Development of a quantitative PCR method to differentiate between viable and nonviable bacteria in environmental water samples.

ABSTRACT: Ethidium monoazide bromide (EMA) treatment of pure culture and environmental waters at low concentrations (1.0-7.5 microg/ml) indicated effective enumeration of viable and viable but nonculturable Escherichia coli in pure cultures, creek waters, and secondary activated sludge effluent samples by quantitative polymerase chain reaction (qPCR) amplification of the uidA and fliC gene targets at turbidity values < 10 NTU. However, EMA treatment was not effective in primary clarifier and secondary trickling filter effluents where turbidities were > or = 10 NTU. In viable pure cultures, rapidly dividing and senescent cells were most affected by increasing EMA concentrations. Amplification of heat-killed pure bacterial cultures decreased 4 to 6 logs depending on EMA concentration and culture age.

SUBMITTER: Gedalanga PB 

PROVIDER: S-EPMC7419450 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature