Forty-nine typical and atypical enteropathogenic Escherichia coli (EPEC) strains belonging to different serotypes and isolated from humans, pets (cats and dogs), farm animals (bovines, sheep, and rabbits), and wild animals (monkeys) were investigated for virulence markers and clonal similarity by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The virulence markers analyzed revealed that atypical EPEC strains isolated from animals have the potential to cause diarrhea in humans. A close clonal relationship between human and animal isolates was found by MLST and PFGE. These results indicate that these animals act as atypical EPEC reservoirs and may represent sources of infection for humans. Since humans also act as a reservoir of atypical EPEC strains, the cycle of mutual infection of atypical EPEC between animals and humans, mainly pets and their owners, cannot be ruled out since the transmission dynamics between the reservoirs are not yet clearly understood.Enteropathogenic Escherichia coli (EPEC) strains are among the major causes of infantile diarrhea in developing countries (71) and can be classified as typical and atypical, depending on the presence or absence of the E. coli adherence factor plasmid (pEAF), respectively (39).The pathogenesis of EPEC resides in the ability to cause the attaching and effacing (A/E) lesion in the gut mucosa of human or animal hosts, leading to diarrheal illness (40). The genes responsible for the A/E lesion formation are located in a chromosomal pathogenicity island of ϳ35 kb, known as the locus of enterocyte effacement (LEE) (23,47). LEE encodes an adhesin called intimin (38), its translocated receptor (Tir) (42), components of a type III secretion system (36), and effector molecules, named E. coli-secreted proteins (Esp proteins) (41). These virulence factors have a crucial role in A/E lesion formation, and their detection in EPEC strains is an indicator of their potential to produce these lesions (19,56).Atypical EPEC strains have been associated with diarrhea outbreaks in developed countries (31,73,77) and with sporadic cases of diarrhea in developing and developed countries (1,12,26,52,55). At present, the prevalence of atypical EPEC is higher than that of typical EPEC in several countries (1,12,26,52,55,65).Different from the situation in developed countries, where atypical EPEC outbreaks and sporadic infections are associated with children and adults, atypical EPEC infection in Brazil is mainly associated with children's illnesses (32, 71).Typical EPEC strains are rarely isolated from animals, and humans are the major natural reservoir for these pathogens (14,32,53,71). In contrast, atypical EPEC strains are present in both healthy and diseased animals (dog, monkey, cats, and bovines) and humans (4,6,18,28,71). Some studies have associated pets and farm and wild animals as reservoirs and infection sources of atypical EPEC strains for humans (32). However, these studies did not compare atypical EPEC strains isolated from humans and animals b...

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Feces of 70 diarrhoeic and 230 non-diarrhoeic domestic cats from Sao Paulo, Brazil were investigated for enteropathogenic (EPEC), enterohaemorrhagic (EHEC) and enterotoxigenic (ETEC) Escherichia coli types. While ETEC and EHEC strains were not found, 15 EPEC strains were isolated from 14 cats, of which 13 were non-diarrhoeic, and one diarrhoeic. None of 15 EPEC strains carried the bfpA gene or the EPEC adherence factor plasmid, indicating atypical EPEC types. The EPEC strains were heterogeneous with regard to intimin types, such as eae-theta (three strains), eae-kappa (n = 3), eae-alpha1 (n = 2), eae-iota (n = 2), one eae-alpha2, eae-beta1 and eae-eta each, and two were not typeable. The majority of the EPEC isolates adhered to HEp-2 cells in a localized adherence-like pattern and were positive for fluorescence actin staining. The EPEC strains belonged to 12 different serotypes, including O111:H25 and O125:H6, which are known to be pathogens in humans. Multi locus sequence typing revealed a close genetic similarity between the O111:H25 and O125:H6 strains from cats, dogs and humans. Our results show that domestic cats are colonized by EPEC, including serotypes previously described as human pathogens. As these EPEC strains are also isolated from humans, a cycle of mutual infection by EPEC between cats and its households cannot be ruled out, though the transmission dynamics among the reservoirs are not yet understood clearly.

The release of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae to the environment is a public health issue worldwide. The aim of this study was to investigate the genetic background of genes encoding ESBLs in wastewater treatment plants (WWTPs) in São Paulo, southeastern Brazil. In 2009, during a local surveillance study, seven ESBL-producing Enterobacteriaceae strains were recovered from five WWTPs and screened for ESBL genes and mobile genetic elements. Multilocus sequence typing (MLST) was carried out, and wild plasmids were transformed into electrocompetent Escherichia coli. S1-PFGE technique was used to verify the presence of high molecular weight plasmids in wild-type strains and in bla ESBL-containing E. coli transformants. Strains harbored bla CTX-M-8, bla CTX-M-15, and/or bla SHV-28. Sequencing results showed that bla CTX-M-8 and bla CTX-M-15 genes were associated with IS26. MLST revealed new sequence types for E. coli (ST4401, ST4402, ST4403, and ST4445) and Klebsiella pneumoniae (ST1574), except for one K. pneumoniae from ST307 and Enterobacter cloacae from ST131. PCR and S1-PFGE results showed CTX-M-producing E. coli transformants carried heavy plasmids sizing 48.5-209 kb, which belonged to IncI1, IncF, and IncM1 incompatibility groups. This is the first report of CTX-M-8 and SHV-28 enzymes in environmental samples, and the present results demonstrate the plasmid-mediated spread of CTX-M-encoding genes through five WWTPs in São Paulo, Brazil, suggesting WWTPs are hotspots for the transfer of ESBL genes and confirming the urgent need to improve the management of sewage in order to minimize the dissemination of resistance genes to the environment.

This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s.) in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.

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Bacterial resistance is a primary public health concern worldwide. Within this context, pets and breeding animals act as reservoirs for multidrug-resistant bacteria (MR), such as those producing extended spectrum beta-lactamases (ESBL) and those presenting plasmid-mediated quinolone resistance (PMQR). The aim of this study was to detect the presence of ESBL and PMQR in members of the Enterobacteriaceae family, isolated from healthy sheep and dogs from non-intense farming rural properties in the Umuarama region of Paraná, Brazil. A total of 81 oral and rectal swabs from dogs and sheep from 11 small rural properties were analyzed. These swabs were inoculated in tubes containing brain heart infusion broth (BHI), and the resulting cultures were inoculated on MacConkey agar (MAC) supplemented with 10 μg/mL cefotaxime for the selection of ESBL producers. The cells were also plated on MAC supplemented with 50 μg/mL nalidixic acid for selecting quinolone-resistant enterobacteria. The bacterial isolates were subjected to biochemical identification tests, antibiograms, double-disk synergic tests, and polymerase chain reaction analysis for resistance-inducing genes (bla ESBL , qnr, and genes encoding efflux pump and acetylases). Four (5.00%) bacterial isolates (3 Escherichia coli and 1 Morganella morganii) resistant to cephalosporins and/or quinolones were identified; of these, three (75%) isolates were from sheep and one (25%) from a dog. These findings indicate the presence of MR bacteria in the normal microbiota of the animals studied. Animals colonized with such bacteria can ResumoResistência bacteriana é considerado o maior problema de saúde pública mundial da atualidade, sendo os relatos de infecções e surtos causados por bactérias multirresistentes cada vez mais frequentes na clínica veterinária e humana. Neste contexto, animais de companhia e criação podem atuar como reservatório de bactérias multirresistentes, como as produtoras de beta-lactamases de espectro estendido (ESBL) e as que apresentam resistência as quinolonas mediada por plasmídeos (PMQR). O objetivo deste trabalho foi detectar beta-lactamases de espectro estendido e resistência as quinolonas mediada por plasmídeos em membros da família Enterobacteriaceae isolados de ovinos e cães de propriedades rurais não tecnificadas da região de Umuarama, Paraná, Brasil. Foram analisados 81 swabs de cães e ovinos provenientes de 11 pequenas propriedades rurais da região de Umuarama (PR). Os swabs foram inoculados em caldo Brain Heart Infusion (BHI) e o crescimento obtivo foi em seguida semeado em placas de Petri contendo ágar MacConkey (MC) acrescido de cefotaxima 10 μg/mL para seleção de bactérias gram-negativas produtoras de ESBL; e placas contendo MC acrescido de ácido nalidíxico 50 g/mL para seleção de bactérias gram-negativas resistentes as quinolonas. Os isolados bacterianos obtidos foram submetidos a testes de antibiograma pelo método de disco-difusão em ágar, teste sinérgico do duplo-disco e reação em cadeia da polimerase (PCR) para genes que confe...

The aim of this study was to determine the occurrence of B. burgdorferi sensu lato s.l. antibodies in stray dogs of the urban area of Umuarama Town, localized in the northwest region of Parana state, Brazil. Serum samples from 168 dogs were tested using indirect immunofluorescence assay (IFA) and Western Blot (WB) in order to detect anti-Borrelia burgdorferi s.l. antibodies. The IFI analysis was used as screening test and positive results were confirmed employing the WB technique. Sixty-five (38.69%) of 168 serum samples were positive in the IFI; 54 of them (83.07%) were confirmed by the WB. The overall data analysis confirmed the presence of anti-B. burgdorferi s.l. antibodies in 32.14% (54/168) dogs, what suggests that northwest region of Parana State may constitute a risk area for Lyme disease. Further studies are necessary to determine the epidemiological characteristics of this disease in the region studied. Key words: Borreliosis, canids, diagnostic, IFA, western blot ResumoO objetivo deste estudo foi determinar a ocorrência de anticorpos anti-B. burgdorferi sensu lato sl em cães errantes da área urbana da cidade de Umuarama, localizado na região noroeste do estado do Paraná, Brasil. Amostras de soro de 168 cães foram testados utilizando imunofluorescência indireta (IFI) e Western Blot (WB), a fim de detectar anticorpos anti-Borrelia burgdorferi sl. A IFI foi utilizada como teste de triagem e os resultados positivos foram confirmados empregando a técnica de WB. Sessenta e cinco (38,69%) de 168 amostras de soro foram positivas na IFI; 54 deles (83,07%) foram confirmados pela WB. A análise geral dos dados confirmaram a presença de anticorpos anti-B. burgdorferi sl em