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


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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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.


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