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Comparative persistence of human sewage-specific enterococcal bacteriophages in freshwater and seawater

Abstract

Enterococcus faecalis bacteria have been recently reported for their ability to host bacteriophages that are specifically from human sewage, suggesting their application to track human fecal contamination in water resources. However, little is known about the survivability of sewage-specific enterococcal bacteriophages in various water matrices under ambient and storage conditions. In this study, bacteriophages that were derived from the Thailand-isolated E. faecalis strains AIM06 and SR14 exhibited morphologies consistent with the Siphoviridae, Podoviridae, and Myoviridae families. Four representative bacteriophages were separately spiked into environmental water samples (n = 7) comprising freshwater and seawater with low- and high-pollution (LF, HF, LS, and HS, respectively) levels, defined according to Thailand Water Quality Standards. All bacteriophages decayed fastest in HS or HF samples at 30 °C, reaching a 5-log10 reduction in 2.2 to 9.8 days, and slowest in LS samples, requiring 8.8 to 23.5 days. The decay rates were 5 to 53 times lower at a storage temperature of 5 °C. HF samples could be stored for as little as 2.5 days to prevent the decay of 50% of the phages. Myoviridae phages decayed faster than Siphoviridae phages and Podoviridae phages in most water matrices at 30 °C. Moreover, the decay rates were 1.8 to 92 times slower in filtered samples, emphasizing a strong role for water constituents, i.e., suspended solids and natural microorganisms, in phage persistence. This study emphasized that differential enterococcal bacteriophage persistence should be considered when planning the monitoring and interpreting of fecal sources by microbial source tracking.

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References

  1. American Public Health Association, American Water Works Association, Water Environment Federation (2017a) 2540 D. Total suspended solids dried at 103 – 105°C. In: Standard methods for the examination of water & wastewater. Online edition. Access date 16 January, vol 2017, pp 4–5

  2. American Public Health Association, American Water Works Association, Water Environment Federation (2017b) 5210 B. 5-day BOD test. In: Standard methods for the examination of water & wastewater. Online edition. Access Date 16 January 2017:1–6

  3. Bachrach G, Leizerovici-Zigmond M, Zlotkin A, Naor R, Steinberg D (2003) Bacteriophage isolation from human saliva. Lett Appl Microbiol 36:50–53. https://doi.org/10.1046/j.1472-765X.2003.01262.x

  4. Bonilla N, Santiago T, Marcos P, Urdaneta M, Santo Domingo J, Toranzos GA (2010) Enterophages, a group of phages infecting Enterococcus faecalis, and their potential as alternate indicators of human faecal contamination. Water Sci Technol 61:293–300. https://doi.org/10.2166/wst.2010.815

  5. Durán AE, Muniesa M, Méndez X, Valero F, Lucena F, Jofre J (2002) Removal and inactivation of indicator bacteriophages in fresh waters. J Appl Microbiol 92:338–347. https://doi.org/10.1046/j.1365-2672.2002.01536.x

  6. Durán AE, Muniesa M, Mocé-Llivina L, Campos C, Jofre J, Lucena F (2003) Usefulness of different groups of bacteriophages as model micro-organisms for evaluating chlorination. J Appl Microbiol 95:29–37. https://doi.org/10.1046/j.1365-2672.2003.t01-1-01948.x

  7. Fard RMN, Barton MD, Heuzenroeder MW (2010) Novel bacteriophages in enterococcus spp. Curr Microbiol 60:400–406. https://doi.org/10.1007/s00284-009-9555-z

  8. Fujioka R, Solo-Gabriele H, Byappanahalli M, Kirs M (2015) U.S. recreational water quality criteria: a vision for the future. Int J Environ Res Public Health 12:7752–7776. https://doi.org/10.3390/ijerph120707752

  9. Griffith JF, Layton B a, Boehm AB, Holden P a, Jay J a, Hagedorn C, McGee CD, Weisberg SB (2013) The California microbial source identification manual: a tiered approach to identifying fecal pollution sources to beaches

  10. Hamdi S, Rousseau GM, Labrie SJ, Tremblay DM, Kourda RS, Ben Slama K, Moineau S (2017) Characterization of two polyvalent phages infecting Enterobacteriaceae. Sci Rep 7:40349. https://doi.org/10.1038/srep40349

  11. Jensen EC, Schrader HS, Rieland B, Thompson TL, Lee KW, Nickerson KW, Kokjohn TA (1998) Prevalence of broad-host-range lytic bacteriophages of Sphaerotilus natans, Escherichia coli, and Pseudomonas aeruginosa. Appl Environ Microbiol 64:575–580

  12. Jofre J (2009) Is the replication of somatic coliphages in water environments significant? J Appl Microbiol 106:1059–1069

  13. Jofre J, Blanch AR, Lucena F, Muniesa M (2014) Bacteriophages infecting Bacteroides as a marker for microbial source tracking. Water Res 55:1–11. https://doi.org/10.1016/j.watres.2014.02.006

  14. Jofre J, Lucena F, Blanch AR, Muniesa M (2016) Coliphages as model organisms in the characterization and management of water resources. Water (Switzerland) 8:1–21. https://doi.org/10.3390/w8050199

  15. Lasobras J, Muniesa M, Frias J, Lucena F, Jofre J (1997) Relationship between the morphology of bacteriophages and their persistence in the environment. Water Sci Technol 35:129–132

  16. Lefkowitz EJ, Adams, Michael J, Davison AJ, Siddell SG, Simmonds P (2017) Virus taxonomy: classification and nomenclature of viruses-online (10th) Report of the International Committee on Taxonomy of Viruses

  17. Leknoi Y, Mongkolsuk S, Sirikanchana K (2016) Assessment of swine-specific bacteriophages of Bacteroides fragilis in swine farms with different antibiotic practices. J Water Health 15:251–261. https://doi.org/10.2166/wh.2016.069

  18. McMinn BR, Ashbolt NJ, Korajkic A (2017) Bacteriophages as indicators of fecal pollution and enteric virus removal. Lett Appl Microbiol 65:11–26. https://doi.org/10.1111/lam.12736

  19. Moldovan R, Chapman-McQuiston E, Wu XL (2007) On kinetics of phage adsorption. Biophys J 93:303–315. https://doi.org/10.1529/biophysj.106.102962

  20. Muniesa M, Lucena F, Jofre J (1999) Study of the potential relationship between the morphology of infectious somatic coliphages and their persistence in the environment. J Appl Microbiol 87:402–409. https://doi.org/10.1046/j.1365-2672.1999.00833.x

  21. Muniesa M, Jofre J (2004) Factors influencing the replication of somatic coliphages in the water environment. Antonie Leeuwenhoek 86:65–76

  22. Muniesa M, Jofre J (2007) The contribution of induction of temperate phages to the numbers of free somatic coliphages in waters is not significant. FEMS Microbiol Lett 270:272–276

  23. National Environment Board (1994) Notification of the National Environmental Board: Surface Water Quality Standards, No. 8, B.E. 2537 (1994), issued under the Enhancement and Conservation of National Environmental Quality Act B.E.2535 (1992)

  24. National Environment Board (2006) Notification of the National Environmental Board, No. 27, B.E. 2549 (2006), entitled Coastal Water Quality Standards

  25. Noble RT, Lee IM, Schiff KC (2004) Inactivation of indicator micro-organisms from various sources of faecal contamination in seawater and freshwater. J Appl Microbiol 96:464–472. https://doi.org/10.1111/j.1365-2672.2004.02155.x

  26. Payan A, Ebdon J, Taylor H, Gantzer C, Ottoson J, Papageorgiou GT, Blanch AR, Lucena F, Jofre J, Muniesa M (2005) Method for isolation of Bacteroides bacteriophage host strains suitable for tracking sources of fecal pollution in water. Appl Environ Microbiol 71:5659–5662. https://doi.org/10.1128/AEM.71.9.5659

  27. Payment P, Locas A (2011) Pathogens in water: value and limits of correlation with microbial indicators. Ground Water 49:4–11. https://doi.org/10.1111/j.1745-6584.2010.00710.x

  28. Pollution Control Department (PCD) (2012a) Thailand State of Pollution Report 2012

  29. Pollution Control Department (PCD) (2012b) Coastal water quality values nationwide in 2012

  30. Purnell SE, Ebdon JE, Taylor HD (2011) Bacteriophage lysis of Enterococcus host strains: a tool for microbial source tracking? Environ Sci Technol 45:10699–10705. https://doi.org/10.1021/es202141x

  31. Santiago-Rodríguez TM, Dávila C, González J, Bonilla N, Marcos P, Urdaneta M, Cadete M, Monteiro S, Santos R, Domingo JS, G a T (2010) Characterization of Enterococcus faecalis-infecting phages (enterophages) as markers of human fecal pollution in recreational waters. Water Res 44:4716–4725. https://doi.org/10.1016/j.watres.2010.07.078

  32. Santiago-Rodriguez TM, Marcos P, Monteiro S, Urdaneta M, Santos R, Toranzos GA (2013) Evaluation of Enterococcus-infecting phages as indices of fecal pollution. J Water Health 11:51–63. https://doi.org/10.2166/wh.2012.100

  33. Savichtcheva O, Okabe S (2006) Alternative indicators of fecal pollution: relations with pathogens and conventional indicators, current methodologies for direct pathogen monitoring and future application perspectives. Water Res 40:2463–2476. https://doi.org/10.1016/j.watres.2006.04.040

  34. Silverman AI, Nguyen MT, Schilling IE, Wenk J, Nelson KL (2015) Sunlight inactivation of viruses in open-water unit process treatment wetlands: modeling endogenous and exogenous inactivation rates. Environ Sci Technol 49:2757–2766. https://doi.org/10.1021/es5049754

  35. Silverman AI, Peterson BM, Boehm AB, McNeill K, Nelson KL (2013) Sunlight inactivation of human viruses and bacteriophages in coastal waters containing natural photosensitizers. Environ Sci Technol 47:1870–1878. https://doi.org/10.1021/es3036913

  36. Sinton LW, Finlay RK, Lynch P a (1999) Sunlight inactivation of fecal bacteriophages and bacteria in sewage-polluted seawater. Appl Environ Microbiol 65:3605–3613

  37. Sinton LW, Hall CH, Lynch P a., Davies-Colley RJ (2002) Sunlight inactivation of fecal indicator bacteria and bacteriophages from waste stabilization pond effluent in fresh and saline waters. Appl Environ Microbiol 68:1122–1131. https://doi.org/10.1128/AEM.68.3.1122-1131.2002

  38. Sirikanchana K, Wangkahad B, Mongkolsuk S (2014) The capability of non-native strains of Bacteroides bacteria to detect bacteriophages as faecal indicators in a tropical area. J Appl Microbiol 117:1820–1829. https://doi.org/10.1111/jam.12646

  39. Soller J a., Schoen ME, Bartrand T, Ravenscroft JE, Ashbolt NJ (2010) Estimated human health risks from exposure to recreational waters impacted by human and non-human sources of faecal contamination. Water Res 44:4674–4691. https://doi.org/10.1016/j.watres.2010.06.049

  40. Soller J, Bartrand T, Ravenscroft J, Molina M, Whelan G, Schoen M, Ashbolt N (2015) Estimated human health risks from recreational exposures to stormwater runoff containing animal faecal material. Environ Model Softw 72:21–32. https://doi.org/10.1016/j.envsoft.2015.05.018

  41. Souza KA, Ginoza HS, Haight RD (1972) Isolation of a polyvalent bacteriophage for Escherichia coli, Klebsiella pneumoniae, and Aerobacter aerogenes. J Virol 9:851–856

  42. Uchiyama J, Rashel M, Maeda Y, Takemura I, Sugihara S, Akechi K, Muraoka A, Wakiguchi H, Matsuzaki S (2008) Isolation and characterization of a novel Enterococcus faecalis bacteriophage øEF24C as a therapeutic candidate. FEMS Microbiol Lett 278:200–206. https://doi.org/10.1111/j.1574-6968.2007.00996.x

  43. US EPA (2015) Review of coliphages as possible indicators of fecal contamination for ambient water quality

  44. US EPA (2011) Using microbial source tracking to support TMDL development and implementation. Seattle, Washington, USA

  45. US EPA (2002a) Method 1604: Total coliforms and Escherichia coli in water by membrane filtration using a simultaneous detection technique (MI medium). EPA 821-R-02-024

  46. US EPA (2002b) Method 1600: enterococci in water by membrane filtration using membrane-enterococcus indoxyl-beta-D-glucoside agar (mEI). EPA 821-R-02-022

  47. Vijayavel K, Byappanahalli MN, Ebdon J, Taylor H, Whitman RL, Kashian DR (2014) Enterococcus phages as potential tool for identifying sewage inputs in the Great Lakes region. J Great Lakes Res 40:989–993. https://doi.org/10.1016/j.jglr.2014.09.011

  48. Wangkahad B, Bosup S, Mongkolsuk S, Sirikanchana K (2015) Occurrence of bacteriophages infecting Aeromonas, Enterobacter, and Klebsiella in water and association with contamination sources in Thailand. J Water Health 13:613–624. https://doi.org/10.2166/hydro.2013.124

  49. Wangkahad B, Mongkolsuk S, Sirikanchana K (2017) Integrated multivariate analysis with nondetects for the development of human sewage source-tracking tools using bacteriophages of Enterococcus faecalis. Environ Sci Technol 51:2235–2245. https://doi.org/10.1021/acs.est.6b04714

  50. Yang Y, Griffiths MW (2013) Comparative persistence of subgroups of F-specific RNA phages in river water. Appl Environ Microbiol 79:4564–4567. https://doi.org/10.1128/AEM.00612-13

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Acknowledgements

Professor Juan Jofre from the University of Barcelona, Spain, is thanked for his kind provision of B. thetaiotaomicron strains HB13 and GA17 and bacteriophages. We would like to also thank Mr. Akechai Kongprajug for his assistance in statistical analysis.

Funding

This study was funded by the Chulabhorn Research Institute (grant number BT2015-02), the Chulabhorn Graduate Institute, and the Center of Excellence on Environmental Health and Toxicology, Thailand.

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Correspondence to Kwanrawee Sirikanchana.

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The authors declare that they have no conflicts of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Booncharoen, N., Mongkolsuk, S. & Sirikanchana, K. Comparative persistence of human sewage-specific enterococcal bacteriophages in freshwater and seawater. Appl Microbiol Biotechnol 102, 6235–6246 (2018) doi:10.1007/s00253-018-9079-1

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Keywords

  • Bacteriophages
  • Enterococci
  • Decay
  • Microbial source tracking
  • Fecal indicator