Abstract
Monitoring of environmental water is crucial to protecting humans and animals from possible health risks. Although numerous human-specific viral markers have been designed to track the presence of human fecal contamination in water, they lack adequate sensitivity and specificity in different geographical regions. We evaluated the performances of six human-specific viral markers [Aichi virus 1 (AiV-1), human adenoviruses (HAdVs), BK and JC polyomaviruses (BKPyVs and JCPyVs), pepper mild mottle virus (PMMoV), and crAssphage] using 122 fecal-source samples collected from humans and five animal hosts in the Kathmandu Valley, Nepal. PMMoV and crAssphage showed high sensitivity (90–100%) with concentrations of 4.5–9.1 and 6.2–7.0 log10 copies/g wet feces (n = 10), respectively, whereas BKPyVs, JCPyVs, HAdVs, and AiV-1 showed poor performances with sensitivities of 30–40%. PMMoV and crAssphage were detected in 40–100% and 8–90%, respectively, of all types of animal fecal sources and showed no significantly different concentrations among most of the fecal sources (Kruskal–Wallis test, P > 0.05), suggesting their applicability as general fecal pollution markers. Furthermore, a total of 115 environmental water samples were tested for PMMoV and crAssphage to identify fecal pollution. PMMoV and crAssphage were successfully detected in 62% (71/115) and 73% (84/115) of water samples, respectively. The greater abundance and higher mean concentration of crAssphage (4.1 ± 0.9 log10 copies/L) compared with PMMoV (3.3 ± 1.4 log10 copies/L) indicated greater chance of detection of crAssphage in water samples, suggesting that crAssphage could be preferred to PMMoV as a marker of fecal pollution.
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Acknowledgements
The authors thank Prof. Suresh Das Shrestha (Tribhuvan University, Nepal), Prof. Kei Nishida, Dr. Takashi Nakamura, and Mr. Bijay Man Shakya, (University of Yamanashi, Japan) for their support in water sampling; Mr. Ocean Thakali, Ms. Niva Sthapit (University of Yamanashi), and Mr. Dinesh Bhandari (Tribhuvan University) for their support in laboratory analysis; and Dr. Sadhana Shrestha (University of Yamanashi) for her support in statistical analysis.
Funding
This study was financially supported by the Science and Technology Research Partnership for Sustainable Development (SATREPS) Program entitled “Hydro-microbiological Approach for Water Security in Kathmandu Valley, Nepal,” which is jointly funded by the Japan International Cooperation Agency (JICA) and the Japan Science and Technology Agency (JST). It is also supported by the Japan Society for the Promotion of Science (JSPS) through Grant-in-Aid for Scientific Research (B) (JP17H03332).
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Malla, B., Ghaju Shrestha, R., Tandukar, S. et al. Performance Evaluation of Human-Specific Viral Markers and Application of Pepper Mild Mottle Virus and CrAssphage to Environmental Water Samples as Fecal Pollution Markers in the Kathmandu Valley, Nepal. Food Environ Virol 11, 274–287 (2019). https://doi.org/10.1007/s12560-019-09389-x
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DOI: https://doi.org/10.1007/s12560-019-09389-x