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Isolation and Identification of Enteroviruses from Sewage and Sewage-Contaminated Water in Lagos, Nigeria

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Abstract

Studies have confirmed silent circulation of enteroviruses in the environment even in the absence of associated clinical conditions in the community. In this light, 26 samples of sewage and sewage-contaminated water serving selected high-risk communities in Lagos Nigeria were examined between June and September 2010. To concentrate virus particles in the sample, 480 μL of each sample was centrifuged at 3,000 rpm for 1 h at 4 °C. Subsequently, pellets were pooled, chloroform treated and further centrifuged at 1,500 rpm for 20 min at 4 °C. The water phase (concentrate) was then collected and stored at −20 °C. The concentrates were subsequently inoculated into RD and L20B cell lines. Recovered isolates were identified by real-time RT-PCR (rRT-PCR), serotyping, VP1 amplification, sequencing and phylogenetic analysis. Overall, 9 (34.6 %) of the samples showed characteristic enterovirus cytopathic effect in RD cell line and were subsequently confirmed by pan-enterovirus rRT-PCR. The isolates were further identified by serotyping to include three E7, one E11 and one E13 isolates whilst four isolates were untypable. Further characterisation by VP1 sequencing confirmed the results of serotyping and rRT-PCR for all but isolate E13. Also, the four previously untypable isolates were identified to include two E19, one E20 and one E7 by VP1 sequencing. Results of the study confirmed circulation of Sub-Saharan Africa-specific enterovirus clades in the region, provide information on their molecular epidemiology and emphasise the need to combine methods of identification to enhance enterovirus surveillance.

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Acknowledgments

We thank Dr. Adewumi MO for thoughtful critique of our manuscript, the entire staff of the WHO National Polio Laboratory in Ibadan Nigeria for their valuable technical assistance and RIVM, The Netherlands, for the enterovirus antisera panel. FTOC is a PhD student studying the evolutionary dynamics of enteric viruses in Nigeria. This study is a part of his PhD work.

Conflict of interest

The authors declare no conflict of interests.

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Authors and Affiliations

  1. Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria

    Johnson Adekunle Adeniji & Temitope Oluwasegun Cephas Faleye

  2. WHO National Polio Laboratory, University of Ibadan, Ibadan, Oyo State, Nigeria

    Johnson Adekunle Adeniji

  3. Department of Microbiology, Faculty of Science, Ekiti State University, Ado Ekiti, Ekiti State, Nigeria

    Temitope Oluwasegun Cephas Faleye

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  1. Johnson Adekunle Adeniji
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Correspondence to Temitope Oluwasegun Cephas Faleye.

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Adeniji, J.A., Faleye, T.O.C. Isolation and Identification of Enteroviruses from Sewage and Sewage-Contaminated Water in Lagos, Nigeria. Food Environ Virol 6, 75–86 (2014). https://doi.org/10.1007/s12560-014-9137-5

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  • DOI: https://doi.org/10.1007/s12560-014-9137-5

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