RESUMO
Enteropathogenic Escherichia coli (EPEC) is a pathogen associated with acute diarrhoea in humans. To determine whether EPEC isolated from healthy food-producing animals possesses the same virulence gene repertoire as EPEC isolated from human with diarrhoea, we compared six typical EPEC (tEPEC) and 20 atypical EPEC (aEPEC) from humans with diarrhoea and 42 aEPEC from healthy animals (swine, sheep and buffaloes), using pulsed-field gel electrophoresis (PFGE), virulence markers, serotyping and subtyping of eae and tir genes. We found that human and animal isolates shared virulence genes, including nleB, nleE and nleF, which were associated with human diarrhoea. Serogroups and serotypes identified in isolates of food-producing animals such as O26:H11, O128:H2, O76:H7, O103, O108, O111 and O145, have previously been implicated in human disease. The subtypes eae and tir were also shared between human and animal isolates, being eae-γ1 and eae-ß1 the most prevalent in both groups, while the most common tir subtypes were α and ß. Despite PFGE analysis demonstrating that EPEC strains are heterogeneous and there was no prevalent clone identified, EPEC isolated from humans and food-producing animals shared some characteristics, such as virulence genes associated with human diarrhoea, indicating that food-producing animals could play a role as reservoirs for those genes.
Assuntos
Escherichia coli Enteropatogênica , Infecções por Escherichia coli , Proteínas de Escherichia coli , Doenças dos Ovinos , Doenças dos Suínos , Humanos , Animais , Suínos , Ovinos , Escherichia coli Enteropatogênica/genética , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Adesinas Bacterianas/genética , Diarreia/veterinária , Sorotipagem/veterináriaRESUMO
Buffaloes and pigs play an important epidemiological roll in the Salmonella infection cycle, and asymptomatic animals can act as key component in the dissemination of the disease by horizontal, vertical, and cross-species transmission. Our study aimed and was able to confirm evidences of a cross-species transmission of Salmonella Agona between asymptomatic buffaloes and pigs. Also, we described Salmonella infection within the pig production phases, involving serotypes Agona, Senftenberg and Schwarzengrund. Rectal samples were collected from Jafarabadi buffaloes (n = 25) and Piau pigs (n = 32), located on a single farm. Salmonella Agona was isolated from lactating buffaloes, gilts, pregnant sows, and weaned pigs, Salmonella Schwarzengrund from lactating sows and Salmonella Senftenberg from gilts, pregnant sows, lactating sows, and weaned pigs. Pulsed-field Gel Electrophoresis protocol (PFGE) was performed and revealed four different profiles. Profile 1 (Salmonella Agona), isolated from a pregnant sow, a gilt and two lactating buffaloes, revealed a indistinguishable PFGE pattern, confirming evidences of potential cross-species transmission. Profile 2 (Salmonella Agona), 3 (Salmonella Senftenberg), and 4 (Salmonella Schwarzengrund), isolated from pigs, revealed important indistinguishable PFGE patterns, evidencing Salmonella infection within the pig production phases. Considering the epidemiological relevance of buffaloes and pigs in the cycle of Salmonella infection, confirmation of a potential cross-species transmission of Salmonella Agona and potential Salmonella infection within the pig production phases highlights the importance of the correct establishment of preventive health strategies in farms, in special the importance of avoiding contact between buffaloes and pigs, since cross-species transmission can occur, increasing the risk of spreading the disease.
RESUMO
Queijos tipo Minas frescal podem veicular microrganismos patogênicos. Este estudo objetivou isolar Listeria spp. e identificar as espécies L. innocua, L. seeligeri, L. ivanovii e L. monocytogenes na obtenção do leite e na elaboração de queijos tipo Minas frescal e detectar a presença de genes de virulência. Foram realizadas coletas em cinco pequenas propriedades rurais produtoras deste tipo de queijo em Jaboticabal-São Paulo. Foram coletadas amostras de suabes de fezes bovinas, amostras de mãos de ordenhador, balde de ordenha, leite, água, superfície de elaboração de queijos, mãos de manipulador do queijo, peneiras, bandejas, fôrmas e escumadeiras. O gênero Listeria spp. teve alta prevalência nas amostras, entretanto, nenhuma das espécies pesquisadas foi identificada. Assim, conclui-se que a presença de Listeria spp. em alta percentagem representa potencial risco de contaminação de Queijos tipo Minas frescal e exige uma vigilância contínua para a presença deste gênero.
Assuntos
Laticínios/análise , Laticínios/microbiologia , Leite/microbiologia , Listeria/isolamento & purificação , Listeria/patogenicidade , Manipulação de Alimentos , Microbiologia de Alimentos , Queijo/análise , Queijo/microbiologiaRESUMO
Queijos tipo Minas frescal podem veicular microrganismos patogênicos. Este estudo objetivou isolar Listeria spp. e identificar as espécies L. innocua, L. seeligeri, L. ivanovii e L. monocytogenes na obtenção do leite e na elaboração de queijos tipo Minas frescal e detectar a presença de genes de virulência. Foram realizadas coletas em cinco pequenas propriedades rurais produtoras deste tipo de queijo em Jaboticabal-São Paulo. Foram coletadas amostras de suabes de fezes bovinas, amostras de mãos de ordenhador, balde de ordenha, leite, água, superfície de elaboração de queijos, mãos de manipulador do queijo, peneiras, bandejas, fôrmas e escumadeiras. O gênero Listeria spp. teve alta prevalência nas amostras, entretanto, nenhuma das espécies pesquisadas foi identificada. Assim, conclui-se que a presença de Listeria spp. em alta percentagem representa potencial risco de contaminação de Queijos tipo Minas frescal e exige uma vigilância contínua para a presença deste gênero.(AU)
Assuntos
Laticínios/análise , Laticínios/microbiologia , Queijo/análise , Queijo/microbiologia , Leite/microbiologia , Microbiologia de Alimentos , Manipulação de Alimentos , Listeria/isolamento & purificação , Listeria/patogenicidadeRESUMO
ABSTRACT Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.
Assuntos
Humanos , Animais , Escherichia coli Enteropatogênica/isolamento & purificação , Escherichia coli Shiga Toxigênica/isolamento & purificação , Peixes/microbiologia , Filogenia , Contaminação de Alimentos/análise , Qualidade de Produtos para o Consumidor , Proteínas de Escherichia coli/genética , Escherichia coli Enteropatogênica/classificação , Escherichia coli Enteropatogênica/genética , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Carne/microbiologiaRESUMO
Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.(AU)
Assuntos
Humanos , Animais , Escherichia coli Enteropatogênica/isolamento & purificação , Ciclídeos/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Virulência/genética , Zoonoses/prevenção & controle , Reação em Cadeia da Polimerase/veterinária , FilogeniaRESUMO
Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.
Assuntos
Escherichia coli Enteropatogênica/isolamento & purificação , Peixes/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Qualidade de Produtos para o Consumidor , Escherichia coli Enteropatogênica/classificação , Escherichia coli Enteropatogênica/genética , Proteínas de Escherichia coli/genética , Contaminação de Alimentos/análise , Humanos , Carne/microbiologia , Filogenia , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genéticaRESUMO
The life cycle of synanthropic flies and their behavior, allows them to serve as mechanical vectors of several pathogens. Given that flies can carry multidrug-resistant (MDR) bacteria, this study aimed to investigate the spread of genes of antimicrobial resistance in Escherichia coli isolated from flies collected in two dairy farms in Brazil. Besides antimicrobial resistance determinants, the presence of virulence genes related to bovine colibacillosis was also assessed. Of 94 flies collected, Musca domestica was the most frequently found in the two farms. We isolated 198 E. coli strains (farm A=135 and farm B=63), and >30% were MDR E. coli. We found an association between blaTEM and phenotypical resistance to ampicillin, or chloramphenicol, or tetracycline; and blaCTX-M and resistance to cefoperazone. A high frequency (86%) of phylogenetic group B1 among MDR strains and the lack of association between multidrug resistance and virulence factors suggest that antimicrobial resistance possibly is associated with the commensal bacteria. Clonal relatedness of MDR E. coli performed by Pulsed-Field Gel Electrophoresis showed wide genomic diversity. Different flies can carry clones, but with distinct antimicrobial resistance pattern. Sanger sequencing showed that the same class 1 integron arrangement is displayed by apparently unrelated strains, carried by different flies. Our conjugation results indicate class 1 integron transfer associated with tetracycline resistance. We report for the first time, in Brazil, that MDR E. coli is carried by flies in the milking environment. Therefore, flies can act as carriers for MDR strains and contribute to dissemination routes of antimicrobial resistance.
Assuntos
Indústria de Laticínios , Dípteros/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Monitoramento Ambiental , Escherichia coli/fisiologia , Animais , Brasil , Bovinos , FazendasRESUMO
Cow raw milk cheese is widely eaten in Brazil. These products may be contaminated with pathogenic bacteria. In this work, we investigated the presence of Escherichia coli in raw milk cheese from different States in Brazil. From 147 "Minas" cheese samples, 28 cheeses were positive for E. coli. Among 39 E. coli isolates of the cheeses, one was positive for eae and negative for bpfA and efa1/lifA using PCR, and so was classified as atypical Enteropathogenic E. coli (aEPEC). Two other isolates were positive for extraintestinal pathogenic E. coli (ExPEC) genes. The aEPEC isolate belongs to serogroup O127 and was classified in A phylogenetic group, and ExPEC isolates were found in O73:H12 (EC-2 strain) and O64474:H8 (EC-9 strain) serotype. This ExPEC belongs to A and C phylogenetic group, respectively. Most of E. coli strains belonged to Clermont phylogenetic groups A (28.2%), C, and E (23.1%). Six strains (15.4%) of E. coli were positive for group B1 and two (5.1%) for B2. E. coli isolates presented an aggregative (46.0%) and diffuse (12.6%) adherence pattern to HeLa cells, and the other isolates did not show adhesion (41.4%). Four E. coli isolates (10.3%) were shown to produce moderate biofilm. The antimicrobial resistance rate was tetracycline (25.6%), followed by ampicillin (17.9%), cefoxitin (7.7%), nalidixic acid (5.1%), and amoxicillin-clavulanic acid (2.6%). One strain was resistant to three antimicrobials (tetracycline, ampicillin, and nalidixic acid). The presence of these microorganisms, the O127 strain, and a new serogroup in Brazil is a potential risk for public health.
Assuntos
Queijo/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli Enteropatogênica/genética , Leite/microbiologia , Combinação Amoxicilina e Clavulanato de Potássio/farmacologia , Ampicilina/farmacologia , Animais , Antibacterianos/farmacologia , Aderência Bacteriana , Brasil , Cefoxitina/farmacologia , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Escherichia coli Enteropatogênica/efeitos dos fármacos , Escherichia coli Enteropatogênica/isolamento & purificação , Contaminação de Alimentos , Microbiologia de Alimentos , Células HeLa , Humanos , Testes de Sensibilidade Microbiana , Ácido Nalidíxico/farmacologia , Pasteurização , Filogenia , Sorogrupo , Sorotipagem , Tetraciclina/farmacologiaRESUMO
The ability to obtain bacterial genomes from the same host has allowed for comparative studies that help in the understanding of the molecular evolution of specific pathotypes. Avian pathogenic Escherichia coli (APEC) is a group of extraintestinal strains responsible for causing colibacillosis in birds. APEC is also suggested to possess a role as a zoonotic agent. Despite its importance, APEC pathogenesis still has several cryptic pathogenic processes that need to be better understood. In this work, a genome-wide survey of eight APEC strains for genes with evidence of recombination revealed that â¼14% of the homologous groups evaluated present signs of recombination. Enrichment analyses revealed that nine Gene Ontology (GO) terms were significantly more represented in recombinant genes. Among these GO terms, several were noted to be ATP-related categories. The search for positive selection in these APEC genomes revealed 32 groups of homologous genes with evidence of positive selection. Among these groups, we found several related to cell metabolism, as well as several uncharacterized genes, beyond the well-known virulence factors ompC, lamB, waaW, waaL, and fliC. A GO term enrichment test showed a prevalence of terms related to bacterial cell contact with the external environment (e.g., viral entry into host cell, detection of virus, pore complex, bacterial-type flagellum filament C, and porin activity). Finally, the genes with evidence of positive selection were retrieved from genomes of non-APEC strains and tested as were done for APEC strains. The result revealed that none of the groups of genes presented evidence of positive selection, confirming that the analysis was effective in inferring positive selection for APEC and not for E. coli in general, which means that the study of the genes with evidence of positive selection identified in this study can contribute for the better understanding of APEC pathogenesis processes.