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United States Department of Agriculture

Agricultural Research Service

Title: Simultaneous Concentration of Pathogenic Viruses, Bacteria and Protozoa from Water Using Sodocalcic Glass Wool Filters

Authors
item Marrero-Ortiz, R - MARSHFIELD CLINIC
item Bertz, P - MARSHFIELD CLINIC
item Spencer, S - MARSHFIELD CLINIC
item Volenec, M - MARSHFIELD CLINIC
item Jokela, William
item Borchardt, M - MARSHFIELD CLINIC

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: January 15, 2008
Publication Date: June 1, 2008
Citation: Marrero-Ortiz, R., Bertz, P.D., Spencer, S.K., Volenec, M.J., Jokela, W.E., Borchardt, M.A. 2008. Simultaneous Concentration of Pathogenic Viruses, Bacteria and Protozoa from Water Using Sodocalcic Glass Wool Filters. American Society for Microbiology General Meeting Abstracts.

Technical Abstract: Background: Methods for simultaneous concentration of multiple waterborne pathogens would be helpful for ensuring water quality and public health safety. The major technical issue of a reliable concentration method is the widely varied physical and chemical characteristics of pathogenic viruses, bacteria and protozoa, and the fact that these microorganisms occur at low concentrations in water. Previously it was demonstrated that sodocalcic glass wool filters can concentrate human enteric viruses from a large volumes of water. This study evaluated the ability of glass wool filters for simultaneous concentration of agricultural zoonotic pathogens expected to occur in runoff from fields with bovine manure applications, namely Cryptosporidium parvum, Salmonella typhimurium, Campylobacter jejuni, as well as bovine enteroviruses. In early tests, Sabin type III poliovirus was used as a surrogate for bovine enteroviruses. Methods: Ten liters dechlorinated tap water (pH = 7) were seeded with 6.5x103 C. parvum oocysts, 1x103 - 5x106 CFU S. typhimurium or 1.5x103 PFU poliovirus. The seeded poliovirus volumes included 5.8 g dry/mass agricultural soil to simulate runoff. Seeded water was passed through a glass wool filter using a peristaltic pump (flow rate = 1 liter min-1). Retained pathogens were eluted from the filters by passing beef extract-glycine buffer (pH 9.5) in the direction opposite of sample flow. C. parvum, S. typhimurium and enteroviruses were enumerated by immunofluorescent microscopy, culture on XLD media and quantitative RT-PCR, respectively. Results: The mean recovery efficiencies for C. parvum, S. typhimurium, and poliovirus were 40% (n=12), 91% (n=12), and 77% (n=4), respectively, with corresponding coefficients of variation of 33%, 34%, and 106%. Campylobacter jejuni seeded at 8x105 CFU into 10 liters water and enumerated by CVA media was present in the eluent, but could not be enumerated. Conclusion: Glass wool filtration appears to be a promising cost-effective method for concentrating several waterborne pathogens simultaneously, although recovery may be low for some specific genera such as Campylobacter.

Last Modified: 4/17/2014
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