The use of multiple indices of physiological activity to access viability in chlorine disinfected Escherichia coli O157:H7.
A suite of fluorescent intracellular stains and probes was used, in conjunction with viable plate counts, to assess the effect of chlorine disinfection on membrane potential (rhodamine 123; Rh123 and bis-(1,3-dibutylbarbituric acid) trimethine oxonol; DiBAC4(3)), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride; CTC) and substrate responsiveness (direct viable counts; DVC) in the commensal pathogen Escherichia coli O157:H7. After a 5 min exposure to the disinfectant, physiological indices were affected in the following order: viable plate counts > substrate responsiveness > membrane potential > respiratory activity > membrane integrity. In situ assessment of physiological activity by examining multiple targets, as demonstrated in this study, permits a more comprehensive determination of the site and extent of injury in bacterial cells following sublethal disinfection with chlorine. This approach to assessing altered bacterial physiology has application in various fields where detection of stressed bacteria is of interest.
Lisle
J T
JT
Montana State University, Department of Microbiology, Bozeman 59717, USA. umbjl@gemini.oscs.montana.edu
Pyle
B H
BH
McFeters
G A
GA
D002713:Chlorine / Q000494:pharmacology*; D015169:Colony Count, Microbial; D004203:Disinfection / Q000379:methods*; D019453:Escherichia coli O157 / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials; D010101:Oxygen Consumption
D002713:Chlorine