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Food and Environmental Virology

, Volume 11, Issue 1, pp 32–39 | Cite as

Comparative Assessment of BGM and PLC/PRF/5 Cell Lines for Enteric Virus Detection in Biosolids

  • Sherif Abd-Elmaksoud
  • Nohelia Castro-del Campo
  • Charles P. Gerba
  • Ian L. Pepper
  • Kelly R. BrightEmail author
Original Paper
  • 27 Downloads

Abstract

The buffalo green monkey (BGM) cell line is required for the detection of enteric viruses in biosolids through a total culturable viral assay (TCVA) by the United States Environmental Protection Agency. In the present study, BGM and PLC/PRF/5 cell lines were evaluated for TCVA and for their use in determining the incidence of adenoviruses and enteroviruses in raw sludge and Class B biosolids. Six raw sludge and 17 Class B biosolid samples were collected from 13 wastewater treatment plants from seven U.S. states. Samples were processed via organic flocculation and concentrate volumes equivalent to 4 g total solids were assayed on BGM and PLC/PRF/5 cells. Cell monolayers were observed for cytopathic effect (CPE) after two 14-days passages. Cell lysates were tested for the presence of adenoviruses and enteroviruses by PCR or RT-PCR. The PLC/PRF/5 cells detected more culturable viruses than the BGM cells by CPE (73.9% vs. 56.5%, respectively). 52% of the samples were positive for CPE using both cell lines. No viruses were detected in either cell line by PCR in flasks in which CPE was not observed. No adenoviruses were detected in 13 CPE-positive samples from BGM lysates. In contrast, of the 17 samples exhibiting CPE on PLC/PRF/5 cells, 14 were positive for adenoviruses (82.4%). In conclusion, PLC/PRF/5 cells were superior for the detection of adenoviruses in both raw sludge and Class B biosolids. Thus, the use of BGM cells alone for TCVA may underestimate the viral concentration in sludge/biosolid samples.

Keywords

Biosolids ICC-PCR Enteric viruses Adenovirus BGM cell line PLC cell line 

Notes

Acknowledgements

The authors would like to thank Dr. Masaaki Kitajima who assisted with the PCR and RT-PCR protocols used during the study and helped with troubleshooting and optimization of the methods.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Sherif Abd-Elmaksoud
    • 1
    • 2
  • Nohelia Castro-del Campo
    • 1
    • 3
  • Charles P. Gerba
    • 1
  • Ian L. Pepper
    • 1
  • Kelly R. Bright
    • 1
    Email author
  1. 1.Department of Soil, Water and Environmental Science, Water & Energy Sustainable Technology (WEST) CenterThe University of ArizonaTucsonUSA
  2. 2.Environmental Virology Laboratory, Department of Water Pollution ResearchNational Research CentreCairoEgypt
  3. 3.Centro de Investigación en Alimentación y Desarrollo A.C. (Research Center for Food and Development A.C.)CuliacánMexico

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