Phenotypic and phylogenetic identification of coliform bacteria obtained from 12 USEPA approved coliform methods

Handle URI:
http://hdl.handle.net/10754/558683
Title:
Phenotypic and phylogenetic identification of coliform bacteria obtained from 12 USEPA approved coliform methods
Authors:
Zhang, Ya; Hong, Pei-Ying ( 0000-0002-4474-6600 ) ; LeChevallier, Mark W.; Liu, Wen-Tso
Abstract:
The current definition of coliform bacteria is method-dependent, and when different culture-based methods are used, discrepancies in results can occur and affect the accuracy in identifying true coliforms. This study used an alternative approach to identify true coliforms by combing the phenotypic traits of the coliform isolates and the phylogenetic affiliation of 16S rRNA gene sequences together with the use of lacZ and uidA genes. A collection of 1404 isolates from 12 US Environmental Protection Agency approved coliform-testing methods were characterized based on their phylogenetic affiliations and responses to their original isolation medium and Lauryl Tryptose broth, m-Endo and MI agar media. Isolates were phylogenetically classified into 32 true coliform or targeted Enterobacteriaceae (TE) groups, and 14 non-coliform or non-targeted Enterbacteriaceae (NTE) groups. It was statistically shown that detecting true-positive (TP) events is more challenging than detecting true-negative (TN) events. Furthermore, most false-negative (FN) events were associated with four TE groups (i.e., Serratia group I, Providencia, Proteus, and Morganella), and most false-positive (FP) events with two NTE groups, Aeromonas and Plesiomonas. In Escherichia coli testing, 18 out of 145 E. coli isolates identified by those enzymatic methods were validated as FNs. The reasons behind the FP and FN reactions could be explained through the analysis of the lacZ and uidA gene. Overall, combining the analyses of 16S rRNA, lacZ and uidA genes with the growth responses of TE and NTE on culture-based media is an effective way to evaluate the performance of coliform detection methods.
KAUST Department:
Water Desalination and Reuse Research Center (WDRC); Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
Phenotypic and phylogenetic identification of coliform bacteria obtained from 12 USEPA approved coliform methods 2015:AEM.01510-15 Applied and Environmental Microbiology
Journal:
Applied and Environmental Microbiology
Issue Date:
26-Jun-2015
DOI:
10.1128/AEM.01510-15
Type:
Article
ISSN:
0099-2240; 1098-5336
Additional Links:
http://aem.asm.org/lookup/doi/10.1128/AEM.01510-15
Appears in Collections:
Articles; Water Desalination and Reuse Research Center (WDRC); Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorZhang, Yaen
dc.contributor.authorHong, Pei-Yingen
dc.contributor.authorLeChevallier, Mark W.en
dc.contributor.authorLiu, Wen-Tsoen
dc.date.accessioned2015-06-30T14:31:38Zen
dc.date.available2015-06-30T14:31:38Zen
dc.date.issued2015-06-26en
dc.identifier.citationPhenotypic and phylogenetic identification of coliform bacteria obtained from 12 USEPA approved coliform methods 2015:AEM.01510-15 Applied and Environmental Microbiologyen
dc.identifier.issn0099-2240en
dc.identifier.issn1098-5336en
dc.identifier.doi10.1128/AEM.01510-15en
dc.identifier.urihttp://hdl.handle.net/10754/558683en
dc.description.abstractThe current definition of coliform bacteria is method-dependent, and when different culture-based methods are used, discrepancies in results can occur and affect the accuracy in identifying true coliforms. This study used an alternative approach to identify true coliforms by combing the phenotypic traits of the coliform isolates and the phylogenetic affiliation of 16S rRNA gene sequences together with the use of lacZ and uidA genes. A collection of 1404 isolates from 12 US Environmental Protection Agency approved coliform-testing methods were characterized based on their phylogenetic affiliations and responses to their original isolation medium and Lauryl Tryptose broth, m-Endo and MI agar media. Isolates were phylogenetically classified into 32 true coliform or targeted Enterobacteriaceae (TE) groups, and 14 non-coliform or non-targeted Enterbacteriaceae (NTE) groups. It was statistically shown that detecting true-positive (TP) events is more challenging than detecting true-negative (TN) events. Furthermore, most false-negative (FN) events were associated with four TE groups (i.e., Serratia group I, Providencia, Proteus, and Morganella), and most false-positive (FP) events with two NTE groups, Aeromonas and Plesiomonas. In Escherichia coli testing, 18 out of 145 E. coli isolates identified by those enzymatic methods were validated as FNs. The reasons behind the FP and FN reactions could be explained through the analysis of the lacZ and uidA gene. Overall, combining the analyses of 16S rRNA, lacZ and uidA genes with the growth responses of TE and NTE on culture-based media is an effective way to evaluate the performance of coliform detection methods.en
dc.relation.urlhttp://aem.asm.org/lookup/doi/10.1128/AEM.01510-15en
dc.rightsArchived with thanks to Applied and Environmental MicrobiologyCopyright © 2015, American Society for Microbiology. All Rights Reserved.en
dc.titlePhenotypic and phylogenetic identification of coliform bacteria obtained from 12 USEPA approved coliform methodsen
dc.typeArticleen
dc.contributor.departmentWater Desalination and Reuse Research Center (WDRC)en
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalApplied and Environmental Microbiologyen
dc.eprint.versionPost-printen
dc.contributor.institutionDepartment of Civil and Environmental Engineering, University of Illinois Urbana-Champaign, 205 N. Mathews Ave., Urbana, IL 61810, USAen
dc.contributor.institutionAmerican Water, 1025 Laurel Oak Rd., Voorhees, NJ 08043, USAen
kaust.authorHong, Pei-Yingen
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