Aims: To utilize microbial source tracking to detect and differentiate sources of fecal bacteria in Texas, addressing the limitations of dated culture-based Enterolert tests, which quantify fecal indicator bacteria (FIB) but fail to indicate the source of the pollution. Study Design: This study involved quantification of FIB using some DNA-based tests validated by the United States Environmental Protection Agency (EPA). Place and Duration of Study: Water samples were collected from two counties along the Texas coast from February 2022 through June 2023. Methodology: EPA Method 1696 was conducted on 198 water samples collected for the detection of human-associated Bacteroidales by HF183/BacR287 quantitative polymerase chain reaction (qPCR) assay. A human-associated Enterococcus qPCR assay was also performed on a subset of Enterococcus isolates subcultured from Enterolert IDEXX trays to further test for the presence of human fecal contamination. A general Bacteroidales qPCR assay was also conducted to detect fecal contamination from various endothermic animals. These additional qPCR assays were used to detect FIB from avian, equine, ruminant, bovine, swine, and canine sources. Results: Although no samples tested positive for human-associated Bacteroidales, 7.6% of the subcultured samples tested positive for human-associated Enterococcus. All samples were positive for general Bacteroidales markers, and most samples were positive for avian FIB, while FIB from other animal sources were absent or detected in less than 5% of samples. Conclusion: This study provides insight into human and non-human contributions to high FIB counts in recreational waters along the Texas coast. Understanding these sources may improve water quality management and public health efforts.
References
[1]
González-Fernández, A., Symonds, E.M., Gallard-Gongora, J.F., Mull, B., Lukasik, J.O., Rivera Navarro, P., et al. (2021) Relationships among Microbial Indicators of Fecal Pollution, Microbial Source Tracking Markers, and Pathogens in Costa Rican Coastal Waters. Water Research, 188, Article ID: 116507. https://doi.org/10.1016/j.watres.2020.116507
[2]
Xiang, R., Xu, Y., Liu, Y., Lei, G., Liu, J. and Huang, Q. (2019) Isolation Distance between Municipal Solid Waste Landfills and Drinking Water Wells for Bacteria Attenuation and Safe Drinking. Scientific Reports, 9, Article No. 17881. https://doi.org/10.1038/s41598-019-54506-2
[3]
Noble, R.T., Blackwood, A.D., Griffith, J.F., McGee, C.D. and Weisberg, S.B. (2010) Comparison of Rapid Quantitative Pcr-Based and Conventional Culture-Based Methods for Enumeration of Enterococcus Spp. And Escherichia coli in Recreational Waters. Applied and Environmental Microbiology, 76, 7437-7443. https://doi.org/10.1128/aem.00651-10
[4]
Parker, J.K., McIntyre, D. and Noble, R.T. (2010) Characterizing Fecal Contamination in Stormwater Runoff in Coastal North Carolina, Usa. Water Research, 44, 4186-4194. https://doi.org/10.1016/j.watres.2010.05.018
[5]
Powers, N.C., Pinchback, J., Flores, L., Huang, Y., Wetz, M.S. and Turner, J.W. (2021) Long-term Water Quality Analysis Reveals Correlation between Bacterial Pollution and Sea Level Rise in the Northwestern Gulf of Mexico. Marine Pollution Bulletin, 166, Article ID: 112231. https://doi.org/10.1016/j.marpolbul.2021.112231
[6]
EPA (2015) Method 1609.1: Enterococci in Water by TaqMan Quantitative Polymerase Chain Reaction (qPCR) with Internal Amplification Control (IAC) Assay. EPA-820-R-15-099. https://www.epa.gov/sites/default/files/2015-08/documents/method_1609-1-enterococcus-iac_2015_3.pdf
[7]
Brooks, Y.M., Spirito, C.M., Bae, J.S., Hong, A., Mosier, E.M., Sausele, D.J., et al. (2020) Fecal Indicator Bacteria, Fecal Source Tracking Markers, and Pathogens Detected in Two Hudson River Tributaries. Water Research, 171, Article ID: 115342. https://doi.org/10.1016/j.watres.2019.115342
[8]
Ahmed, W., Hughes, B. and Harwood, V. (2016) Current Status of Marker Genes of Bacteroides and Related Taxa for Identifying Sewage Pollution in Environmental Waters. Water, 8, Article 231. https://doi.org/10.3390/w8060231
[9]
EPA (2019) Method 1696: Characterization of Human Fecal Pollution in Water by HF183/BacR287 TaqMan Quantitative Polymerase Chain Reaction (qPCR) Assay. EPA-820-R-19-002. https://www.epa.gov/sites/default/files/2019-03/documents/method_1696_draft_2019.pdf
[10]
Nshimyimana, J.P., Cruz, M.C., Thompson, R.J. and Wuertz, S. (2017) Bacteroidales Markers for Microbial Source Tracking in Southeast Asia. Water Research, 118, 239-248. https://doi.org/10.1016/j.watres.2017.04.027
[11]
McMinn, B.R., Korajkic, A., Kelleher, J., Diedrich, A., Pemberton, A., Willis, J.R., et al. (2024) Quantitative Fecal Pollution Assessment with Bacterial, Viral, and Molecular Methods in Small Stream Tributaries. Science of the Total Environment, 951, Article ID: 175740. https://doi.org/10.1016/j.scitotenv.2024.175740
[12]
Green, H.C., Dick, L.K., Gilpin, B., Samadpour, M. and Field, K.G. (2012) Genetic Markers for Rapid PCR-Based Identification of Gull, Canada Goose, Duck, and Chicken Fecal Contamination in Water. Applied and Environmental Microbiology, 78, 503-510. https://doi.org/10.1128/aem.05734-11
[13]
EPA (2009) Method 1600: Enterococci in Water by Membrane Filtration Using Mem-brane-Enterococcus Indoxyl-β-D-Glucoside Agar (mEl). EPA-821-R-09-016. https://www.epa.gov/sites/default/files/2015-08/documents/method_1600_2009.pdf
[14]
Brady, J.A., Faske, J.B., Castañeda-Gill, J.M., King, J.L. and Mitchell, F.L. (2011) High-throughput DNA Isolation Method for Detection of Xylella Fastidiosa in Plant and Insect Samples. Journal of Microbiological Methods, 86, 310-312. https://doi.org/10.1016/j.mimet.2011.06.007
[15]
Ahmed, W., Stewart, J., Gardner, T. and Powell, D. (2008) A Real‐time Polymerase Chain Reaction Assay for Quantitative Detection of the Human‐Specific Enterococci Surface Protein Marker in Sewage and Environmental Waters. Environmental Microbiology, 10, 3255-3264. https://doi.org/10.1111/j.1462-2920.2008.01715.x
[16]
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. Applied and Environmental Microbiology, 72, 4214-4224. https://doi.org/10.1128/aem.01036-05
[17]
Mieszkin, S., Yala, J., Joubrel, R. and Gourmelon, M. (2010) Phylogenetic Analysis of Bacteroidales 16S rRNA Gene Sequences from Human and Animal Effluents and Assessment of Ruminant Faecal Pollution by Real‐Time PCR. Journal of Applied Microbiology, 108, 974-984. https://doi.org/10.1111/j.1365-2672.2009.04499.x
[18]
Kildare, B.J., Leutenegger, C.M., McSwain, B.S., Bambic, D.G., Rajal, V.B. and Wuertz, S. (2007) 16S rRNA-Based Assays for Quantitative Detection of Universal, Human-, Cow-, and Dog-Specific Fecal Bacteroidales: A Bayesian Approach. Water Research, 41, 3701-3715. https://doi.org/10.1016/j.watres.2007.06.037
[19]
Tambalo, D.D., Boa, T., Liljebjelke, K. and Yost, C.K. (2012) Evaluation of Two Quantitative PCR Assays Using Bacteroidales and Mitochondrial DNA Markers for Tracking Dog Fecal Contamination in Waterbodies. Journal of Microbiological Methods, 91, 459-467. https://doi.org/10.1016/j.mimet.2012.09.029
[20]
Mieszkin, S., Furet, J., Corthier, G. and Gourmelon, M. (2009) Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers. Applied and Environmental Microbiology, 75, 3045-3054. https://doi.org/10.1128/aem.02343-08
[21]
R Core Team (2022) R: A Language and Environment for Statistical Computing (Version 4.2.1). R Foundation for Statistical Computing. https://www.R-project.org/
[22]
Shrestha, A., Kelty, C.A., Sivaganesan, M., Shanks, O.C. and Dorevitch, S. (2020) Fecal Pollution Source Characterization at Non-Point Source Impacted Beaches under Dry and Wet Weather Conditions. Water Research, 182, Article ID: 116014. https://doi.org/10.1016/j.watres.2020.116014
[23]
Hachad, M., Lanoue, M., Vo Duy, S., Villemur, R., Sauvé, S., Prévost, M., et al. (2022) Locating Illicit Discharges in Storm Sewers in Urban Areas Using Multi-Parameter Source Tracking: Field Validation of a Toolbox Composite Index to Prioritize High Risk Areas. Science of the Total Environment, 811, Article ID: 152060. https://doi.org/10.1016/j.scitotenv.2021.152060
[24]
Walters, S.P., Yamahara, K.M. and Boehm, A.B. (2009) Persistence of Nucleic Acid Markers of Health-Relevant Organisms in Seawater Microcosms: Implications for Their Use in Assessing Risk in Recreational Waters. Water Research, 43, 4929-4939. https://doi.org/10.1016/j.watres.2009.05.047
[25]
Staley, C., Reckhow, K.H., Lukasik, J. and Harwood, V.J. (2012) Assessment of Sources of Human Pathogens and Fecal Contamination in a Florida Freshwater Lake. Water Research, 46, 5799-5812. https://doi.org/10.1016/j.watres.2012.08.012
[26]
Ballesté, E., Demeter, K., Masterson, B., Timoneda, N., Sala-Comorera, L. and Meijer, W.G. (2020) Implementation and Integration of Microbial Source Tracking in a River Watershed Monitoring Plan. Science of the Total Environment, 736, Article ID: 139573. https://doi.org/10.1016/j.scitotenv.2020.139573
[27]
Inglis, J.M., Hood, R.E., Brown, B.A. and DeYoung, C.A. (1979) Home Range of White-Tailed Deer in Texas Coastal Prairie Brushland. Journal of Mammalogy, 60, 377-389. https://doi.org/10.2307/1379810
[28]
Fogarty, L.R., Haack, S.K., Wolcott, M.J. and Whitman, R.L. (2003) Abundance and Characteristics of the Recreational Water Quality Indicator Bacteria Escherichia coli and Enterococci in Gull Faeces. Journal of Applied Microbiology, 94, 865-878. https://doi.org/10.1046/j.1365-2672.2003.01910.x