Molecular epidemiology of Pseudomonas aeruginosa isolated from lower respiratory tract of ICU patients / Epidemiologia molecular de Pseudomonas aeruginosa isolada em pacientes com infecções do trato respiratório inferior admitidos em UTI

Lower respiratory tract infections (LRTIs) caused by Pseudomonas aeruginosa are the most common infection among hospitalized patients, associated with increased levels of morbidity, mortality and attributable health care costs. Increased resistant Pseudomonas worldwide has been quite meaningful to patients, especially in intensive care unit (ICUs). Different species of Pseudomonas exhibit different genetic profile and varied drug resistance. The present study determines the molecular epidemiology through DNA fingerprinting method and drug resistance of P. aeruginosa isolated from patients with LTRIs admitted in ICU. A total of 79 P. aeruginosa isolated from patients with LRTIs admitted in ICU were characterized by Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD) and Repetitive Extrapalindromic PCR (REP-PCR). Antibiotic resistance was determined by minimum inhibitory concentration (MIC) assay while MDR genes, viz, blaTEM, blaOXA, blaVIM, blaCTX-M-15 were detected by polymerase chain reaction (PCR). Of the 137 Pseudomonas sp isolated from ICU patients, 57.7% of the isolates were reported to be P. aeruginosa. The overall prevalence of P. aeruginosa among the all included patients was 34.5%. The RAPD analysis yielded 45 different patterns with 72 clusters with 57% to 100% similarity level. The RFLP analysis yielded 8 different patterns with 14 clusters with 76% to 100% similarity level. The REP PCR analysis yielded 37 different patterns with 65 clusters with 56% to 100% similarity level. There was no correlation among the different DNA patterns observed between the three different methods. Predominant of the isolates (46.8%) were resistant to amikacin. Of the 79 isolates, 60.8% were positive for blaTEM gene and 39.2% were positive for blaOXA gene. P. aeruginosa was predominantly isolated from patients with LRTIs admitted in ICU. The difference in the similarity level observed between the three DNA fingerprinting methods indicates that there is high inter-strain variability. The high genetic variability and resistance patterns indicates that we should continuously monitor the trend in the prevalence and antibiotic resistance of P. aeruginosa especially in patients with LRTIs admitted in ICU.(AU)

Infecções do trato respiratório inferior (ITRIs) são as infecções mais comuns entre pacientes internados em unidade de terapia intensiva (UTI). Pseudomonas aeruginosa é a causa mais comum de ITRIs e está associada ao aumento da mortalidade. Diferentes espécies de Pseudomonas exibem diferentes perfis genéticos e resistência variada as drogas. O presente estudo determina a epidemiologia molecular através do método de fingerprinting de DNA e resistência as drogas de P. aeruginosa isoladas de pacientes com LTRIs internados em UTI. Um total de 79 P. aeruginosa isoladas de pacientes com ITRIs internados em UTI foram caracterizados por Polimorfismo de Comprimento de Fragmentos de Restrição (RFLP), DNA Polimórfico Amplificado ao Acaso (RAPD) e PCR Extrapalindrômico Repetitivo (REP-PCR). A resistência aos antibióticos foram determinadas pelos ensaios de concentrações inibitória mínima (MIC), enquanto os genes MDR, blaTEM, blaOXA, blaVIM, blaCTX-M-15 foram detectados pela reação em cadeia da polimerase (PCR). Das 137 Pseudomonas sp isoladas de pacientes de UTI, 57,7% dos isolados foram relatados como P. aeruginosa. A prevalência geral de P. aeruginosa entre os pacientes incluídos foram de 34,5%. A análise RAPD renderam 45 padrões diferentes com 72 clusters com nível de similaridade de 57% a 100%. A análise RFLP renderam 8 padrões diferentes com 14 clusters com 76% a 100% de similaridade. A análise de PCR do REP produziram 37 padrões diferentes com 65 clusters com nível de similaridade de 56% a 100%. Não houveram correlações entre os diferentes padrões de DNA observados entre os três diferentes métodos. Predominantes dos isolados (46,8%) eram resistentes à amicacina. Dos 79 isolados, 60,8% foram positivos para o gene blaTEM e 39,2% foram positivos para o gene blaOXA. P. aeruginosa foi predominantemente isolado de pacientes com ITRIs internados em UTI. A diferença no nível de similaridade observado entre os três métodos de fingerprinting do DNA indica que há alta variabilidade inter-strain. A alta variabilidade genética e os padrões de resistência indicam que devemos monitorar continuamente a tendência na prevalência e resistência a antibióticos de P. aeruginosa, especialmente em pacientes com ITRIs internados em UTI.(AU)

Molecular epidemiology of Pseudomonas aeruginosa isolated from lower respiratory tract of ICU patients / Epidemiologia molecular de Pseudomonas aeruginosa isolada em pacientes com infecções do trato respiratório inferior admitidos em UTI

Lower respiratory tract infections (LRTIs) caused by Pseudomonas aeruginosa are the most common infection among hospitalized patients, associated with increased levels of morbidity, mortality and attributable health care costs. Increased resistant Pseudomonas worldwide has been quite meaningful to patients, especially in intensive care unit (ICUs). Different species of Pseudomonas exhibit different genetic profile and varied drug resistance. The present study determines the molecular epidemiology through DNA fingerprinting method and drug resistance of P. aeruginosa isolated from patients with LTRIs admitted in ICU. A total of 79 P. aeruginosa isolated from patients with LRTIs admitted in ICU were characterized by Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD) and Repetitive Extrapalindromic PCR (REP-PCR). Antibiotic resistance was determined by minimum inhibitory concentration (MIC) assay while MDR genes, viz, blaTEM, blaOXA, blaVIM, blaCTX-M-15 were detected by polymerase chain reaction (PCR). Of the 137 Pseudomonas sp isolated from ICU patients, 57.7% of the isolates were reported to be P. aeruginosa. The overall prevalence of P. aeruginosa among the all included patients was 34.5%. The RAPD analysis yielded 45 different patterns with 72 clusters with 57% to 100% similarity level. The RFLP analysis yielded 8 different patterns with 14 clusters with 76% to 100% similarity level. The REP PCR analysis yielded 37 different patterns with 65 clusters with 56% to 100% similarity level. There was no correlation among the different DNA patterns observed between the three different methods. Predominant of the isolates (46.8%) were resistant to amikacin. Of the 79 isolates, 60.8% were positive for blaTEM gene and 39.2% were positive for blaOXA gene. P. aeruginosa was predominantly isolated from patients with LRTIs admitted in ICU. The difference in the similarity level observed between the three DNA fingerprinting methods indicates that there is high inter-strain variability. The high genetic variability and resistance patterns indicates that we should continuously monitor the trend in the prevalence and antibiotic resistance of P. aeruginosa especially in patients with LRTIs admitted in ICU.

Infecções do trato respiratório inferior (ITRIs) são as infecções mais comuns entre pacientes internados em unidade de terapia intensiva (UTI). Pseudomonas aeruginosa é a causa mais comum de ITRIs e está associada ao aumento da mortalidade. Diferentes espécies de Pseudomonas exibem diferentes perfis genéticos e resistência variada as drogas. O presente estudo determina a epidemiologia molecular através do método de fingerprinting de DNA e resistência as drogas de P. aeruginosa isoladas de pacientes com LTRIs internados em UTI. Um total de 79 P. aeruginosa isoladas de pacientes com ITRIs internados em UTI foram caracterizados por Polimorfismo de Comprimento de Fragmentos de Restrição (RFLP), DNA Polimórfico Amplificado ao Acaso (RAPD) e PCR Extrapalindrômico Repetitivo (REP-PCR). A resistência aos antibióticos foram determinadas pelos ensaios de concentrações inibitória mínima (MIC), enquanto os genes MDR, blaTEM, blaOXA, blaVIM, blaCTX-M-15 foram detectados pela reação em cadeia da polimerase (PCR). Das 137 Pseudomonas sp isoladas de pacientes de UTI, 57,7% dos isolados foram relatados como P. aeruginosa. A prevalência geral de P. aeruginosa entre os pacientes incluídos foram de 34,5%. A análise RAPD renderam 45 padrões diferentes com 72 clusters com nível de similaridade de 57% a 100%. A análise RFLP renderam 8 padrões diferentes com 14 clusters com 76% a 100% de similaridade. A análise de PCR do REP produziram 37 padrões diferentes com 65 clusters com nível de similaridade de 56% a 100%. Não houveram correlações entre os diferentes padrões de DNA observados entre os três diferentes métodos. Predominantes dos isolados (46,8%) eram resistentes à amicacina. Dos 79 isolados, 60,8% foram positivos para o gene blaTEM e 39,2% foram positivos para o gene blaOXA. P. aeruginosa foi predominantemente isolado de pacientes com ITRIs internados em UTI. A diferença no nível de similaridade observado entre os três métodos de fingerprinting do DNA indica que há alta variabilidade inter-strain. A alta variabilidade genética e os padrões de resistência indicam que devemos monitorar continuamente a tendência na prevalência e resistência a antibióticos de P. aeruginosa, especialmente em pacientes com ITRIs internados em UTI.

Genetic traceability of Staphylococcus aureus strains isolated from primiparous dairy cows mastitis, humans and environment in the Northeast region of Brazil / Rastreabilidade genética de cepas de Staphylococcus aureus isoladas de mastite de vacas primíparas, ordenhadores e ambiente de ordenha na região Nordeste do Brasil

This research aimed to investigate the genotypic relatedness of 18 Staphylococcus aureus strains isolated from intramammary infections in primiparous cows and extramammary sites on five dairy herds by rep-PCR using RW3A primers, and by PFGE using the endonuclease SmaI. The isolates were also evaluated in vitro for the susceptibility against beta-lactam antimicrobials drugs (penicillin and oxacillin), considering that beta-lactams are frequently used for treating staphylococcal intrammamary infections. The rep-PCR typing was highly discriminatory (D value= 0.9804) and a total of 15 patterns were detected. The PFGE method was also highly discriminatory (D value= 0.9667) and a total of 13 patterns were observed. A total of 15 out of 18 (83%) isolates were resistant to penicillin and one out of 18 (6%) to oxacillin. In conclusion, these findings confirmed the occurrence of a high genetic diversity of S. aureus strains at the herds and the presence of clonally-related strains only at the same herd, emphasizing a variety of genotypic profiles among the isolates.(AU)

Objetivou-se com este estudo investigar a correlação genética de 18 cepas de Staphylococcus aureus isoladas de infecções intramamárias em vacas primíparas e de locais extramamários em cinco propriedades leiteiras através das técnicas de PCR por sequências palindrômicas extragênicas repetitivas (rep-PCR), usando iniciadores RW3A, e de eletroforese em gel de campo pulsado (PFGE), usando a endonuclease SmaI. Os isolados também foram avaliados in vitro quanto à suscetibilidade aos antimicrobianos beta-lactâmicos (penicilina e oxacilina). A tipagem por rep-PCR foi altamente discriminatória (valor D = 0,9804) e um total de 15 padrões foram detectados. Os isolados de S. aureus foram agrupados em três grupos diferentes (A a C), com 80% de similaridade. A técnica de PFGE também foi altamente discriminatória (valor D = 0,9667) e um total de 13 padrões foi observado. A análise do dendrograma com um coeficiente de similaridade de 80% gerou dois grupos diferentes (A e B). Além disso, cepas clonais isoladas do leite foram identificadas na mesma propriedade pelos dois métodos de tipificação e, apesar da presença de cepas dominantes, nossos resultados sugerem uma alta diversidade genética dentre as cepas de S. aureus analisadas. Um total de 15, dos 18 (83%) isolados, eram resistentes à penicilina e um dos 18 (6%) à oxacilina. Assim, esses achados confirmam a ocorrência de uma alta diversidade genética de cepas de S. aureus nas propriedades e a presença de cepas clonalmente relacionadas apenas na mesma propriedade, enfatizando uma variedade de perfis genotípicos entre os isolados.(AU)

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.(AU)

Molecular epidemiology of Pseudomonas aeruginosa isolated from lower respiratory tract of ICU patients

Abstract Lower respiratory tract infections (LRTIs) caused by Pseudomonas aeruginosa are the most common infection among hospitalized patients, associated with increased levels of morbidity, mortality and attributable health care costs. Increased resistant Pseudomonas worldwide has been quite meaningful to patients, especially in intensive care unit (ICUs). Different species of Pseudomonas exhibit different genetic profile and varied drug resistance. The present study determines the molecular epidemiology through DNA fingerprinting method and drug resistance of P. aeruginosa isolated from patients with LTRIs admitted in ICU. A total of 79 P. aeruginosa isolated from patients with LRTIs admitted in ICU were characterized by Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD) and Repetitive Extrapalindromic PCR (REP-PCR). Antibiotic resistance was determined by minimum inhibitory concentration (MIC) assay while MDR genes, viz, blaTEM, blaOXA, blaVIM, blaCTX-M-15 were detected by polymerase chain reaction (PCR). Of the 137 Pseudomonas sp isolated from ICU patients, 57.7% of the isolates were reported to be P. aeruginosa. The overall prevalence of P. aeruginosa among the all included patients was 34.5%. The RAPD analysis yielded 45 different patterns with 72 clusters with 57% to 100% similarity level. The RFLP analysis yielded 8 different patterns with 14 clusters with 76% to 100% similarity level. The REP PCR analysis yielded 37 different patterns with 65 clusters with 56% to 100% similarity level. There was no correlation among the different DNA patterns observed between the three different methods. Predominant of the isolates (46.8%) were resistant to amikacin. Of the 79 isolates, 60.8% were positive for blaTEM gene and 39.2% were positive for blaOXA gene. P. aeruginosa was predominantly isolated from patients with LRTIs admitted in ICU. The difference in the similarity level observed between the three DNA fingerprinting methods indicates that there is high inter-strain variability. The high genetic variability and resistance patterns indicates that we should continuously monitor the trend in the prevalence and antibiotic resistance of P. aeruginosa especially in patients with LRTIs admitted in ICU.

Resumo Infecções do trato respiratório inferior (ITRIs) são as infecções mais comuns entre pacientes internados em unidade de terapia intensiva (UTI). Pseudomonas aeruginosa é a causa mais comum de ITRIs e está associada ao aumento da mortalidade. Diferentes espécies de Pseudomonas exibem diferentes perfis genéticos e resistência variada as drogas. O presente estudo determina a epidemiologia molecular através do método de fingerprinting de DNA e resistência as drogas de P. aeruginosa isoladas de pacientes com LTRIs internados em UTI. Um total de 79 P. aeruginosa isoladas de pacientes com ITRIs internados em UTI foram caracterizados por Polimorfismo de Comprimento de Fragmentos de Restrição (RFLP), DNA Polimórfico Amplificado ao Acaso (RAPD) e PCR Extrapalindrômico Repetitivo (REP-PCR). A resistência aos antibióticos foram determinadas pelos ensaios de concentrações inibitória mínima (MIC), enquanto os genes MDR, blaTEM, blaOXA, blaVIM, blaCTX-M-15 foram detectados pela reação em cadeia da polimerase (PCR). Das 137 Pseudomonas sp isoladas de pacientes de UTI, 57,7% dos isolados foram relatados como P. aeruginosa. A prevalência geral de P. aeruginosa entre os pacientes incluídos foram de 34,5%. A análise RAPD renderam 45 padrões diferentes com 72 clusters com nível de similaridade de 57% a 100%. A análise RFLP renderam 8 padrões diferentes com 14 clusters com 76% a 100% de similaridade. A análise de PCR do REP produziram 37 padrões diferentes com 65 clusters com nível de similaridade de 56% a 100%. Não houveram correlações entre os diferentes padrões de DNA observados entre os três diferentes métodos. Predominantes dos isolados (46,8%) eram resistentes à amicacina. Dos 79 isolados, 60,8% foram positivos para o gene blaTEM e 39,2% foram positivos para o gene blaOXA. P. aeruginosa foi predominantemente isolado de pacientes com ITRIs internados em UTI. A diferença no nível de similaridade observado entre os três métodos de fingerprinting do DNA indica que há alta variabilidade inter-strain. A alta variabilidade genética e os padrões de resistência indicam que devemos monitorar continuamente a tendência na prevalência e resistência a antibióticos de P. aeruginosa, especialmente em pacientes com ITRIs internados em UTI.

Contamination of pigs by Yersinia enterocolitica in the abattoir flowchart and its relation to the farm / Contaminação de suínos por Yersinia enterocolitica no fluxograma de abate e sua relação com a granja

The research intends to detect sources of contamination by Yersinia enterocolitica in the abattoir flowchart and endeavors to study its relation with the contamination in the farm. For this purpose, sixty pigs were followed up. In order to carry out the study, samples of faeces were collected from the animal farm, where the animals were originally kept and from the abattoir, directly from the animals rectum, after desensitization. Additionally, samples were also collected from the carcass, after passage into the hair removal machine, after evisceration, prior to entry into the cold chambre, from the jowls, and water of the scald tank, before the commencement of the abattoir as well as after the passage of the animals. Further, the isolates were obtained through microbiological analyzes, upon being identified by PCR and compared via rep-PCR. Basically, Yersinia enterocolitica was isolated from three bays in the original farm (20 %) and from 20 samples (6.67 %), obtained in the abattoir flowchart. Comparison made via rep-PCR revealed that the contaminated pigs on the farm could carry the microorganism to different points in the abattoir flowchart. However, apart from the farm, other sources of the contamination were reported to be more frequent and diverse. Indeed, the chins and the carcass at the entrance of the cold chamber were identified as the most critical points. Therefore, we concluded that Y. enterocolitica present in the gastrointestinal tract of pigs on the farm, cannot be eliminated throughout theabattoir flowchart and remain in the chambers intended for the cold room.(AU)

O objetivo deste estudo foi detectar fontes de contaminação por Yersinia enterocolitica no fluxograma de abate e sua relação com a contaminação na granja. Sessenta suínos foram acompanhados. Foram coletadas amostras de fezes dos animais na granja de origem e durante o abate, diretamente do reto, após a insensibilização. Também foram coletadas amostras da carcaça após a passagem na depiladeira, após a evisceração, antes da entrada na câmara fria, da papada e da água do tanque de escaldagem antes de iniciar o abate e após a passagem dos animais. Os isolados foram obtidos através de análises microbiológicas, identificados por PCR e comparados através de rep-PCR. Yersinia enterocolitica foi isolada de três baias na granja de origem (20%) e de 20 amostras (6,67%) obtidas no fluxograma de abate. Após a rep-PCR, observou-se que os suínos contaminados na granja podem carrear o micro-organismo para diferentes pontos do fluxograma de abate. No entanto, outras fontes de contaminação que não a granja são mais frequentes e diversas. A papada e a carcaça na entrada da câmara fria são os pontos mais críticos. Conclui-se que Y. enterocolitica presente no trato gastrointestinal de suínos na granja pode não ser eliminada ao longo de todo o fluxograma de abate e permanecer na carcaça destinada à câmara fria.(AU)

Molecular methods for diversity assessment among xanthomonads of Bulgarian and Macedonian pepper

ABSTRACT Bacterial spot is an important disease of pepper in Bulgaria and Macedonia. For characterization of Xanthomonas species associated with bacterial spot, 161 strains were collected from various field pepper-growing regions. Among them, 131 strains were identified as Xanthomonas euvesicatoria and 30 as Xanthomonas vesicatoria using species-specific primers and polymerase chain reaction followed by restriction fragment length polymorphism analysis. To assess the genetic diversity of the strains, two methods (Random Amplified Polymorphic DNA and Repetitive Element Palindromic-Polymerase Chain Reaction) were applied. Discriminatory index was calculated and analysis of molecular variance was carried out.Combined random amplified polymorphic DNA analysis of the X. euvesicatoria strains with primers CUGEA-4 and CUGEA-6 had greater discriminative power (0.60) than repetitive element palindromic-polymerase chain reaction with ERIC and BOX A1R primers, which makes this method applicable for strain diversity evaluation. Discrimination among the X. vesicatoria strains was achieved by the use of ERIC primers and only for the Bulgarian strains. The results demonstrated that X. euvesicatoria was more diverse than X. vesicatoria and heterogeneity was observed mainly in the Bulgarian populations. According to the analysis of molecular variance, genetic variations in X. euvesicatoria were observed among and within populations from different regions, while the differences between the two countries were minor. Following the principal coordinates analysis, a relation between the climatic conditions of the regions and a genetic distance of the populations may be suggested.

Molecular methods for diversity assessment among xanthomonads of Bulgarian and Macedonian pepper

Bacterial spot is an important disease of pepper in Bulgaria and Macedonia. For characterization of Xanthomonas species associated with bacterial spot, 161 strains were collected from various field pepper-growing regions. Among them, 131 strains were identified as Xanthomonas euvesicatoria and 30 as Xanthomonas vesicatoria using species-specific primers and polymerase chain reaction followed by restriction fragment length polymorphism analysis. To assess the genetic diversity of the strains, two methods (Random Amplified Polymorphic DNA and Repetitive Element Palindromic-Polymerase Chain Reaction) were applied. Discriminatory index was calculated and analysis of molecular variance was carried out.Combined random amplified polymorphic DNA analysis of the X. euvesicatoria strains with primers CUGEA-4 and CUGEA-6 had greater discriminative power (0.60) than repetitive element palindromic-polymerase chain reaction with ERIC and BOX A1R primers, which makes this method applicable for strain diversity evaluation. Discrimination among the X. vesicatoria strains was achieved by the use of ERIC primers and only for the Bulgarian strains. The results demonstrated that X. euvesicatoria was more diverse than X. vesicatoria and heterogeneity was observed mainly in the Bulgarian populations. According to the analysis of molecular variance, genetic variations in X. euvesicatoria were observed among and within populations from different regions, while the differences between the two countries were minor. Following the principal coordinates analysis, a relation between the climatic conditions of the regions and a genetic distance of the populations may be suggested.(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)

Formação de biofilme por Staphylococcus aureus isolados de queijo mussarela elaborado com leite de búfala e seu efeito sobre a sensibilidade a sanitizantes / Biofilm formation by coagulase-positive staphylococcus aureus isolated from mozzarella cheese elaborated with buffalo milk and its effect on sensitivity to sanitizers

Background: The buffalo milk mozzarella cheese is a new product in the market, with high consumer acceptance and excellent prospects for trade. The cheese is rich in nutrients, which favors the proliferation of microorganisms that can cause food-borne diseases in the consumer. Staphylococcus aureus can cause gastro-enteritis in humans by the production of enterotoxins in food. One problem that may hinder the elimination of undesirable microorganisms in the food industry is the formation of biofilms. The objective of this study was to determine the effect of biofilm formation by Staphylococcus aureus isolated from buffalo mozzarella cheese on sensitivity to sanitizers.Materials, Methods & Results: Fifty samples of buffalo mozzarella cheese were analyzed to investigate the presence of S. aureus. The isolates were obtained through microbiological analysis and identified by PCR. The similarity of the strains was compared through rep-PCR. The distinct strains were tested for biofilm formation in microtiter plates. Soy Tripticase Broth (TSB) was placed in each well of the microtiter plate and overnight cultures of each strain was added. Wells without bacterial culture were used as controls. A villous cap was then placed on the plate and incubated for 48 h at 37°C. During incubation, the biofilms formed on the surface of the villi of the caps. For quantification of biofilm formation, material that remained attached to the cap was stained with crystal violet, the stained biofilm was extracted and the OD570 of each well was measured. Each strain was classified as non-biofilm forming, weak forming, moderately formed or formative strong. Strong forming and non-biofilm forming strains were tested on high density polyethylene, stainless steel and glass surfaces. Plates of 4 cm² of the different materials were placed in TSB where the culture of each isolate was inoculated separately.[...]

Formação de biofilme por Staphylococcus aureus isolados de queijo mussarela elaborado com leite de búfala e seu efeito sobre a sensibilidade a sanitizantes / Biofilm formation by coagulase-positive staphylococcus aureus isolated from mozzarella cheese elaborated with buffalo milk and its effect on sensitivity to sanitizers

Background: The buffalo milk mozzarella cheese is a new product in the market, with high consumer acceptance and excellent prospects for trade. The cheese is rich in nutrients, which favors the proliferation of microorganisms that can cause food-borne diseases in the consumer. Staphylococcus aureus can cause gastro-enteritis in humans by the production of enterotoxins in food. One problem that may hinder the elimination of undesirable microorganisms in the food industry is the formation of biofilms. The objective of this study was to determine the effect of biofilm formation by Staphylococcus aureus isolated from buffalo mozzarella cheese on sensitivity to sanitizers.Materials, Methods & Results: Fifty samples of buffalo mozzarella cheese were analyzed to investigate the presence of S. aureus. The isolates were obtained through microbiological analysis and identified by PCR. The similarity of the strains was compared through rep-PCR. The distinct strains were tested for biofilm formation in microtiter plates. Soy Tripticase Broth (TSB) was placed in each well of the microtiter plate and overnight cultures of each strain was added. Wells without bacterial culture were used as controls. A villous cap was then placed on the plate and incubated for 48 h at 37°C. During incubation, the biofilms formed on the surface of the villi of the caps. For quantification of biofilm formation, material that remained attached to the cap was stained with crystal violet, the stained biofilm was extracted and the OD570 of each well was measured. Each strain was classified as non-biofilm forming, weak forming, moderately formed or formative strong. Strong forming and non-biofilm forming strains were tested on high density polyethylene, stainless steel and glass surfaces. Plates of 4 cm² of the different materials were placed in TSB where the culture of each isolate was inoculated separately.[...](AU)

Pocilgas de espera como fonte de contaminação por Staphylococcus coagulase positiva de suínos destinados ao abate / Holding pens as sources of contamination of coagulase-positive Staphylococcus to pigs waiting for slaughter

Background: Coagulase-Positive Staphylococcus (SCP) are important pathogens related to foodborne illness associated with pork consumption. The isolation of SCP from pork products has been reported in several countries, including Brazil. Therefore, the identification of the sources of contamination of the pork products is fundamental to ensure the food safety. Although the animals remain in the holding pens during the pre-slaughter, these facilities have not been studied as a possible source of contamination for pigs. The aim of this study was to determine the importance of holding pens as sources of contamination of SCP to pigs and to identify other sources in the slaughter flowchart.Materials, Methods & Results: It was followed four pigs from ten different lots sent to slaughter. Prior to slaughter, samples were collected from the floors of the holding pens in the slaughterhouse. During slaughter, samples from seven different points were collected: 1) stool from the rectum immediately after stunning; 2) external surface of the carcass after dehairing; 3) internal surface of the carcass after evisceration; 4) external surface of the half-carcass prior to entry into the cold chamber; 5) tongue surface; 6) jowls; and 7) mesenteric lymph nodes. The strains were obtained through microbiological analysis. To compare the similarity between the strains, rep-PCR was performed. Of the ten samples collected in the holding pens, four (40%) were contaminated with SCP. At slaughter, 280 samples were collected and 56 (20%) SCP isolates were obtained. The lymph nodes were the point of greatest isolation (19.6%), followed by the surface of the carcass at the entrance to the cold chamber (17.8%), the rectum after desensitization (16.1%), carcass surface after opening of the abdominal cavity (16.1%), jowls (12.5%), carcass surface after dehairing (8.9%) and tongue surface (8.9%).[...]

Pocilgas de espera como fonte de contaminação por Staphylococcus coagulase positiva de suínos destinados ao abate / Holding pens as sources of contamination of coagulase-positive Staphylococcus to pigs waiting for slaughter

Background: Coagulase-Positive Staphylococcus (SCP) are important pathogens related to foodborne illness associated with pork consumption. The isolation of SCP from pork products has been reported in several countries, including Brazil. Therefore, the identification of the sources of contamination of the pork products is fundamental to ensure the food safety. Although the animals remain in the holding pens during the pre-slaughter, these facilities have not been studied as a possible source of contamination for pigs. The aim of this study was to determine the importance of holding pens as sources of contamination of SCP to pigs and to identify other sources in the slaughter flowchart.Materials, Methods & Results: It was followed four pigs from ten different lots sent to slaughter. Prior to slaughter, samples were collected from the floors of the holding pens in the slaughterhouse. During slaughter, samples from seven different points were collected: 1) stool from the rectum immediately after stunning; 2) external surface of the carcass after dehairing; 3) internal surface of the carcass after evisceration; 4) external surface of the half-carcass prior to entry into the cold chamber; 5) tongue surface; 6) jowls; and 7) mesenteric lymph nodes. The strains were obtained through microbiological analysis. To compare the similarity between the strains, rep-PCR was performed. Of the ten samples collected in the holding pens, four (40%) were contaminated with SCP. At slaughter, 280 samples were collected and 56 (20%) SCP isolates were obtained. The lymph nodes were the point of greatest isolation (19.6%), followed by the surface of the carcass at the entrance to the cold chamber (17.8%), the rectum after desensitization (16.1%), carcass surface after opening of the abdominal cavity (16.1%), jowls (12.5%), carcass surface after dehairing (8.9%) and tongue surface (8.9%).[...](AU)

Molecular epidemiology of Pseudomonas aeruginosa isolated from lower respiratory tract of ICU patients

Abstract Lower respiratory tract infections (LRTIs) caused by Pseudomonas aeruginosa are the most common infection among hospitalized patients, associated with increased levels of morbidity, mortality and attributable health care costs. Increased resistant Pseudomonas worldwide has been quite meaningful to patients, especially in intensive care unit (ICUs). Different species of Pseudomonas exhibit different genetic profile and varied drug resistance. The present study determines the molecular epidemiology through DNA fingerprinting method and drug resistance of P. aeruginosa isolated from patients with LTRIs admitted in ICU. A total of 79 P. aeruginosa isolated from patients with LRTIs admitted in ICU were characterized by Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD) and Repetitive Extrapalindromic PCR (REP-PCR). Antibiotic resistance was determined by minimum inhibitory concentration (MIC) assay while MDR genes, viz, blaTEM, blaOXA, blaVIM, blaCTX-M-15 were detected by polymerase chain reaction (PCR). Of the 137 Pseudomonas sp isolated from ICU patients, 57.7% of the isolates were reported to be P. aeruginosa. The overall prevalence of P. aeruginosa among the all included patients was 34.5%. The RAPD analysis yielded 45 different patterns with 72 clusters with 57% to 100% similarity level. The RFLP analysis yielded 8 different patterns with 14 clusters with 76% to 100% similarity level. The REP PCR analysis yielded 37 different patterns with 65 clusters with 56% to 100% similarity level. There was no correlation among the different DNA patterns observed between the three different methods. Predominant of the isolates (46.8%) were resistant to amikacin. Of the 79 isolates, 60.8% were positive for blaTEM gene and 39.2% were positive for blaOXA gene. P. aeruginosa was predominantly isolated from patients with LRTIs admitted in ICU. The difference in the similarity level observed between the three DNA fingerprinting methods indicates that there is high inter-strain variability. The high genetic variability and resistance patterns indicates that we should continuously monitor the trend in the prevalence and antibiotic resistance of P. aeruginosa especially in patients with LRTIs admitted in ICU.

Resumo Infecções do trato respiratório inferior (ITRIs) são as infecções mais comuns entre pacientes internados em unidade de terapia intensiva (UTI). Pseudomonas aeruginosa é a causa mais comum de ITRIs e está associada ao aumento da mortalidade. Diferentes espécies de Pseudomonas exibem diferentes perfis genéticos e resistência variada as drogas. O presente estudo determina a epidemiologia molecular através do método de fingerprinting de DNA e resistência as drogas de P. aeruginosa isoladas de pacientes com LTRIs internados em UTI. Um total de 79 P. aeruginosa isoladas de pacientes com ITRIs internados em UTI foram caracterizados por Polimorfismo de Comprimento de Fragmentos de Restrição (RFLP), DNA Polimórfico Amplificado ao Acaso (RAPD) e PCR Extrapalindrômico Repetitivo (REP-PCR). A resistência aos antibióticos foram determinadas pelos ensaios de concentrações inibitória mínima (MIC), enquanto os genes MDR, blaTEM, blaOXA, blaVIM, blaCTX-M-15 foram detectados pela reação em cadeia da polimerase (PCR). Das 137 Pseudomonas sp isoladas de pacientes de UTI, 57,7% dos isolados foram relatados como P. aeruginosa. A prevalência geral de P. aeruginosa entre os pacientes incluídos foram de 34,5%. A análise RAPD renderam 45 padrões diferentes com 72 clusters com nível de similaridade de 57% a 100%. A análise RFLP renderam 8 padrões diferentes com 14 clusters com 76% a 100% de similaridade. A análise de PCR do REP produziram 37 padrões diferentes com 65 clusters com nível de similaridade de 56% a 100%. Não houveram correlações entre os diferentes padrões de DNA observados entre os três diferentes métodos. Predominantes dos isolados (46,8%) eram resistentes à amicacina. Dos 79 isolados, 60,8% foram positivos para o gene blaTEM e 39,2% foram positivos para o gene blaOXA. P. aeruginosa foi predominantemente isolado de pacientes com ITRIs internados em UTI. A diferença no nível de similaridade observado entre os três métodos de fingerprinting do DNA indica que há alta variabilidade inter-strain. A alta variabilidade genética e os padrões de resistência indicam que devemos monitorar continuamente a tendência na prevalência e resistência a antibióticos de P. aeruginosa, especialmente em pacientes com ITRIs internados em UTI.

Fontes de contaminação de Yersinia enterocolitica durante a produção de leite / Contamination sources of Yersinia enterocolitica during milk production

Objetivou-se determinar as possíveis fontes de contaminação de Yersinia enterocolitica em diferentes pontos do processo de ordenha de vacas leiteiras em oito propriedades da região de Pelotas, RS, ao longo de um ano. Foram analisadas amostras de leite cru de conjunto logo após a ordenha, água de estábulo leiteiro, mão de ordenhador, balde de recolhimento do leite e insuflador de teteiras. As amostras de leite cru e água foram coletadas em frascos estéreis, e as amostras de mão, balde e teteiras com zaragatoas estéreis. As amostras de leite cru foram submetidas a um pré-enriquecimento em água peptonada, sendo posteriormente incubadas em caldo PSTA, adicionado de ampicilina. As amostras de água foram filtradas em membrana de éster de celulose e incubadas em caldo TSB. As amostras de leite após incubação em PSTA, as membranas utilizadas na filtragem da água incubadas em TSB, bem como o material de mãos, balde e teteiras coletadas nas zaragatoas, foram semeados em ágar MacConkey e incubados para a obtenção de colônias. Colônias características foram analisadas por meio de duplex PCR para confirmação da espécie. Os perfis moleculares dos isolados de Y. enterocolitica foram comparados utilizando-se a técnica de rep-PCR. Y. enterocolitica foi isolada de 9,37% das amostras de leite, 6,25% das amostras de água e 12,5% das amostras de mão. Não houve similaridade no perfil de bandas dos isolados encontrados, entretanto foi identificada a presença de cepas diferentes na mesma amostra, demonstrando uma variedade grande de cepas distribuídas no ambiente. A presença de Y. enterocolitica em leite cru no Brasil é preocupante, já que uma quantidade considerável do produto ainda é comercializada de forma clandestina, expondo o consumidor ao risco de infecção pela bactéria, ao consumi-lo sem tratamento térmico adequado.(AU)

This work was performed in order to determine the possible Yersinia enterocolitica contamination sources at different points of the dairy cows milking process in eight properties of Pelotas, RS, in a year. Raw milk samples were analyzed immediately after milking, as well as water from milking parlor, milkers' hands, milk collection bucket, and inflator liners. The samples of raw milk and water were collected in sterile bottles and hand samples, and sterile swabs were used for the buckets and liners. The raw milk samples were subjected to a pre-enrichment peptone water buffered and subsequently incubated in PSTA broth with added ampicillin. Water samples were filtered through cellulose ester membrane and incubated in TSB medium. The milk samples after incubation in PSTA, the membranes used in water filtration were incubated in TSB and the material of the hands material, bucket and liners collected in the swabs were plated on MacConkey agar to obtain colonies. Characteristics of colonies were analyzed by duplex PCR to confirm the species. The molecular profiles of Y. enterocolitica isolates were compared using rep-PCR. Y. enterocolitica was isolated from 9,37% of milk samples, 6,25% of water samples and 12,5% of hand samples. There weren't similarities in the band profile of the isolates found; however, the presence of different strains was found in the same sample, demonstrating a variety of strains distributed in the environment. The presence of Y. enterocolitica in raw milk in Brazil is dangerous, considering that the product is sold clandestinely, exposing consumers to the risk of infection by the bacterium, when consuming it without proper heat treatment.(AU)

Fontes de contaminação de Yersinia enterocolitica durante a produção de leite / Contamination sources of Yersinia enterocolitica during milk production

Objetivou-se determinar as possíveis fontes de contaminação de Yersinia enterocolitica em diferentes pontos do processo de ordenha de vacas leiteiras em oito propriedades da região de Pelotas, RS, ao longo de um ano. Foram analisadas amostras de leite cru de conjunto logo após a ordenha, água de estábulo leiteiro, mão de ordenhador, balde de recolhimento do leite e insuflador de teteiras. As amostras de leite cru e água foram coletadas em frascos estéreis, e as amostras de mão, balde e teteiras com zaragatoas estéreis. As amostras de leite cru foram submetidas a um pré-enriquecimento em água peptonada, sendo posteriormente incubadas em caldo PSTA, adicionado de ampicilina. As amostras de água foram filtradas em membrana de éster de celulose e incubadas em caldo TSB. As amostras de leite após incubação em PSTA, as membranas utilizadas na filtragem da água incubadas em TSB, bem como o material de mãos, balde e teteiras coletadas nas zaragatoas, foram semeados em ágar MacConkey e incubados para a obtenção de colônias. Colônias características foram analisadas por meio de duplex PCR para confirmação da espécie. Os perfis moleculares dos isolados de Y. enterocolitica foram comparados utilizando-se a técnica de rep-PCR. Y. enterocolitica foi isolada de 9,37% das amostras de leite, 6,25% das amostras de água e 12,5% das amostras de mão. Não houve similaridade no perfil de bandas dos isolados encontrados, entretanto foi identificada a presença de cepas diferentes na mesma amostra, demonstrando uma variedade grande de cepas distribuídas no ambiente. A presença de Y. enterocolitica em leite cru no Brasil é preocupante, já que uma quantidade considerável do produto ainda é comercializada de forma clandestina, expondo o consumidor ao risco de infecção pela bactéria, ao consumi-lo sem tratamento térmico adequado.(AU)

This work was performed in order to determine the possible Yersinia enterocolitica contamination sources at different points of the dairy cows milking process in eight properties of Pelotas, RS, in a year. Raw milk samples were analyzed immediately after milking, as well as water from milking parlor, milkers' hands, milk collection bucket, and inflator liners. The samples of raw milk and water were collected in sterile bottles and hand samples, and sterile swabs were used for the buckets and liners. The raw milk samples were subjected to a pre-enrichment peptone water buffered and subsequently incubated in PSTA broth with added ampicillin. Water samples were filtered through cellulose ester membrane and incubated in TSB medium. The milk samples after incubation in PSTA, the membranes used in water filtration were incubated in TSB and the material of the hands material, bucket and liners collected in the swabs were plated on MacConkey agar to obtain colonies. Characteristics of colonies were analyzed by duplex PCR to confirm the species. The molecular profiles of Y. enterocolitica isolates were compared using rep-PCR. Y. enterocolitica was isolated from 9,37% of milk samples, 6,25% of water samples and 12,5% of hand samples. There weren't similarities in the band profile of the isolates found; however, the presence of different strains was found in the same sample, demonstrating a variety of strains distributed in the environment. The presence of Y. enterocolitica in raw milk in Brazil is dangerous, considering that the product is sold clandestinely, exposing consumers to the risk of infection by the bacterium, when consuming it without proper heat treatment.(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)