970 resultados para endogenous nucleases : pulsed field gel electrophoresis : hybridisation :


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Microbiologia Agropecuária - FCAV

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Although vancomycin-resistant enterococci (VRE) are reported in Brazil since 1996, data on their impact over settings of different complexity are scarce. We performed a study aimed at identifying determinants ofVRE emergence and spread in a public hospital consortium (comprising 2 hospitals, with 318 and 57 beds) in inner Brazil. Molecular typing and case-control studies (addressing predictors of acquisition or clonality) were performed. Among 122 authocthonous isolates, 106 were Enterococcus faecium (22 clones), and 16, Enterococcus faecalis (5 clones). Incidence was greater in the small-sized hospital, and a previous admission to this hospital was associated with greater risk of VRE colonization or infection during admission to the larger one. Overall risk factors included comorbidities, procedures, and antimicrobials (piperacillin-tazobactam, cefepime, and imipenem). Risk factors varied among different hospitals, species, and clones. Our findings demonstrate that VRE can spread within low-complexity facilities and from these to larger hospitals. (C) 2015 Elsevier Inc. All rights reserved.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Erysipelothrix rhusiopathiae causes a variety of diseases in many animal species, including human beings. Most human infections caused by this pathogen are related to occupational exposure, and swine are considered to be the most important reservoir of E. rhusiopathiae. The white-lipped peccary (Tayassu pecari) is an ungulate that has some genetic relationship to swine, and since the demand for T. pecari meat has recently increased in Brazil and nothing is known about the relationship of this peccary with the occupational zoonotic agent, E. rhusiopathiae, an investigation on the matter was conducted. Tonsils from 21 T. pecari slaughtered in southern Brazil were examined, and one animal was positive for E. rhusiopathiae isolation. Five colonies of this positive specimen had their species identity confirmed by PCR, and were characterized by serotyping, broth microdilution susceptibility test, and pulsed field gel electrophoresis (PFGE). All colonies belonged to serotype 2b, and presented identical susceptibility profiles. Nevertheless, the five colonies showed three different PFGE profiles, demonstrating the occurrence of infection by different E. rhusiopathiae genotypes. This is the first report of E. rhusiopathiae infection in T. pecari as well as the first description of animals carrying different E. rhusiopathiae genotypes.

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Carbapenem resistance amongst Acinetobacter spp. has been increasing in the last decade. This study evaluated the outer membrane protein (OMP) profile and production of carbapenemases in 50 carbapenem-resistant Acinetobacter spp. isolates from bloodstream infections. Isolates were identified by API20NE. Minimum inhibitory concentrations (MICs) for carbapenems were determined by broth microdilution. Carbapenemases were studied by phenotypic tests, detection of their encoding gene by polymerase chain reaction (PCR) amplification, and imipenem hydrolysis. Nucleotide sequencing confirming the enzyme gene type was performed using MegaBACE 1000. The presence of OMPs was studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and PCR. Molecular typing was performed using pulsed-field gel electrophoresis (PFGE). All isolates were resistant to carbapenems. Moreover, 98% of the isolates were positive for the gene encoding the enzyme OXA-51-like, 18% were positive for OXA-23-like (only one isolate did not show the presence of the insertion sequence ISAba1 adjacent to this gene) and 76% were positive for OXA-143 enzyme. Five isolates (10%) showed the presence of the IMP-1 gene. Imipenem hydrolysing activity was detected in only three strains containing carbapenemase genes, comprising two isolates containing the bla(IMP) gene and one containing the bla(OXA-51/OXA-23-like) gene. The OMP of 43 kDa was altered in 17 of 25 strains studied, and this alteration was associated with a high meropenem MIC (256 mu g/mL) in 5 of 7 strains without 43 kDa OMP. On the other hand, decreased OMP 33-36 kDa was found in five strains. The high prevalence of OXA-143 and alteration of OMPs might have been associated with a high level of carbapenem resistance. (C) 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

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Clostridium perfringens is an anaerobic Gram-positive bacterium known as common pathogen for humans, for domestic and wildlife animals. Although infections caused by C. perfringens type C and A in swine are well studied, just a few reports describe the genetic relationship among strains in the epidemiological chain of swine clostridioses, as well as the presence of the microorganism in the slaughterhouses. The aim of the present study was to isolate C. perfringens from feces and carcasses from swine slaughterhouses, characterize the strains in relation to the presence of enterotoxin, alpha, beta, epsilon, iota and beta-2 toxins genes. using polymerase chain reaction (PCR) and comparing strains by means of Pulsed field gel electrophoresis (PFGE). Clostridium perfringens isolation frequencies in carcasses and finishing pig intestines were of 58.8% in both types of samples. According to the polymerase chain reaction assay, only, alfa toxin was detected, being all isolates also negative to enterotoxin and beta2 toxin. Through PFGE technique, the strains were characterized in 35 pulsotypes. In only one pulsotype, the isolate from carcass sample was grouped with fecal isolate of the same animal, suggesting that the risk of cross-contamination was low. Despite the high prevalence of C. perfringens in swine carcasses from the slaughterhouses assessed, the risk of food poisoning to Brazilian pork consumers is low, since all strains were negative to cpe-gene, codifying enterotoxin.

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Bordetella avium is an opportunistic pathogen that presents tropism for ciliated epithelia, leading to upper respiratory tract disease in turkeys. This agent has also been associated with Lockjaw Syndrome in psittacine birds, but literatures describing the importance of this agent in such species are rare. The purpose of the present study was to report the first outbreak of B. avium infection in juvenile cockatiels demonstrating the Lockjaw Syndrome in Brazil and to investigate the antimicrobial resistance profile and phenotypic and genotypic characteristics of these strains. Surprising, the strains obtained from five infected cockatiel chicks from three different breeders from different Brazilian states showed a clonal relationship using the Pulsed Field Gel Electrophoresis and Single Enzyme Amplified Fragment Length Polymorphism techniques. The virulence potentials of the B. avium strains were assessed using tracheal adherence and cytotoxic effects on a VERO cell monolayer. (C) 2012 Elsevier B.V. All rights reserved.