title: Comparison of ZetaPlus 60S and nitrocellulose membrane filters for the simultaneous concentration of F-RNA coliphages, porcine teschovirus and porcine adenovirus from river water.
abstract: Increasing attention is being paid to the impact of agricultural activities on water quality to understand the impact on public health. F-RNA coliphages have been proposed as viral indicators of fecal contamination while porcine teschovirus (PTV) and porcine adenovirus (PAdV) are proposed indicators of fecal contamination of swine origin. Viruses and coliphages are present in water in very low concentrations and must be concentrated to permit their detection. There is little information comparing the effectiveness of the methods for concentrating F-RNA coliphages with concentration methods for other viruses and vice versa. The objective of this study was to compare 5 current published methods for recovering F-RNA coliphages, PTV and PAdV from river water samples concentrated by electronegative nitrocellulose membrane filters (methods A and B) or electropositive Zeta Plus 60S filters (methods C-E). Method A is used routinely for the detection of coliphages (Méndez et al., 2004) and method C (Brassard et al., 2005) is the official method in Health Canada's compendium for the detection of viruses in bottled mineral or spring water. When river water was inoculated with stocks of F-RNA MS2, PAdV, and PTV to final concentrations of 1×10(6) PFU/100 mL, 1×10(5) gc/100 mL and 3×10(5) gc/100 mL, respectively, a significantly higher recovery for each virus was consistently obtained for method A with recoveries of 52% for MS2, 95% for PAdV, and 1.5% for PTV. When method A was compared with method C for the detection of F-coliphages, PAdV and PTV in river water samples, viruses were detected with higher frequencies and at higher mean numbers with method A than with method C. With method A, F-coliphages were detected in 11/12 samples (5-154 PFU/100 mL), PTV in 12/12 samples (397-10,951 gc/100 mL), PAdV in 1/12 samples (15 gc/100 mL), and F-RNA GIII in 1/12 samples (750 gc/100 mL) while F-RNA genotypes I, II, and IV were not detected by qRT-PCR.
authors.affiliation: Agriculture and Agri-Food Canada, Lacombe Research Centre, 6000 C & E Trail, Lacombe, Alberta, Canada T4L 1W1. Electronic address: Tineke.Jones@agr.gc.ca.
authors.forename: T H
authors.initials: TH
authors.lastname: Jones
authors.affiliation: Agriculture and Agri-Food Canada, Lacombe Research Centre, 6000 C & E Trail, Lacombe, Alberta, Canada T4L 1W1.
authors.forename: V
authors.initials: V
authors.lastname: Muehlhauser
authors.affiliation: Agriculture and Agri-Food Canada, Soils and Crops Research and Development Centre, 2560 Hochelaga Boulevard, Quebec, Quebec, Canada G1V 2J3.
authors.forename: G
authors.initials: G
authors.lastname: Thériault
mesh_terms: D025662:Adenoviruses, Porcine / Q000235:genetics / Q000302:isolation & purification*; D002170:Canada; D005374:Filtration / Q000379:methods; D017909:Levivirus / Q000235:genetics / Q000302:isolation & purification*; D045483:Rivers / Q000821:virology*; D012680:Sensitivity and Specificity; D052700:Teschovirus / Q000235:genetics / Q000302:isolation & purification*; D053585:Virus Attachment; D014876:Water Pollution; D060753:Water Quality
keywords: F-RNA coliphage; Nitrocellulose membrane filter; Porcine adenovirus; Porcine teschovirus; River water