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Applied and Environmental Microbiology, December 2007, p. 7891-7897, Vol. 73, No. 24
0099-2240/07/$08.00+0     doi:10.1128/AEM.01428-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Removal of Noroviruses during Wastewater Treatment, Using Real-Time Reverse Transcription-PCR: Different Behaviors of Genogroups I and II{triangledown}

Allegra Kyria da Silva,1 Jean-Claude Le Saux,2 Sylvain Parnaudeau,2 Monique Pommepuy,2 Menachem Elimelech,1 and Françoise S. Le Guyader2*

Department of Chemical Engineering, Environmental Engineering Program, Yale University, P.O. Box 208286, New Haven, Connecticut 06520-8286,1 Laboratoire de Microbiologie, IFREMER, 44311 Nantes Cedex 03, France2

Received 27 June 2007/ Accepted 2 October 2007

Noroviruses, an important cause of gastroenteritis, are excreted by infected individuals and are therefore present in wastewater. We quantified norovirus genogroup I (GI) and GII in wastewater at different locations in France and evaluated removal by a range of treatment types, including basic (waste stabilization pond), current industry standard (activated sludge), and state-of-the-art (submerged membrane bioreactor) treatments. Noroviruses were quantified using real-time reverse transcription-PCR (rRT-PCR). Mengovirus was used as a virus extraction control, and internal controls were used to verify the level of GI and GII rRT-PCR inhibition. A total of 161 (81 influent and 79 effluent) samples were examined; GI and GII were detected in 43 and 88% of the influent samples, respectively, and in 24 and 14% of the effluent samples, respectively. Physicians in France report far more cases of GII than GI during outbreaks; thus, the frequent presence of GI was unexpected. The GI influent concentrations were more variable, the peak GI influent concentrations were higher than the peak GII influent concentrations at all four sites (up to 1 x 109 and 6 x 107 genome copies/liter, respectively), and the average positive influent concentrations of GI were higher than the average positive influent concentrations of GII. The maximum effluent breakthrough concentrations were 6 x 106 and 3 x 106 genome copies/liter for GI and GII, respectively, indicating that the four treatment systems studied decreased the norovirus contamination load in receiving waters.

* Corresponding author. Mailing address: Microbiology Laboratory. IFREMER BP 21105, 44311 Nantes cedex 03, France. Phone: (33) 2 40 37 40 52. Fax: (33) 2 40 37 40 73. E-mail: sleguyad{at}ifremer.fr

{triangledown} Published ahead of print on 12 October 2007.

Applied and Environmental Microbiology, December 2007, p. 7891-7897, Vol. 73, No. 24
0099-2240/07/$08.00+0     doi:10.1128/AEM.01428-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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