ABSTRACT
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.
Subject(s)
Bacteria/classification , Bacteria/isolation & purification , Caves/microbiology , Fresh Water/microbiology , Bacteria/genetics , Environment , Phylogeny , Polymerase Chain Reaction , SlovakiaABSTRACT
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.