Zoonotic potential of atypical enteropathogenic Escherichia coli (aEPEC) isolated from puppies with diarrhoea in Brazil.

Recent studies point atypical enteropathogenic Escherichia coli (aEPEC) to be an important agent in childhood diarrhoea in Brazil. aEPEC are commonly found in various animal species, including dogs. Although the true zoonotic risk remains unknown, some strains recovered from dogs present the same serotypes and carry the same virulence genes implicated in human disease. In this study, we compared the virulence and genetic relationship among a set of aEPEC strains previously isolated from diarrheic faeces from companion dogs and humans. A total of 17 strains, 12 from puppies and five from children, were studied. The strains were assessed for: (i) presence of virulence-associated genes (a total of 31 genes) using PCR assays; (ii) genetic relationship by Random Amplified Polymorphic DNA (RAPD), Multilocus Sequence Typing (MLST) and Pulsed-field Gel Electrophoresis (PFGE); and (iii) adherence pattern in intestinal Caco-2 cells. The occurrence of virulence genes was similar between the canine and human isolates presenting the same serotype. The fimbrial genes ecpA and fimH were the most frequently detected, followed by hcpA, tccP, tccP2, lpfA1, lpfA2, astA and toxB genes. Several nle genes were also detected, with one canine strain (O156:H- / ST327) showing all PAI O-122 genes investigated (efa-1, nleB, nleE and ent/espL2). Canine and human strains of the same serotype were grouped into a single cluster by RAPD and PFGE, in which the ST10 and ST206 were identified. Additionally, most of the strains exhibited a localized adherence-like phenotype when interacting with Caco-2 cells. The results showed that some canine aEPEC strains share virulence genes commonly found in human pathogenic strains. Moreover, strains of the same serotype, isolated from dogs and children, share virulence genes and are phylogenetically close, suggesting a potential zoonotic risk.

Major globally disseminated clonal complexes of antimicrobial resistant enterococci associated with infections in cancer patients in Brazil.

Cancer and hematological malignancies constitute major comorbidities in enterococcal infections, but little is known about the characteristics of enterococci affecting cancer patients. The aim of this study was to characterize 132 enterococcal clinical isolates obtained from cancer patients attending a Cancer Reference Center in Brazil between April 2013 and March 2014. Susceptibility to 17 antimicrobial agents was assessed by disk diffusion method. Resistance and virulence genes were investigated by PCR. Multilocus sequence typing (MLST) was performed for selected Enterococcus faecalis and Enterococcus faecium isolates. The predominant species was E. faecalis (108 isolates), followed by E. faecium (18), Enterococcus gallinarum (3), Enterococcus avium (2) and Enterococcus durans (1). Multidrug-resistant (MDR) isolates made up 44.7%, but all isolates were susceptible to fosfomycin, linezolid and glycopeptides. The most prevalent genes associated with erythromycin- and tetracycline-non susceptible isolates were erm(B) (47/71; 66.2%) and tet(M) (24/68; 35.3%), respectively. High-level resistance (HLR) to gentamicin was found in 22 (16.7%) isolates and 13 (59.1%) of them carried the aac(6')-Ie-aph(2″)-Ia gene. HLR to streptomycin was detected in 34 (25.8%) isolates, of which 15 (44.1%) isolates had the ant(6')-Ia gene. The most common virulence genes were gelE (48.9%), esp (30.5%) and asa1 (29.8%). MLST performed for 26 E. faecalis isolates revealed 18 different sequence-types (STs), with seven corresponding to novel STs (625, 626, 627, 628, 629, 630, and 635). On the other hand, nine of 10 E. faecium isolates analyzed by MLST belonged to a single clonal complex, comprised of mostly ST412, which emerged worldwide after mid-2000s, but also two novel STs (963 and 964). We detected major globally disseminated E. faecalis and E. faecium clonal complexes along with novel closely related STs, indicating the fitness and continuous evolution of these hospital-adapted lineages.

Antimicrobial resistance, integron carriage, and gyrA and gyrB mutations in Pseudomonas aeruginosa isolated from dogs with otitis externa and pyoderma in Brazil.

BACKGROUND: Pseudomonas aeruginosa is associated with otitis and pyoderma in dogs and is frequently resistant to several antimicrobial drugs. Resistance genes can be carried by integrons with quinolone resistance mainly due to mutations in DNA topoisomerases II and IV. OBJECTIVE: To evaluate the antimicrobial susceptibility, integron carriage, and gyrA and gyrB mutations in P. aeruginosa isolates from canine otitis and pyoderma. ANIMALS: One hundred and four P. aeruginosa strains isolated from dogs with otitis externa (n = 93) and pyoderma (n = 11). METHODS: Antimicrobial susceptibility against 16 antibacterial agents was evaluated through agar diffusion tests. Integron carriage, class and gyrA and gyrB mutations were analysed by PCR, restriction fragment length polymorphism (RFLP)-PCR and genetic sequencing assays. RESULTS: Isolates were mostly resistant to enrofloxacin (72.2%) and ticarcillin (59.7%). Lower resistance to ciprofloxacin (7.7%), tobramycin (3.8%) and polymixin B (0.0%) was detected. Ten (9.6%) multidrug-resistant (MDR) strains were detected. Eight (7.7%) strains carried class 1 integrons and this was associated with MDR (three isolates, P ≤ 0.05). Five of the integron-carrying strains exhibited aminoglycoside resistance genes. Mutations of gyrA and gyrB were observed in 10 isolates, seven of them resistant to all fluoroquinolones tested. CONCLUSIONS AND CLINICAL IMPORTANCE: Enrofloxacin and ticarcilin resistance was widespread in P. aeruginosa isolated from dogs in Brazil. Pseudomonas aeruginosa carrying integrons may present a significant challenge for treatment.