RESUMO
The diversity and abundance of coliform bacteria (taxonomically enterobacterias), an important quality water indicator, were determined for four representative caves in Slovak Karst: Domica Cave, Gombasecká Cave, Milada Cave and Krásnohorská Cave. Three hundred and fifty-two enterobacterial isolates were successfully identified by biochemical testing (commercial ENTEROtest 24) and selected isolates confirmed by molecular techniques (PCR, 16S rDNA sequence analysis). A total of 39 enterobacterial species were isolated from cave waters, with predominance of Escherichia coli, Serratia spp. and Enterobacter spp. PCR amplification of lacZ gene is not specific enough to provide a reliable detection of coliform bacteria isolated from the environment. Sequence analysis of 16S rDNA confirmed that all of the selected isolates belong to the family Enterobacteriaceae. In general, physical and chemical parameters of cave waters in Slovak Karst corresponded to national drinking water quality standards.
Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Cavernas/microbiologia , Água Doce/microbiologia , Bactérias/genética , Meio Ambiente , Filogenia , Reação em Cadeia da Polimerase , EslováquiaRESUMO
The occurrence of Vibrio cholerae, an important aquatic pathogen, was assessed in the surface water of the Danube River near Bratislava. The isolates were distinguished by biochemical tests and grouped by ARDRA to three clusters corresponding to three species (V. cholerae, Vibrio metschnikovii, and Aeromonas spp.). The identification of V. cholerae was confirmed by multiplex PCR using primer pairs targeted to ompW gene (membrane protein), ctxA gene (toxicity gene), and toxR gene (regulatory gene). None from the isolated V. cholerae from surface water contained ctxA gene; seven of them possessed toxR gene. Serotyping of V. cholerae isolates with polyvalent O antiserum and O/139 antiserum was negative. All isolates of V. cholerae were susceptible to chloramphenicol, rifampicin, tetracycline, variable to ampicillin, and resistant to kanamycin and streptomycin.
Assuntos
Água Doce/microbiologia , Vibrio cholerae/classificação , Vibrio cholerae/isolamento & purificação , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Toxina da Cólera/genética , Primers do DNA/genética , DNA Bacteriano/genética , Testes de Sensibilidade Microbiana , Tipagem Molecular , Reação em Cadeia da Polimerase Multiplex/métodos , Rios , Sorotipagem , Eslováquia , Vibrio cholerae/genética , Vibrio cholerae/metabolismoRESUMO
A set of reference strains and a group of previously unidentified enterococci were analysed by rep-PCR with the (GTG)(5) primer to evaluate the discriminatory power and suitability of this method for typing and identification of enterococcal species. A total of 49 strains representing all validly described species were obtained from bacterial collections. For more extensive evaluation of this identification approach 112 well-defined and identified enterococci isolated from bryndza cheese were tested. The (GTG)(5)-PCR fingerprinting assigned all strains into well-differentiated clusters representing individual species. Subsequently, a group including 44 unidentified enterococci isolated from surface waters was analysed to evaluate this method for identification of unknown isolates. Obtained band patterns allowed us to identify all the strains clearly to the species level. This study proved that rep-PCR with (GTG)(5) primer is a reliable and fast method for species identification of enterococci.
