Detection and Persistence of Clinical Escherichia coli in Drinking Water Evaluated by a Rapid Enzyme Assay and qPCR

Annette S. Bukh, Nina E. Hansen, Peter Roslev

Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Aalborg, Denmark.

DOI: 10.4236/aim.2012.23030                

Abstract

The aims of this study were to evaluate two methods, qPCR and a chemiluminescent assay (ColiLight II), for rapid detection of E. coli in water, and to examine the survival and persistence of clinical E. coli in drinking water and biofilm using qPCR and ColiLight II. qPCR and ColiLight II were compared with a cultivation-based method (MPN), and survival and persistence of four clinical E. coli strains in water and biofilms on stainless steel (SS) and polyethylene (PE) surfaces were studied in a flow-through reactor with non-disinfected drinking water using ColiLight II, qPCR, ATP bioluminescence, and MPN. ColiLight II and qPCR correlated well with MPN. In drinking water, some clinical E. coli strains showed prolonged survival in drinking water flow-through systems, and persisted 3 - 3.4 times longer than the theoretical washout due to incorporation into biofilms. Strain specific attributes can significantly affect detection and persistence of E. coli in drinking water matrices.

Keywords

Biofilm; Clinical E. coli; Chemiluminescent Assay; Lab-Scale Drinking Water Reactor System; Drinking Water; qPCR

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	A. Rompré, P. Servais, J. Baudart, M. de-Roubin and P. Laurent, “Detection and Enumeration of Coliforms in Drinking Water: Current Methods and Emerging Approaches,” Journal of Microbiological Methods, Vol. 49, No. 1, 2002, pp. 31-54. doi:10.1016/S0167-7012(01)00351-7
[2]	L. Fiksdal and I. Tryland, “Application of Rapid Enzyme Assay Techniques for Monitoring of Microbial Water Quality,” Current Opinion in Biotechnology, Vol. 19, No. 3, 2008, pp. 289-294. doi:10.1016/j.copbio.2008.03.004
[3]	J. D. Berg and L. Fiksdal, “Rapid Detection of Total and Fecal Coliforms in Water by Enzymatic Hydrolysis of 4-Methylumbelliferone-beta-D-galactoside,” Applied and Environmental Microbiology, Vol. 54, 1988, pp. 2118- 2122.
[4]	L. Fiksdal, M. Pommepuy, M. P. Caprais and I. Midttun, “Monitoring of Fecal Pollution in Coastal Waters by Use of Rapid Enzymatic Techniques,” Applied and Environmental Microbiology, Vol. 60, 1994, pp. 1581-1584.
[5]	I. George, M. Petit and P. Servais, “Use of Enzymatic Methods for Rapid Enumeration of Coliforms in Freshwaters,” Journal of Applied Microbiology, Vol. 88, No. 3, 2000, pp. 404-413. doi:10.1046/j.1365-2672.2000.00977.x
[6]	T. Garcia-Armisen, P. Lebaron and P. Servais, “β-D-Glucuronidase Activity Assay to Assess Viable Escherichia coli Abundance in Freshwaters,” Letters in Applied Microbiology, Vol. 40, No. 4, 2005, pp. 278-282. doi:10.1111/j.1472-765X.2005.01670.x
[7]	P. Lebaron, A. Henry, A. Lepeuple, G. Pena and P. Servais, “An Operational Method for the Real-Time Monitoring of E. coli Numbers in Bathing Waters,” Marine Pollution Bulletin, Vol. 50, No. 6, 2005, pp. 652-659. doi:10.1016/j.marpolbul.2005.01.016
[8]	S. O. Van Poucke and H. J. Nelis, “Rapid Detection of Fluorescent and Chemiluminescent Total Coliforms and Escherichia coli on Membrane Filters,” Journal of Microbiological Methods, Vol. 42, No. 3, 2000, pp. 233- 244. doi:10.1016/S0167-7012(00)00193-7
[9]	A. S. Bukh and P. Roslev, “Characterization and Validation of a Chemiluminescent Assay Based on 1,2-Dioxetanes for Rapid Detection of Viable Escherichia coli,” Applied Microbiology and Biotechnology, Vol. 86, No. 6, 2010, pp. 1947-1957. doi:10.1007/s00253-010-2514-6
[10]	K. L. Cook and C. H. Bolster, “Survival of Campylobacter jejuni and Escherichia coli in Groundwater during Prolonged Starvation at Low Temperatures,” Journal of Applied Microbiology, Vol. 103, No. 3, 2007, pp. 573- 583. doi:10.1111/j.1365-2672.2006.03285.x
[11]	W. Ahmed, F. Huygens, A. Goonetilleke and T. Gardner, “Real-Time PCR Detection of Pathogenic Microorganisms in Roof-Harvested Rainwater in Southeast Queensland, Australia,” Applied and Environmental Microbiology, Vol. 74, No. 17, 2008, pp. 5490-5496. doi:10.1128/AEM.00331-08
[12]	?. Lothigius, ?. Sj?ling, A. Svennerholm and I. B?lin, “Survival and Gene Expression of Enterotoxigenic Escherichia coli during Long-Term Incubation in Sea Water and Freshwater,” Journal of Applied Microbiology, Vol. 108, No. 4, 2010, pp. 1441-1449. doi:10.1111/j.1365-2672.2009.04548.x
[13]	C. Lee, J. Kim, S. G. Shin and S. Hwang, “Absolute and Relative QPCR Quantification of Plasmid Copy Number in Escherichia coli,” Journal of Biotechnology, Vol. 123, No. 3, 2006, pp. 273-280. doi:10.1016/j.jbiotec.2005.11.014
[14]	O. Clermont, M. Lescat, C. L. O’Brien, D. M. Gordon, O. Tenaillon and E. Denamur, “Evidence for a Human-Specific Escherichia coli Clone,” Environmental Microbiology, Vol. 10, No. 4, 2008, pp. 1000-1006. doi:10.1111/j.1462-2920.2007.01520.x
[15]	DS 6222-1, “Water Quality—Enumeration of Culturable Micro-Organisms—Colony Count by Inoculation in a Nutrient Agar Culture Medium,” Danish Standards Association, Copenhagen, 2002.
[16]	W. E. Garthright and R. J. Blodgett, “FDA’s Preferred MPN Methods for Standard, Large or Unusual Tests, with a Spreadsheet,” Food Microbiology, Vol. 20, No. 4, 2003, pp. 439-445. doi:10.1016/S0740-0020(02)00144-2
[17]	I. Tryland and L. Fiksdal, “Enzyme Characteristics of β-D-Galactosidase- and β-D-Glucuronidase-Positive Bacteria and Their Interference in Rapid Methods for Detection of Waterborne Coliforms and Escherichia coli,” Applied and Environmental Microbiology, Vol. 64, 1998, pp. 1018-1023.
[18]	C. Johansen, A. Verheul, L. Gram, T. Gill and T. Abee, “Protamine-Induced Permeabilization of Cell Envelopes of Gram-Positive and Gram-Negative Bacteria,” Applied and Environmental Microbiology, Vol. 63, 1997, pp. 1155- 1159.
[19]	I. Bronstein, B. Edwards and J. C. Voyta, “1,2-Dioxetanes: Novel Chemiluminescent Enzyme Substrates. Applications to Immunoassays,” Journal of Bioluminescence and Chemiluminescence, Vol. 4, No. 1, 1989, pp. 99-111. doi:10.1002/bio.1170040116
[20]	S. O. Van Poucke and H. J. Nelis, “Development of a Sensitive Chemiluminometric Assay for the Detection of β-Galactosidase in Permeabilized Coliform Bacteria and Comparison with Fluorometry and Colorimetry,” Applied and Environmental Microbiology, Vol. 61, 1995, pp. 4505-4509.
[21]	M. Raymaekers, R. Smets, B. Maes and R. Cartuyvels, “Checklist for Optimization and Validation of Real-Time PCR Assays,” Journal of Clinical Laboratory Analysis, Vol. 23, No. 3, 2009, pp. 145-151. doi:10.1002/jcla.20307
[22]	A. Al-Turki and M. G. El-Ziney, “Evaluation of Commercial Colilert18-Quantitray? Method by ISO Techniques for Enumeration and Quantification of Total Coliforms and Escherichia coli in Drinking-Water of Buraidah, Saudi Arabia,” Journal of Applied Sciences, Vol. 9, No. 18, 2009, pp. 3357-3363. doi:10.3923/jas.2009.3357.3363
[23]	J. Yu, D. Kim and T. Lee, “Microbial Diversity in Biofilms on Water Distribution Pipes of Different Materials,” Water Science and Technology, Vol. 61, No. 1, 2010, pp. 163-171. doi:10.2166/wst.2010.813
[24]	J. Ryu and L. R. Beuchat, “Biofilm Formation by Escherichia coli O157:H7 on Stainless Steel: Effect of Exopolysaccharide and Curli Production on Its Resistance to Chlorine,” Applied and Environmental Microbiology, Vol. 71, No. 1, 2005, 247-254. doi:10.1128/AEM.71.1.247-254.2005
[25]	O. Vidal, R. Longin, C. Prigent-Combaret, C. Dorel, M. Hooreman and P. Lejeune, “Isolation of an Escherichia coli K-12 Mutant Strain Able to Form Biofilms on Inert Surfaces: Involvement of a New ompR Allele That Increases Curli Expression,” Journal of Bacteriology, Vol. 180, 1998, pp. 2442-2449.
[26]	Z. Salda?a, J. Xicohtencatl-Cortes, F. Avelino, A. D. Phillips, J. B. Kaper, J. L. Puente and J. A. Girón, “Synergistic Role of Curli and Cellulose in Cell Adherence and Biofilm Formation of Attaching and Effacing Escherichia coli and Identification of Fis as a Negative Regulator of Curli,” Environmental Microbiology, Vol. 11, No. 4, 2009, pp. 992-1006. doi:10.1111/j.1462-2920.2008.01824.x
[27]	K. L. Anderson, J. E. Whitlock and V. J. Harwood, “Persistence and Differential Survival of Fecal Indicator Bacteria in Subtropical Waters and Sediments,” Applied and Environmental Microbiology, Vol. 71, No. 6, 2005, pp. 3041-3048. doi:10.1128/AEM.71.6.3041-3048.2005
[28]	L. Watterworth, B. Rosa, H. Schraft, E. Topp and K. T. Leung, “Survival of Various ERIC-Genotypes of Shiga Toxin-Producing Escherichia coli in Well Water,” Water, Air, & Soil Pollution, Vol. 177, No. 1-4, 2006, pp. 367-382. doi:10.1007/s11270-006-9179-x