Evaluation of methodology for detection of human adenoviruses in wastewater, drinking water, stream water and recreational waters.
OBJECTIVE
This study evaluates dialysis filtration and a range of PCR detection methods for identification and quantification of human adenoviruses in a range of environmental waters.


RESULTS
Adenovirus was concentrated from large volumes (50-200 l) of environmental and potable water by hollow fibre microfiltration using commercial dialysis filters. By this method, an acceptable recovery of a seeded control bacteriophage MS2 from seawater (median 95.5%, range 36-98%, n=5), stream water (median 84.7%, range 23-94%, n=5) and storm water (median 59.5%, range 6.3-112%, n=5) was achieved. Adenovirus detection using integrated cell culture PCR (ICC-PCR), direct PCR, nested PCR, real-time quantitative PCR (qPCR) and adenovirus group F-specific direct PCR was tested with PCR products sequenced for confirmation. Adenovirus was routinely detected from all water types by most methods, with ICC-PCR more sensitive than direct-nested PCR or qPCR. Group F adenovirus dominated in wastewater samples but was detected very infrequently in environmental waters.


CONCLUSIONS
Human adenoviruses (HAdv) proved relatively common in environmental and potable waters when assessed using an efficient concentration method and sensitive detection method. ICC-PCR proved most sensitive, could be used semiquantitatively and demonstrated virus infectivity but was time consuming and expensive. qPCR provided quantitative results but was c. ten-fold less sensitive than the best methods.
Dong
Y
Y
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Kim
J
J
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Lewis
G D
GD
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
D000260:Adenoviruses, Human / Q000302:isolation & purification*; D001493:Bathing Beaches; D002460:Cell Line; D004279:DNA, Viral / Q000302:isolation & purification; D004784:Environmental Monitoring / Q000379:methods*; D005374:Filtration / Q000379:methods; D006801:Humans; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods; D011998:Recreation; D045483:Rivers / Q000821:virology*; D012623:Seawater / Q000821:virology; D012680:Sensitivity and Specificity; D014865:Waste Disposal, Fluid; D014871:Water Microbiology; D014873:Water Pollutants / Q000302:isolation & purification; D014876:Water Pollution / Q000032:analysis; D014881:Water Supply
D004279:DNA, Viral; D014873:Water Pollutants
APHA, (2005) Standard Methods for the Examination of Water and Wastewater, 21st edn, ed. Eaton, A.D., Clesceri, L.S. and Rice, E.W. Washington, DC: American Public Health Association.
Bosch, A., Lucena, F., Diez, J.M., Gajardo, R., Blasi, M. and Jofre, J. (1991) Human enteric viruses and indicator microorganisms in a water supply associated with an outbreak of infectious hepatitis. J Am Water Works Assoc 83, 80-83.
Castignolles, N., Petit, F., Mendel, I., Simon, L., Cattolico, L. and Buffet-Janvresse, C. (1998) Detection of adenovirus in the waters of the Seine River estuary by nested-PCR. Mol Cell Probes 12, 175-180.
Choi, S. and Jiang, S.C. (2005) Real-time PCR quantification of human adenoviruses in urban rivers indicates genome prevalence but low infectivity. Appl Environ Microbiol 71, 7426-7433.
Elliott, M.A., Stauber, C.E., Koksal, F., DiGiano, F.A. and Sobsey, M.D. (2008) Reductions of E. coli, echovirus type 12 and bacteriophages in an intermittently operated household-scale slow sand filter. Water Res 42, 2662-2670.
Genthe, B., Gericke, M., Bateman, B., Mjoli, N. and Kfir, R. (1995) Detection of enteric adenoviruses in South African waters using gene probes. Water Sci Technol 23, 2091-2098.
Gerba, C.P. (1984) Applied and theoretical aspects of virus adsorption to surfaces. Adv Appl Microbiol 30, 133-168.
Greening, G., Hewitt, J. and Lewis, G. (2002) Evaluation of integrated cell culture-PCR (C-PCR) for virological analysis of environmental samples. J Appl Microbiol 93, 745-750.
Haramoto, E., Katayama, H., Oguma, K. and Ohgaki, S. (2007) Quantitative analysis of human enteric adenoviruses in aquatic environments. J Appl Microbiol 103, 2153-2159.
Havelaar, A., Van Olphen, M. and Dorst, Y. (1993) F-Specific RNA phage are adequate model organisms for enteric viruses in freshwater. Appl Environ Microbiol 59, 2956-2962.
Heim, A., Ebnet, C., Harste, G. and Pring-Akerblom, P. (2003) Rapid and quantitative detection of human adenovirus DNA by real-time PCR. J Med Virol 70, 228-239.
Horwitz, M.S. (2001) Adenoviruses. In Field’s Virology ed. Knipe, D.M. and Howley, P.M. pp. 2327-2359. Philadelphia, PA: Lippincott Williams & Wilkins.
Huang, M.L., Nguy, L., Ferrenberg, J., Boeckh, M., Cent, A. and Corey, L. (2008) Development of multiplexed real-time quantitative polymerase chain reaction assay for detecting human adenoviruses. Diagn Microbiol Infect Dis 62, 263-271.
Jiang, S., Noble, R. and Chu, W. (2001) Human adenoviruses and coliphages in urban runoff-impacted coastal waters of Southern California. Appl Environ Microbiol 67, 179-184.
Juliano, N. and Sobsey, M.D. (1997) Simultaneous concentration of Cryptosporidium bacteria and viruses from water by hollow fiber ultrafiltration. Water Quality Technology Conference. Denver, Colorado: American Water Works Association.
Keswick, B.H., Gerba, C.P., DuPont, H.L. and Rose, J.B. (1984) Detection of enteric viruses in treated drinking water. Appl Environ Microbiol 47, 1290-1294.
Ko, G., Jothikumar, N., Hill, V.R. and Sobsey, M.D. (2005) Rapid detection of infectious adenoviruses by mRNA real-time RT-PCR. J Virol Methods 127, 148-153.
Krikelis, V., Markoulatos, P., Spyron, N. and Serie, C. (1985) Detection of indigenous enteric viruses in raw sewage effluents of the city of Athens, Greece, during a two-year survey. Water Sci Technol 17, 159-164.
Labelle, R.L. and Gerba, C.P. (1980) Influence of Estuarine sediments on virus survival under field conditions. Appl Environ Microbiol 39, 749-755.
Lambertini, E., Spencer, S.K., Bertz, P.D., Loge, F.J., Kieke, B.A. and Borchardt, M.A. (2008) Concentration of enteroviruses, adenoviruses, and noroviruses from drinking water by use of glass wool filters. Appl Environ Microbiol 74, 2990-2996.
Langlet, J., Gaboriaud, F., Duval, J. and Gantzer, C. (2008) Aggregation and surface properties of F-specific RNA phages: implication for membrane filtration processes. Water Res 42, 2769-2777.
Lee, S.H. and Kim, S.J. (2002) Detection of infectious enteroviruses and adenoviruses in tap water in urban areas in Korea. Water Res 36, 248-256.
Lee, S.H., Lee, C., Lee, K.W., Cho, H.B. and Kim, S.J. (2005) The simultaneous detection of both enteroviruses and adenoviruses in environmental water samples including tap water with an integrated cell culture-multiplex-nested PCR procedure. Appl Microbiol 98, 1020-1029.
Lewis, G.D. and Metcalf, T.G. (1988) Polyethylene Glycol precipitation for recovery of pathogenic viruses including hepatitis a virus and human rotavirus from oyster water and sediment samples. Appl Environ Microbiol 54, 1983-1988.
Lin, K., Lin, Y., Chen, H., Ke, G., Chiang, C., Hwang, K., Chu, P., Lin, J. et al. (2004) A two decade survey of respiratory adenovirus in Taiwan: the reemergence of adenovirus types 7 and 4. J Med Virol 73, 274-279.
Lund, E. (1982) Waterborne Virus Disease. Ecol Dis 1, 27-35.
Mei, Y.F., Lindman, K. and Wadell, G. (2002) Human adenoviruses of subgenera B, C, and E with various tropisms differ in both binding to, and replication in, the epithelial A549 and 293 cells. Virol 295, 30-43.
Metcalf, T.G., Melnick, J.L. and Estes, M.K. (1995) Environmental virology: from detection of virus in sewage and water by isolation to identification by molecular biology - a trip of 50 years. Annu Rev Microbiol 49, 461-487.
Muniain-Mujika, J., Girones, R. and Lucena, F. (2000) Viral contamination of shellfish: evaluation of methods and analysis of bacteriophages and human viruses. J Virol Methods 89, 109-118.
Muscillo, M., Pourshaban, M., Iaconelli, M., Fontana, S., Di Grazia, A., Manzara, S., Fadda, G., Santangelo, R. et al. (2008) Detection and quantification of human adenoviruses in surface waters by nested PCR, TaqMan real-time PCR and cell culture assays. Water Air Soil Pollut 191, 1-11.
Papapetropoulou, M. and Vantarakis, A.C. (1998) Detection of adenovirus outbreak at a municipal swimming pool by nested PCR amplification. J Infect 36, 101-103.
Payment, P., Trudel, M. and Plante, R. (1985) Elimination of viruses and indicator bacteria at each step of treatment during preparation of drinking water at seven water treatment plants. Appl Environ Microbiol 99, 1418-1428.
Pina, S., Puig, M., Lucena, F., Jofre, J. and Girones, R. (1998) Viral pollution in the environment and in shellfish: human Adenovirus detection by PCR as an index of human viruses. Appl Environ Microbiol 64, 3376-3382.
Pinto, R.M., Gajardo, R., Abad, F.X. and Bosch, A. (1995) Detection of fastidious infectious enteric viruses in water. Environ Sci Technol 29, 2636-2638.
Puig, M., Jofre, J., Lucena, F., Allard, A., Gorgan, W. and Girones, R. (1994) Detection of adenoviruses and enteroviruses in polluted waters by nested PCR amplification. Appl Env Microbiol 60, 2963-2970.
Reynolds, K.A. (2004) Integrated cell culture/PCR for detection of enteric viruses in environmental samples. Methods Mol Biol 268, 69-78.
Rigotto, C., Sincero, T.C.M., Simoes, C.M.O. and Barardi, C.R.M. (2005) Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR). Water Res 39, 297-304.
Sobsey, M.D., Yates, M.V., Hsu, F.-C., Lovelace, G., Battigelli, D., Margolin, A., Pillai, S.D. and Nwachuku, N. (2004) Development and evaluation of methods to detect coliphages in large volumes of waterWater Sci Technol 50, 211-217.
Uhnoo, I., Svensson, L. and Wadell, G. (1990) Enteric adenoviruses. In Balliere’s Clinical Gastroenterology ed. Farthing, M.J.G. pp. 627-642. London: Balliere Tindall.
Van Heerden, J., Ehlers, M.M., Heim, A. and Grabow, W.O.K. (2005) Prevalence, quantification and typing of adenoviruses detected in river and treated drinking water in South Africa. J Appl Microbiol 99, 234-242.
WHO. (2006) Microbial aspects. In: Guidelines for Drinking-Water Quality, 1st ad., 3rded. pp. 121-144. Geneva: World Health Organization.
Williams, F.P. Jr and Hurst, C.J. (1988) Detection of environmental viruses in sludge: enhancement of enterovirus plaque assay titers with 5-IODO-2′-Deoxyuridine and comparison to adenovirus and coliphage titers. Water Res 22, 847-851.
Xu, W., McDonough, M.C. and Erdman, D.D. (2000) Species-specific identification of human adenoviruses by a multiplex PCR assay. J Clin Microbiol 38, 4114-4120.