Prevalence and Survival of Enterococcus faecium Populations Carrying the esp Gene as a Source-Tracking Marker
Publication: Journal of Environmental Engineering
Volume 137, Issue 5
Abstract
Microbial marking for human fecal pollution with Enterococcus faecium relies on cultivation followed by the detection of the enterococcal surface protein gene (). This study examined the prevalence of the populations carrying the gene among the enterococci populations cultured out of domestic wastewater. Through year-long monitoring at a wastewater treatment plant in Michigan, the proportion of in the enterococci population cultivated from sewage was 0.99%. A more sensitive detection method was developed for disinfected sewage and some impacted ambient waters by using larger membrane filters to increase the volume sampled, and preliminary application of the new protocol showed the presence of in the Grand River, where enterococcal concentrations were low. An strain isolated from wastewater in Michigan was very stable in phosphate buffer and river water at 4 and 25°C, suggesting that it can survive for months after being released into environmental waters. Among 44 samples from human and nonhuman sources, was detected only from samples with human fecal pollution. These results provide evidence that the with large volume sampling is useful as a microbial source tracking marker for disinfected wastewaters.
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Acknowledgments
This study was in part supported by Grant-in-Aid for Young Scientists (Start-up, UNSPECIFIED20860010), Grant-in-Aid for JSPS Fellows (UNSPECIFIED17-04617) from the Japan Society for the Promotion of Science (JSPS), and by a grant from the National Oceanic and Atmospheric Administration (NOAANA04OAR4600199).
References
Ahmed, W., et al. (2007). “Sourcing faecal pollution: A combination of library-dependent and library-independent methods to identify human faecal pollution in non-sewered catchments.” Water Res., 41(16), 3771–3779.
Ballesté, E., Bonjoch, X., Belanche, L. A., and Blanch, A. R. (2010). “Molecular indicators used in the development of predictive models for microbial source tracking.” Appl. Environ. Microbiol., 76(6), 1789–1795.
Bernhard, A. E., and Field, K. G. (2000). “A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-prevotella genes encoding 16S rRNA.” Appl. Environ. Microbiol., 66(10), 4571–4574.
Cheng, S., et al. (1997). “A PCR assay for identification of Enterococcus faecium.” J. Clin. Microbiol., 35(5), 1248–1250.
Chou, C. C., Lin, Y. C., and Su, J. J. (2004). “Microbial indicators for differentiation of human- and pig-sourced fecal pollution.” J. Environ. Sci. Health A, Toxic/Hazard. Subst. Environ. Eng., 39(6), 1415–1421.
Dombek, P. E., Johnson, L. K., Zimmerley, S. T., and Sadowsky, M. J. (2000). “Use of repetitive DNA sequences and the PCR to differentiate Escherichia coli isolates from human and animal sources.” Appl. Environ. Microbiol., 66(6), 2572–2577.
Eaton, T. J., and Gasson, M. J. (2002). “A variant enterococcal surface protein Esp(fm) in Enterococcus faecium; distribution among food, commensal, medical, and environmental isolates.” FEMS Microbiol. Lett., 216(2), 269–275.
Fong, T. T., Griffin, D. W., and Lipp, E. K. (2005). “Molecular assays for targeting human and bovine enteric viruses in coastal waters and their application for library-independent source tracking.” Appl. Environ. Microbiol., 71(4), 2070–2078.
Harada, T., Tsuji, N., Otsuki, K., and Murase, T. (2005). “Detection of the esp gene in high-level gentamicin resistant Enterococcus faecalis strains from pet animals in Japan.” Vet. Microbiol., 106(1-2), 139–143.
Hartel, P. G., Summer, J. D., Hill, J. L., Collins, J. V., Entry, J. A., and Segars, W. I. (2002). “Geographic variability of Escherichia coli ribotypes from animals in Idaho and Georgia.” J. Environ. Qual., 31(4), 1273–1278.
Hartel, P. G., Summer, J. D., and Segars, W. I. (2003). “Deer diet affects ribotype diversity of Escherichia coli for bacterial source tracking.” Water Res., 37(13), 3263–3268.
Hassan, W. M., Wang, S. Y., and Ellender, R. D. (2005). “Methods to increase fidelity of repetitive extragenic palindromic PCR fingerprint-based bacterial source tracking efforts.” Appl. Environ. Microbiol., 71(1), 512–518.
Jackson, C. R., Fedorka-Cray, P. J., and Barrett, J. B. (2004). “Use of a genus- and species-specific multiplex PCR for identification of enterococci.” J. Clin. Microbiol., 42(8), 3558–3565.
Jenkins, M. B., Hartel, P. G., Olexa, T. J., and Stuedemann, J. A. (2003). “Putative temporal variability of Escherichia coli ribotypes from yearling steers.” J. Environ. Qual., 32(1), 305–309.
Khatib, L. A., Tsai, Y. L., and Olson, B. H. (2002). “A biomarker for the identification of cattle fecal pollution in water using the LTIIa toxin gene from enterotoxigenic Escherichia coli.” Appl. Microbiol. Biotechnol., 59(1), 97–104.
Kim, S.-Y., Lee, J. E., Lee, S., Lee, H. T., Hur, H.-G., and Ko, G. (2010). “Characterization of Enterococcus spp. from human and animal feces using 16S rRNA sequences, the esp gene, and PFGE for microbial source tracking in Korea.” Environ. Sci. Technol., 44(9), 3423–3428.
Kühn, I., et al. (2003). “Comparison of enterococcal populations in animals, humans, and the environment—a European study.” Int. J. Food Microbiol., 88(2-3), 133–145.
Layton, A., McKay, L., Williams, D., Garrett, V., Gentry, R., and Sayler, G. (2006). “Development of Bacteroides 16S rRNA gene TaqMan-based real-time PCR assays for estimation of total, human, and bovine fecal pollution in water.” Appl. Environ. Microbiol., 72(6), 4214–4224.
Layton, B. A., Walters, S. P., and Boehm, A. B. (2009). “Distribution and diversity of the enterococcal surface protein (esp) gene in animal hosts and the Pacific coast environment.” J. Appl. Microbiol., 106(5), 1521–1531.
Leavis, H. L., et al. (2003). “Epidemic and nonepidemic multidrug-resistant Enterococcus faecium.” Emerg. Infect. Dis., 9(9), 1108–1115.
Liu, L., et al. (2006). “Modeling the transport and inactivation of E. coli and enterococci in the near-shore region of Lake Michigan.” Environ. Sci. Technol., 40(16), 5022–5028.
Lund, B., Billstrom, H., and Edlund, C. (2006). “Increased conjugation frequencies in clinical Enterococcus faecium strains harbouring the enterococcal surface protein gene esp.” Clin. Microbiol. Infect., 12(6), 588–591.
McDonald, J. L., et al. (2006). “Identifying sources of fecal contamination inexpensively with targeted sampling and bacterial source tracking.” J. Environ. Qual., 35(3), 889–897.
Meays, C. L., Broersma, K., Nordin, R., and Mazumder, A. (2004). “Source tracking fecal bacteria in water: A critical review of current methods.” J. Environ. Manage., 73(1), 71–79.
Payan, A., et al. (2005). “Method for isolation of Bacteroides bacteriophage host strains suitable for tracking sources of fecal pollution in water.” Appl. Environ. Microbiol., 71(9), 5659–5662.
Pope, J. (2009). “Tracking the sources of fecal contamination in the Wissahickon Creek Watershed using phenotypic and genotypic analytical methods.” Ph.D. thesis, Civil, Architectural and Environmental Engineering, Drexel Univ., Philadelphia.
Scott, T. M., et al. (2003). “Geographical variation in ribotype profiles of Escherichia coli isolates from humans, swine, poultry, beef, and dairy cattle in Florida.” Appl. Environ. Microbiol., 69(2), 1089–1092.
Scott, T. M., Jenkins, T. M., Lukasik, J., and Rose, J. B. (2005). “Potential use of a host associated molecular marker in Enterococcus faecium as an index of human fecal pollution.” Environ. Sci. Technol., 39(1), 283–287.
Scott, T. M., Rose, J. B., Jenkins, T. M., Farrah, S. R., and Lukasik, J. (2002). “Microbial source tracking: Current methodology and future directions.” Appl. Environ. Microbiol., 68(12), 5796–5803.
Silva, J. A., et al. (2005). “Prevalence of antibiotic resistant Enterococcus spp in waste waters in the north of Chile.” Rev. Med. Chil., 133, 1201–1210.
Simpson, J. M., Santo Domingo, J. W., and Reasoner, D. J. (2002). “Microbial source tracking: State of the science.” Environ. Sci. Technol., 36(24), 5279–5288.
Singh, S. (2007). “Investigation of bacterial fecal indicators and coliphage virus in sediment and surface water of parks and beaches along the Grand River (MI) and Lake Michigan (MI).” M.S. thesis, Dept. of Fisheries and Wildlife, Michigan State Univ., East Lansing, MI.
Stoeckel, D. M., and Harwood, V. J. (2007). “Performance, design, and analysis in microbial source tracking studies.” Appl. Environ. Microbiol., 73(8), 2405–2415.
U.S. Environmental Protection Agency (USEPA). (1986). “Ambient water quality criteria for bacteria-1986.” EPA-440/5-84-002, Office of Water Regulations and Standards, Criteria and Standards Div., Washington DC.
U.S. Environmental Protection Agency (USEPA). (2002). “Method 1600: Enterococci in water by membrane filtration using membrane-Enterococcus Indoxyl-beta-D-Glucoside agar (mEI).” EPA-821-R-02-022, Office of Water, Washington DC.
Whitman, R. L., Przybyla-Kelly, K., Shively, D. A., and Byappanahalli, M. N. (2007). “Incidence of the enterococcal surface protein (esp) gene in human and animal fecal sources.” Environ. Sci. Technol., 41(17), 6090–6095.
Willems, R. J. L., et al. (2001). “Variant esp gene as a marker of a distinct genetic lineage of vancomycin-resistant Enterococcus faecium spreading in hospitals.” Lancet, 357(9259), 853–855.
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Information
Published In
Journal of Environmental Engineering
Volume 137 • Issue 5 • May 2011
Pages: 315 - 321
Copyright
© 2011 American Society of Civil Engineers.
History
Received: May 6, 2010
Accepted: Oct 24, 2010
Published online: Apr 15, 2011
Published in print: May 1, 2011
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