the use of antibiotics and biocides, and also being mobilized to be transferred horizontally. Some studies have focused on integrons outside of clinical environments. They have characterized many gene cassettes, without however reaching the diversity, from bacteria or bacterial communities coming from environments with different levels of contamination. However, the diversity of integrase has been little studied, because the majority of studies are limited to clinical integron sequences. Thus, environmental integrons are still poorly characterized and their diversity are little understood. The objectives of my thesis were to characterize the diversity of environmental integrate with a particular focus on integrase, from environments with varying chemical contamination, to evaluate the possible role of integrating in adapting bacteria face environmental disturbances. In this work, approximately 800 different integrase sequences, mostly unknown, were obtained from various freshwater and coastal water sediments. Field studies and microcosms of freshwater or coastal environments, with different types and levels of pollutants allowed to demonstrate an impact of environmental contaminations on the integron diversity, whether on the integrase or the gene cassettes, independently to the bacterial community structure. Finally, in this thesis the characterization of a potentially adaptive integron facing an oil pollution were performed. This integron carrying the intIOPS sequence highlighted and named by Lionel Huang during his thesis. Finally, the results obtained in this thesis provide further elements which support our main hypothesis that environmental integrons would be involved in the adaptation of bacterial communities in response to the presence of contaminants in non- clinical settings.
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Compartmentalization of class 1 integrons and IncP-1 plasmids in the Orne river (France), an aquatic ecosystem impacted by urban and industrial anthropogenic pressures / Compartimentation des intégrons de classe 1 et des plasmides IncP-1 dans la rivière Orne (France), un écosystème aquatique soumis à des pressions anthropiques urbaines et industriellesCruz Barrón, Magali de la 20 December 2018 (has links)
Les éléments génétiques mobiles (EGM) sont des structures génétiques fréquemment associées à la dissémination de gènes de résistance aux antibiotiques (GRA). Dans ce travail, nous avons utilisé deux EGM comme « proxies », les intégrons de classe 1 et les plasmides IncP-1, afin de mieux comprendre (i) le devenir possible des GRA une fois relargués dans un écosystème fluvial (l’Orne, France), ainsi que (ii) l’effet des pressions anthropiques sur leur persistance. À partir d'analyses de l'eau des rivières, nous avons pu montrer que les deux EGM ne se comportaient pas de la même manière. L'entrée des intégrons de classe 1 dans le système fluvial semblait être diffuse plutôt que ponctuelle, tandis que l'abondance du plasmide IncP-1 est relativement stable le long de la section de la rivière étudiée (23 km), indiquant ainsi une origine plutôt indigène. Les intrants anthropiques tels que les stations d’épuration des eaux usées ne semblent pas affecter l’abondance des EGM en raison d’un niveau trop élevé de dilution des effluents. Par ailleurs, il est intéressant de noter que les bactéries porteuses d’EGM semblaient être enrichies sur les matières en suspension, susceptibles de servir de véhicule pour amener des communautés de bactéries plus riches en EGM vers les sédiments. L'analyse de deux carottes de sédiment indique clairement que seules les couches supérieures présentent un niveau élevé de bactéries porteuses d’EGM. Ces abondances diminuent dans les couches plus profondes où seules des zones ponctuelles présentent des microréservoirs avec des abondances d’EGM plus élevées. Pour une carotte sédimentaire au moins, nous avons pu montrer que l'abondance relative d’EGM corrèle négativement la présence de polluants tel que le plomb ou certains HAP / Mobile genetic elements (MGEs) are genetic structures frequently associated to the dissemination of antibiotic resistance genes (ARGs). In this work, we used two of them as proxies, class 1 integrons and IncP-1 plasmids, to better understand (i) the possible fate of ARGs once released in a river ecosystem (Orne, France), as well as (ii) the effect of anthropogenic pressures on their persistence. From river water analyses, we could show that the two MGEs do not behave the same way. The entry of class 1 integrons in the river system appeared to be diffuse rather than punctual, while the abundance of IncP-1 plasmid is relatively stable along the river section studied (23 km) thus indicating a rather indigenous origin. Anthropic inputs such as wastewater treatment plant did not seem to affect the abundance of MGEs because a too high level of effluent dilution. Interestingly, MGE-bearing bacteria appeared to be enriched on suspended material, which is likely to serve as a vehicle to drive MGE-richer communities of bacteria toward the sediments. The analysis of two sediment cores clearly indicates that only the top layers displayed an elevated level of MGE-bearing bacteria. These abundances decrease in deeper layers where only localized zones display micro-reservoirs of elevated MGE abundances. For one sediment core at least, we could show that the relative abundance of MGE negatively correlates with pollutants such as lead or certain PAHs
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