Possible interference of lactose-fermenting marine vibrios in coliform beta-D-galactosidase assays

An investigation into possible interferences in beta-D-galactosidase-based assays for coliform bacteria in marine waters was carried out. A rapid instrumental fluorescence assay for beta-D-galactosidase activity, using 4-methylumbelliferyl-beta-D-galactoside as a substrate, was used to investigate activities of this enzyme in non-coliform bacterial isolates from coastal waters. Only 2% of isolates showed slight enzyme activity after a 1-h incubation period at 44.5 degrees C. At a lower incubation temperature of 20 degrees C, 51% and 94% of the isolates showed some enzyme activity within 6 h and 48 h, respectively. Fifty-nine out of 67 of these isolates were identified as Vibrio species. A lac+ strain of Vibrio vulnificus was found to produce beta-D-galactosidase which caused significant false-positive reactions in the Colilert-Marine Water assay when present at concentrations of 10 cfu ml-1 or greater. This interference could be overcome by addition of the vibriostatic agent O/129. The high fluorescence of this reagent, however, precluded the simultaneous determination of Escherichia coli in the Colilert test and also its use in instrumental fluorescence assays. It was concluded that in assays employing high temperatures and short incubation times, Vibrio species are unlikely to cause significant interferences.