Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates

Detalhes bibliográficos
Autor(a) principal: Dias, Diana Patrícia Pires
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10773/14979
Resumo: Antimicrobial resistance is as an emerging global problem in both human and veterinary medicine. In theory, wild animals are rarely exposed to antimicrobial agents and therefore low levels of AMR are to be expected. However, the growing interaction of these animals with anthropogenic activities can have a huge impact in their bacterial flora. Escherichia coli is commonly found in the intestinal tract of a wide variety of animals and humans. This intestinal bacterium can be easily disseminated in different ecosystems. Therefore, it can be an useful indicator of the selective pressure exerted by the use of antimicrobials. Salmonella is a pathogenic bacterium, commonly found in the intestine of healthy birds and mammals that can cause salmonellosis in humans. In the European Union, over 90,000 salmonellosis cases are reported every year to EFSA. This study was conducted in wild ungulates from three distinct geographical areas in Portugal (Montesinho, Idanha-a-Nova and Lousã) and aimed to: i) access the levels of antibacterial resistance occurring in E. coli strains ii) determine the occurrence levels of Salmonella spp. and iii) determine the occurrence levels of shiga toxin-producing E. coli (STEC). To that purpose, a total of 67 faecal samples from red deer (n=41), wild boar (n=21) and roe deer (n=4) were collected. Before antibacterial susceptibility testing (according to the EUCAST guidelines), the E. coli isolates obtained were typed by BOX-PCR to select for genetically different strains for each sample (n=152). The detection of Salmonella was performed according to ISO 6579:2002 Annex D. Results revealed that in E. coli resistance was observed to ampicillin (10%), tetracycline (9%), streptomycin (5%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%) and cefoxitin (1%). A total of 3.3% of the isolates exhibited a multiresistant phenotype, all from Lousã. The results were also analyzed according to ECOFFs. Non-wildtype phenotypes were obtained to ampicillin (10%), ceftazidime (6%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%), aztreonam (1%) and cefotxitin (1%). A low incidence of Salmonella spp. (1.5%) was observed and it was only identified in wild boar from Lousã. The isolate was susceptible to all the tested antimicrobials. Regarding the presence of STEC, it was possible to establish that red and roe deer from the three sampling sites carry this bacterium. The stx variants detected in the STEC isolates included stx1c, stx2d and stx2g. Moreover, the hemolysin gene ehxA was identified in a strain possessing the stx2g variant. Overall, our results reveal that these populations of wild ungulates are reservoirs of antibiotic resistant and potential pathogenic bacteria. Therefore, these animals can act as dissemination vehicles between wildlife-livestock-human interfaces.
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spelling Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulatesMicrobiologiaBactérias patogénicasResistência a antibióticosUngulados - PortugalAntimicrobial resistance is as an emerging global problem in both human and veterinary medicine. In theory, wild animals are rarely exposed to antimicrobial agents and therefore low levels of AMR are to be expected. However, the growing interaction of these animals with anthropogenic activities can have a huge impact in their bacterial flora. Escherichia coli is commonly found in the intestinal tract of a wide variety of animals and humans. This intestinal bacterium can be easily disseminated in different ecosystems. Therefore, it can be an useful indicator of the selective pressure exerted by the use of antimicrobials. Salmonella is a pathogenic bacterium, commonly found in the intestine of healthy birds and mammals that can cause salmonellosis in humans. In the European Union, over 90,000 salmonellosis cases are reported every year to EFSA. This study was conducted in wild ungulates from three distinct geographical areas in Portugal (Montesinho, Idanha-a-Nova and Lousã) and aimed to: i) access the levels of antibacterial resistance occurring in E. coli strains ii) determine the occurrence levels of Salmonella spp. and iii) determine the occurrence levels of shiga toxin-producing E. coli (STEC). To that purpose, a total of 67 faecal samples from red deer (n=41), wild boar (n=21) and roe deer (n=4) were collected. Before antibacterial susceptibility testing (according to the EUCAST guidelines), the E. coli isolates obtained were typed by BOX-PCR to select for genetically different strains for each sample (n=152). The detection of Salmonella was performed according to ISO 6579:2002 Annex D. Results revealed that in E. coli resistance was observed to ampicillin (10%), tetracycline (9%), streptomycin (5%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%) and cefoxitin (1%). A total of 3.3% of the isolates exhibited a multiresistant phenotype, all from Lousã. The results were also analyzed according to ECOFFs. Non-wildtype phenotypes were obtained to ampicillin (10%), ceftazidime (6%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%), aztreonam (1%) and cefotxitin (1%). A low incidence of Salmonella spp. (1.5%) was observed and it was only identified in wild boar from Lousã. The isolate was susceptible to all the tested antimicrobials. Regarding the presence of STEC, it was possible to establish that red and roe deer from the three sampling sites carry this bacterium. The stx variants detected in the STEC isolates included stx1c, stx2d and stx2g. Moreover, the hemolysin gene ehxA was identified in a strain possessing the stx2g variant. Overall, our results reveal that these populations of wild ungulates are reservoirs of antibiotic resistant and potential pathogenic bacteria. Therefore, these animals can act as dissemination vehicles between wildlife-livestock-human interfaces.A resistência antimicrobiana é um problema emergente e global, tanto a nível clínico como veterinário. Em teoria, os animais selvagens raramente estão expostos a agentes antimicrobianos, e deste modo espera-se que a sua flora bacteriana apresente baixos níveis de resistência. Contudo, a crescente interação destes animais com atividades antropogénicas pode influenciar a aquisição de uma flora bacteriana resistente. Escherichia coli faz parte do trato intestinal de uma grande variedade de animais, incluindo o Homem. Esta bactéria pode disseminar-se facilmente em diferentes ecossistemas, sendo também um importante indicador da pressão seletiva exercida pela utilização de antimicrobianos. Salmonella spp. é uma bactéria patogénica, normalmente encontrada no intestino de diversos animais. Anualmente, na União Europeia são reportados à EFSA mais de 90,000 casos de salmoneloses. O presente estudo foi realizado em três espécies de ungulados selvagens que habitam três localizações geográficas distintas em Portugal (Montesinho, Idanha-a-Nova e Lousã) e teve como objetivos: i) avaliar os níveis de resistência de isolados de E. coli ii) determinar o nível de ocorrência de Salmonella spp. e iii) determinar o nível de ocorrência de E. coli produtora da toxina shiga (STEC). Para tal foram recolhidas 67 amostras fecais de veado (n=41), javali (n=21) e corço (n=4). Numa primeira fase os isolados recolhidos foram tipados por BOX-PCR para selecionar estirpes geneticamente diferentes em cada amostra (n=152). Posteriormente realizou-se o teste de suscetibilidade a antimicrobianos (de acordo com o EUCAST). A deteção de Salmonella foi realizada de acordo com a norma ISO 6579:2002 Anexo D. Os resultados obtidos revelaram que para E. coli se verificou resistência aos antibióticos ampicilina (10%), tetraciclina (9%), streptomicina (5%), cotrimoxazol (4%), amoxicilina/ácido clavulânico (2%) e cofoxitina (1%). Um fenótipo de multirresistência foi encontrado em 3.3% dos isolados, todos provenientes da região da Lousã. Os resultados foram também analisados de acordo com os valores de ECOFFs, tendo sido encontrados fenótipos do tipo não-selvagem para a ampicilina (10%), ceftazidima (6%), cotrimoxazol (4%), amoxicilina/ácido clavulânico (2%), aztreonam (1%) e cefoxitina (1%). No que se refere à pesquisa de Salmonella, os resultados revelaram uma baixa incidência na população estudada (1.5%). Esta estirpe revelou-se suscetível a todos os antimicrobianos testados. Relativamente à presença de STEC, foi possível determinar que veados e corços dos três locais estudados são portadores deste tipo de estirpes. Detetaram-se três variantes do gene stx nos isolados STEC, incluindo stx1c, stx2d e stx2g. Foi ainda identificado o gene ehxA, que codifica para uma hemolisina, num isolado contendo a variante stx2g. No seu conjunto, os resultados obtidos mostram que as populações de ungulados selvagens estudados são reservatórios de bactérias resistentes, assim como de bactérias potencialmente patogénicas e podem, por isso, atuar como veículo de transmissão entre a vida selvagem, o gado e o Homem.Universidade de Aveiro2018-07-20T14:00:51Z2014-12-19T00:00:00Z2014-12-192017-12-19T12:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/14979TID:201572524engDias, Diana Patrícia Piresinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:27:42Zoai:ria.ua.pt:10773/14979Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:50:28.720932Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
title Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
spellingShingle Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
Dias, Diana Patrícia Pires
Microbiologia
Bactérias patogénicas
Resistência a antibióticos
Ungulados - Portugal
title_short Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
title_full Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
title_fullStr Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
title_full_unstemmed Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
title_sort Presence of pathogenic bacteria and antimicrobial resistance in Portuguese wild ungulates
author Dias, Diana Patrícia Pires
author_facet Dias, Diana Patrícia Pires
author_role author
dc.contributor.author.fl_str_mv Dias, Diana Patrícia Pires
dc.subject.por.fl_str_mv Microbiologia
Bactérias patogénicas
Resistência a antibióticos
Ungulados - Portugal
topic Microbiologia
Bactérias patogénicas
Resistência a antibióticos
Ungulados - Portugal
description Antimicrobial resistance is as an emerging global problem in both human and veterinary medicine. In theory, wild animals are rarely exposed to antimicrobial agents and therefore low levels of AMR are to be expected. However, the growing interaction of these animals with anthropogenic activities can have a huge impact in their bacterial flora. Escherichia coli is commonly found in the intestinal tract of a wide variety of animals and humans. This intestinal bacterium can be easily disseminated in different ecosystems. Therefore, it can be an useful indicator of the selective pressure exerted by the use of antimicrobials. Salmonella is a pathogenic bacterium, commonly found in the intestine of healthy birds and mammals that can cause salmonellosis in humans. In the European Union, over 90,000 salmonellosis cases are reported every year to EFSA. This study was conducted in wild ungulates from three distinct geographical areas in Portugal (Montesinho, Idanha-a-Nova and Lousã) and aimed to: i) access the levels of antibacterial resistance occurring in E. coli strains ii) determine the occurrence levels of Salmonella spp. and iii) determine the occurrence levels of shiga toxin-producing E. coli (STEC). To that purpose, a total of 67 faecal samples from red deer (n=41), wild boar (n=21) and roe deer (n=4) were collected. Before antibacterial susceptibility testing (according to the EUCAST guidelines), the E. coli isolates obtained were typed by BOX-PCR to select for genetically different strains for each sample (n=152). The detection of Salmonella was performed according to ISO 6579:2002 Annex D. Results revealed that in E. coli resistance was observed to ampicillin (10%), tetracycline (9%), streptomycin (5%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%) and cefoxitin (1%). A total of 3.3% of the isolates exhibited a multiresistant phenotype, all from Lousã. The results were also analyzed according to ECOFFs. Non-wildtype phenotypes were obtained to ampicillin (10%), ceftazidime (6%), co-trimoxazole (4%), amoxicillin/clavulanic acid (2%), aztreonam (1%) and cefotxitin (1%). A low incidence of Salmonella spp. (1.5%) was observed and it was only identified in wild boar from Lousã. The isolate was susceptible to all the tested antimicrobials. Regarding the presence of STEC, it was possible to establish that red and roe deer from the three sampling sites carry this bacterium. The stx variants detected in the STEC isolates included stx1c, stx2d and stx2g. Moreover, the hemolysin gene ehxA was identified in a strain possessing the stx2g variant. Overall, our results reveal that these populations of wild ungulates are reservoirs of antibiotic resistant and potential pathogenic bacteria. Therefore, these animals can act as dissemination vehicles between wildlife-livestock-human interfaces.
publishDate 2014
dc.date.none.fl_str_mv 2014-12-19T00:00:00Z
2014-12-19
2017-12-19T12:00:00Z
2018-07-20T14:00:51Z
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dc.publisher.none.fl_str_mv Universidade de Aveiro
publisher.none.fl_str_mv Universidade de Aveiro
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