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Applied and Environmental Microbiology, November 1998, p. 4307-4312, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Detection of Infectious Enteroviruses, Enterovirus Genomes, Somatic Coliphages, and Bacteroides fragilis Phages in Treated Wastewater

C. Gantzer,1 A. Maul,2 J. M. Audic,3 and L. Schwartzbrod1,*

Faculté de Pharmacie, Laboratoire de Virologie, 54000 Nancy,1 I.U.T. de Metz, 57000 Metz,2 and Lyonnaise des Eaux, CIRSEE, 78230 Le Pecq,3 France

Received 13 January 1998/Accepted 13 July 1998

In this study, three types of treated wastewater were tested for infectious enteroviruses, the enterovirus genome, somatic coliphages, and Bacteroides fragilis phages. The aim of this work was to determine whether the presence of the two types of bacteriophages or of the enterovirus genome was a good indicator of infectious enterovirus contamination. The enterovirus genome was detected by reverse transcription-polymerase chain reaction. Infectious enteroviruses were quantified by cell culturing (BGM cells), and the bacteriophages were quantified by plaque formation on the host bacterium (Escherichia coli or B. fragilis) in agar medium. Forty-eight samples of treated wastewater were analyzed. Sixteen samples had been subjected to a secondary treatment for 8 to 12 h (A), 16 had been subjected to a secondary treatment for 30 h (B1), and 16 had been subjected to both secondary and tertiary treatments (B2). The mean concentrations of somatic coliphages were 4.9 × 104 PFU · liter-1 for treatment line A, 9.8 × 103 PFU · liter-1 for B1, and 1.4 × 103 PFU · liter-1 for B2, with all the samples testing positive (100%). The mean concentrations of B. fragilis phages were 1.7 × 103 PFU · liter-1 for A (100% positive samples), 17 to 24 PFU · liter-1 for B1 (44% positive samples), and 0.8 to 13 PFU · liter-1 for B2 (6% positive samples). The mean concentrations of infectious enteroviruses were 4 most probable number of cytopathogenic units (MPNCU) · liter-1 for A (31% positive samples) and <1 MPNCU · liter-1 for B1 and B2 (0% positive samples). The percentages of samples testing positive for the enterovirus genome were 100% for A, 56% for B1, and 19% for B2. The percentages of samples testing positive for the enterovirus genome were significantly higher than those for infectious enteroviruses. This finding may have been due to the presence of noninfectious enteroviruses or to the presence of infectious enteroviruses that do not multiply in BGM cell cultures. However, under our experimental conditions, nondetection of the genome implies the absence of infectious viruses. There was a significant correlation between the concentration of somatic coliphages or B. fragilis phages and the presence of infectious enteroviruses or the presence of the enterovirus genome. However, the somatic coliphage concentration did not lead to fluctuations in the infectious enterovirus concentration, whereas the B. fragilis phage concentration did.

* Corresponding author. Mailing address: Faculté de Pharmacie, Laboratoire de Virologie, 5 rue Albert Lebrun, 54000 Nancy, France. Phone: 33.3.83.17.88.25. Fax: 33.3.83.17.88.79. E-mail: lschwart{at}pharma.u-nancy.fr.

Applied and Environmental Microbiology, November 1998, p. 4307-4312, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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