Water, Air, & Soil Pollution

, 230:223 | Cite as

Seasonality of E. coli and Enterococci Concentrations in Creek Water, Sediment, and Periphyton

  • Matthew Daniel StockerEmail author
  • Jaclyn Elizabeth Smith
  • Cesar Hernandez
  • Dumitru Macarisin
  • Yakov Pachepsky


Environmental reservoirs of fecal indicator bacteria (FIB) are attracting increasing attention because of the ambiguity they present when assessing the microbial quality of water. FIB can survive and even grow in various environmental reservoirs which means FIB measured in the water column may not have originated directly from a fecal source. Sediment and periphyton, i.e., aquatic biofilms growing on submerged rocks, have been shown to harbor large populations of FIB in the environment. However, little is known about the spatial and temporal dynamics of FIB in periphyton. The objective of this work was to determine levels of the common FIB, Escherichia coli and enterococci, in creek water, sediment, and periphyton during the summer and winter. FIB were measured during two summer and winter sampling dates at five locations along a 2.8-km stretch of creek in Beltsville, Maryland. Significant differences in FIB by location were only observed for E. coli in water at one time point. Levels of FIB significantly declined from summer to winter in all media. FIB concentrations in periphyton ranged from 102 to 104 gdw−1 in the summer and from 100 to 104 CFU gdw−1 in the winter. When compared on a dry weight basis, periphyton contained higher concentrations of FIB than the sediment. Variability of FIB was in the order of water < sediment < periphyton. Levels of E. coli and enterococci measured in the same sample showed significant positive correlation in all media (rs = 0.87, 0.48, 0.70, for water, sediment, and periphyton, respectively). Results from this work show that fecal bacteria can persist in creek periphyton which may act as both a reservoir for fecal pathogens as well as a probable source of fecal bacteria to the water column.


Fecal indicator bacteria FIB Periphyton Sediments Water quality Food safety 



The authors would like to acknowledge the help of the USDA’s Agricultural Research Learning Experience (ARLE) program along with the Hispanic Serving Institutions initiative for supporting researchers to help plan and conduct the work.

Funding Information

This work was supported through the USDA’s Agricultural Research Service project number 8042-12630-011-00D.

Supplementary material

11270_2019_4263_MOESM1_ESM.jpg (485 kb)
Supplemental Figure 1. Flow and precipitation data recorded during the observation period. S1, S2, W1, and W2 represent sampling dates of 7/21/2017, 7/27/2017, 1/31/2018, and 2/6/2018, respectively. Panels a and b represent summer and winter sampling periods, respectively. (JPG 485 kb)
11270_2019_4263_MOESM2_ESM.jpg (798 kb)
Supplemental Figure 2. Cumulative probability distributions of E. coli and enterococci concentrations along the stream reach for the four observation periods. E. coli concentrations in water, sediment, and periphyton are presented in panels a, b, and c while enterococci in the same media are presented in d, e and f. (JPG 797 kb)
11270_2019_4263_MOESM3_ESM.jpg (367 kb)
Supplemental Figure 3. Comparison of FIB concentrations in periphyton as reported using different units. White circles and triangles represent E. coli concentrations in CFU gdw-1 and CFU cm-2, respectively. Black circles and triangles represent enterococci concentrations in CFU gdw-1 and CFU cm-2, respectively. (JPG 366 kb)
11270_2019_4263_MOESM4_ESM.docx (70 kb)
ESM 4 (DOCX 69 kb)


  1. Allende, A., & Monaghan, J. (2015). Irrigation water quality for leafy crops: a perspective of risks and potential solutions. International Journal of Environmental Research and Public Health, 12(7), 7457–7477.Google Scholar
  2. Balzer, M., Witt, N., Flemming, H. C., & Wingender, J. (2010). Faecal indicator bacteria in river biofilms. Water Science and Technology, 61(5), 1105–1111.Google Scholar
  3. Blaustein, R. A., Pachepsky, Y., Hill, R. L., Shelton, D. R., & Whelan, G. (2013). Escherichia coli survival in waters: temperature dependence. Water Research, 47(2), 569–578.Google Scholar
  4. Byappanahalli, M. N., Sawdey, R., Ishii, S., Shively, D. A., Ferguson, J. A., Whitman, R. L., & Sadowsky, M. J. (2009). Seasonal stability of Cladophora-associated Salmonella in Lake Michigan watersheds. Water Research, 43(3), 806–814.Google Scholar
  5. Cantonati, M., & Lowe, R. L. (2014). Lake benthic algae: toward an understanding of their ecology. Freshwater Science, 33(2), 475–486.Google Scholar
  6. Carr, G. M., Morin, A., & Chambers, P. A. (2005). Bacteria and algae in stream periphyton along a nutrient gradient. Freshwater Biology, 50(8), 1337–1350.Google Scholar
  7. Cho, K. H., Pachepsky, Y. A., Kim, J. H., Guber, A. K., Shelton, D. R., & Rowland, R. (2010). Release of Escherichia coli from the bottom sediment in a first-order creek: experiment and reach-specific modeling. Journal of Hydrology, 391(3–4), 322–332.Google Scholar
  8. Cinotto, P. J. (2005). Occurrence of fecal-indicator bacteria and protocols for identification of fecal-contamination sources in selected reaches of the West Branch Brandywine Creek, Chester County, Pennsylvania. Reston: US Department of the Interior, US Geological Survey.Google Scholar
  9. Cooper, I. R., Taylor, H. D., & Hanlon, G. W. (2007). Virulence traits associated with verocytotoxigenic Escherichia coli O157 recovered from freshwater biofilms. Journal of Applied Microbiology, 102(5), 1293–1299.Google Scholar
  10. Davies, C. M., Long, J. A., Donald, M., & Ashbolt, N. J. (1995). Survival of fecal microorganisms in marine and freshwater sediments. Applied and Environmental Microbiology, 61(5), 1888–1896.Google Scholar
  11. de Souza, M. L., Pellegrini, B. G., & Ferragut, C. (2015). Periphytic algal community structure in relation to seasonal variation and macrophyte richness in a shallow tropical reservoir. Hydrobiologia, 755(1), 183–196.Google Scholar
  12. Derlet, R. W., Richards, J. R., Tanaka, L. L., Hayden, C., Ger, K. A., & Goldman, C. R. (2012). Impact of summer cattle grazing on the Sierra Nevada watershed: aquatic algae and bacteria. Journal of Environmental and Public Health, 2012. Google Scholar
  13. Englebert, E. T., McDermott, C., & Kleinheinz, G. T. (2008). Effects of the nuisance algae, Cladophora, on Escherichia coli at recreational beaches in Wisconsin. The Science of the Total Environment, 404(1), 10–17.Google Scholar
  14. Ford, R. M., & Harvey, R. W. (2007). Role of chemotaxis in the transport of bacteria through saturated porous media. Advances in Water Resources, 30(6–7), 1608–1617.Google Scholar
  15. Gaertner, J. P., Mendoza, J. A., Forstner, M. R., & Hahn, D. (2011). Recovery of Salmonella from biofilms in a headwater spring ecosystem. Journal of Water and Health, 9(3), 458–466.Google Scholar
  16. Garzio-Hadzick, A., Shelton, D. R., Hill, R. L., Pachepsky, Y. A., Guber, A. K., & Rowland, R. (2010). Survival of manure-borne E. coli in streambed sediment: effects of temperature and sediment properties. Water Research, 44(9), 2753–2762.Google Scholar
  17. Grant, S. B., Litton-Mueller, R. M., & Ahn, J. H. (2011). Measuring and modeling the flux of fecal bacteria across the sediment-water interface in a turbulent stream. Water Resources Research, 47(5).
  18. Haglund, A. L., & Hillebrand, H. (2005). The effect of grazing and nutrient supply on periphyton associated bacteria. FEMS Microbiology Ecology, 52(1), 31–41.Google Scholar
  19. Haller, L., Amedegnato, E., Poté, J., & Wildi, W. (2009). Influence of freshwater sediment characteristics on persistence of fecal indicator bacteria. Water, Air, and Soil Pollution, 203(1–4), 217–227.Google Scholar
  20. Hammer, Ø., Harper, D. A., & Ryan, P. D. (2001). PAST: paleontological statistics software package for education and data analysis. Palaeontologia Electronica, 4(1), 9.Google Scholar
  21. 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.Google Scholar
  22. Ishii, S., & Sadowsky, M. J. (2008). Escherichia coli in the environment: implications for water quality and human health. Microbes and Environments, 23(2), 101–108.Google Scholar
  23. Jamieson, R., Joy, D. M., Lee, H., Kostaschuk, R., & Gordon, R. (2005a). Transport and deposition of sediment-associated Escherichia coli in natural streams. Water Research, 39(12), 2665–2675.Google Scholar
  24. Jamieson, R. C., Joy, D. M., Lee, H., Kostaschuk, R., & Gordon, R. J. (2005b). Resuspension of sediment-associated Escherichia coli in a natural stream. Journal of Environmental Quality, 34(2), 581–589.Google Scholar
  25. Kim, J. W., Pachepsky, Y. A., Shelton, D. R., & Coppock, C. (2010). Effect of streambed bacteria release on E. coli concentrations: monitoring and modeling with the modified SWAT. Ecological Modelling, 221(12), 1592–1604.Google Scholar
  26. Koirala, S. R., Gentry, R. W., Perfect, E., Schwartz, J. S., & Sayler, G. S. (2008). Temporal variation and persistence of bacteria in streams. Journal of Environmental Quality, 37(4), 1559–1566.Google Scholar
  27. Kovačić, A., Tripković, I., Galov, A., & Žitko, T. (2011). Distribution of microbiological indicators of fecal pollution in the riverine substrates. Environmental Monitoring and Assessment, 172(1–4), 623–630.Google Scholar
  28. Ksoll, W. B., Ishii, S., Sadowsky, M. J., & Hicks, R. E. (2007). Presence and sources of fecal coliform bacteria in epilithic periphyton communities of Lake Superior. Applied and Environmental Microbiology, 73(12), 3771–3778.Google Scholar
  29. Latorre, A. A., Van Kessel, J. S., Karns, J. S., Zurakowski, M. J., Pradhan, A. K., Boor, K. J., et al. (2010). Biofilm in milking equipment on a dairy farm as a potential source of bulk tank milk contamination with Listeria monocytogenes. Journal of Dairy Science, 93(6), 2792–2802.Google Scholar
  30. Li, X., Peed, L., Sivaganesan, M., Kelty, C.A., Neitch, C., Shanks, O.C. (2017). Simultaneous measurement of genetic fecal indicators in water column and periphyton biofilms in artificial streams. Environmental Protection Agency. Poster presentation at Water Microbiology Conference, Chapel Hill, N.C, May 15 – 19th, 2017.Google Scholar
  31. Mackowiak, M., Leifels, M., Hamza, I. A., Jurzik, L., & Wingender, J. (2018). Distribution of Escherichia coli, coliphages and enteric viruses in water, epilithic biofilms and sediments of an urban river in Germany. Science of the Total Environment, 626, 650–659.Google Scholar
  32. Mathai, P. P., Dunn, H. M., Magnone, P., Zhang, Q., Ishii, S., Chun, C. L., & Sadowsky, M. J. (2019). Association between submerged aquatic vegetation and elevated levels of Escherichia coli and potential bacterial pathogens in freshwater lakes. Science of the Total Environment, 657, 319-324.Google Scholar
  33. Moreira, S., Brown, A., Ha, R., Iserhoff, K., Yim, M., Yang, J., et al. (2012). Persistence of E scherichia coli in freshwater periphyton: biofilm-forming capacity as a selective advantage. FEMS Microbiology Ecology, 79(3), 608–618.Google Scholar
  34. Muirhead, R. W., Davies-Colley, R. J., Donnison, A. M., & Nagels, J. W. (2004). Faecal bacteria yields in artificial flood events: quantifying in-stream stores. Water Research, 38(5), 1215–1224.Google Scholar
  35. Mulling, B. T. M., van den Boomen, R. M., van der Geest, H. G., Kappelhof, J. W. N. M., & Admiraal, W. (2013). Suspended particle and pathogen peak discharge buffering by a surface-flow constructed wetland. Water Research, 47, 1091–1100.Google Scholar
  36. Naranjo, R. C., Niswonger, R. G., Smith, D., Rosenberry, D., & Chandra, S. (2019). Linkages between hydrology and seasonal variations of nutrients and periphyton in a large oligotrophic subalpine lake. Journal of Hydrology, 568, 877–890.Google Scholar
  37. Pachepsky, Y. A., Allende, A., Boithias, L., Cho, K., Jamieson, R., Hofstra, N., & Molina, M. (2018). Microbial water quality: monitoring and modeling. Journal of Environmental Quality, 47(5), 931–938.Google Scholar
  38. Pachepsky, Y. A., & Shelton, D. R. (2011). Escherichia coli and fecal coliforms in freshwater and estuarine sediments. Critical Reviews in Environmental Science and Technology, 41(12), 1067–1110.Google Scholar
  39. Pachepsky, Y., Stocker, M., Saldaña, M. O., & Shelton, D. (2017). Enrichment of stream water with fecal indicator organisms during baseflow periods. Environmental Monitoring and Assessment, 189(2), 51.Google Scholar
  40. Pandey, P. K., Soupir, M. L., Ikenberry, C. D., & Rehmann, C. R. (2016). Predicting streambed sediment and water column Escherichia coli levels at watershed scale. JAWRA Journal of the American Water Resources Association, 52(1), 184–197.Google Scholar
  41. Park, Y., Pachepsky, Y., Hong, E. M., Shelton, D., & Coppock, C. (2017). Escherichia coli release from streambed to water column during baseflow periods: a modeling study. Journal of Environmental Quality, 46(1), 219–226.Google Scholar
  42. Phillips, M. C., Solo-Gabriele, H. M., Reniers, A. J., Wang, J. D., Kiger, R. T., & Abdel-Mottaleb, N. (2011). Pore water transport of enterococci out of beach sediments. Marine Pollution Bulletin, 62(11), 2293–2298.Google Scholar
  43. Piorkowski, G., Jamieson, R., Bezanson, G., Hansen, L. T., & Yost, C. (2014). Reach specificity in sediment E. coli population turnover and interaction with waterborne populations. Science of the Total Environment, 496, 402–413.Google Scholar
  44. Piorkowski, G. S., Jamieson, R. C., Hansen, L. T., Bezanson, G. S., & Yost, C. K. (2014). Characterizing spatial structure of sediment E. coli populations to inform sampling design. Environmental Monitoring and Assessment, 186(1), 277–291.Google Scholar
  45. Romani, H. G. A. M. (Ed.). (2016). Aquatic biofilms. Caister Academic Press, Wymondham.Google Scholar
  46. Rosemond, A. D. (1994). Multiple factors limit seasonal variation in periphyton in a forest stream. Journal of the North American Benthological Society, 13(3), 333–344.Google Scholar
  47. Sha, Q. (2012). Distribution, diversity, and fate of salmonella in natural biofilms. (Doctoral dissertation). Retrieved from
  48. Smith, J.E., Stocker, M.D., Pachepsky, Y. (2019). The effect of temperature oscillations and sediment texture on fecal indicator bacteria survival in sediments. In Review. Journal of Environmental Quality.Google Scholar
  49. Stocker, M. D., Penrose, M., & Pachepsky, Y. A. (2018). Spatial patterns of Escherichia coli concentrations in sediment before and after high-flow events in a first-order creek. Journal of Environmental Quality, 47(5), 958–966.Google Scholar
  50. Stocker, M. D., Rodriguez-Valentin, J. G., Pachepsky, Y. A., & Shelton, D. R. (2016). Spatial and temporal variation of fecal indicator organisms in two creeks in Beltsville, Maryland. Water Quality Research Journal, 51(2), 167–179.Google Scholar
  51. Traister, E., & Anisfeld, S. C. (2006). Variability of indicator bacteria at different time scales in the upper Hoosic River watershed. Environmental Science & Technology, 40(16), 4990–4995.Google Scholar
  52. Tyrrel, S. F., & Quinton, J. N. (2003). Overland flow transport of pathogens from agricultural land receiving faecal wastes. Journal of Applied Microbiology, 94, 87–93.Google Scholar
  53. USEPA.2012. Recreational water quality criteria; Office of Water, United States Environmental Potection Agency: Washington, D.C.Google Scholar
  54. van Dam, A. A., Beveridge, M. C., Azim, M. E., & Verdegem, M. C. (2002). The potential of fish production based on periphyton. Reviews in Fish Biology and Fisheries, 12(1), 1–31.Google Scholar
  55. Vogeleer, P., Tremblay, Y. D., Mafu, A. A., Jacques, M., & Harel, J. (2014). Life on the outside: role of biofilms in environmental persistence of Shiga-toxin producing Escherichia coli. Frontiers in Microbiology, 5, 317.Google Scholar
  56. Whitman, R. L., Shively, D. A., Pawlik, H., Nevers, M. B., & Byappanahalli, M. N. (2003). Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan. Applied and Environmental Microbiology, 69(8), 4714–4719.Google Scholar

Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2019

Authors and Affiliations

  1. 1.Oak Ridge Institute for Science and EducationOak RidgeUSA
  2. 2.USDA-ARS Environmental Microbial and Food Safety LaboratoryBeltsvilleUSA
  3. 3.Department of Chemistry and BiochemistryUniversity of Texas at El PasoEl PasoUSA
  4. 4.Office of Regulatory Science, Center for Food Safety and Applied NutritionFood and Drug AdministrationCollege ParkUSA

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