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Estimation of Contamination Sources of Human Enteroviruses in a Wastewater Treatment and Reclamation System by PCR–DGGE

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Abstract

A polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE) method was employed to estimate the contamination sources of human enteroviruses and understand how their dominant strains vary in a wastewater treatment and reclamation system consisting of sewage collection, wastewater treatment with membrane bioreactor and open lakes for reclaimed water storage and reuse. After PCR–DGGE using a selected primer set targeting enteroviruses, phylogenetic analysis of acquired enterovirus gene sequences was performed. Enteroviruses identified from the septic tank were much more diverse than those from grey water and kitchen wastewater. Several unique types of enterovirus different from those in wastewater samples were dominant in a biological wastewater treatment unit. Membrane filtration followed by chlorination was proved effective for physically eliminating enteroviruses; however, secondary contamination likely occurred as the reclaimed water was stored in artificial lakes. Enterovirus 71 (EV71), a hand-foot-and-mouth disease (HFMD) viral pathogen, was detected mainly from the artificial lakes, implying that wastewater effluent was not the contamination source of EV71 and that there were unidentified non-point sources of the contamination with the HFMD viral pathogen in the reclaimed water stored in the artificial lakes. The PCR–DGGE targeting enteroviruses provided robust evidence about viral contamination sources in the wastewater treatment and reclamation system.

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Acknowledgments

We appreciate the support of Dr. Hiroyuki Shimizu, National Institute of Infectious Diseases, Japan, and Dr. Kazuyoshi Yano, Tokyo Metropolitan Institute of Public Health, Japan for kindly providing enterovirus strains. We also thank Dr. Yoshifumi Masago (Tohoku University, Japan), Dr. Chaofeng Ma (Xi’an Center for Disease Control and Prevention, China) and Dr. Takayuki Miura (Hokkaido University, Japan) for their kind suggestions and assistance. This study was supported by the Strategic China-Japan Joint Research Program on ‘S&T for Environmental Conservation and Construction of a Society with Less Environmental Burden’ (NSFC Grant No. 51021140002).

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  1. Key Laboratory of Northwest Water Resource, Ecology and Environment, Ministry of Education, Xi’an University of Architecture and Technology, Xi’an, 710055, Shaanxi, China

    Zheng Ji, Xiaochang C. Wang, Limei Xu & Chongmiao Zhang

  2. Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, North 13, West 8, Kita-ku, Sapporo, Hokkaido, 060-8628, Japan

    Naoyuki Funamizu, Satoshi Okabe & Daisuke Sano

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  1. Zheng Ji
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  2. Xiaochang C. Wang
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Ji, Z., Wang, X.C., Xu, L. et al. Estimation of Contamination Sources of Human Enteroviruses in a Wastewater Treatment and Reclamation System by PCR–DGGE. Food Environ Virol 6, 99–109 (2014). https://doi.org/10.1007/s12560-014-9140-x

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