Phenotypic and molecular characterization of erythromycin resistance in Campylobacter jejuni and Campylobacter coli strains isolated from swine and broiler chickens / Caracterização fenotípica e molecular da resistência à eritromicina em cepas de Campylobacter jejuni e Campylobacter coli isoladas de suínos e frangos de corte

Campylobacter spp. is a bacterial agent that causes gastroenteritis in humans and may trigger Guillain-Barré Syndrome (GBS) and is also considered one of the main foodborne diseases in developed countries. Poultry and pigs are considered reservoirs of these microorganisms, as well as raw or undercooked by-products are often incriminated as a source of human infection. Treatment in human cases is with macrolide, such erythromycin, that inhibits the protein synthesis of the microorganism. This study aimed to isolate Campylobacter jejuni and Campylobacter coli from intestinal content samples of broiler chickens (n=20) and swine (n=30) to characterize the erythromycin resistance profile of the strains and to detect molecular mechanisms involved in this resistance. The minimum inhibitory concentration was determined by agar dilution. The Mismatch Amplification Mutation Assay-Polymerase Chain Reaction (MAMA-PCR) was performed to detect mutations at positions 2074 and 2075 of 23S rRNA region, in addition to PCR test to detect the erm(B) gene. From the intestinal content of broiler chickens, 18 strains of C. jejuni and two strains of C. coli were isolated, whereas, from swine samples, no C. jejuni strain and 14 strains of C. coli were isolated. All C. coli strains were resistant, and three C. jejuni strains from broilers chickens were characterized with intermediate resistance to erythromycin. The MIC of the strains ranged from ≤0.5mg/μL to ≥128mg/μL. All resistant strains had the A2075G mutation, and one strain with intermediate resistance had the A2075G mutation. However, the A2074C mutation and the erm(B) gene were not detected. High resistance levels were detected in C. coli strains isolated from swine. The MAMA-PCR is a practical tool for detecting the erythromycin resistance in Campylobacter strains.(AU)

Campylobacter spp. é um agente bacteriano causador de gastroenterite em humanos e associado à síndrome de Guillain-Barré, sendo a campilobacteriose considerada uma das principais enfermidades de origem alimentar. Aves e suínos são importantes reservatórios desses microrganismos e seus produtos derivados crus ou mal cozidos são muitas vezes incriminados como fonte de infecção humana. A primeira escolha para o tratamento em casos humanos são os antimicrobianos da classe dos macrolídeos como à eritromicina. Dentro desse contexto, o objetivo deste estudo foi isolar Campylobacter jejuni e C. coli a partir de 20 amostras de conteúdo intestinal de frangos de corte e de 30 de suínos ao abate e investigar a resistência à eritromicina das estirpes obtidas e os possíveis mecanismos moleculares envolvidos nesta resistência. A concentração inibitória mínima foi determinada pela diluição em ágar e a técnica MAMA-PCR foi utilizada para detecção de mutações nas posições 2074 e 2075 da região 23s rRNA, foi pesquisado também a presença do gene erm(B) pela PCR. A partir do conteúdo intestinal de frangos de corte foram isoladas 18 estirpes de C. jejuni e duas de C. coli, enquanto de suínos foram obtidas 14 estirpes de C. coli e nenhuma estirpe de C. jejuni. Todas as estirpes de C. coli de suínos foram identificadas como resistentes e três estirpes de C. jejuni de frangos foram caracterizadas com resistência intermediária. A CIM das estirpes variou de ≤0,5mg/μL a ≥128mg/μL. Todas as estirpes resistentes tinham a mutação A2075G e uma cepa com resistência intermediária também apresentou a mutação A2075G. Não foi detectada a mutação A2074C ou a presença do gene erm(B) em nenhuma das estirpes obtidas. Os resultados revelam um alto nível de resistência em estirpes de C. coli isoladas de suínos frente a eritromicina. A técnica MAMA PCR utilizada se constitui em uma ferramenta prática para detecção da resistência à eritromicina em estirpes de C. jejuni e C. coli.(AU)

Phenotypic and molecular characterization of erythromycin resistance in Campylobacter jejuni and Campylobacter coli strains isolated from swine and broiler chickens / Caracterização fenotípica e molecular da resistência à eritromicina em cepas de Campylobacter jejuni e Campylobacter coli isoladas de suínos e frangos de corte

Campylobacter spp. is a bacterial agent that causes gastroenteritis in humans and may trigger Guillain-Barré Syndrome (GBS) and is also considered one of the main foodborne diseases in developed countries. Poultry and pigs are considered reservoirs of these microorganisms, as well as raw or undercooked by-products are often incriminated as a source of human infection. Treatment in human cases is with macrolide, such erythromycin, that inhibits the protein synthesis of the microorganism. This study aimed to isolate Campylobacter jejuni and Campylobacter coli from intestinal content samples of broiler chickens (n=20) and swine (n=30) to characterize the erythromycin resistance profile of the strains and to detect molecular mechanisms involved in this resistance. The minimum inhibitory concentration was determined by agar dilution. The Mismatch Amplification Mutation Assay-Polymerase Chain Reaction (MAMA-PCR) was performed to detect mutations at positions 2074 and 2075 of 23S rRNA region, in addition to PCR test to detect the erm(B) gene. From the intestinal content of broiler chickens, 18 strains of C. jejuni and two strains of C. coli were isolated, whereas, from swine samples, no C. jejuni strain and 14 strains of C. coli were isolated. All C. coli strains were resistant, and three C. jejuni strains from broilers chickens were characterized with intermediate resistance to erythromycin. The MIC of the strains ranged from ≤0.5mg/μL to ≥128mg/μL. All resistant strains had the A2075G mutation, and one strain with intermediate resistance had the A2075G mutation. However, the A2074C mutation and the erm(B) gene were not detected. High resistance levels were detected in C. coli strains isolated from swine. The MAMA-PCR is a practical tool for detecting the erythromycin resistance in Campylobacter strains.(AU)

Campylobacter spp. é um agente bacteriano causador de gastroenterite em humanos e associado à síndrome de Guillain-Barré, sendo a campilobacteriose considerada uma das principais enfermidades de origem alimentar. Aves e suínos são importantes reservatórios desses microrganismos e seus produtos derivados crus ou mal cozidos são muitas vezes incriminados como fonte de infecção humana. A primeira escolha para o tratamento em casos humanos são os antimicrobianos da classe dos macrolídeos como à eritromicina. Dentro desse contexto, o objetivo deste estudo foi isolar Campylobacter jejuni e C. coli a partir de 20 amostras de conteúdo intestinal de frangos de corte e de 30 de suínos ao abate e investigar a resistência à eritromicina das estirpes obtidas e os possíveis mecanismos moleculares envolvidos nesta resistência. A concentração inibitória mínima foi determinada pela diluição em ágar e a técnica MAMA-PCR foi utilizada para detecção de mutações nas posições 2074 e 2075 da região 23s rRNA, foi pesquisado também a presença do gene erm(B) pela PCR. A partir do conteúdo intestinal de frangos de corte foram isoladas 18 estirpes de C. jejuni e duas de C. coli, enquanto de suínos foram obtidas 14 estirpes de C. coli e nenhuma estirpe de C. jejuni. Todas as estirpes de C. coli de suínos foram identificadas como resistentes e três estirpes de C. jejuni de frangos foram caracterizadas com resistência intermediária. A CIM das estirpes variou de ≤0,5mg/μL a ≥128mg/μL. Todas as estirpes resistentes tinham a mutação A2075G e uma cepa com resistência intermediária também apresentou a mutação A2075G. Não foi detectada a mutação A2074C ou a presença do gene erm(B) em nenhuma das estirpes obtidas. Os resultados revelam um alto nível de resistência em estirpes de C. coli isoladas de suínos frente a eritromicina. A técnica MAMA PCR utilizada se constitui em uma ferramenta prática para detecção da resistência à eritromicina em estirpes de C. jejuni e C. coli.(AU)

Campylobacter jejuni and Campylobacter coli originated from chicken carcasses modulate their transcriptome to translate virulence genes in human cells / Campylobacter jejuni e Campylobacter coli originadas de carcaças de frango modulam seu transcriptoma para traduzir genes de virulência em células humanas

The aim was to determine the spread of genetically similar profiles of Campylobacter in chicken carcasses and evaluate their ability to produce transcripts for ciaB, dnaJ, p19 and sodB genes, before and after cultivation in Caco-2 cells. The strains used were isolated from 420 samples of chicken carcasses chilled and frozen ready for marketing. The species were identified by PCR-multiplex, the phylogeny was determined by RAPD-PCR and the presence of transcripts was performed by RT-PCR. We identified 74 (17.6%) of Campylobacter strains, being 55 (74.3%) C. jejuni and 19 (25.7%) C. coli. The phylogenetic relationship demonstrated heterogeneity between isolates of the same species, with absence of clones, indicating the high level of diversity of circulating genotypes. The gene transcription showed conflicting results before and after the culture in Caco-2 cell, so that before cultivation isolates showed greater capacity to transcribe genes related to survival and after the interaction with human cells, the strains showed higher potential to transcribe genes associated with virulence. The result of this study contributes to the understanding of how these seemingly fragile microorganisms are the most prevalent bacterial agents in human gastroenteritis.(AU)

O objetivo foi determinar a disseminação de perfis geneticamente semelhantes de Campylobacter em carcaças de frango e avaliar sua capacidade de produzir transcritos para os genes ciaB, dnaJ, p19 e sodB, antes e após o cultivo em células Caco-2. As cepas utilizadas foram isoladas de 420 amostras de carcaças de frango resfriadas e congeladas prontas para comercialização. As espécies foram identificadas por PCR-multiplex, a filogenia foi determinada por RAPD-PCR e a presença de transcritos foi realizada por RT-PCR. Identificamos 74 (17,6%) das cepas de Campylobacter, sendo 55 (74,3%) C. jejuni e 19 (25,7%) C. coli. A relação filogenética demonstrou heterogeneidade entre isolados da mesma espécie, com ausência de clones, indicando o alto nível de diversidade dos genótipos circulantes. A transcrição gênica mostrou resultados conflitantes antes e após a cultura em células Caco-2, de modo que, antes do cultivo, os isolados apresentaram maior capacidade de transcrever genes relacionados à sobrevivência e após a interação com células humanas, as linhagens apresentaram maior potencial para transcrever genes associados à virulência. O resultado deste estudo contribui para a compreensão de como esses microrganismos aparentemente frágeis são os agentes bacterianos mais prevalentes na gastroenterite humana.(AU)

Campylobacter jejuni and Campylobacter coli originated from chicken carcasses modulate their transcriptome to translate virulence genes in human cells / Campylobacter jejuni e Campylobacter coli originadas de carcaças de frango modulam seu transcriptoma para traduzir genes de virulência em células humanas

The aim was to determine the spread of genetically similar profiles of Campylobacter in chicken carcasses and evaluate their ability to produce transcripts for ciaB, dnaJ, p19 and sodB genes, before and after cultivation in Caco-2 cells. The strains used were isolated from 420 samples of chicken carcasses chilled and frozen ready for marketing. The species were identified by PCR-multiplex, the phylogeny was determined by RAPD-PCR and the presence of transcripts was performed by RT-PCR. We identified 74 (17.6%) of Campylobacter strains, being 55 (74.3%) C. jejuni and 19 (25.7%) C. coli. The phylogenetic relationship demonstrated heterogeneity between isolates of the same species, with absence of clones, indicating the high level of diversity of circulating genotypes. The gene transcription showed conflicting results before and after the culture in Caco-2 cell, so that before cultivation isolates showed greater capacity to transcribe genes related to survival and after the interaction with human cells, the strains showed higher potential to transcribe genes associated with virulence. The result of this study contributes to the understanding of how these seemingly fragile microorganisms are the most prevalent bacterial agents in human gastroenteritis.(AU)

O objetivo foi determinar a disseminação de perfis geneticamente semelhantes de Campylobacter em carcaças de frango e avaliar sua capacidade de produzir transcritos para os genes ciaB, dnaJ, p19 e sodB, antes e após o cultivo em células Caco-2. As cepas utilizadas foram isoladas de 420 amostras de carcaças de frango resfriadas e congeladas prontas para comercialização. As espécies foram identificadas por PCR-multiplex, a filogenia foi determinada por RAPD-PCR e a presença de transcritos foi realizada por RT-PCR. Identificamos 74 (17,6%) das cepas de Campylobacter, sendo 55 (74,3%) C. jejuni e 19 (25,7%) C. coli. A relação filogenética demonstrou heterogeneidade entre isolados da mesma espécie, com ausência de clones, indicando o alto nível de diversidade dos genótipos circulantes. A transcrição gênica mostrou resultados conflitantes antes e após a cultura em células Caco-2, de modo que, antes do cultivo, os isolados apresentaram maior capacidade de transcrever genes relacionados à sobrevivência e após a interação com células humanas, as linhagens apresentaram maior potencial para transcrever genes associados à virulência. O resultado deste estudo contribui para a compreensão de como esses microrganismos aparentemente frágeis são os agentes bacterianos mais prevalentes na gastroenterite humana.(AU)

Isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli found in chilled chicken carcasses in the Federal District Region and surrounding areas / Isolamento e resistência antimicrobiana de Campylobacter jejuni e Campylobacter coli encontradas em carcaças de frango resfriadas na região do Distrito Federal e entorno

This study aimed to isolate Campylobacter jejuni and Campylobacter coli from chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as to detect the occurrence of antimicrobial resistance and genes responsible for the same. A total of 105 chilled chicken carcasses were collected, of which 7 (6.67%) were positive for C. jejuni and 4 (3.81%) were positive for C. coli. These results were obtained using both the conventional microbiological isolation method and polymerase chain reaction assays. All of the positive strains were subjected to antimicrobial susceptibility testing for seven antimicrobials. The resistance incidences found in the C. jejuni strains were as follows: 71.43% for tetracycline and nalidixic acid, 42.86% for streptomycin and gentamicin, 57.14% for ciprofloxacin and erythromycin, and 28.57% for chloramphenicol. Among the C. coli strains, 100% were resistant to tetracycline and streptomycin, 75% were resistant to erythromycin, 50% were resistant to ciprofloxacin, gentamicin, and nalidixic acid, and no strains were resistant to chloramphenicol. While analyzing the presence of antimicrobial resistance genes in the isolated C. jejuni strains, the aph3-1 (resistance to aminoglycosides), aadE (resistance to streptomycin), and tet(O) (resistance to tetracycline) genes were identified, with occurrence rates of 57.14%, 28.57%, and 42.86%, respectively, whereas in the C. coli strains, there was a 25% occurrence rate for both the aph3-1 and tet(O) genes. The aadE gene was not found in the C. coli isolates. The results of this study demonstrated the presence of C. jejuni and C. coli in chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as the antimicrobial resistance and the presence of resistance genes in these bacteria, which may pose threats to public health.

O objetivo deste trabalho foi isolar Campylobacter jejuni e Campylobacter coli de carcaças de frango resfriadas comercializadas no Distrito Federal e entorno, bem como detectar a ocorrência de resistência antimicrobiana e genes responsáveis pela resistência antimicrobiana. Foram coletadas um total de 105 carcaças de frango resfriadas, das quais 7 (6,67%) foram positivas para C. jejuni e 4 (3,81%) para C. coli. Estes resultados foram obtidos usando tanto o método convencional de isolamento microbiológico quanto os ensaios de reação em cadeia da polymerase (PCR). Todas as cepas positivas foram submetidas ao teste de susceptibilidade antimicrobiana para sete antimicrobianos. As incidências de resistência encontradas nas cepas de C. jejuni foram as seguintes: 71,43% para tetraciclina e ácido nalidíxico, 42,86% para estreptomicina e gentamicina, 57,14% para ciprofloxacina e eritromicina e 28,57% para cloranfenicol. Entre as cepas de C. coli, 100% foram resistentes à tetraciclina e à estreptomicina, 75% eram resistentes à eritromicina, 50% eram resistentes à ciprofloxacina, gentamicina e ácido nalidíxico, e nenhuma cepa eram resistentes ao cloranfenicol. Ao analisar a presença de genes de resistência antimicrobiana nas cepas isoladas de C. jejuni, foram identificados os genes aph3-1 (resistência a aminoglicosídeos), aadE (resistência à estreptomicina) e tet (O) (resistência à tetraciclina) genes foram identificados, com taxas de ocorrência de 57,14%, 28,57% e 42,86%, respectivamente, enquanto que nas cepas de C. coli, houve uma taxa de ocorrência de 25% para os genes aph3-1 e tet (O). O gene aadE não foi encontrado nos isolados de C. coli. Os resultados deste estudo demonstraram a presença de C. jejuni e C. coli em carcaças de frango resfriadas comercializadas na Região do Distrito Federal e entorno, além da resistência antimicrobiana e a presença de genes de resistência nestas cepas, que podem se tornar uma possível ameaça a saúde pública.

Isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli found in chilled chicken carcasses in the Federal District Region and surrounding areas / Isolamento e resistência antimicrobiana de Campylobacter jejuni e Campylobacter coli encontradas em carcaças de frango resfriadas na região do Distrito Federal e entorno

This study aimed to isolate Campylobacter jejuni and Campylobacter coli from chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as to detect the occurrence of antimicrobial resistance and genes responsible for the same. A total of 105 chilled chicken carcasses were collected, of which 7 (6.67%) were positive for C. jejuni and 4 (3.81%) were positive for C. coli. These results were obtained using both the conventional microbiological isolation method and polymerase chain reaction assays. All of the positive strains were subjected to antimicrobial susceptibility testing for seven antimicrobials. The resistance incidences found in the C. jejuni strains were as follows: 71.43% for tetracycline and nalidixic acid, 42.86% for streptomycin and gentamicin, 57.14% for ciprofloxacin and erythromycin, and 28.57% for chloramphenicol. Among the C. coli strains, 100% were resistant to tetracycline and streptomycin, 75% were resistant to erythromycin, 50% were resistant to ciprofloxacin, gentamicin, and nalidixic acid, and no strains were resistant to chloramphenicol. While analyzing the presence of antimicrobial resistance genes in the isolated C. jejuni strains, the aph3-1 (resistance to aminoglycosides), aadE (resistance to streptomycin), and tet(O) (resistance to tetracycline) genes were identified, with occurrence rates of 57.14%, 28.57%, and 42.86%, respectively, whereas in the C. coli strains, there was a 25% occurrence rate for both the aph3-1 and tet(O) genes. The aadE gene was not found in the C. coli isolates. The results of this study demonstrated the presence of C. jejuni and C. coli in chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as the antimicrobial resistance and the presence of resistance genes in these bacteria, which may pose threats to public health.(AU)

O objetivo deste trabalho foi isolar Campylobacter jejuni e Campylobacter coli de carcaças de frango resfriadas comercializadas no Distrito Federal e entorno, bem como detectar a ocorrência de resistência antimicrobiana e genes responsáveis pela resistência antimicrobiana. Foram coletadas um total de 105 carcaças de frango resfriadas, das quais 7 (6,67%) foram positivas para C. jejuni e 4 (3,81%) para C. coli. Estes resultados foram obtidos usando tanto o método convencional de isolamento microbiológico quanto os ensaios de reação em cadeia da polymerase (PCR). Todas as cepas positivas foram submetidas ao teste de susceptibilidade antimicrobiana para sete antimicrobianos. As incidências de resistência encontradas nas cepas de C. jejuni foram as seguintes: 71,43% para tetraciclina e ácido nalidíxico, 42,86% para estreptomicina e gentamicina, 57,14% para ciprofloxacina e eritromicina e 28,57% para cloranfenicol. Entre as cepas de C. coli, 100% foram resistentes à tetraciclina e à estreptomicina, 75% eram resistentes à eritromicina, 50% eram resistentes à ciprofloxacina, gentamicina e ácido nalidíxico, e nenhuma cepa eram resistentes ao cloranfenicol. Ao analisar a presença de genes de resistência antimicrobiana nas cepas isoladas de C. jejuni, foram identificados os genes aph3-1 (resistência a aminoglicosídeos), aadE (resistência à estreptomicina) e tet (O) (resistência à tetraciclina) genes foram identificados, com taxas de ocorrência de 57,14%, 28,57% e 42,86%, respectivamente, enquanto que nas cepas de C. coli, houve uma taxa de ocorrência de 25% para os genes aph3-1 e tet (O). O gene aadE não foi encontrado nos isolados de C. coli. Os resultados deste estudo demonstraram a presença de C. jejuni e C. coli em carcaças de frango resfriadas comercializadas na Região do Distrito Federal e entorno, além da resistência antimicrobiana e a presença de genes de resistência nestas cepas, que podem se tornar uma possível ameaça a saúde pública.(AU)

Campylobacter jejuni, Campylobacter coli, Salmonella spp. e Yersinia enterocolitica isoladas de animais silvestres em um centro de reabilitação / Campylobacter jejuni, Campylobacter coli, Salmonella spp. and Yersinia enterocolitica isolated from wildlife animals of a reabilitation center

Muitas espécies de animais silvestres de vida livre servem como reservatório de bactérias patogênicas que ameaçam a saúde humana e dos animais domésticos. Algumas bactérias, como Campylobacter jejuni, Campylobacter coli, Yersinia enterocolitica e Salmonella enterica, causam enfermidades em humanos e podem contaminar os animais domésticos e silvestres. O Núcleo de Reabilitação da Fauna Silvestre da Universidade Federal de Pelotas (NURFS-UFPel) soluciona uma demanda regional específica de atenção à fauna silvestre brasileira. O objetivo desse trabalho foi identificar a presença de Campylobacter jejuni, Campylobacter coli, Salmonella spp. e Yersinia enterocolitica em animais silvestres que se encontravam em processo de reabilitação. Foram coletadas amostras de fezes, com uso de zaragatoas estéreis, de 34 aves, 16 mamíferos e 23 répteis. Dos 73 animais amostrados, quatro (5,48%) albergavam Y. enterocolitica, sendo duas aves, um mamífero e um réptil. Salmonella e Campylobacter não foram isolados. Os perfis de bandas dos isolados de Y. enterocolitica analisados pela rep-PCR foram diferentes entre si. Esses resultados indicam que as cepas isoladas não estão relacionadas entre si, não possuindo uma origem comum recente. Vanellus chilensis, Turdus rufiventris, Didelphis albiventris e Pantherophis guttatus podem albergar Y. enterocolitica e eliminá-la nas fezes, oferecendo risco de disseminação desse micro-organismo no ambiente, além de constituírem possíveis fontes de contaminação para humanos e outros animais.(AU)

Wild animals can transmit pathogenic bacteria to human and domestic animal's health. Some bacteria, such as Campylobacter jejuni, Campylobacter coli, Yersinia enterocolitica and Salmonella enterica, cause diseases in humans and can contaminate domestic and wild animais. The Núcleo de Reabilitação da Fauna Silvestre of Universidade Federal de Pelotas (Nurfs-UFPel) attend a specific regional demand of wildlife in Brazil. The aim of this paper was to identify the presence of these pathogenic bacteria in wild animals in rehabilitation. Stool samples were collected using sterile swabs from 34 birds, 16 mammals and 23 reptilian that were housed at Nurfs. Of the 73 collections, Y. enterocolitica was isolated from four (5.48%) of two birds, one mammal and one reptile. Salmonella and Campylobacter were not isolated. The molecular profile of bands of Y. enterocolitica identified in rep-PCR had differences. These results indicated that the isolates did not have a recent common origin. Pantherophis guttatus, Didelphis albiventris, Turdus rufiventris and Vanellus chilensis could shelt Y. enterocolitica and eliminate the bacteria in stool, offering risk of dissemination of these microorganisms in the environment with possible contamination of humans and other animals.(AU)

Campylobacter jejuni, Campylobacter coli, Salmonella spp. e Yersinia enterocolitica isoladas de animais silvestres em um centro de reabilitação / Campylobacter jejuni, Campylobacter coli, Salmonella spp. and Yersinia enterocolitica isolated from wildlife animals of a reabilitation center

Muitas espécies de animais silvestres de vida livre servem como reservatório de bactérias patogênicas que ameaçam a saúde humana e dos animais domésticos. Algumas bactérias, como Campylobacter jejuni, Campylobacter coli, Yersinia enterocolitica e Salmonella enterica, causam enfermidades em humanos e podem contaminar os animais domésticos e silvestres. O Núcleo de Reabilitação da Fauna Silvestre da Universidade Federal de Pelotas (NURFS-UFPel) soluciona uma demanda regional específica de atenção à fauna silvestre brasileira. O objetivo desse trabalho foi identificar a presença de Campylobacter jejuni, Campylobacter coli, Salmonella spp. e Yersinia enterocolitica em animais silvestres que se encontravam em processo de reabilitação. Foram coletadas amostras de fezes, com uso de zaragatoas estéreis, de 34 aves, 16 mamíferos e 23 répteis. Dos 73 animais amostrados, quatro (5,48%) albergavam Y. enterocolitica, sendo duas aves, um mamífero e um réptil. Salmonella e Campylobacter não foram isolados. Os perfis de bandas dos isolados de Y. enterocolitica analisados pela rep-PCR foram diferentes entre si. Esses resultados indicam que as cepas isoladas não estão relacionadas entre si, não possuindo uma origem comum recente. Vanellus chilensis, Turdus rufiventris, Didelphis albiventris e Pantherophis guttatus podem albergar Y. enterocolitica e eliminá-la nas fezes, oferecendo risco de disseminação desse micro-organismo no ambiente, além de constituírem possíveis fontes de contaminação para humanos e outros animais.(AU)

Wild animals can transmit pathogenic bacteria to human and domestic animal's health. Some bacteria, such as Campylobacter jejuni, Campylobacter coli, Yersinia enterocolitica and Salmonella enterica, cause diseases in humans and can contaminate domestic and wild animais. The Núcleo de Reabilitação da Fauna Silvestre of Universidade Federal de Pelotas (Nurfs-UFPel) attend a specific regional demand of wildlife in Brazil. The aim of this paper was to identify the presence of these pathogenic bacteria in wild animals in rehabilitation. Stool samples were collected using sterile swabs from 34 birds, 16 mammals and 23 reptilian that were housed at Nurfs. Of the 73 collections, Y. enterocolitica was isolated from four (5.48%) of two birds, one mammal and one reptile. Salmonella and Campylobacter were not isolated. The molecular profile of bands of Y. enterocolitica identified in rep-PCR had differences. These results indicated that the isolates did not have a recent common origin. Pantherophis guttatus, Didelphis albiventris, Turdus rufiventris and Vanellus chilensis could shelt Y. enterocolitica and eliminate the bacteria in stool, offering risk of dissemination of these microorganisms in the environment with possible contamination of humans and other animals.(AU)

Campylobacter coli no fluxograma de abate de suínos e pesquisa dos genes cdt / Campylobacter coli in swine slaughtering flowchart and research of cdt genes

Background: Campylobacter spp. are among the microorganisms most commonly associated with foodborne disease. Campylobacter spp. isolation from pigs during the slaughter and final products have been reported in several countries, including Brazil. However, very little is known about the sources of contamination in the slaughtering flowchart and how these microorganisms are spread in processing plants. Considering the possibility of the pigs carry Campylobacter spp. since the farm or its products are contaminated in the slaughterhouse, this study had as aim to track Campylobacter spp. in pig slaughtering flowchart to understand the behavior of these pathogens in the production line.Materials, Methods & Results: Forty animals of 10 lots, four from each lot, were followed during slaughter. Stool samples were collected from the floor of each enclosure where the pigs were housed on the farm and immediately after stunning on slaughterhouse. Samples from carcass surface were collected after removal of the animals from scrap machine, after evisceration and before the refrigeration chamber. It was also collected surface samples from jowls and samples from the scalding tank water before and after the passage of animals. The swabs containing samples were plated onto Columbia agar supplemented with activated charcoal, oxygen reduction solution and antibiotics supplement, and incubated at 42°C for 48 h under microaerobic conditions. The colonies which presented with a shiny and moist appearance were analyzed by Gram staining for identification of Campylobacter by morphology, and then tested for catalase and oxidase. The Campylobacter isolates were identified for species C. jejuni or C. coli by PCR. Bands profiles were determined by rep-PCR and used to compare the strains. Campylobacter was isolated from 19 (9.5%) of the 200 pig samples analyzed, seven (36.8%) of the rectum, seven (36.8%) after evisceration and five (26.3%) before the refrigeration chamber.[...]

Campylobacter coli no fluxograma de abate de suínos e pesquisa dos genes cdt / Campylobacter coli in swine slaughtering flowchart and research of cdt genes

Background: Campylobacter spp. are among the microorganisms most commonly associated with foodborne disease. Campylobacter spp. isolation from pigs during the slaughter and final products have been reported in several countries, including Brazil. However, very little is known about the sources of contamination in the slaughtering flowchart and how these microorganisms are spread in processing plants. Considering the possibility of the pigs carry Campylobacter spp. since the farm or its products are contaminated in the slaughterhouse, this study had as aim to track Campylobacter spp. in pig slaughtering flowchart to understand the behavior of these pathogens in the production line.Materials, Methods & Results: Forty animals of 10 lots, four from each lot, were followed during slaughter. Stool samples were collected from the floor of each enclosure where the pigs were housed on the farm and immediately after stunning on slaughterhouse. Samples from carcass surface were collected after removal of the animals from scrap machine, after evisceration and before the refrigeration chamber. It was also collected surface samples from jowls and samples from the scalding tank water before and after the passage of animals. The swabs containing samples were plated onto Columbia agar supplemented with activated charcoal, oxygen reduction solution and antibiotics supplement, and incubated at 42°C for 48 h under microaerobic conditions. The colonies which presented with a shiny and moist appearance were analyzed by Gram staining for identification of Campylobacter by morphology, and then tested for catalase and oxidase. The Campylobacter isolates were identified for species C. jejuni or C. coli by PCR. Bands profiles were determined by rep-PCR and used to compare the strains. Campylobacter was isolated from 19 (9.5%) of the 200 pig samples analyzed, seven (36.8%) of the rectum, seven (36.8%) after evisceration and five (26.3%) before the refrigeration chamber.[...](AU)

Caracterização de estirpes de Campylobacter coli isoladas de carcaças de ovinos e de efluentes de abatedouro do estado de São Paulo / Characterization of Campylobacter coli strains isolated from the carcasses of sheep and of wastewater of abattoir in the state of São Paulo

Doença bacteriana zoonótica, a campilobacteriose é responsável mundialmente por frequentes casos de gastroenterite humana. Campylobacter spp. apresenta fator de virulência associado à diarreia, denominado toxina citoletal distensiva (CDT), sendo codificado pelos genes do complexo cdt. Os objetivos do presente estudo foram: 1) isolar e identificar estirpes de Campylobacter spp. de 102 suabes de carcaças e 102 suabes retais de ovinos (Ovis aries) e de sete amostras de água dos efluentes, antes e depois do tratamento de desinfecção de abatedouro localizado no estado de São Paulo; e 2) detectar, pela técnica de multiplex-PCR, a presença do complexo de genes cdt. Foram isoladas e identificadas, por métodos fenotípicos e genotípicos, sete estirpes de Campylobacter coli provenientes de 4/102 (3,92%) das amostras de suabes retais, 1/102 (0,98%) de suabes de carcaças e 2/7 (28,5%) das águas dos efluentes. Dos isolados de suabes retais, em 2/7 (28,6%) estirpes foi detectada a presença dos genes cdt. Trata-se do primeiro relato de isolamento de estirpes de Campylobacter coli provenientes de abatedouro de ovinos e das estirpes portadoras do complexo de genes cdt nessa espécie animal no Brasil.(AU)

A zoonosis and bacterial disease, campylobacteriosis is responsible for frequent cases of human gastroenteritis worldwide. Campylobacter spp. presents the virulence factor called cytolethal distensive toxine (CDT), responsible for diarrhea and codified by the cdt gene. The aims of this study were: 1) to isolate and identify Campylobacter spp. strains from 102 carcass swabs and 102 rectal swabs of sheep (Ovis aries) and seven samples of wastewater, before and after the disinfection treatment, collected from the abattoir of the state of São Paulo; and 2) to detect the presence of cdt gene complex by Multiplex-PCR in strains of Campylobacter spp. Seven strains of Campylobacter coli were isolated and identified by phenotypic and genotypic methods: 4/102 (3.92%) from rectal swabs, 1/102 (0.98%) from carcass swabs and 2/7 (28.5%) from wastewater. From the rectal swab samples 2/7 (28.6%) strains were detected with the cdt gene. This is the first report on the isolation of Campylobacter coli from sheep abattoir, and of strains carrying the cdt gene complex in this animal species in Brazil.(AU)

Occurrence of Campylobacter jejuni and C. coli on broiler carcasses after chilling in southern Brazil / Campylobacter jejuni e C. coli em carcaças de frangos após a refrigeração por imersão no sul do Brasil

Campylobacter jejuni and C. coli have been associated with gastrointestinal disorders in human beings, due mainly to the consumption of chicken meat. Despite control measures for reducing contamination by these bacteria, the detection of Campylobacter in carcasses after chilling remains high. A total of 105 carcasses were assessed by the horizontal detection method in five federally inspected slaughterhouses in southern Brazil in 2012 and in the first three months of 2013. Campylobacterwas isolated in 37.1% of the carcasses, of which 97.5% contained C. jejuni and 2.5% were infected by C. coli. The rate of positive carcasses across the slaughterhouses ranged from 0 to 71.4%. Determining the occurrence of Campylobacter among flocks is crucial for estimating the microbial load at specific points along the slaughtering process and for minimizing the risk of contamination of end products by Campylobacter.(AU)

Campylobacter jejuni e C. coli têm sido associados a problemas gastroentéricos em seres humanos principalmente devido ao consumo de carne de frango. Embora medidas de controle sejam realizadas para reduzir a contaminação por estas bactérias, a identificação de Campylobacterem carcaças após a refrigeração por imersão é alto. Foram analisadas 105 carcaças pelo método de detecção horizontal em cinco abatedouros sob Inspeção Federal no sul do Brasil em 2012 e nos três primeiros meses de 2013. Campylobacter foi isolada em 37,1% das carcaças analisadas, as quais 97,5% foram identificados como C. jejuni e 2,5% como C. coli. A ocorrência de carcaças positivas entre matadouros variou de zero a 71,4%. O conhecimento sobre a ocorrência de Campylobacter entre os lotes é fundamental para estimar a extensão da carga microbiana em pontos específicos do abate e consequentemente minimizar o risco de contaminação por Campylobacter em produtos finais de frangos.(AU)

Occurrence and antimicrobial susceptibility of Campylobacter jejuni and C. coli in dog feces from public parks in Southern Ecuador

Background: Campylobacter jejuni and C. coli are important agents of diarrhea in human beings. Several animals are reservoirs for these bacteria and they may contribute to environmental contamination through fecal shedding. The presence of dog feces in urban settings could be a risk of Campylobacter environmental contamination and a problem of public health concern. Besides environmental contamination, the increasing antimicrobial resistance among Campylobacter is another issue of public health significance. The aims of this study were to determine the prevalence of C. jejuni and C. coli in dog feces collected from public parks of Loja city, Ecuador, and establishing their susceptibility to six antibiotics.Materials, Methods & Results: A total of 70 fresh-looking non-diarrheal dog fecal samples were collected from public parks of Loja city. An amount (c. a. 1 g) of each fecal specimen was collected with a swab, placed in a tube containing Bolton enrichment broth and processed within the next six hours. Campylobacter spp. were isolated by direct plating onto Butzler selective agar and into Bolton enrichment broth, which were incubated at 42°C for 48 h in microaerophilic conditions. From Bolton enrichment broth subcultures to Butzler selective agar were made and also to blood agar, the latter using the passive filtration method in order to isolate C. upsaliensis. Identification was made firstly using the traditional phenotypic test (catalase, oxidase, oxygen tolerance, growth at 25°C, hippurate and indoxyl acetate hydrolysis, and cephalothin and nalidixic acid susceptibility) and corroborated by the genus-specific polymerase chain reaction (PCR) and by the multiplex PCR (m-PCR) method proposed for the characterization of the six most common clinical Campylobacter species.[...](AU)

Occurrence and antimicrobial susceptibility of Campylobacter jejuni and C. coli in dog feces from public parks in Southern Ecuador

Background: Campylobacter jejuni and C. coli are important agents of diarrhea in human beings. Several animals are reservoirs for these bacteria and they may contribute to environmental contamination through fecal shedding. The presence of dog feces in urban settings could be a risk of Campylobacter environmental contamination and a problem of public health concern. Besides environmental contamination, the increasing antimicrobial resistance among Campylobacter is another issue of public health significance. The aims of this study were to determine the prevalence of C. jejuni and C. coli in dog feces collected from public parks of Loja city, Ecuador, and establishing their susceptibility to six antibiotics.Materials, Methods & Results: A total of 70 fresh-looking non-diarrheal dog fecal samples were collected from public parks of Loja city. An amount (c. a. 1 g) of each fecal specimen was collected with a swab, placed in a tube containing Bolton enrichment broth and processed within the next six hours. Campylobacter spp. were isolated by direct plating onto Butzler selective agar and into Bolton enrichment broth, which were incubated at 42°C for 48 h in microaerophilic conditions. From Bolton enrichment broth subcultures to Butzler selective agar were made and also to blood agar, the latter using the passive filtration method in order to isolate C. upsaliensis. Identification was made firstly using the traditional phenotypic test (catalase, oxidase, oxygen tolerance, growth at 25°C, hippurate and indoxyl acetate hydrolysis, and cephalothin and nalidixic acid susceptibility) and corroborated by the genus-specific polymerase chain reaction (PCR) and by the multiplex PCR (m-PCR) method proposed for the characterization of the six most common clinical Campylobacter species.[...]

Occurrence of Campylobacter jejuni and Campylobacter coli biotypes and antimicrobial susceptibility in healthy dogs in Southern Chile

Background: The thermotolerant species of Campylobacter, mainly C. jejuni and C. coli, are as important agents of human gastroenteritis worldwide, being a serious public health problem. They are widely distributed in the environment and the animal kingdom having as reservoirs a wide variety of animals, including dogs, which in turn can act as a source of infection for humans. The Campylobacter isolation rates found in dogs are heterogeneous with few available data in Latin American countries. However Campylobacter diarrhea in humans is a self-limited clinical process being antimicrobial treatment not always necessary, it is necessary to know their antimicrobial susceptibility/resistance profiles in clinical and reservoirs isolates, especially in C. jejuni and C. coli strains isolated from animals close to man as dogs are. The aims of this work were to determine the isolation frequency of C. jejuni and C. coli and their biotypes in healthy dogs and the susceptibility/resistance profiles of the isolated strains to six antimicrobial drugs. Materials, Methods & Results: Fecal samples from 141 healthy dogs were obtained, seeded into the TEC transport medium. In the laboratory, each sample was plated out on modified Skirrow medium and incubated at 42°C for 48 h under microaerobic conditions. The isolated strains were identified to species and biotype levels through their phenotypic characteristics using the API Campy® procedure (bioMérieux, Marcy/Etoile, France) and the method described by Lior, respectively. The antimicrobial susceptibility patterns to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin and tetracycline were determined by means of the E-test method, considering as the minimal inhibitory concentrations (MICs) the lowest antibiotic concentrations yielding no growth. Ampicillin resistant strains were tested for ß-lactamase production with the chromogenic cephalosporin test and the disc diffusion susceptibility test for ampicillin-sulbactam. The recovery rate of Campylobacter was 31.2% (C. jejuni 22.7%, C. coli 8.5%). Three of the four biotypes described for C. jejuni and the two described for C. coli were found being C. jejuni biotype I the most frequent one (53.1%). None of the strains showed resistance to chloramphenicol, erythromycin, gentamicin and tetracycline. Resistant strains were found for ciprofloxacin (6 strains, 13.9%) and ampicillin (4 strains, 9.1%). All the ampicillin resistant strains were found to be ß-lactamase producers. Discussion: The Campylobacter isolation rate (31.2%) found is not negligible. It may provide an epidemiological insight about the risk of zoonotic infection for individuals maintaining contact with dogs, especially for children. Direct contact with pets is a risk factor for acquiring campylobacteriosis, which was also observed in Latin-American countries. Ampicillin resistance is related to ß-lactamase production and was reported in different countries, including Chile. Campylobacter resistance to quinolones has been increasingly reported in strains of human and animal origin, especially from animals related to husbandry. The ciprofloxacin resistant strains found in this study probably could be a reflection of their spread to human pets. Because dogs have been identified as a potential source of human Campylobacter infections, exposure to animals carrying fluoroquinolone-resistant microorganisms could be a risk factor for acquiring this kind of strains.