Resumo
Enterococci have been used as sentinel organisms for monitoring antimicrobial resistance in food, humans, and other animals. In this sense, the present study evaluated the antimicrobial susceptibility profile and the presence of genes associated with resistance to erythromycin (msrC and ermB) and tetracycline [tet(M) and/or tet(L)] in enterococci isolated from raw sheep's milk and cheeses (colonial, feta-, and pecorino-type) from South region of Brazil. A total of 156 enterococci were isolated from milk (n=80) and cheese (n=76) samples, identified by MALDI-TOF. Enterococcus faecalis (50.6%; n=79) was the most frequent species isolated from both samples. According to in vitro susceptibility tests, enterococci strains were not susceptible to the most commonly antimicrobial agents used in human and veterinary medicine. The frequency of MDR strains in enterococci isolated from milk (53.7%) was higher than those from cheese (24.2%). The tet(M) gene was the most commonly detected among tetracycline not-susceptible strains. The present study provided the first evidence of antimicrobial not-susceptible enterococci in raw sheep's milk and cheeses in South Brazil. Drug-resistant strains, particularly those that are MDR, constitute a One Health issue.
Os enterococos têm sido usados como organismos sentinela para monitorar o padrão de suscetibilidade a antimicrobianos em alimentos, humanos e outros animais. Neste sentido, o presente estudo objetivou avaliar o perfil de susceptibilidade a antimicrobianos e os genes associados com a resistência a eritromicina (msrC and ermB) e à tetraciclina [tet(M) and/or tet(L)] em enterococos isolados de leite cru de ovelha e queijos (colonial, tipo-feta e tipo-pecorino) do Sul do Brasil. Um total de 156 enterococos foram isolados de leite (n=80) e queijo (n=76), identificados por MALDI-TOF. Enterococcus faecalis (50,6%; n=79) foi a espécie mais frequentemente isolada de ambas as amostras. De acordo com o teste de suscetibilidade in vitro, as cepas de enterococos não foram susceptíveis aos agentes antimicrobianos mais comumente utilizados na clínica humana e veterinária. A frequência de cepas de enterococos MDR isoladas do leite (53,7%) foi superior à do queijo (24,2%). O gene tet(M) foi o mais comumente detectado entre as cepas não susceptíveis à tetraciclina. O presente estudo fornece as primeiras evidências de enterococos não susceptíveis aos antimicrobianos em leite cru de ovelha e queijos no Sul do Brasil. Cepas resistentes a drogas, particularmente as que são MDR, representam uma preocupação de Saúde Única.
Assuntos
Ovinos , Queijo/parasitologia , Enterococcus , Laticínios/parasitologia , Abastecimento de AlimentosResumo
The aim of present study is to characterize the resistance and virulence profile of enterococci isolated from aquaculture excavated ponds and masonry tanks (6 samples) in southern Brazil. Samples were cultured in selective medium, 10 colonies were randomly selected from each sample, which were identified by MALDI-TOF and tested against 13 antimicrobials. The presence of resistance (tetL, tetM, tetS, ermB and msrC) and virulence (ace, esp, agg, cylA and gelE) genes were determined by PCR. A total of 79 enterococci were identified, and Entecococcus faecalis (44.3%) and E. casseliflavus (36.7%) were the most prevalent species isolated. Sixty-five strains (82.3%) were resistant to at least one of the antimicrobials tested, whereas 27 (34.2%) strains were multiresistant. The overall percentages of antimicrobial resistant isolates were: 58.2% to rifampicin, 40.5% to fluoroquinolones, 36.7% to erythromycin and 30.4% to tetracycline. The tetL and tetM genes were found in 57.7% of the tetracycline-resistant strains; and msrC in 31.01% of erythromycin-resistant strains. The most frequently detected virulence factors were ace and gelE genes. Although limited to a single farm, these data suggest that aquaculture may be a reservoir of resistant and virulent enterococci. This study is the first step towards enhancing our understandingof distribution, resistance and virulence profile in enterococci isolated from fish farming environments in the south Brazil.(AU)
O objetivo do estudo apresentado é caracterizar o perfil de resistência e virulência de enterococos isolados de viveiros escavados e tanques de alvenaria (6 amostras) de uma pisicultura no Sul do Brasil. As amostras foram cultivadas em meio seletivo, 10 colônias foram selecionadas aleatoriamente de cada amostra, que foram identificadas por MALDI-TOF e testadas contra 13 antimicrobianos. A presença de genes de resistência (tetL, tetM, tetS, ermB e msrC) e virulência (ace, esp, agg, cylA e gelE) foi determinada por PCR. Foram identificados 79 enterococos, sendo Entecococcus faecalis (44,3%) e E. casseliflavus (36,7%) as espécies mais frequentes isoladas. Sessenta e cinco cepas (82,3%) eram resistentes a pelo menos um dos antimicrobianos testados, enquanto 27 (34,2%) eram multirresistentes. As porcentagens gerais de isolados resistentes a antimicrobianos foram: 58,2% para rifampicina, 40,5% para fluoroquinolonas, 36,7% para eritromicina e 30,4% para tetraciclina. Os genes tetL e tetM foram encontrados em 57,7% das cepas resistentes à tetraciclina; e msrC em 31,01% das cepas resistentes à eritromicina. Os fatores de virulência mais comumente detectados foram ace e gelE. Embora limitados a uma única fazenda, esses dados indicam que a aquicultura pode ser uma fonte de enterococos resistentes e virulentos. Este estudo é o primeiro passo para melhorar nosso entendimento da distribuição, resistência e perfil de virulência em enterococos isolados de ambientes de piscicultura no sul do Brasil.(AU)
Assuntos
Animais , Enterococcus/isolamento & purificação , Pesqueiros , Fatores de Virulência , BrasilResumo
Abstract The aim of present study is to characterize the resistance and virulence profile of enterococci isolated from aquaculture excavated ponds and masonry tanks (6 samples) in southern Brazil. Samples were cultured in selective medium, 10 colonies were randomly selected from each sample, which were identified by MALDI-TOF and tested against 13 antimicrobials. The presence of resistance (tetL, tetM, tetS, ermB and msrC) and virulence (ace, esp, agg, cylA and gelE) genes were determined by PCR. A total of 79 enterococci were identified, and Entecococcus faecalis (44.3%) and E. casseliflavus (36.7%) were the most prevalent species isolated. Sixty-five strains (82.3%) were resistant to at least one of the antimicrobials tested, whereas 27 (34.2%) strains were multiresistant. The overall percentages of antimicrobial resistant isolates were: 58.2% to rifampicin, 40.5% to fluoroquinolones, 36.7% to erythromycin and 30.4% to tetracycline. The tetL and tetM genes were found in 57.7% of the tetracycline-resistant strains; and msrC in 31.01% of erythromycin-resistant strains. The most frequently detected virulence factors were ace and gelE genes. Although limited to a single farm, these data suggest that aquaculture may be a reservoir of resistant and virulent enterococci. This study is the first step towards enhancing our understandingof distribution, resistance and virulence profile in enterococci isolated from fish farming environments in the south Brazil.
Resumo O objetivo do estudo apresentado é caracterizar o perfil de resistência e virulência de enterococos isolados de viveiros escavados e tanques de alvenaria (6 amostras) de uma pisicultura no Sul do Brasil. As amostras foram cultivadas em meio seletivo, 10 colônias foram selecionadas aleatoriamente de cada amostra, que foram identificadas por MALDI-TOF e testadas contra 13 antimicrobianos. A presença de genes de resistência (tetL, tetM, tetS, ermB e msrC) e virulência (ace, esp, agg, cylA e gelE) foi determinada por PCR. Foram identificados 79 enterococos, sendo Entecococcus faecalis (44,3%) e E. casseliflavus (36,7%) as espécies mais frequentes isoladas. Sessenta e cinco cepas (82,3%) eram resistentes a pelo menos um dos antimicrobianos testados, enquanto 27 (34,2%) eram multirresistentes. As porcentagens gerais de isolados resistentes a antimicrobianos foram: 58,2% para rifampicina, 40,5% para fluoroquinolonas, 36,7% para eritromicina e 30,4% para tetraciclina. Os genes tetL e tetM foram encontrados em 57,7% das cepas resistentes à tetraciclina; e msrC em 31,01% das cepas resistentes à eritromicina. Os fatores de virulência mais comumente detectados foram ace e gelE. Embora limitados a uma única fazenda, esses dados indicam que a aquicultura pode ser uma fonte de enterococos resistentes e virulentos. Este estudo é o primeiro passo para melhorar nosso entendimento da distribuição, resistência e perfil de virulência em enterococos isolados de ambientes de piscicultura no sul do Brasil.
Resumo
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.
Assuntos
Alimentos Crus/análise , Carne , Enterococos Resistentes à Vancomicina/genética , Resistência a Tetraciclina/genética , Bovinos , Galinhas , Reação em Cadeia da Polimerase , SuínosResumo
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)
Assuntos
Enterococos Resistentes à Vancomicina/genética , Resistência a Tetraciclina/genética , Alimentos Crus/análise , Carne , Reação em Cadeia da Polimerase , Bovinos , Galinhas , SuínosResumo
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)
Assuntos
Animais , Papagaios/microbiologia , Reservatórios de Doenças/veterinária , Gentamicinas , Resistência a Vancomicina , Farmacorresistência Bacteriana , Animais de Estimação/microbiologia , Enterococcus/isolamento & purificaçãoResumo
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)
Assuntos
Animais , Papagaios/microbiologia , Reservatórios de Doenças/veterinária , Gentamicinas , Resistência a Vancomicina , Farmacorresistência Bacteriana , Animais de Estimação/microbiologia , Enterococcus/isolamento & purificaçãoResumo
The fidelity of the genomes is defended by mechanism known as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems. Three Type II CRISPR systems (CRISPR1- cas, CRISPR2 and CRISPR3-cas) have been identified in enterococci isolates from clinical and environmental samples. The aim of this study was to observe the distribution of CRISPR1-cas, CRISPR2 and CRISPR3-cas in non-clinical strains of Enterococcus faecalis and Enterococcus faecium isolates from food and fecal samples, including wild marine animals. The presence of CRISPRs was evaluated by PCR in 120 enterococci strains, 67 E. faecalis and 53 E. faecium. It is the first report of the presence of the CRISPRs system in E. faecalis and E. faecium strains isolated from wild marine animal fecal samples. The results showed that in non-clinical strains, the CRISPRs were more frequently detected in E. faecalis than in E. faecium. And the frequencies of CRISPR1-cas and CRISPR2 were higher (60%) in E. faecalis strains isolated from animal feces, compared to food samples. Both strains showed low frequencies of CRISPR3-cas (8.95% and 1.88%). In conclusion, the differences in the habitats of enterococcal species may be related with the results observe in distribution of CRISPRs systems.(AU)
A fidelidade dos genomas é defendida por mecanismos conhecidos como sistemas de repetições palindrômicas curtas agrupadas e regularmente interespaçadas (CRISPRs). Três tipos de sistemas CRISPR II (CRISPR1-cas, CRISPR2 e CRISPR3-cas) têm sido identificados em cepas de enterococos isolados de amostras clínicas e ambientais. O objetivo deste estudo foi observar a distribuição dos CRISPR1-cas, CRISPR2 e CRISPR3-cas em cepas não-clínicas de Enterococcus faecalis e Enterococcus faecium isoladas de amostras alimentícias e fecais, incluindo animais marinhos selvagens. A presenca dos CRISPRs foi determinada por PCR em 120 cepas de enterococos, sendo 67 E. faecalis e 53 E. faecium. É o primeiro relato da presença do sistema CRISPRs nas estirpes E. faecalis e E. faecium isoladas de amostras fecais de animais marinhos selvagens. Os resultados mostraram que em cepas não-clínicas, os CRISPRs foram mais frequentemente detectados em E. faecalis do que em E. faecium. E as frequências de CRISPR1-cas e CRISPR2 foram maiores (60%) em cepas de E. faecalis isoladas de fezes de animais, quando comparadas à amostras de alimentos. Ambas as cepas apresentaram baixas freqüências de CRISPR3-cas (8,95% e 1,88%). Em conclusão, as diferenças nos habitats das espécies de enterococos podem estar relacionadas com os resultados observados na distribuição dos sistemas CRISPRs.(AU)
Resumo
Abstract The fidelity of the genomes is defended by mechanism known as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems. Three Type II CRISPR systems (CRISPR1- cas, CRISPR2 and CRISPR3-cas) have been identified in enterococci isolates from clinical and environmental samples. The aim of this study was to observe the distribution of CRISPR1-cas, CRISPR2 and CRISPR3-cas in non-clinical strains of Enterococcus faecalis and Enterococcus faecium isolates from food and fecal samples, including wild marine animals. The presence of CRISPRs was evaluated by PCR in 120 enterococci strains, 67 E. faecalis and 53 E. faecium. It is the first report of the presence of the CRISPRs system in E. faecalis and E. faecium strains isolated from wild marine animal fecal samples. The results showed that in non-clinical strains, the CRISPRs were more frequently detected in E. faecalis than in E. faecium. And the frequencies of CRISPR1-cas and CRISPR2 were higher (60%) in E. faecalis strains isolated from animal feces, compared to food samples. Both strains showed low frequencies of CRISPR3-cas (8.95% and 1.88%). In conclusion, the differences in the habitats of enterococcal species may be related with the results observe in distribution of CRISPRs systems.
Resumo A fidelidade dos genomas é defendida por mecanismos conhecidos como sistemas de repetições palindrômicas curtas agrupadas e regularmente interespaçadas (CRISPRs). Três tipos de sistemas CRISPR II (CRISPR1-cas, CRISPR2 e CRISPR3-cas) têm sido identificados em cepas de enterococos isolados de amostras clínicas e ambientais. O objetivo deste estudo foi observar a distribuição dos CRISPR1-cas, CRISPR2 e CRISPR3-cas em cepas não-clínicas de Enterococcus faecalis e Enterococcus faecium isoladas de amostras alimentícias e fecais, incluindo animais marinhos selvagens. A presenca dos CRISPRs foi determinada por PCR em 120 cepas de enterococos, sendo 67 E. faecalis e 53 E. faecium. É o primeiro relato da presença do sistema CRISPRs nas estirpes E. faecalis e E. faecium isoladas de amostras fecais de animais marinhos selvagens. Os resultados mostraram que em cepas não-clínicas, os CRISPRs foram mais frequentemente detectados em E. faecalis do que em E. faecium. E as frequências de CRISPR1-cas e CRISPR2 foram maiores (60%) em cepas de E. faecalis isoladas de fezes de animais, quando comparadas à amostras de alimentos. Ambas as cepas apresentaram baixas freqüências de CRISPR3-cas (8,95% e 1,88%). Em conclusão, as diferenças nos habitats das espécies de enterococos podem estar relacionadas com os resultados observados na distribuição dos sistemas CRISPRs.
Resumo
ABSTRACT Rapid identification of vancomycin-resistant enterococci (VRE) can assist in choosing the appropriate treatment and preventing VRE spread. The performance of chromIDTM VRE agar was evaluated using 184 clinical isolates of Enterococcus spp. and reference strains. The test had a sensitivity of 95.52% but a low specificity of 30%.(AU)
Assuntos
Compostos Cromogênicos/análise , Compostos Cromogênicos/química , Enterococos Resistentes à Vancomicina/química , Enterococos Resistentes à Vancomicina/isolamento & purificaçãoResumo
Abstract The aim of present study is to characterize the resistance and virulence profile of enterococci isolated from aquaculture excavated ponds and masonry tanks (6 samples) in southern Brazil. Samples were cultured in selective medium, 10 colonies were randomly selected from each sample, which were identified by MALDI-TOF and tested against 13 antimicrobials. The presence of resistance (tetL, tetM, tetS, ermB and msrC) and virulence (ace, esp, agg, cylA and gelE) genes were determined by PCR. A total of 79 enterococci were identified, and Entecococcus faecalis (44.3%) and E. casseliflavus (36.7%) were the most prevalent species isolated. Sixty-five strains (82.3%) were resistant to at least one of the antimicrobials tested, whereas 27 (34.2%) strains were multiresistant. The overall percentages of antimicrobial resistant isolates were: 58.2% to rifampicin, 40.5% to fluoroquinolones, 36.7% to erythromycin and 30.4% to tetracycline. The tetL and tetM genes were found in 57.7% of the tetracycline-resistant strains; and msrC in 31.01% of erythromycin-resistant strains. The most frequently detected virulence factors were ace and gelE genes. Although limited to a single farm, these data suggest that aquaculture may be a reservoir of resistant and virulent enterococci. This study is the first step towards enhancing our understandingof distribution, resistance and virulence profile in enterococci isolated from fish farming environments in the south Brazil.
Resumo O objetivo do estudo apresentado é caracterizar o perfil de resistência e virulência de enterococos isolados de viveiros escavados e tanques de alvenaria (6 amostras) de uma pisicultura no Sul do Brasil. As amostras foram cultivadas em meio seletivo, 10 colônias foram selecionadas aleatoriamente de cada amostra, que foram identificadas por MALDI-TOF e testadas contra 13 antimicrobianos. A presença de genes de resistência (tetL, tetM, tetS, ermB e msrC) e virulência (ace, esp, agg, cylA e gelE) foi determinada por PCR. Foram identificados 79 enterococos, sendo Entecococcus faecalis (44,3%) e E. casseliflavus (36,7%) as espécies mais frequentes isoladas. Sessenta e cinco cepas (82,3%) eram resistentes a pelo menos um dos antimicrobianos testados, enquanto 27 (34,2%) eram multirresistentes. As porcentagens gerais de isolados resistentes a antimicrobianos foram: 58,2% para rifampicina, 40,5% para fluoroquinolonas, 36,7% para eritromicina e 30,4% para tetraciclina. Os genes tetL e tetM foram encontrados em 57,7% das cepas resistentes à tetraciclina; e msrC em 31,01% das cepas resistentes à eritromicina. Os fatores de virulência mais comumente detectados foram ace e gelE. Embora limitados a uma única fazenda, esses dados indicam que a aquicultura pode ser uma fonte de enterococos resistentes e virulentos. Este estudo é o primeiro passo para melhorar nosso entendimento da distribuição, resistência e perfil de virulência em enterococos isolados de ambientes de piscicultura no sul do Brasil.
Resumo
Abstract The fidelity of the genomes is defended by mechanism known as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems. Three Type II CRISPR systems (CRISPR1- cas, CRISPR2 and CRISPR3-cas) have been identified in enterococci isolates from clinical and environmental samples. The aim of this study was to observe the distribution of CRISPR1-cas, CRISPR2 and CRISPR3-cas in non-clinical strains of Enterococcus faecalis and Enterococcus faecium isolates from food and fecal samples, including wild marine animals. The presence of CRISPRs was evaluated by PCR in 120 enterococci strains, 67 E. faecalis and 53 E. faecium. It is the first report of the presence of the CRISPRs system in E. faecalis and E. faecium strains isolated from wild marine animal fecal samples. The results showed that in non-clinical strains, the CRISPRs were more frequently detected in E. faecalis than in E. faecium. And the frequencies of CRISPR1-cas and CRISPR2 were higher (60%) in E. faecalis strains isolated from animal feces, compared to food samples. Both strains showed low frequencies of CRISPR3-cas (8.95% and 1.88%). In conclusion, the differences in the habitats of enterococcal species may be related with the results observe in distribution of CRISPRs systems.
Resumo A fidelidade dos genomas é defendida por mecanismos conhecidos como sistemas de repetições palindrômicas curtas agrupadas e regularmente interespaçadas (CRISPRs). Três tipos de sistemas CRISPR II (CRISPR1-cas, CRISPR2 e CRISPR3-cas) têm sido identificados em cepas de enterococos isolados de amostras clínicas e ambientais. O objetivo deste estudo foi observar a distribuição dos CRISPR1-cas, CRISPR2 e CRISPR3-cas em cepas não-clínicas de Enterococcus faecalis e Enterococcus faecium isoladas de amostras alimentícias e fecais, incluindo animais marinhos selvagens. A presenca dos CRISPRs foi determinada por PCR em 120 cepas de enterococos, sendo 67 E. faecalis e 53 E. faecium. É o primeiro relato da presença do sistema CRISPRs nas estirpes E. faecalis e E. faecium isoladas de amostras fecais de animais marinhos selvagens. Os resultados mostraram que em cepas não-clínicas, os CRISPRs foram mais frequentemente detectados em E. faecalis do que em E. faecium. E as frequências de CRISPR1-cas e CRISPR2 foram maiores (60%) em cepas de E. faecalis isoladas de fezes de animais, quando comparadas à amostras de alimentos. Ambas as cepas apresentaram baixas freqüências de CRISPR3-cas (8,95% e 1,88%). Em conclusão, as diferenças nos habitats das espécies de enterococos podem estar relacionadas com os resultados observados na distribuição dos sistemas CRISPRs.
Resumo
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)
Assuntos
Clostridioides difficile , Enterococos Resistentes à Vancomicina , Técnicas Imunoenzimáticas , Infecção HospitalarResumo
This study highlights the prevalence of aminoglycoside-modifying enzyme genes and virulence determinants among clinical enterococci with high-level aminoglycoside resistance in Inner Mongolia, China. Screening for high-level aminoglycoside resistance against 117 enterococcal clinical isolates was performed using the agar-screening method. Out of the 117 enterococcal isolates, 46 were selected for further detection and determination of the distribution of aminoglycoside-modifying enzyme-encoding genes and virulence determinants using polymerase chain reaction -based methods. Enterococcus faecium and Enterococcus faecalis were identified as the species of greatest clinical importance. The aac(6')-Ie-aph(2")-Ia and ant(6')-Ia genes were found to be the most common aminoglycoside-modifying enzyme genes among high-level gentamicin resistance and high-level streptomycin resistance isolates, respectively. Moreover, gelE was the most common virulence gene among high-level aminoglycoside resistance isolates. Compared to Enterococcus faecium, Enterococcus faecalis harbored multiple virulence determinants. The results further indicated no correlation between aminoglycoside-modifying enzyme gene profiles and the distribution of virulence genes among the enterococcal isolates with high-level gentamicin resistance or high-level streptomycin resistance evaluated in our study.(AU)
Assuntos
Aminoglicosídeos/análise , Aminoglicosídeos/síntese química , Aminoglicosídeos/genética , Enterococcus/imunologiaResumo
Background: As a result of the extensive use of antimicrobials in agriculture, animals have been implicated as possible reservoirs of resistant strains of bacteria. Enterococci are members of the normal flora of the gastrointestinaltract of human and animals. Because of their ubiquity, enterococci have been introduced in programs to evaluate thehazard of transmission of resistant bacterial strains through the food chain. The aim of this study was to assess theantimicrobial resistance profile of Enterococcus isolated from swine carcasses at the pre-chill step of processing.Material, Methods & Results: Pig carcasses were sampled at three commercial slaughterhouses (A, B and C). Oneach of two sampling occasions swabs of 100 cm2 areas were taken from each ham, back, belly and jowl of a total of14 pre-chill carcasses. Enterococci were isolated and counted in KF Streptococcus Agar, and typical colonies wereconfirmed by PCR assay targeting the tuf gene. Enterococcus isolates were tested for susceptibility to nine differentantimicrobial agents by agar disc diffusion. From a total of 252 carcasses sampled, 240 (95.2%) presented presumptivecolonies of Enterococcus in counts ranging from 0.02 log CFU.cm-2 to 2.9 log CFU.cm-2. All isolates were confirmedas belonging to the genus Enterococcus, and the great majority was identified as E. faecalis (218/240; 90.83%). Half(125/240; 52.1%) of the Enterococcus isolates were susceptible to all tested antimicrobials. No resistance to ampicillin, vancomycin or teicoplanin was found. The most frequent resistance was to tetracycline (42.5%), followed byerythromycin (26.7%), high level (HLR) streptomycin (20.4%), ciprofloxacin (13.8%), chloramphenicol (12.1%)and HLR-gentamicin (10.4%). Among the 115 resistant Enterococcus isolates, 55 (47.8%) were multi-resistant, andthe distribution of the most common profiles was related to the slaughterhouse...(AU)
Assuntos
Animais , Enterococcus , Suínos/microbiologia , Resistência Microbiana a Medicamentos , Carne/microbiologia , MatadourosResumo
Background: As a result of the extensive use of antimicrobials in agriculture, animals have been implicated as possible reservoirs of resistant strains of bacteria. Enterococci are members of the normal flora of the gastrointestinaltract of human and animals. Because of their ubiquity, enterococci have been introduced in programs to evaluate thehazard of transmission of resistant bacterial strains through the food chain. The aim of this study was to assess theantimicrobial resistance profile of Enterococcus isolated from swine carcasses at the pre-chill step of processing.Material, Methods & Results: Pig carcasses were sampled at three commercial slaughterhouses (A, B and C). Oneach of two sampling occasions swabs of 100 cm2 areas were taken from each ham, back, belly and jowl of a total of14 pre-chill carcasses. Enterococci were isolated and counted in KF Streptococcus Agar, and typical colonies wereconfirmed by PCR assay targeting the tuf gene. Enterococcus isolates were tested for susceptibility to nine differentantimicrobial agents by agar disc diffusion. From a total of 252 carcasses sampled, 240 (95.2%) presented presumptivecolonies of Enterococcus in counts ranging from 0.02 log CFU.cm-2 to 2.9 log CFU.cm-2. All isolates were confirmedas belonging to the genus Enterococcus, and the great majority was identified as E. faecalis (218/240; 90.83%). Half(125/240; 52.1%) of the Enterococcus isolates were susceptible to all tested antimicrobials. No resistance to ampicillin, vancomycin or teicoplanin was found. The most frequent resistance was to tetracycline (42.5%), followed byerythromycin (26.7%), high level (HLR) streptomycin (20.4%), ciprofloxacin (13.8%), chloramphenicol (12.1%)and HLR-gentamicin (10.4%). Among the 115 resistant Enterococcus isolates, 55 (47.8%) were multi-resistant, andthe distribution of the most common profiles was related to the slaughterhouse...
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Animais , Carne/microbiologia , Enterococcus , Resistência Microbiana a Medicamentos , Suínos/microbiologia , MatadourosResumo
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)
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Humanos , Proteínas de Bactérias/genética , Carbono-Oxigênio Ligases/genética , Enterococos Resistentes à Vancomicina , Enterococos Resistentes à Vancomicina/genética , Antibacterianos/farmacologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Irã (Geográfico) , Reação em Cadeia da Polimerase em Tempo RealResumo
Among multiresistant Gram-positive microorganisms, stands out methicillin-resistant Staphylococcus (MRS), an opportunistic pathogen associated with hospital acquired and community infections reported in medicine and large increase in reports of veterinary medicine. In veterinary medicine, numerous reports regarding several species of animals have been described. MRS is intrinsically resistant to all ?-lactam drugs. In veterinary medicine, numerous reports regarding several species of animals have been described, but Staphylococcus aureus with intermediate resistance and resistant to vancomycin (VISA/VRSA) has not yet been reported in veterinary medicine, still need further study. Staphylococcus spp. are also related to antimicrobial resistance of macrolides, lincosamides, and streptogramin B (MLSB) group, that has the same mechanism of action, although the drugs belong to different classes. In veterinary medicine, clindamycin (lincosamide class) is widely used for skin infections, wounds, bone infections, pneumonia, infections of the oral cavity, and infections caused by anaerobic bacteria, besides being used for treatments of MRS infections. Enterococcus is another resistant Gram-positive microorganism, from which vancomycin-resistant enterococci (VREs) are the most important strains. There are several reports of VREs in veterinary medicine due the use of a similar...(AU)
Dentre os micro-organismos gram positivos multirresistentes destacam-se, principalmente os Staphylococcus spp. meticilina resistente (MRS), patógenos considerados oportunistas e relacionados tanto a infecções hospitalares como infecções comunitárias, tendo inúmeros relatos na medicina e um grande aumento de relatos na medicina veterinária, em diversas espécies de animais. MRS são intrinsicamente resistentes a todas as drogas beta-lactâmicas. Os Staphylococcus aureus com resistência intermediária e os resistentes à vancomicina (VISA/VRSA) ainda não foram reportados em animais, porém são necessários estudos mais aprofundados. Os Staphylococcus spp. também estão relacionados com resistência aos antimicrobianos do grupo dos Macrolídeos, Lincosamidas e Streptogramineas B (MLSb), que apesar de serem de classes diferentes, possuem o mesmo mecanismo de ação. Na medicina veterinária, a clindamicina (antimicrobiano da classe da Lincosamida) é amplamente utilizada para tratamentos de infecções de pele, feridas, infecções ósseas, pneumonia, infecção da cavidade oral e infecções causadas por bactérias anaeróbicas, além de ser utilizada em infecções causadas por MRS. Outro gênero de micro-organismos gram positivos resistente é o Enterococcus, sendo os Enterococcus vancomicina resistente (VRE) os de maior importância. Após vários relatos de VRE na medicina veterinária, devido ao grande uso...(AU)
Assuntos
Farmacorresistência Bacteriana , Bactérias Gram-Positivas , Staphylococcus , Saúde Pública , Medicina VeterináriaResumo
Vancomycin resistant
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Humanos , Antibacterianos/uso terapêutico , Infecção Hospitalar/epidemiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Resistência a Vancomicina/fisiologia , Enterococos Resistentes à Vancomicina/isolamento & purificação , Vancomicina/uso terapêutico , Cefalosporinas/uso terapêutico , Ciprofloxacina/uso terapêutico , Infecção Hospitalar/microbiologia , DNA Bacteriano/genética , Egito/epidemiologia , Enterococcus faecium/isolamento & purificação , Gentamicinas/uso terapêutico , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/microbiologia , Controle de Infecções/métodos , Unidades de Terapia Intensiva , Estudos Prospectivos , Insuficiência Renal , Fatores de Risco , Enterococos Resistentes à VancomicinaResumo
Enterococcus spp., principalmente E. faecalis e E. faecium, são importantes patógenos de 62 infecções associadas à assistência à saúde (IRAS) devido a sua capacidade de resistir à 63 terapia antimicrobiana. São amplamente distribuídos no ambiente e abundantes na 64 microbiota intestinal de humanos e de outros mamíferos. O leite bovino contaminado por 65 enterococos pode ter implicações na qualidade dos produtos lácteos e na saúde humana. 66 Deste modo, os objetivos deste estudo foram investigar os perfis fenotípicos e genotípicos 67 de resistência aos antimicrobianos e analisar a população clonal de Enterococcus spp. 68 isolados de fezes e leite de bovinos armazenado em latões ou tanques de refrigeração em 69 propriedades leiteiras. Foram coletados 126 swabs retais de vacas leiteiras e 16 amostras 70 de leite de 16 rebanhos do estado do Acre. Até três colônias foram selecionadas por 71 espécime para identificação bacteriana por técnica de espectrometria de massa (MALDI- 72 TOF). O teste de difusão em disco foi realizado para determinar os perfis fenotípicos de 73 resistência aos antimicrobianos. Genes de resistência à vancomicina, macrolídeos, 74 tetraciclina e alto nível de aminoglicosídeos foram investigados entre as amostras 75 resistentes. A população clonal de amostras de E. faecalis e E. faecium foi analisada por 76 eletroforese em gel de campo pulsado (PFGE). Cento e sessenta e sete amostras (33,4%) 77 foram identificadas como espécies de Enterococcus, incluindo E. casseliflavus (27,0%), 78 E. gallinarum (17,5%), E. faecalis (13,5%), E. faecium (6,3%), E mundtii (1,4%) e E. 79 durans (1,4%). E. casseliflavus e E. faecalis foram as espécies mais frequentemente 80 isoladas de swab retal e leite, respectivamente. A maior frequência de resistência para E. 81 faecalis foi para ciprofloxacina (47,4%) enquanto para E. faecium foi para penicilina 82 (36,8%). Amostras resistentes à vancomicina não foram detectadas e resistentes à 83 ampicilina e a alto nível de aminoglicosídeos foram raramente. Cepas MDR 84 (multirresistentes) foram recuperadas de swab retal e leite de cinco rebanhos, sendo E. 85 faecium (32%), E. faecalis (2,6%) e E. casseliflavus (1,8%). Apenas os genes de 86 resistência à tetraciclina [tet(L) e tet(K)] foram detectados. Foi observada baixa 87 diversidade clonal tanto para E. faecalis quanto para E. faecium dentro de cada rebanho. 88 No entanto, a população bacteriana era diversa quando considerados os diferentes 89 rebanhos. Não foi observada associação entre perfil de resistência e genótipos de PFGE. 90 Os mesmos genótipos de E. faecalis foram detectadas nos swabs retais e no leite dos 91 animais em vários rebanhos, apontando a necessidade de usar boas práticas de higiene em 92 pequenas propriedades. Embora tenha sido observada baixa frequência de resistência a 93 importantes antimicrobianos prescritos para infecções enterocócicas entre as amostras de 94 origem bovina, programas para controlar a qualidade do leite são essenciais para 95 monitorar o surgimento de resistência aos antimicrobianos entre patógenos de origem 96 alimentar no Brasil.
Enterococcus spp., mainly E. faecalis and E. faecium, are important pathogens of 107 healthcare-associated infections (HAI) due their ability to resist antimicrobial therapy. 108 They are broadly distributed in environment and abundant in the intestinal microbiota of 109 humans and animals, including cows. Bovine milk contaminated by enterococci may have 110 implications for both dairy product quality and human health. Then, the objectives of this 111 study were survey the phenotypic and genotypic antimicrobial resistance profiles and 112 clonal populations of Enterococcus spp. recovered from bovine feces and milk stored in 113 cans or refrigeration tanks in dairy farms. A total of 126 rectal swabs and 16 milk samples 114 were collected from16 dairy herds in the Acre state. Until three colonies were selected 115 per sample to bacterial identification by MALDI-TOF. Disk diffusion test was performed 116 to determinate the phenotypic antimicrobial resistance profiles. Vancomycin, macrolide, 117 tetracycline, and high-level aminoglycoside resistance genes were investigated among 118 resistant isolates. The clonal population of E. faecalis and E. faecium isolates was 119 analyzed by PFGE. One hundred sixty-seven (33.4%) isolates were identified as 120 Enterococcus species, including E. casseliflavus (27.0%), E. gallinarum (17.5%), E. 121 faecalis (13.5%), E. faecium (6.3%), E. mundtii (1.4%), and E. durans (1.4%). E. 122 casseliflavus and E. faecalis were the species most frequently isolated from rectal swabs 123 and milk samples, respectively. E. faecalis and E. faecium isolates showed the highest 124 resistance frequency to ciprofloxacin (47.4%) and penicillin (36.8%), respectively. 125 Vancomycin-resistant isolates were not detected and ampicillin-, and high-level 126 aminoglycoside-resistant isolates were rarely detected. MDR (multidrug resistant) 127 isolates were recovered from rectal swabs and milk from five dairy farms: E. faecium 128 (32.0%), E. faecalis (2.6%), and E. casseliflavus (1.8%). Only tetracycline resistance 129 genes [tet(L) and tet(K)] were detected among the isolates investigated. Low clonal 130 diversity was observed among the E. faecalis and E. faecium isolates obtained within the 131 investigated herds. However, the bacterial population was diverse between different 132 herds. It was not observed an association between antimicrobial resistance profiles and 133 PFGE genotypes. The same E. faecalis genotypes were detected in rectal swabs and milk 134 in several herds, pointing out the need to use good hygiene practices in small farms. 135 Although low frequency of resistance to important antimicrobial prescribed to 136 enterococcal infections has been observed among isolates from bovine origin, programs 137 to control milk quality are essential to monitor the emergence of antimicrobial resistance 138 among foodborne pathogens in Brazil. 139