Enterococos resistentes à vancomicina e tetraciclina em carnes cruas e processadas: características fenotípicas e genotípicas / Vancomycin and tetracycline-resistant enterococci from from raw and processed meats: phenotypic and genotypic characteristics of isolates

The ubiquitous nature of enterococci and their ability to colonize different habitats account for their easy spread throughout the food chain. Here, we evaluated the distribution and antimicrobial susceptibility of Enterococcus isolates from meats obtained from different supermarkets. We acquired and cultured 100 products (raw chicken meat, raw pork, and boiled meats) to screen for the presence of Enterococcus spp. In total, 194 isolates were recovered from the samples, with contamination rates of 63.6% in the chicken samples, 31% in the raw pork meat, and 1.4% in the boiled meat samples. PCR amplification with specific primers was performed to screen the DNA of Enterococcus spp. (95/96), E. faecalis (66/96), E. faecium (30/96), and E. casseliflavus/E. flavescens (3/96). The antimicrobial susceptibility tests showed that all the isolates were resistant to at least one of the antibiotics. All E. faecium isolates were resistant to vancomycin, streptomycin, ciprofloxacin, norfloxacin, erythromycin, and tetracycline. The E. casseliflavus/E. flavescens isolates were resistant to gentamicin, streptomycin, ciprofloxacin, norfloxacin, erythromycin, and tetracycline. E. faecalis isolates were resistant to ciprofloxacin, tetracycline, and erythromycin (92%), norfloxacin (83%), vancomycin, and streptomycin (50%). The resistance genes tetL and vanB were detected by genotyping. The presence of these antimicrobial-resistant microorganisms in food might pose problems for public health.

A natureza ubíqua dos enterococos e sua capacidade de colonizar diferentes habitats são responsáveis pela sua fácil disseminação pela cadeia alimentar. No presente estudo, avaliamos a distribuição e a susceptibilidade antimicrobiana de isolados de Enterococcus provenientes de produtos cárneos. Cem produtos (carne de frango cru, carne de porco crua e carne cozida) foram adquiridos e cultivados para a presença de Enterococcus spp. No total, 194 amostras foram avaliadas, com taxas de contaminação de 63,6% nas amostras de frango, 31% na carne de porco crua e 1,4% nas amostras de carne cozida. A amplificação por PCR foi realizada para confirmar a presença de Enterococcus spp. (95/96), E. faecalis (66/96), E. faecium (30/96) E. casseliflavus/E. flavescens (3/96). Resultados de susceptibilidade mostraram que 100% dos isolados foram resistentes a pelo menos um antibiótico, sendo 100% de E. faecium resistentes a vancomicina, estreptomicina, ciprofloxacina, norfloxacina, eritromicina e tetraciclina. E. casseliflavus / E. flavescens resistentes a gentamicina, estreptomicina, ciprofloxacina, norfloxacina, eritromicina e tetraciclina. E. faecalis foram resistentes a ciprofloxacina, tetraciclina e eritromicina (92%), norfloxacina (83%), vancomicina e estreptomicina (50%). Na genotipagem, foram detectados os genes tetL e vanB. A presença desses microrganismos resistentes aos antimicrobianos nos alimentos pode causar problemas para a saúde pública.

Enterococos resistentes à vancomicina e tetraciclina em carnes cruas e processadas: características fenotípicas e genotípicas / Vancomycin and tetracycline-resistant enterococci from from raw and processed meats: phenotypic and genotypic characteristics of isolates

The ubiquitous nature of enterococci and their ability to colonize different habitats account for their easy spread throughout the food chain. Here, we evaluated the distribution and antimicrobial susceptibility of Enterococcus isolates from meats obtained from different supermarkets. We acquired and cultured 100 products (raw chicken meat, raw pork, and boiled meats) to screen for the presence of Enterococcus spp. In total, 194 isolates were recovered from the samples, with contamination rates of 63.6% in the chicken samples, 31% in the raw pork meat, and 1.4% in the boiled meat samples. PCR amplification with specific primers was performed to screen the DNA of Enterococcus spp. (95/96), E. faecalis (66/96), E. faecium (30/96), and E. casseliflavus/E. flavescens (3/96). The antimicrobial susceptibility tests showed that all the isolates were resistant to at least one of the antibiotics. All E. faecium isolates were resistant to vancomycin, streptomycin, ciprofloxacin, norfloxacin, erythromycin, and tetracycline. The E. casseliflavus/E. flavescens isolates were resistant to gentamicin, streptomycin, ciprofloxacin, norfloxacin, erythromycin, and tetracycline. E. faecalis isolates were resistant to ciprofloxacin, tetracycline, and erythromycin (92%), norfloxacin (83%), vancomycin, and streptomycin (50%). The resistance genes tetL and vanB were detected by genotyping. The presence of these antimicrobial-resistant microorganisms in food might pose problems for public health.(AU)

A natureza ubíqua dos enterococos e sua capacidade de colonizar diferentes habitats são responsáveis pela sua fácil disseminação pela cadeia alimentar. No presente estudo, avaliamos a distribuição e a susceptibilidade antimicrobiana de isolados de Enterococcus provenientes de produtos cárneos. Cem produtos (carne de frango cru, carne de porco crua e carne cozida) foram adquiridos e cultivados para a presença de Enterococcus spp. No total, 194 amostras foram avaliadas, com taxas de contaminação de 63,6% nas amostras de frango, 31% na carne de porco crua e 1,4% nas amostras de carne cozida. A amplificação por PCR foi realizada para confirmar a presença de Enterococcus spp. (95/96), E. faecalis (66/96), E. faecium (30/96) E. casseliflavus/E. flavescens (3/96). Resultados de susceptibilidade mostraram que 100% dos isolados foram resistentes a pelo menos um antibiótico, sendo 100% de E. faecium resistentes a vancomicina, estreptomicina, ciprofloxacina, norfloxacina, eritromicina e tetraciclina. E. casseliflavus / E. flavescens resistentes a gentamicina, estreptomicina, ciprofloxacina, norfloxacina, eritromicina e tetraciclina. E. faecalis foram resistentes a ciprofloxacina, tetraciclina e eritromicina (92%), norfloxacina (83%), vancomicina e estreptomicina (50%). Na genotipagem, foram detectados os genes tetL e vanB. A presença desses microrganismos resistentes aos antimicrobianos nos alimentos pode causar problemas para a saúde pública.(AU)

Vancomycin susceptibility profiles of Staphylococcus spp. isolates from domestic and wild animals / Perfil de suscetibilidade à Vancomicina de isolados de Staphylococcus spp. de animais domésticos e silvestres

Staphylococcus spp. are bacteria involved in human and animal infections. They are resistant to antimicrobials and have become a major public health concern. In recent years, there has been a significant increase in methicillin-resistant Staphylococcus strains and vancomycin is the drug of choice for the treatment of such isolates. However, the minimum inhibitory concentration (MIC) of vancomycin necessary to combat this microorganism has been showing an increase. The aim of the present study was to determine the susceptibility profile of the Staphylococcus spp. of domestic and wild animals to vancomycin, using the microdilution in broth and E-test® techniques, as well as comparing the results of both tests. Of the 50 isolates tested, 47 (94 %) were sensitive to vancomycin in the microdilution and 43 (86 %) were sensitive to vancomycin in the E-test®. Seven (14 %) isolates had an intermediate result showing a risk to public health since the detection of these isolates may precede the occurrence of isolates resistant to vancomycin. In addition, the mecA gene was detected in 78 % of the tested samples. Six of the seven isolates with intermediate resistance to vancomycin were carriers of the mecA gene, showing that these isolates had a potential risk of becoming resistant. Thus, control measures must be taken to prevent the spread of these isolates with intermediate resistance and preserve the effectiveness of this antimicrobial for the treatment of infections caused by multiresistant Staphylococcus spp.(AU)

Staphylococcus spp. são bactérias envolvidas em infecções de humanos e animais, resistentes a antimicrobianos e tem se tornado uma grande preocupação em saúde pública. Nos últimos anos houve um aumento significativo de Staphylococcus resistentes à meticilina e a vancomicina é a droga de escolha para o tratamento desses isolados, porém vem apresentando elevação nos valores de Concentração Inibitória Mínima (CIM) necessários para combater este microrganismo. O objetivo do presente trabalho foi determinar o perfil de suscetibilidade à vancomicina para isolados de Staphylococcus spp. de animais domésticos e silvestres pelas técnicas de Microdiluição em caldo e E-test®, bem como comparar os resultados de ambos os testes. Dos 50 isolados testados 47 (94%) foram sensíveis à vancomicina na Microdiluição e 43 (86%) foram sensíveis à vancomicina no E-test®. Sete (14%) isolados tiveram resultado intermediário demonstrando um risco à saúde pública visto que a detecção destes isolados pode preceder a ocorrência de isolados resistentes à vancomicina. Ademais o gene mecA foi detectado em 78% das amostras testadas, sendo que dos sete isolados com resistência intermediária à vancomicina, seis eram portadores do gene mecA, evidenciando que esses isolados possuem potencial risco de se tornarem resistentes. Dessa forma medidas de controle devem ser tomadas para evitar a propagação destes isolados com resistência intermediária e preservar a eficácia deste antimicrobiano para o tratamento de infecções causadas por Staphylococcus multirresitentes.(AU)

Companion psittacine birds as reservoir of gentamicin and vancomycin-resistant Enterococcus spp / Psitacídeos de companhia como reservatórios de Enterococcus spp. gentamicina e vancomicina-resistentes

Enterococcus are recognized worldwide as significant nosocomial agents that have been continuously envolving to adapt to different niches and acquire resistance to several antibiotic classes. Vancomycin and gentamicin-resistant strains of E. faecalis and E. faecium have been associated with nosocomial human infections. Some epidemiological studies suggest the participation of pets as reservoirs of vancomycin and gentamicin-resistant Enterococcus strains. However, the role of companion birds as reservoirs of these strains has been poorly studied. In this study, 126 psittacine birds were evaluated and 26.9% carried Enterococcus spp., including the species E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum and E. casseliflavus. The antibiotic resistance profile showed four high-level gentamicin-resistance (HLGR) strains. In addition, two strains presented intermediate levels of vancomycin resistance. Resistant strains were isolated from fecal and oropharynx samples of sick and clinically healthy birds, suggesting that psittacine birds may act as reservoirs of HLGR Enterococcus spp. However, sick birds appear to be more implicated in the enterococci transmission than healthy birds.(AU)

Enterococcus são reconhecidos mundialmente como significantes agentes nosocomiais, que têm continuamente se adaptado a diferentes nichos e adquirido resistência a várias classes de antibióticos. Cepas de E. faecalis e E. faecium vancomicina e gantamicina-resistentes têm sido associadas a infecções nosocomiais em humanos. Alguns estudos epidemiológicos sugerem a participação de aves como reservatórios de cepas de Enterococcus vancomicina e gentamicina-resistentes. Entretanto, a relação das aves de companhia como reservatórios destas cepas tem sido pouco estudada. Neste estudo, 126 psitacídeos foram avaliados, e 26,9% destes eram portadores de Enterococcus spp., incluindo as espécies E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum e E. casseliflavus. O perfil de resistência antibiótica mostrou quatro cepas com alto nível de resistência a gentamicina (ANRG). Além de duas cepas com nível intermediário de resistência a vancomicina. As cepas resistentes foram isoladas de amostras fecais e de orofaringe de aves doentes e clinicamente saudáveis, sugerindo que psitacídeos podem estar atuando como reservatórios para Enterococcus spp. com ANRG. Contudo, Aves doentes parecem estar mais relacionadas à transmissão de enterococcus, do que aves saudáveis.(AU)

Companion psittacine birds as reservoir of gentamicin and vancomycin-resistant Enterococcus spp / Psitacídeos de companhia como reservatórios de Enterococcus spp. gentamicina e vancomicina-resistentes

Enterococcus are recognized worldwide as significant nosocomial agents that have been continuously envolving to adapt to different niches and acquire resistance to several antibiotic classes. Vancomycin and gentamicin-resistant strains of E. faecalis and E. faecium have been associated with nosocomial human infections. Some epidemiological studies suggest the participation of pets as reservoirs of vancomycin and gentamicin-resistant Enterococcus strains. However, the role of companion birds as reservoirs of these strains has been poorly studied. In this study, 126 psittacine birds were evaluated and 26.9% carried Enterococcus spp., including the species E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum and E. casseliflavus. The antibiotic resistance profile showed four high-level gentamicin-resistance (HLGR) strains. In addition, two strains presented intermediate levels of vancomycin resistance. Resistant strains were isolated from fecal and oropharynx samples of sick and clinically healthy birds, suggesting that psittacine birds may act as reservoirs of HLGR Enterococcus spp. However, sick birds appear to be more implicated in the enterococci transmission than healthy birds.(AU)

Enterococcus são reconhecidos mundialmente como significantes agentes nosocomiais, que têm continuamente se adaptado a diferentes nichos e adquirido resistência a várias classes de antibióticos. Cepas de E. faecalis e E. faecium vancomicina e gantamicina-resistentes têm sido associadas a infecções nosocomiais em humanos. Alguns estudos epidemiológicos sugerem a participação de aves como reservatórios de cepas de Enterococcus vancomicina e gentamicina-resistentes. Entretanto, a relação das aves de companhia como reservatórios destas cepas tem sido pouco estudada. Neste estudo, 126 psitacídeos foram avaliados, e 26,9% destes eram portadores de Enterococcus spp., incluindo as espécies E. faecalis, E. faecium, E. hirae, E. phoeniculicola, E. gallinarum e E. casseliflavus. O perfil de resistência antibiótica mostrou quatro cepas com alto nível de resistência a gentamicina (ANRG). Além de duas cepas com nível intermediário de resistência a vancomicina. As cepas resistentes foram isoladas de amostras fecais e de orofaringe de aves doentes e clinicamente saudáveis, sugerindo que psitacídeos podem estar atuando como reservatórios para Enterococcus spp. com ANRG. Contudo, Aves doentes parecem estar mais relacionadas à transmissão de enterococcus, do que aves saudáveis.(AU)

The effects of ozone on bacterial growth and thiol-disulphide homeostasis in vascular graft infection caused by MRSA in rats

Purpose: To investigate the microbiological, inflammatory and oxidant effects of adjuvant ozone administration in experimental rat vascular graft infection model which has not been previously investigated. Methods: Forty adult Wistar rats were divided into Sham, Control, Vancomycin, Ozone, Vancomycin+Ozone groups. Grafts were inoculated with Methicillin-resistant Staphylococcus aureus (MRSA) strain and implanted subcutaneously. Rats were treated intraperitoneally with ozone and /or intramuscularly with vancomycin for 10 days. Grafts were evaluated by quantitative bacterial cultures. Blood samples were harvested for determination of thiol-disulphide and cytokine profiles. Results: There was no significant difference in bacterial counts between Control and Ozone Groups. In the Ozone Group median colony count was significantly higher than the Vancomycin and Vancomycin+Ozone Groups. Total thiol and disulphide levels increased and disulphide/native thiol and disulphide/total thiol ratios decreased in Ozone Group significantly. Albumin levels decreased significantly in Vancomycin and Vancomycin+Ozone Groups compared to the Sham Group. IL-1 and TNF-alpha levels significantly increased in infected rats. Decreased levels of VEGF due to infection reversed by ozone therapy in control and vancomycin groups. Conclusions: We didn't observe any benefit of the agent on MRSA elimination in our model. Likewise, effects of ozone on thiol-disulphide homeostasis and inflammatory cytokines were contradictory.(AU)

Detection of vancomycin-resistant enterococci (VRE) in stool specimens submitted for Clostridium difficile toxin testing

The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24 h at 37 °C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p 0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p 0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.(AU)

Perfil de sensibilidade antimicrobiana de Clostridioides (Clostridium) difficile ao metronidazol, vancomicina e moxifloxacina isolados de animais e seres humanos em Minas Gerais, Brasil

Clostridioides difficile é um enteropatógeno bacteriano que infecta animais e seres humanos, capaz de promover distúrbios intestinais que podem levar o seu hospedeiro ao óbito. A infecção por C. difficile (ICD) representa atualmente uma das principais causas de infecção hospitalar, sendo a antibioticoterapia o principal fator predisponente. Na década de 2000, surtos ocasionados por C. difficile estiveram intimamente relacionados ao uso indiscriminado de antibióticos da classe das fluoroquinolonas. No Brasil, o tratamento da ICD é baseado no uso do metronidazol e da vancomicina. O uso de antimicrobianos parece ter relação direta com a seleção de determinadas estirpes de C. difficile. Nessa perspectiva, estudos epidemiológicos relacionados à resistência antimicrobiana desse patógeno são fundamentais para evitar a emergência de estirpes que podem representar um problema de saúde pública. Entretanto, o teste de sensibilidade antimicrobiana padrão-ouro recomendado para C. difficile é laborioso e de custo elevado. Dessa forma, o teste de disco difusão torna-se uma opção mais atrativa, uma vez que o método é simples de executar, mais barato e eficaz. O estudo teve como objetivo avaliar o perfil de sensibilidade ao metronidazol, à vancomicina e à moxifloxacina através do método de disco difusão de 120 estirpes de C. difficile isoladas de animais e seres humanos no Brasil. De acordo com pontos de corte já estabelecidos em estudos anteriores, as estirpes consideradas resistentes nesse teste foram submetidas ao método E-test para confirmação do fenótipo através da concentração inibitória mínima (CIM), mediante os critérios preconizados pelos órgãos de referência. Adicionalmente, três isolados classificados como resistentes à moxifloxacina foram sequenciados (gene gyrA) para detectar mutações que poderiam indicar o mecanismo genotípico de resistência à droga. Nenhum isolado foi resistente à vancomicina e somente uma estirpe (0,8%), considerada não toxigênica (RT010, clado 1), apresentou resistência ao metronidazol (CIM > 256 g/mL). Em relação à moxifloxacina, 14% (n = 17) das estirpes avaliadas foram resistentes, as quais obtiveram CIM > 32 g/mL. O ribotipo RT126 (clado 5) foi o único a apresentar associação estatística positiva de resistência à moxifloxacina (P < 0,0001), bem como os isolados das espécies equina e suína (P = 0,0139 e P = 0,0228, respectivamente). A mutação do tipo Thr82Ile foi encontrada nos três isolados sequenciados. O estudo revelou uma baixa frequência de resistência ao metronidazol e à vancomicina, enquanto a resistência à moxifloxacina foi superior a trabalhos anteriores realizados no Brasil

Clostridioides difficile is a bacterial enteropathogen that infects animals and humans, able to promote intestinal disorders that can lead to its host's death. Infection with C. difficile (CDI) currently represents one of the main causes of nosocomial infection, with antibiotic therapy being the main predisposing factor. In the 2000s, outbreaks caused by C. difficile were closely related to the indiscriminate use of antibiotics of the fluoroquinolone class. In Brazil, the treatment of CDI is based on the use of metronidazole and vancomycin. The use of antimicrobials seems to have a direct relationship with the selection of certain strains of C. difficile. In this perspective, epidemiological studies related to the antimicrobial resistance of this pathogen are essential to avoid the emergence of strains that may represent a public health problem. However, the gold standard antimicrobial susceptibility test recommended for C. difficile is laborious and expensive. Thus, the diffusion disk test becomes a more attractive option, since the method is simple to perform, cheaper and more effective. The study aimed to evaluate the sensitivity profile to metronidazole, vancomycin and moxifloxacin using the disk diffusion method from 120 strains of C. difficile from animals and humans isolated in Brazil. According to cutoff points already established in previous studies, strains considered resistant in this test were submitted to the E-test method to confirm the phenotype through the minimum inhibitory concentration (MIC), according to the criteria recommended by the reference institution. In addition, three isolates classified as resistant to moxifloxacin were sequenced (gyrA gene) to detect mutations that could indicate the genotypic mechanism of drug resistance. There was no vancomycin-resistant isolate and one strain (0.8%), non-toxigenic (RT010, clade 1), was resistant to metronidazole (MIC > 256 g/mL). Regarding moxifloxacin, 14% (n = 17) of the strains evaluated were resistant, which obtained MIC > 32 g/mL. The ribotype RT126 (clade 5) was the only one to present a positive statistical association of resistance to moxifloxacin (P < 0.0001), as well as isolates of the equine and swine species (P = 0.0139 and P = 0.0228, respectively). The Thr82Ile mutation was found in the three sequenced isolates. The study revealed a low frequency of resistance to metronidazole and vancomycin, while resistance to moxifloxacin was superior to previous studies carried out in Brazil.

Detection of both vanA & vanB genes in vanA phenotypes of Enterococci by Taq Man RT-PCR

Twenty seven isolates of vancomycin resistant Enterococci based on the disk diffusion and E- test have been screened; being found eight (0.3%) clinical isolates of vanA & vanB through Taq Man Real Time PCR assay. This study shows the presence of both vanA & vanB genotypes in vanA phenotypes clinical isolates in the three hospitals in Iran.(AU)

In vitro susceptibility to methicillin, vancomycin and linezolid of staphylococci isolated from bloodstream infections in eastern Turkey

Staphylococcus species are one of the major causes of bacterial bloodstream infections. Multi-resistant staphylococci infections are major therapeutic problems. This study was aimed to detect methicillin, linezolid and vancomycin susceptibilities of Staphylococcus isolates. A total of 870 Staphylococcus strains isolated from blood cultures of hospitalized patients with BSI. Antimicrobial susceptibilities of methicillin, linezolid and vancomycin were detected according to the Clinical and Laboratory Standards Institute (CLSI). A total of 771 (88.6%) isolates were coagulase-negative staphylococci (CoNS). 700 (80.5%) isolates were methicillin-resistant (MR) and 170 (19.5%) were methicillin-susceptible (MS). All the MS isolates were also susceptible to linezolid. However 15 (1.7%) of MR strains were resistant to linezolid. The minimum inhibitory concentration range for the linezolid-resistant isolates by Etest was 6-32 µg/mL. The difference between linezolid susceptibilities for MS and MR staphylococci was not quite statistically significant (p = 0.052). There was no statistically significant difference between S. aureus and CoNS isolates for linezolid susceptibility. All of the isolates were susceptible to vancomycin. In conclusion, linezolid is currently an efficient option for the treatment of methicillin-resistant staphylococci infections.

Detection of Staphylococcus aureus with an intermediate profile to vancomycin (VISA) isolate from Santa Maria, RS

Twenty-three isolates of Staphylococcus aureus resistant to methicillin have been analyzed, being found a clinical isolate of VISA through microdilution technique. The others techniques were unable to detect such isolates. This is the first study that shows the presence of VISA in clinical isolates in the city of Santa Maria-RS.

Detection of Staphylococcus aureus with an intermediate profile to vancomycin (VISA) isolate from Santa Maria, RS

Twenty-three isolates of Staphylococcus aureus resistant to methicillin have been analyzed, being found a clinical isolate of VISA through microdilution technique. The others techniques were unable to detect such isolates. This is the first study that shows the presence of VISA in clinical isolates in the city of Santa Maria-RS.

Micro-organismos de importância veterinária e zoonóticos em pombos domésticos (Columba livia domestica) do Distrito Federal, Brasil

Pombos domésticos estão amplamente distribuídos nos centros urbanos e podem constituir um reservatório de patógenos fúngicos e bacterianos. Este trabalho teve como objetivos caracterizar micro-organismos de importância veterinária e zoonóticos em swab cloacal de Columba livia domestica clinicamente saudáveis mantidos em criatórios e em um laboratório de experimentação animal no Distrito Federal. Das 100 amostras de swab cloacal analisadas, 75% foram positivas no cultivo microbiológico em ágar sangue, sendo que Escherichia coli (64%) e Enterococcus spp. (18%) caracterizaram-se como as bactérias mais frequentes. Nenhuma amostra mostrou-se positiva para Salmonella spp. e em apenas uma (1%) foi identificado Cryptococcus spp. Em relação à sensibilidade de E. coli aos antimicrobianos, tetraciclina, estreptomicina e sulfazotrim constituíram as drogas menos eficazes in vitro. Nos isolados de Enterococcus spp. testados, os menores percentuais de sensibilidade observados foram para oxacilina, cefalexina e ácido nalidíxico. A ciprofloxacina destacou-se como a droga mais eficaz in vitro para ambas as espécies. Doze (18,1%) isolados de E. coli e oito (80%) de Enterococcus spp. demonstraram resistência múltipla a três ou mais classes de drogas testadas. A formação de biofilme foi evidenciada em todos os isolados de Enterococcus spp. avaliados e em 61% das cepas de E. coli. A caracterização filogenética de E. coli revelou que a maior parte dos isolados pertenceram ao grupo B2 (46,9%), seguido dos grupos A (29,7%) e D (23,4%). Os pombos domésticos avaliados são portadores de micro-organismos multirresistentes e potencialmente patogênicos para humanos, incluindo Enterococcus spp. resistente à vancomicina.

Domestic pigeons are widely distributed in urban centers and may constitute a reservoir of fungic and bacterial pathogens. This work aimed to characterize microorganisms of veterinary and zoonotic significance in cloacal swabs of healthy Columba livia domestica from pigeons breeders and from na animal experimentation laboratory located in Distrito Federal. Of the 100 samples of cloacal swabs analyzed, 75% were positive in the microbiological culture in blood agar, being Escherichia coli (64%) and Enterococcus spp. (18%) the most frequent bacteria. No samples were positive for Salmonella spp. and only one (1%) was positive to Cryptococcus spp. Regarding the sensitivity of E. coli to antimicrobials, tetracycline, streptomycin and sulfazotrim were the least effective drugs in vitro. For Enterococcus spp. isolates, the lowest percentages of sensitivity were observed for oxacillin, cephalexin and nalidixic acid. Ciprofloxacin was the most effective drug in vitro for both species. Twelve (18.1%) E. coli isolates and eight (80%) Enterococcus spp. showed multiple resistance to three or more classes of drugs tested. Biofilm formation was evidenced in all Enterococcus spp. evaluated and in 61% of E. coli strains. The phylogenetic characterization of E. coli revealed that most of the isolates belonged to group B2 (46.9%), followed by groups A (29.7%) and D (23.4%). The evaluated domestic pigeons are carriers of multiresistant and potentially pathogenic microorganisms, including vancomycin resistant Enterococci.

Detecção de Enterococcus resistentes a vancomicina em criações comerciais de ovinos e caprinos das regiões centro-leste e nordeste do Estado de São Paulo

As exigências para com as condições higiênico-sanitárias na produção de animais de interesse zootécnico vêm aumentando progressivamente dada à necessidade de aliar-se produtividade a produtos de alta qualidade para atender a mercados consumidores cada vez mais exigentes. Nesse sentido, a utilização de antimicrobianos, tanto na profilaxia como na terapêutica, permanece como estratégia de controle para vários microrganismos patogênicos, de importância não apenas para a produção animal como também para a saúde humana, ainda que restrições ao uso indiscriminado desses produtos têm se intensificado. Não obstante, o uso excessivo desses produtos está associado à seleção de microrganismos resistentes nas áreas de produção. Por outro lado, investigações sobre circulação de cepas resistentes em rebanhos animais, até então restritas a populações humanas, ainda permanecem limitadas no Brasil. Bactérias do gênero Enterococcus, integrantes usuais da microbiota gastrointestinal animal e humana, são indicadoras ambientais de contaminação fecal e tem-se tornado objeto de preocupação em saúde pública e veterinária dada a ocorrência de cepas resistentes à vancomicina (VRE). O presente trabalho teve como objetivo isolar, quantificar e caracterizar VRE presentes em amostras fecais de ovinos oriundos de pequenas propriedades das regiões centro-leste e nordeste do estado de São Paulo. Para tanto, 132 amostras fecais foram coletadas diretamente do reto dos animais ou do piso das instalações. As amostras foram semeadas em ágar m-Enterococcus e subcultivadas em Ágar Bile Esculina acrescido de 6 g/mL de Vancomicina (ABEV), para confirmação de Enterococcus spp e detecção de cepas resistentes. Procedeu-se igualmente a observação da morfologia, características tintoriais, bioquímicas e moleculares. O número máximo de Enterococcus spp. encontrado foi de 2,6 × 105 e 1,70 × 105 UFC/g de fezes do ambiente e dos animais, respectivamente. Na caracterização bioquímica espécies mais prevalentes foram: Enterococcus faecalis e Vagococcus fluvialis. No ABEV, houve crescimento de colônias VRE em 33 das 84 amostras de ovinos-caprinos e em 21 das 48 amostras ambientais, representando, respectivamente 46,7% e 29,3% das amostras analisadas. A análise por multiplex PCR das 54 cepas VRE obtidas indicaram que 23 (43%), 22 (41%), 2 (3,5%) e 2 (3,5%) foram positivas, respectivamente, para os genes vanC2/C3, vanC1, vanA e vanB, sendo que para 5,3% dos isolados nenhum produto foi amplificado, sugerindo a possível ocorrência de genes dos demais grupos van conhecidos entre os isolados. Os resultados obtidos indicam, de forma inédita no país, a circulação de VRE em propriedades produtoras de ovinos e caprinos, sem ocorrência de manifestações clínicas aparentes nos animais, porém com possíveis riscos à saúde dos produtores e profissionais envolvidos, bem como a eventuais consumidores.

Demands for sanitary conditions in animal farming have been increasing progressively given the need to combine productivity and high quality products to support increasingly demanding consumer markets. In this context, antimicrobial drugs used in prevention as well as in therapy remain as the control strategy for several pathogenic microorganisms, not important only in animal production but also in human health, although restrictions for the indiscriminate use of these drugs have been intensified. However, the excessive use of these products has been associated to the selection of resistant microorganisms in production areas. On the other hand, investigation on strains of public health importance circulating in animal herds is still limited in Brazil. Enterococcus genus bacteria, usually present in animal and human gastrointestinal microbiota, are environmental indicators of fecal contamination and have become a concerning subject in public and veterinary health given the occurrence of strains resistant to vancomycin (VRE). The present study aimed to isolate, quantify and characterize VRE present in stool samples of sheep and goats from several farms in the center-east and northeast regions of São Paulo State. Swabs collected one hundred and thirty-two stool samples either directly from the animal´s rectum or from the ground. Samples were plated onto m-Enterococcus agar plates and subcultivated in Bile esculin agar with 6 g/mL of vancomycin (BEAV) to confirm Enterococcus spp and detect resistant samples. Colonies were identified by colonial morphology, Gram's staining, biochemical, and molecular profile. The highest colony count was equal to 2.6 105 and 1.7 105 CFU/g of feces from environmental and animal samples, respectively. Regarding biochemical characterization, Enterococcus faecalis e Vagococcus fluvialis were the most prevalent species. VRE was detected on BEAV in 33 out of 84 sheep-goat samples and in 21 out of 48 ambient samples, indicating a positivity rate of 46.7% and 29.3% respectively in the investigated samples. Analysis by multiplex PCR of the obtained 54 VRE strains indicated that 23 (43%), 22 (41%) 2 (3.5%) and 2 (3.5%) were positive, respectively, for the vanC2/C3, vanC1, vanA and vanB genes, and no product was amplified for 5.3% of the isolates, suggesting the possible occurrence of other known van gene groups among the isolates. The results obtained in this study indicate, for the first time in the studied areas, the circulation of VRE in sheep and goat farms, with no occurrence of apparent clinical signs in the animals, but with possible health risks to the farmers and workers involved, as well as potential consumers.

First report of vancomycin-resistant Enterococcus faecalis in Uberaba, Minas Gerais state

In this study we report the first isolation of VanA-type vancomycin-resistant Enterococcus faecalis strains from two different patients hospitalized in the same intensive care unit at the hospital of Universidade Federal do Triângulo Mineiro (UFTM), Uberaba, Minas Gerais, Brazil.

Prevalência de enterococos isolados de frangos caipiras em diferentes regiões do Distrito Federal / Prevalence of Enterococcus spp. isolated from free-range chickens in different regions of Distrito Federal, Brazil

It was studied whether vancomycin-resistant enterococci (VRE) colonize poultry raised without receiving antimicrobial growth promoters (AGP), in non-intensive production systems. A total of 200 cloacal swabs were colleted in farms (n=40) of eight different regions of the Distrito Federal. After selective isolation, the typical enterococcal colonies were submitted to the multiplex PCR to identify enterococcal species (E. faecalis, E. faecium, E. gallinarum, and E. casseliflavus), and genes coding for high-level vancomycin resistance phenotypes. No VRE were found in the examined samples. The prevalence rates were higher for E. gallinarum (n=26; 13.0 percent) and E. casseliflavus (n=11; 5.5 percent). It was found remarkable differences in the prevalence of E. gallinarum and E. casseliflavus among the poultry farms and studied regions, and it seems that poultry raised in non-intensive production systems in the Distrito Federal of Brazil are not reservoirs of VRE.(AU)

Susceptibility of Staphylococcus aureus isolates to vancomycin at a university hospital in southern Brazil

The in vitro susceptibilty of Staphylococcus aureus to vancomycin was evaluated from impatients at a Brazilian University Hospital by the Etest and a screening method. No vancomycin intermediate (VISA) or vancomycin resistant (VRSA) S. aureus isolate was identified. Three patients presented as heteroresistant VISA (h-VISA) isolates but none of them received vancomycin previously.

A suscetibilidade in vitro de Staphylococcus aureus à vancomicina foi avaliada em pacientes internados em um Hospital Universitário Brasileiro pelo Etest e por um método de triagem. Nenhuma amostra de S. aureus resistente (VRSA) ou com resistência intermediária (VISA) à vancomicina foi isolada. Três pacientes tiveram amostras heteroresistentes VISA (h-VISA), mas nenhum destes recebeu vancomicina previamente.

Emergence of VanB phenotype-vanA genotype in vancomycin-resistant enterococci in Brazilian hospital

In Brazil, vancomycin-resistant enterococci (VRE) have been reported as nosocomial pathogens since 1998. Recently, in a VRE surveillance in a hospital, we detected two Enterococcus faecalis isolates with vanA genotype and susceptible to teicoplanin. This is the first report of VanB phenotype-vanA genotype enterococci isolated from humans in Brazil.

No Brasil, enterococos resistente à vancomicina (VRE) têm sido descritos como patógenos hospitalares, desde 1998. Durante um monitoramento de VRE em um hospital, foram detectadas duas cepas de Enterococcus faecalis com genótipo vanA, e sensibilidade à teicoplanina. Este é o primeiro relato do isolamento de enterococo fenótipo VanB e genótipo vanA de amostra clínica no Brasil.

Enterococcus spp. resistentes à vancomicina isolados de fezes de frangos, pombos, gamos e ratos / Vancomycin-resistant Enterococcus isolated from feces of poultry, pigeons, deers and rats

The occurrence of vancomycin resistant enterococci (VRE) was investigated in feces of poultry, pigeons, deers and rats. Feces samples were collected from 25 broilers growing in poultry production farms that never used in the diets avaporcine as growth promoter, 6 broiler from different poultry production farms that used regularly avaporcine in the formulated diets until 1997, 8 deers from natural reservoirs, 15 pigeons, and 20 rats arrested from field conditions. The samples of Enterococcus gallinarum isolated from poultry feces were resistant to vancomycin independently of the use of avaporcine in the formulated diet.(AU)

Study on the susceptibility of Enterococcus faecalis from infectious processes to ciprofloxacin and vancomycin

Enterococcus faecalis is considered a pathogen responsible for hospital infections and, due to its frequent multi-resistant profile, has caused preoccupations among many medical authorities. The objective of this study was to determine the antimicrobial susceptibility of 74 strains isolated from blood cultures and purulent secretions to vancomycin and ciprofloxacin through the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) by using the Microdilution test. The results showed a greater efficacy of vancomycin compared to ciprofloxacin (98.6% of the strains were inhibited by vancomycin at lower concentrations: 0.06 - 1 µg/ml). However, in the MBC analysis 73% of the strains showed a MBC of vancomycin only at high concentrations (equal to or higher than 64 µg/ml). For ciprofloxacin, the strains showed a broad sensitivity with MICs and MBCs distributed along all the MIC classes. Results also revealed a probability that some strains are tolerant to vancomycin, which indicates the need of other tests to confirm this characteristic.