Noncoliform enumeration and identification in potable water, and their senstivity to commonly used disinfectants

Tap water collected according to standard methods was examined for microbial presence. Epifluorescent diagnoses using redox probe 5-cyano-2,3ditolyl tetrazolium chloride (CTC), 4',6-diamidino-2-phenylindole (DAPI), and acridine orange (AO) were employed for direct evidence of microorganisms. Evidence of total (DAPI or AO), respiring (CTC) bacteria, and heterotrophic plate count (HPC) was determined on multiple occasions during the summer, fall, and winter 1996-1997. Pseudomonas aeruginosa, Acinetobacter sp., Bacillus licheniformis, and Methylobacterium rhodinum were isolated and identified by the API and Biolog system using GN and GP procedures. On the basis of comparisons presented in this study between the CTC method and the standard HPC procedure, it appeared that the number of CTC-reducing bacteria in the tap water samples was typically higher than that determined by HPC, indicating that many respiring bacteria detected by the CTC reduction technique fail to produce visible colonieson the agar media used. In the seasonal data obtained by the CTC method, no difference was shown among respiring bacterial counts obtained from June through January. In the examination of P. aeruginosa viability in presence of chlorine, the number of CTC-positive bacteria exceeded the number of CFU by more than 2 logs after exposure to chlorine, suggesting that reliance on HPC overestimate the efficacy of disinfection treatment. In inactivation assays using the Biolog MT plate, no sensitivity to chlorine or chloramine disinfectants was noted even at high concentration levels (5 mg/liter). Following initial drop, bacterial activities increased as contact time increased. Thus, it appears that the MT microplate provides too low a cell concentration, too great a contact time, and/or too low a concentration of tetrazolium dye within the well for successful analysis of disinfectant capability to selected bacterial strains isolated from distribution water.Department of BiologyThesis (M.S.