Prevalence and identification of Salmonella spp. in water buffaloes from São Paulo State, Brazil / Prevalência e identificação de Salmonella spp. em búfalos do Estado de São Paulo, Brasil

O objetivo do estudo foi investigar a prevalência de Salmonella spp. em amostras de fezes de búfalos do estado de São Paulo, Brasil, e identificar os sorotipos isolados. Foram examinadas 116 amostras de suabes retais de búfalos das raças Jafarabadi e Murrah, coletadas em triplicata, em seis propriedades rurais localizadas nas regiões Central, Centro-Oeste e Nordeste do estado de São Paulo, Brasil. Para avaliar a presença de Salmonella spp., foram utilizados três diferentes caldos de enriquecimento (caldo selenito cistina, caldo tetrationado Muller-Kauffmann e caldo Rappaport-Vassiliadis) e dois diferentes meios de cultura (ágar verde brilhante modificado e ágar XLT4). Das 116 amostras de suabes retais examinadas, oito amostras (6,90%; 8/116) foram positivas para Salmonella spp., incluindo quatro sorotipos: S. Panama (50%; 4/8), S. Agona (25%; 2/8) , S. Newport (12,5%; 1/8) e S. Saintpaul (12,5%; 1/8), todos isolados de búfalos sem sinais clínicos de salmonelose, indicando a importância dos animais assintomáticos como fonte de infecção para outros animais e seres humanos. Das seis propriedades rurais avaliadas, apenas em duas fazendas (33,3%; 2/6) não foi detectada Salmonella spp. O uso de mais de um caldo de enriquecimento seletivo e de mais de um meio de cultura é indicado para o isolamento de Salmonella.(AU)

Short communication: Detection of stx2 and elt genes in bovine milk by using a multiplex PCR system.

The aim of this study was to detect 2 important toxin genes from diarrheagenic Escherichia coli (DEC) in bovine milk using a new multiplex PCR. To standardize the multiplex PCR, the stx2 and elt genes were investigated for the detection of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC), respectively. The DNA template was prepared with a thermal procedure (boiling) and a commercial kit. Samples consisted of UHT and pasteurized milk, both skimmed, and STEC and ETEC were tested in concentrations between 101 and 109 cfu/mL. With the thermal procedure, the multiplex PCR system detected both pathotypes of E. coli at 109 cfu/mL in UHT and pasteurized milk. When the commercial kit was used for template preparation, STEC and ETEC could be detected at concentrations as low as 104 cfu/mL in UHT and pasteurized milk. Negative controls (Listeria monocytogenes, Salmonella Typhimurium, Salmonella Enteritidis, and Escherichia coli strain APEC 13) were not amplified with the multiplex PCR. These results indicate that the multiplex PCR was a rapid (less than 6 h) and efficient method to detect STEC and ETEC in milk using different methods for DNA preparation; however, the commercial kit was more sensitive than the thermal procedure.

Multidrug-resistant pathogenic Escherichia coli isolated from wild birds in a veterinary hospital.

Wild birds are carriers of Escherichia coli. However, little is known about their role as reservoirs for extra-intestinal pathogenic E. coli (ExPEC). In this work we investigated E. coli strains carrying virulence genes related to human and animal ExPEC isolated from free-living wild birds treated in a veterinary hospital. Multidrug resistance was found in 47.4% of the strains, but none of them were extended-spectrum beta-lactamase producers. Not only the virulence genes, but also the serogroups (e.g. O1 and O2) detected in the isolates of E. coli have already been implicated in human and bird diseases. The sequence types detected were also found in wild, companion and food animals, environmental and human clinical isolates in different countries. Furthermore, from the 19 isolates, 17 (89.5%) showed a degree of pathogenicity on an in vivo infection model. The isolates showed high heterogeneity by pulsed-field gel electrophoresis indicating that E. coli from these birds are clonally diverse. Overall, the results showed that wild birds can be reservoirs and/or vectors of highly pathogenic and multidrug-resistant E. coli that have the potential to cause disease in humans and poultry.

Comparison between avian pathogenic (APEC) and avian faecal (AFEC) Escherichia coli isolated from different regions in Brazil.

Detection and analysis of virulence-associated genes (VAGs) of avian pathogenic Escherichia coli (APEC) may be helpful to distinguish pathogenic from commensal faecal strains (AFEC). The aim of this study was to characterise 120 isolates of avian Escherichia coli, comprising 91 APEC (from diseased birds) and 29 AFEC (from healthy chickens), collected in Brazil. Phylogenetic analysis and in vivo pathogenicity testing was performed on 38 VAGs. The VAGs iucD, iutA, iroN, fepC, ompT, cvi and hlyF were statistically associated with medium and high pathogenicity (MP/HP) strains. A minimal group of seven VAGs may be required to accurately discriminate pathogenic and non-pathogenic avian strains of E. coli in Brazil.

Shiga toxigenic and enteropathogenic Escherichia coli in water and fish from pay-to-fish ponds.

UNLABELLED: Escherichia coli is part of the normal microflora of the intestines of mammals. However, among the enteric pathogens, it is one of the leading causes of intestinal diseases, especially Shiga toxigenic E. coli, which can cause diarrhoea, haemorrhagic colitis and complications like haemolytic uraemic syndrome and thrombotic thrombocytopaenic purpura. Escherichia coli is considered a serious public health problem. Water and fish samples were subjected to biochemical tests to confirm the presence of E. coli and by PCR to verify the presence of pathogenic strains (O157, enteropathogenic and shiga toxigenic) in water and fish (skin, gastrointestinal tract and muscles) from pay-to-fish ponds located in the Córrego Rico watershed in the northeastern region of the state of São Paulo, Brazil. Of the 115 E. coli isolates from fish or water, five (4·34%) contained eae and stx2 genes, one had only the eae gene and two had the stx1 gene. An isolate containing the stx2 gene was also found in the water sample. In addition, eight isolates (6·95%) from the fish gastrointestinal tract contained rfbEO157:H7 (O157 gene), and three (2·61%) contained stx2 and eae genes, demonstrating the potential risk to the environment and public health. The results provide useful basic information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population. SIGNIFICANCE AND IMPACT OF THE STUDY: Pay-to-fish ponds are a common commercial activity in Brazil. Samples of water and Oreochromis niloticus were examined by PCR to detect the presence of pathogenic strains of Escherichia coli (O157, enteropathogenic and shiga toxigenic). Several pathogenic strains were detected in this study, providing useful epidemiological information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population.

Fate of non O157 Shigatoxigenic Escherichia coli in ovine manure composting / Eliminação de Escherichia coli Shigatoxigênica não O157 em compostagem de esterco ovino

Livestock manure may contain pathogenic microorganisms which pose a risk to the health of animal or humans if the manure is not adequately treated or disposed of. To determine the fate of Shiga toxigenic Escherichia coli (STEC) non O157 in composted manure from naturally colonized sheep, fresh manure was obtained from animals carrying bacterial cells with stx1/ stx2 genes. Two composting systems were used, aerated and non-aerated, and the experiments were done in Dracena city, São Paulo State. Every week, for seven weeks, one manure sample from six different points in both systems was collected and cultured to determine the presence of E. coli, the presence of the virulence genes in the cells, and also the susceptibility to 10 antimicrobial drugs. The temperature was verified at each sampling. STEC non-O157 survived for 49 days in both composting systems. E. coli non-STEC showing a high degree of antibiotic resistance was recovered all long the composting period. No relationship was established between the presence of virulence genes and antibiotic resistance. The presence of virulence genes and multiple antibiotic resistances in E. coli implicates a potential risk for these genes spread in the human food chain, which is a reason for concern...

Esterco de animais de criação pode conter microrganismos patogênicos, o que representa um risco para a saúde animal e a humana se o esterco não for adequadamente tratado ou descartado. Determinou-se o tempo necessário para a eliminação de Escherichia coli Shiga toxigenica (STEC) não O157 em esterco ovino composto, obtido de fezes frescas de ovelhas naturalmente colonizadas com cepas STEC não O157 que apresentavam os genes stx1/ stx2. Foram utilizados dois sistemas de compostagem, aerado e não aerado, em experimentos realizados na cidade de Dracena, estado de São Paulo. Todas as semanas, durante sete semanas, uma amostra de compostagem proveniente de seis pontos diferentes na leira, nos dois sistemas, foi coletada e semeada para a determinação da presença de E. coli, da presença de genes de virulência nas células, bem como da sensibilidade dessas células a 10 drogas antimicrobianas. Em cada amostragem, a temperatura da leira foi analisada. Células de STEC não O157 sobreviveram por 49 dias nos dois sistemas de compostagem. E. coli não STEC com um alto grau de resistência a antibióticos foi recuperada ao longo de todo o período de compostagem. Não foi possível estabelecer relação entre a presença de genes de virulência e a resistência a antibióticos. A presença de genes de virulência e a resistência a múltiplos antibióticos em E. coli representam um risco potencial para o espalhamento desses genes na cadeia alimentar humana, o que é motivo de grande preocupação...

Virulence genes in isolates of Escherichia coli from samples of milk and feces from dairy cattle.

The aim of this work was to determine if Escherichia coli isolates carrying the virulence genes eae and eltB and exhibiting the Ehly phenotype are present in feces and milk samples from healthy dairy cattle on farms. Isolates from calves showed a statistically higher prevalence of eae and eltB compared with isolates from older animals. The other factors tested (stx(1), stx(2), and Ehly) were not statistically different between the two groups. Two isolates originating from calf feces were identified as serotype O157:H7; one of these isolates carried stx(1) and eae, the other stx(2) and eae. E. coli isolated from milk contained stx(1), stx(2), and eltB. The results show that feces or milk from healthy dairy cattle may contain E. coli pathotypes that express virulence genes, indicating that these materials have zoonotic potential. The results also reinforce the idea that host age can influence the dynamics of virulence genes in E. coli from cattle.

Prevalence of methicillin-resistant Staphylococci on a farm: staff can harbour MRS when animals do not.

The aim of this work was to establish the prevalence of methicillin-resistant Staphylococci (MRS) in the animals and staff of a teaching and research farm. Samples of dairy cattle (36), beef cattle (26), sheep (19), horses (21), pigs (23), goats (23) and humans (13) were collected and screened for the presence of MRS. The detection of mecA gene was performed by PCR to determine the resistance of the samples to methicillin. Antimicrobial-resistance testing to penicillin, meropenem, ceftriaxone, cephalothin, oxacillin, levofloxacin, enrofloxacin, chloramphenicol, ciprofloxacin, gentamicin, clindamycin, erytromycin, linezolid, sulfamethoxazole/trimethoprim, tetracycline, doxycycline and vancomycin was performed on the mecA+ isolates. From the 161 samples, four methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were isolated from human beings (31%), whereas none was isolated from animals (0%). No methicillin-resistant Staphylococcus aureus (MRSA) were isolated. All of the MRCoNS isolates from this work presented different antimicrobial resistance patterns. MRCoNS may be present in humans associated with animals while not present in the animals. Selective pressure outside of the farm and a lack of MRCoNS transmission between humans and animals may be responsible for this lack of correlation.