Seasonal Loading and Concentration Patterns for Fecal Bacteroidales qPCR Markers and Relationships to Water Quality Parameters at Baseflow
Abstract
Bacteria belonging to the Order Bacteroidales predominate the intestines of warm-blooded animals, and monitoring of these bacteria can indicate fecal pollution impacts to a waterbody. Differences in seasonal concentrations and loadings for Bacteroidales and their relationship with physicochemical water parameters were investigated in temperate, inland streams. Seasonal samples (n = 321) were collected during baseflow in three central Tennessee, USA, watersheds. To estimate total fecal bacteria in receiving streams, general Bacteroidales 16S rRNA gene targets were analyzed by quantitative PCR and reported as concentration and loadings for individual and combined watersheds. In most cases, Bacteroidales marker concentrations were highest during spring/summer and loading values were highest in the spring. Bacteroidales concentrations were positively correlated with temperature and total suspended solids and negatively with dissolved oxygen, while no consistent correlations were found between loadings and abiotic factors. Temperature, total suspended solids, and dissolved oxygen are likely drivers influencing seasonal patterns for Bacteroidales concentrations. Researchers and water quality stakeholders should carefully consider measurement type (concentration versus loading), season, and water quality parameters as elements that could impact results when developing fecal monitoring projects.
Keywords
Fecal indicator bacteria Bacteroidales qPCR Season Water quality parametersNotes
Acknowledgments
This project was funded as a special project by the Metro Water Services, Nashville, TN, USA, and supported by the Middle Tennessee State University Molecular Biosciences PhD Program. Expertise and training on qPCR assays were provided by Dr. Alice Layton and Dan Williams, Center for Biotechnology, University of Tennessee, Knoxville, TN, USA.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
References
- American Public Health Association (APHA). (2012). Standard methods for the examination of water and wastewater (22nd ed.). Washington, DC: APHA.Google Scholar
- Bae, S., & Wuertz, S. (2009). Rapid decay of host-specific fecal Bacteroidales cells in seawater as measured by quantitative PCR with propidium monoazide. Water Research, 43(19), 4850–4859.CrossRefGoogle Scholar
- Balleste, E., & Blanch, A. R. (2010). Persistence of Bacteroides species populations in a river as measured by molecular and culture techniques. Applied and Environmental Microbiology, 76(22), 7608–7616.CrossRefGoogle Scholar
- Baughn, A. D., & Malamy, M. H. (2004). The strict anaerobe Bacteroides fragilis grows in and benefits from nanomolar concentrations of oxygen. Nature, 427, 441–444.CrossRefGoogle Scholar
- Bell, A., Layton, A. C., McKay, L., Williams, D., Gentry, R., & Sayler, G. (2009). Factors influencing the persistence of fecal Bacteroides in stream water. Journal of Environmental Quality, 38(3), 1224–1232.CrossRefGoogle Scholar
- Bernhard, A. E., & Field, K. G. (2000a). A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. Applied and Environmental Microbiology, 66(10), 4571–4574.CrossRefGoogle Scholar
- Bernhard, A. E., & Field, K. G. (2000b). Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA genetic markers from fecal anaerobes. Applied and Environmental Microbiology, 66(4), 1587–1594.CrossRefGoogle Scholar
- Bradshaw, J. K., Snyder, B. J., Oladeinde, A., Spidle, D., Berrang, M. E., Meinersmann, R. J., & Molina, M. (2016). Characterizing relationships among fecal indicator bacteria, microbial source tracking markers, and associated waterborne pathogen occurrence in stream water and sediments in a mixed land use watershed. Water Research, 101, 498–509.CrossRefGoogle Scholar
- Byappanahalli, M. N., Whitman, R. L., Shively, D. A., Sadowsky, M. J., & Ishii, S. (2006). Population structure, persistence, and seasonality of autochthonous Escherichia coli in temperate, coastal forest soil from a Great Lakes watershed. Environmental Microbiology, 8(3), 504–513.CrossRefGoogle Scholar
- Converse, R. R., Piehler, M. F., & Noble, R. T. (2011). Contrasts in concentrations and loads of conventional and alternative indicators of fecal contamination in coastal stormwater. Water Research, 45, 5229–5240.CrossRefGoogle Scholar
- Dick, L. K., Bernhard, A. E., Brodeur, T. J., Santo Domingo, J. W., Simpson, J. M., Walters, S. P., & Field, K. G. (2005). Host distributions of uncultivated fecal Bacteroidales bacteria reveal genetic markers for fecal source identification. Applied and Environmental Microbiology, 71(6), 3184–3191.CrossRefGoogle Scholar
- Dick, L. K., Stelzer, E. A., Bertke, E. E., Fong, D. L., & Stoeckel, D. M. (2010). Relative decay of Bacteroidales microbial source tracking markers and cultivated Escherichia coli in freshwater microcosms. Applied and Environmental Microbiology, 76(10), 3255–3262.CrossRefGoogle Scholar
- Fiksdal, L., Maki, J. S., LaCroix, S. J., & Staley, J. T. (1985). Survival and detection of Bacteroides spp., prospective indicator bacteria. Applied and Environmental Microbiology, 49(1), 148–150.Google Scholar
- Fode-Vaughn, K. A., Wimpee, C. F., Remsen, C. C., & Collins, M. L. P. (2001). Detection of bacteria in environmental samples by direct PCR without DNA extraction. BioTechniques, 31(3), 598–607.CrossRefGoogle Scholar
- Gentry, R. W., Layton, A. C., McKay, L. D., McCarthy, J. F., Williams, D. E., Koirala, S. R., & Sayler, G. S. (2007). Efficacy of Bacteroides measurements for reducing the statistical uncertainty associated with hydrologic flow and fecal loads in a mixed use watershed. Journal of Environmental Quality, 36(5), 1324–1330.CrossRefGoogle Scholar
- Gentry, R. W., McCarthy, J., Layton, A. C., McKay, L. D., Williams, D. E., Koirala, S. R., & Sayler, G. S. (2006). Escherichia coli loading at or near base flow in a mixed-use watershed. Journal of Environmental Quality, 35(6), 2244–2249.CrossRefGoogle Scholar
- Ishii, S., Ksoll, W. B., Hicks, R. E., & Sadowsky, M. J. (2006). Presence and growth of naturalized Escherichia coli in temperate soils from Lake Superior watersheds. Applied and Environmental Microbiology, 72(1), 612–621.CrossRefGoogle Scholar
- Kreader, C. A. (1995). Design and evaluation of Bacteroides DNA probes for the specific detection of human fecal pollution. Applied and Environmental Microbiology, 61(4), 1171–1179.Google Scholar
- Kreader, C. A. (1998). Persistence of PCR-detectable Bacteroides distasonis from human feces in river water. Applied and Environmental Microbiology, 64(10), 4103–4105.Google Scholar
- Layton, A., McKay, L., Williams, D., Garrett, V., Gentry, R., & 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. Applied and Environmental Microbiology, 72(6), 4214–4224.CrossRefGoogle Scholar
- Mallin, M. A., & Cahoon, L. B. (2003). Industrialized animal production a major source of nutrient and microbial pollution to aquatic ecosystems. Population and Environment, 24(5), 369–385.CrossRefGoogle Scholar
- Okabe, S., Okayama, N., Savichtcheva, O., & Ito, T. (2007). Quantification of host-specific Bacteroides-Prevotella 16S rRNA genetic markers for assessment of fecal pollution in freshwater. Applied Microbiology and Biotechnology, 74(4), 890–901.CrossRefGoogle Scholar
- Okabe, S., & Shimazu, Y. (2007). Persistence of host-specific Bacteroides-Prevotella 16S rRNA genetic markers in environmental waters: effects of temperature and salinity. Applied Microbiology and Biotechnology, 76(4), 935–944.CrossRefGoogle Scholar
- Prϋss, A. (1998). Review of epidemiological studies on health effects from exposure to recreational water. International Journal of Epidemiology, 27(1), 1–9.CrossRefGoogle Scholar
- Rowny, J. G., & Stewart, J. R. (2012). Characterization of nonpoint source microbial contamination in an urbanizing watershed serving as a municipal water supply. Water Research, 46, 6143–6153.CrossRefGoogle Scholar
- Schriewer, A., Miller, W. A., Byrne, B. A., Miller, M. A., Oates, S., Conrad, P. A., Hardin, D., Yang, H.-H., Chouicha, N., Melli, A., Jessup, D., Dominik, D., & Wuertz, S. (2010). Presence of Bacteroidales as a predictor of pathogens in surface waters of the central California coast. Applied and Environmental Microbiology, 76(17), 5802–5814.CrossRefGoogle Scholar
- Schulz, C. J., & Childers, G. W. (2011). Fecal Bacteroidales diversity and decay in response to variations in temperature and salinity. Applied and Environmental Microbiology, 77(8), 2563–2572.CrossRefGoogle Scholar
- Seurinck, S., Defoirdt, T., Verstraete, W., & Siciliano, S. D. (2005). Detection and quantification of the human-specific HF183 Bacteroides 16S rRNA genetic marker with real-time PCR for assessment of human faecal pollution in freshwater. Environmental Microbiology, 7(2), 249–259.CrossRefGoogle Scholar
- Shanks, O. C., Kelty, C. A., Sivaganesan, M., Varma, M., & Haugland, R. A. (2009). Quantitative PCR for genetic markers of human fecal pollution. Applied and Environmental Microbiology, 75(17), 5507–5513.CrossRefGoogle Scholar
- Stallard, M. A., Otter, R. R., Winesett, S., Barbero, M., Bruce, M., Layton, A. C., & Bailey, F. C. (2016). A watershed analysis of seasonal concentration-and loading-based results for Escherichia coli in inland waters. Bulletin of Environmental Contamination and Toxicology, 97(6), 838–842.CrossRefGoogle Scholar
- Stumpf, C. H., Piehler, M. F., Thompson, S., & Noble, R. T. (2010). Loading of fecal indicator bacteria in North Carolina tidal creek headwaters: hydrographic patterns and terrestrial runoff relationships. Water Research, 44, 4704–4715.CrossRefGoogle Scholar
- Tennessee Department of Environment and Conservation (TDEC). (2000). Tennessee ecoregion project: 1994–1999. Nashville: TDEC Division of Water Pollution Control.Google Scholar
- Tennessee Department of Environment and Conservation (TDEC). (2009). Quality systems for standard operating procedure for chemical and bacteriological sampling of surface water. Nashville: TDEC Division of Water Pollution Control.Google Scholar
- Tennessee Department of Environment and Conservation (TDEC). (2014). Proposed final version, year 2014 303(d) list. Nashville: TDEC Planning and Standards Unit, Division of Water Resources.Google Scholar
- Traister, E., & Anisfeld, S. C. (2006). Variability of indicator bacteria at different time scales in the upper Hoosic River watershed. Environmental Science and Technology, 40(16), 4990–4995.CrossRefGoogle Scholar
- United States Environmental Protection Agency (U.S. EPA). (1986). Bacteriological ambient water quality criteria for marine and fresh recreational waters. Washington, DC: U.S. EPA Office of Water.Google Scholar
- United States Environmental Protection Agency (U.S. EPA). (2001). Protocol for developing pathogen TMDLs. Washington, DC: U.S. EPA Office of Water.Google Scholar
- United States Environmental Protection Agency (U.S. EPA). (2005). Microbial source tracking guide document. Cincinnati: U.S. EPA National Risk Management Research Laboratory.Google Scholar
- Vierheilig, J., Farnleitner, A. H., Kollanur, D., Blöschl, G., & Reischer, G. H. (2012). High abundance of genetic Bacteroidetes markers for total fecal pollution in pristine alpine soils suggest lack of specificity for feces. Journal of Microbiological. Methods, 88, 433–435.CrossRefGoogle Scholar
- Wade, T. J., Calderon, R. L., Brenner, K. P., Sams, E., Beach, M., Haugland, R., Wymer, L., & Dufour, A. (2008). High sensitivity of children to swimming-associated gastrointestinal illness. Epidemiology, 19(3), 375–383.CrossRefGoogle Scholar
- Walters, S. P., & Field, K. G. (2009). Survival and persistence of human and ruminant-specific faecal Bacteroidales in freshwater microcosms. Environmental Microbiology, 11(6), 1410–1421.CrossRefGoogle Scholar
- Weidhaas, J., Mantha, S., Hair, E., Nayak, B., & Harwood, V. J. (2015). Evidence for extraintestinal growth of Bacteroidales originating from poultry litter. Applied and Environmental Microbiology, 81(1), 196–202.CrossRefGoogle Scholar