Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal

Detalhes bibliográficos
Autor(a) principal: Freire, Caroline Fernandes [UNIFESP]
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5456892
http://repositorio.unifesp.br/handle/11600/50655
Resumo: Shiga toxin-producing Escherichia coli (STEC) are food-borne pathogens being ruminant animals their main natural reservoir. STEC is responsible for causing severe human illnesses as hemorragic colitis and the Hemolytic Uremic Syndrome (HUS). Recent studies have pointed out the prevalence of non-O157 STEC serotypes associated with both sporadic cases and outbreaks of severe diarrhea. Moreover, it has been observed that several non-O157 STEC serotypes unable to cause attaching and effacing lesion (A/E) have been implicated as agents of serious human diseases and HUS. Although Shiga toxin hás a central role in the pathogenesis of STEC diseases, other accessory virulence factors may also participate in this process. Therefore, to know the distribution of stx subtypes and accessory virulence genes in non-O157 STEC isolates devoid of A/E from the animal reservoir could indicate a higher pathogenic potential of these isolates and, therefore, constitutes the aimof this study. Gene sequences related to Stx1 and Stx2 subtypes, autotransporter proteins (EspP, EspI, EpeA, Sab), adhesins (Saa), toxins (EhxA) andiron uptake protein (Irp2) were sought by PCR in 125 STEC strains isolated from sheep, goat, buffalo and cattle. STEC isolates most frequently harbor stx2 (52%) and stx1stx2 (38%), being subtypes stx1a (57%), stx1c (47%), stx2a (67%), and stx2d (38%) those most frequently identified. Distribution of subtypes varied according to the reservoir. stx1a prevailed in cattle and buffalo isolates, stx1c predominate in sheep and goat, stx2a in cattle, and stx2d prevailed in goat and buffalo. The saa, espP, ehxA, espI,epeA, sab andirp2 genes were identified in 97%, 58%, 55%, 42%, 30%, 13% and11% of the STEC isolates, respectively, and differences were also observed depending on the reservoir. espP and ehxA molecular characterization showed that variants C (90%) and A (100%) prevailed, respectively. A great diversity of serotypes and virulence profiles wasobserved independently of the isolate origin. However, serotypes O39:H49, O79:H14, O113:H21 and O178:H19 were more representative among cattle and buffalos, and serotype O5:H- among goat isolates. This study made important contributions related to the virulence profile of STEC isolates from the animal reservoir in our settings. In addition, showed that STEC isolates from cattle most frequently carry the stx2 subtypes related to human infections, and harbor virulence factors at higher frequencies.
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spelling Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animalSearch of shiga toxin subtypes and accessory virulence makers in shiga toxin-producing Escherichia coli (STEC) devoid of eae gene isolated from animal reservoirsEscherichia coliDisease reservoirsVirulenceHemolytic-uremic syndromeEscherichia coliReservatórios de doençasVirulênciaSíndrome hemolítico-urêmicaShiga toxin-producing Escherichia coli (STEC) are food-borne pathogens being ruminant animals their main natural reservoir. STEC is responsible for causing severe human illnesses as hemorragic colitis and the Hemolytic Uremic Syndrome (HUS). Recent studies have pointed out the prevalence of non-O157 STEC serotypes associated with both sporadic cases and outbreaks of severe diarrhea. Moreover, it has been observed that several non-O157 STEC serotypes unable to cause attaching and effacing lesion (A/E) have been implicated as agents of serious human diseases and HUS. Although Shiga toxin hás a central role in the pathogenesis of STEC diseases, other accessory virulence factors may also participate in this process. Therefore, to know the distribution of stx subtypes and accessory virulence genes in non-O157 STEC isolates devoid of A/E from the animal reservoir could indicate a higher pathogenic potential of these isolates and, therefore, constitutes the aimof this study. Gene sequences related to Stx1 and Stx2 subtypes, autotransporter proteins (EspP, EspI, EpeA, Sab), adhesins (Saa), toxins (EhxA) andiron uptake protein (Irp2) were sought by PCR in 125 STEC strains isolated from sheep, goat, buffalo and cattle. STEC isolates most frequently harbor stx2 (52%) and stx1stx2 (38%), being subtypes stx1a (57%), stx1c (47%), stx2a (67%), and stx2d (38%) those most frequently identified. Distribution of subtypes varied according to the reservoir. stx1a prevailed in cattle and buffalo isolates, stx1c predominate in sheep and goat, stx2a in cattle, and stx2d prevailed in goat and buffalo. The saa, espP, ehxA, espI,epeA, sab andirp2 genes were identified in 97%, 58%, 55%, 42%, 30%, 13% and11% of the STEC isolates, respectively, and differences were also observed depending on the reservoir. espP and ehxA molecular characterization showed that variants C (90%) and A (100%) prevailed, respectively. A great diversity of serotypes and virulence profiles wasobserved independently of the isolate origin. However, serotypes O39:H49, O79:H14, O113:H21 and O178:H19 were more representative among cattle and buffalos, and serotype O5:H- among goat isolates. This study made important contributions related to the virulence profile of STEC isolates from the animal reservoir in our settings. In addition, showed that STEC isolates from cattle most frequently carry the stx2 subtypes related to human infections, and harbor virulence factors at higher frequencies.Escherichia coli produtora de toxina Shiga (STEC) é um importante patógeno de origem alimentar, sendo os ruminantes o seu principal reservatório natural. STEC é responsável por causar graves infecções humanas como a colite hemorrágica e a Síndrome Hemolítica Urêmica (SHU). Estudos recentes têm sinalizadopara uma prevalência de sorotipos STEC não-O157 associados tanto a casos esporádicos quanto a surtos de diarréia grave. Além disso, tem sido observado quedoenças graves e SHU podem ocorrer associadasa sorotiposincapazes de causar a lesão "attaching and effacing" (A/E).Embora a toxina Shiga tenha um papel central na patogênese das doenças causadas por STEC, é certo que fatores adicionais de virulência também participam deste processo. Conhecer entãoadistribuição dos subtipos de stx e a presença de diversos fatores "acessórios" de virulência, em amostras STEC incapazes de provocar a lesão A/E isoladas do reservatório animal poderia indicar um maior potencial de patogenicidade destas cepas e, portanto, constituiu o objetivo deste estudo. Sequencias genéticas relacionadas aos subtipos de Stx1 and Stx2, as proteínas autotransportadoras (EspP, EspI, EpeA, Sab), adesina (Saa), a enterohemolisina (EhxA) e a proteína ferro-reprimível (Irp2) foram pesquisadas pela técnica de reação em cadeia da polimerase (PCR) em 125 cepas de STEC isoladas de ovinos, caprinos, búfalos e bovinos. As cepas STEC foram mais frequentemente carreadoras de stx2 (52%) ede stx1stx2 (38%), sendo os subtipos stx1a (57%), stx1c (47%), stx2a (67%), estx2d (38%) aqueles mais frequentemente identificados. A distribuição dos subtipos variou de acordo com o reservatório..stx1aprevaleceu entre os isolados de bovinos e búfalos, stx1cpredominou entre ovinos e caprinos, stx2aem bovinos, estx2dentre caprinos e bufalos. Os genes saa, espP, ehxA, espI,epeA, sab eirp2 foram identificadosrespectivamente, em97%, 58%, 55%, 42%, 30%, 13% e11% dos isolados STEC, e diferenças foram observadas de acordo com o reservatório. A caracterização molecular deespPeehxAdemonstrou que as variantes C (90%) e A (100%) prevaleceram, respectivamente. Uma grande diversidade de sorotipos e perfis de virulência foi observada independentemente da origem das amostras. Porém, os sorotipos O39:H49, O79:H14, O113:H21 e O178:H19 foram mais representativos entre bovinos e bufalinos e o sorotipo O5:H- dentre as cepas isoladas de caprinos.Este estudo trouxe contribuições importantes sobre o arsenal de virulência de cepas STEC de origem animal em nosso meio. Além disso, demonstrou que as cepas STEC isoladas de bovinos mais frequentemente apresentam os subtipos de stx2 associados a infecções humanas e são carreadoras em maior frequencia de fatores de virulência.Dados abertos - Sucupira - Teses e dissertações (2017)Universidade Federal de São Paulo (UNIFESP)Guth, Beatriz Ernestina Cabilio [UNIFESP]http://lattes.cnpq.br/5661607777623571http://lattes.cnpq.br/3280477592714250Universidade Federal de São Paulo (UNIFESP)Freire, Caroline Fernandes [UNIFESP]2019-06-19T14:58:13Z2019-06-19T14:58:13Z2017-12-21info:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersion63 f.application/pdfhttps://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5456892http://repositorio.unifesp.br/handle/11600/50655porSão Pauloinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-02T19:21:02Zoai:repositorio.unifesp.br/:11600/50655Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-02T19:21:02Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
Search of shiga toxin subtypes and accessory virulence makers in shiga toxin-producing Escherichia coli (STEC) devoid of eae gene isolated from animal reservoirs
title Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
spellingShingle Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
Freire, Caroline Fernandes [UNIFESP]
Escherichia coli
Disease reservoirs
Virulence
Hemolytic-uremic syndrome
Escherichia coli
Reservatórios de doenças
Virulência
Síndrome hemolítico-urêmica
title_short Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
title_full Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
title_fullStr Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
title_full_unstemmed Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
title_sort Pesquisa dos subtipos de toxina shiga e de marcadores de virulência acessórios em amostras de Escherichia coli produtora de toxina shiga (STEC) desprovidas do gene eae isoladas do reservatório animal
author Freire, Caroline Fernandes [UNIFESP]
author_facet Freire, Caroline Fernandes [UNIFESP]
author_role author
dc.contributor.none.fl_str_mv Guth, Beatriz Ernestina Cabilio [UNIFESP]
http://lattes.cnpq.br/5661607777623571
http://lattes.cnpq.br/3280477592714250
Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Freire, Caroline Fernandes [UNIFESP]
dc.subject.por.fl_str_mv Escherichia coli
Disease reservoirs
Virulence
Hemolytic-uremic syndrome
Escherichia coli
Reservatórios de doenças
Virulência
Síndrome hemolítico-urêmica
topic Escherichia coli
Disease reservoirs
Virulence
Hemolytic-uremic syndrome
Escherichia coli
Reservatórios de doenças
Virulência
Síndrome hemolítico-urêmica
description Shiga toxin-producing Escherichia coli (STEC) are food-borne pathogens being ruminant animals their main natural reservoir. STEC is responsible for causing severe human illnesses as hemorragic colitis and the Hemolytic Uremic Syndrome (HUS). Recent studies have pointed out the prevalence of non-O157 STEC serotypes associated with both sporadic cases and outbreaks of severe diarrhea. Moreover, it has been observed that several non-O157 STEC serotypes unable to cause attaching and effacing lesion (A/E) have been implicated as agents of serious human diseases and HUS. Although Shiga toxin hás a central role in the pathogenesis of STEC diseases, other accessory virulence factors may also participate in this process. Therefore, to know the distribution of stx subtypes and accessory virulence genes in non-O157 STEC isolates devoid of A/E from the animal reservoir could indicate a higher pathogenic potential of these isolates and, therefore, constitutes the aimof this study. Gene sequences related to Stx1 and Stx2 subtypes, autotransporter proteins (EspP, EspI, EpeA, Sab), adhesins (Saa), toxins (EhxA) andiron uptake protein (Irp2) were sought by PCR in 125 STEC strains isolated from sheep, goat, buffalo and cattle. STEC isolates most frequently harbor stx2 (52%) and stx1stx2 (38%), being subtypes stx1a (57%), stx1c (47%), stx2a (67%), and stx2d (38%) those most frequently identified. Distribution of subtypes varied according to the reservoir. stx1a prevailed in cattle and buffalo isolates, stx1c predominate in sheep and goat, stx2a in cattle, and stx2d prevailed in goat and buffalo. The saa, espP, ehxA, espI,epeA, sab andirp2 genes were identified in 97%, 58%, 55%, 42%, 30%, 13% and11% of the STEC isolates, respectively, and differences were also observed depending on the reservoir. espP and ehxA molecular characterization showed that variants C (90%) and A (100%) prevailed, respectively. A great diversity of serotypes and virulence profiles wasobserved independently of the isolate origin. However, serotypes O39:H49, O79:H14, O113:H21 and O178:H19 were more representative among cattle and buffalos, and serotype O5:H- among goat isolates. This study made important contributions related to the virulence profile of STEC isolates from the animal reservoir in our settings. In addition, showed that STEC isolates from cattle most frequently carry the stx2 subtypes related to human infections, and harbor virulence factors at higher frequencies.
publishDate 2017
dc.date.none.fl_str_mv 2017-12-21
2019-06-19T14:58:13Z
2019-06-19T14:58:13Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5456892
http://repositorio.unifesp.br/handle/11600/50655
url https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5456892
http://repositorio.unifesp.br/handle/11600/50655
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 63 f.
application/pdf
dc.coverage.none.fl_str_mv São Paulo
dc.publisher.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
publisher.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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