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Culture-independent techniques for rapid detection of bacteria associated with loss of chloramine residual in a drinking water system

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dc.contributor.authorHoefel, D.
dc.contributor.authorMonis, P.
dc.contributor.authorGrooby, W.
dc.contributor.authorAndrews, S.
dc.contributor.authorSaint, C.P.
dc.date.issued2005
dc.description.abstractChloramination is often the disinfection regimen of choice for extended drinking water systems. However, this process is prone to instability due to the growth of nitrifying bacteria. This is the first study to use alternative approaches for rapid investigation of chloraminated drinking water system instability in which flow cytometric cell sorting of bacteria with intact membranes (membrane-intact fraction) (BacLight kit) or with active esterases (esterase-active fraction) (carboxyfluorescein diacetate) was combined with 16S rRNA gene-directed PCR and denaturing gradient gel electrophoresis (DGGE). No active bacteria were detected when water left the water treatment plant (WTP), but 12 km downstream the chloramine residual had diminished and the level of active bacteria in the bulk water had increased to more than 1 x 10(5) bacteria ml(-1). The bacterial diversity in the system was represented by six major DGGE bands for the membrane-intact fraction and 10 major DGGE bands for the esterase-active fraction. PCR targeting of the 16S rRNA gene of chemolithotrophic ammonia-oxidizing bacteria (AOB) and subsequent DGGE and DNA sequence analysis revealed the presence of an active Nitrosospira-related species and Nitrosomonas cryotolerans in the system, but no AOB were detected in the associated WTP. The abundance of active AOB was then determined by quantitative real-time PCR (qPCR) targeting the amoA gene; 3.43 x 10(3) active AOB ml(-1) were detected in the membrane-intact fraction, and 1.40 x 10(4) active AOB ml(-1) were detected in the esterase-active fraction. These values were several orders of magnitude greater than the 2.5 AOB ml(-1) detected using a routine liquid most-probable-number assay. Culture-independent techniques described here, in combination with existing chemical indicators, should allow the water industry to obtain more comprehensive data with which to make informed decisions regarding remedial action that may be required either prior to or during an instability event.
dc.identifier.citationApplied and Environmental Microbiology, 2005; 71(11):6479-6488
dc.identifier.doi10.1128/AEM.71.11.6479-6488.2005
dc.identifier.issn0099-2240
dc.identifier.issn1098-5336
dc.identifier.urihttps://hdl.handle.net/1959.8/28194
dc.language.isoen
dc.publisherAMER SOC MICROBIOLOGY
dc.rightsCopyright status unknown
dc.source.urihttps://doi.org/10.1128/AEM.71.11.6479-6488.2005
dc.subjectBacteria
dc.subjectChloramines
dc.subjectOxidoreductases
dc.subjectDNA, Bacterial
dc.subjectDNA, Ribosomal
dc.subjectRNA, Ribosomal, 16S
dc.subjectDisinfectants
dc.subjectCulture Media
dc.subjectElectrophoresis
dc.subjectFlow Cytometry
dc.subjectIn Situ Hybridization, Fluorescence
dc.subjectColony Count, Microbial
dc.subjectPolymerase Chain Reaction
dc.subjectWater Microbiology
dc.subjectWater Purification
dc.subjectTime Factors
dc.titleCulture-independent techniques for rapid detection of bacteria associated with loss of chloramine residual in a drinking water system
dc.typeJournal article
pubs.publication-statusPublished
ror.mmsid9915911974301831

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