title: Application of human and animal viral microbial source tracking tools in fresh and marine waters from five different geographical areas.
abstract: Integrated river basin management planning to mitigate the impacts of economic, demographic and climate change is an important issue for the future protection of water resources. Identifying sources of microbial contamination via the emerging science of Microbial Source Tracking (MST) plays a key role in risk assessment and the design of remediation strategies. Following an 18-month surveillance program within the EU-FP7-funded VIROCLIME project, specific MST tools were used to assess human markers such as adenoviruses (HAdV) and JC polyomaviruses (JCPyV) and porcine and bovine markers such as porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) via quantification with real-time PCR to analyze surface water collected from five sites within different climatic zones: the Negro River (Brazil), Glafkos River (Greece), Tisza River (Hungary), Llobregat River (Spain) and Umeälven River (Sweden). The utility of the viral MST tools and the prevalence and abundance of specific human and animal viruses in the five river catchments and adjacent seawater, which is impacted by riverine contributions from the upstream catchments, were examined. In areas where no sanitation systems have been implemented, sewage can directly enter surface waters, and river water exhibited high viral loads; HAdV and JCPyV could be detected at mean concentrations of 10(5) and 10(4) Genome Copies/Liter (GC/L), respectively. In general, river water samples upstream of urban discharges presented lower human viral loads than downstream sampling sites, and those differences appeared to increase with urban populations but decrease in response to high river flow, as the elevated river water volume dilutes microbial loads. During dry seasons, river water flow decreases dramatically, and secondary effluents can represent the bulk of the riverine discharge. We also observed that ice cover that formed over the river during the winter in the studied areas in North Europe could preserve viral stability due to the low temperatures and/or the lack of solar inactivation. Porcine and bovine markers were detected where intensive livestock and agricultural activities were present; mean concentration values of 10(3) GC/L indicated that farms were sometimes unexpected and important sources of fecal contamination in water. During spring and summer, when livestock is outdoors and river flows are low, animal pollution increases due to diffuse contamination and direct voiding of feces onto the catchment surface. The field studies described here demonstrate the dynamics of fecal contamination in all catchments studied, and the data obtained is currently being used to develop dissemination models of fecal contamination in water with respect to future climate change scenarios. The results concerning human and animal targets presented in this study demonstrate the specificity and applicability of the viral quantitative parameters developed to widely divergent geographical areas and their high interest as new indicators of human and animal fecal contamination in water and as MST tools.
authors.affiliation: Laboratory of Virus Contaminants of Water and Food, Department of Microbiology, University of Barcelona, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Rusiñol
authors.affiliation: Laboratory of Virus Contaminants of Water and Food, Department of Microbiology, University of Barcelona, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Fernandez-Cassi
authors.affiliation: Laboratory of Virus Contaminants of Water and Food, Department of Microbiology, University of Barcelona, Barcelona, Spain.
authors.forename: Ayalkibet
authors.initials: A
authors.lastname: Hundesa
authors.affiliation: Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Rio de Janeiro, Brazil.
authors.forename: Carmen
authors.initials: C
authors.lastname: Vieira
authors.affiliation: Department of Water Microbiology, National Institute for Environmental Health, Budapest, Hungary.
authors.forename: Anita
authors.initials: A
authors.lastname: Kern
authors.affiliation: Department of Virology, Umeå University, Umeå, Sweden.
authors.forename: Irene
authors.initials: I
authors.lastname: Eriksson
authors.affiliation: Environmental Microbiology Unit, Department of Public Health, Medical School, University of Patras, Patras, Greece.
authors.forename: Panos
authors.initials: P
authors.lastname: Ziros
authors.affiliation: Institute of Geography and Earth Sciences (IGES), Aberystwyth University, Durham City, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Kay
authors.affiliation: Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Rio de Janeiro, Brazil.
authors.forename: Marize
authors.initials: M
authors.lastname: Miagostovich
authors.affiliation: Department of Water Microbiology, National Institute for Environmental Health, Budapest, Hungary.
authors.forename: Marta
authors.initials: M
authors.lastname: Vargha
authors.affiliation: Department of Virology, Umeå University, Umeå, Sweden.
authors.forename: Annika
authors.initials: A
authors.lastname: Allard
authors.affiliation: Environmental Microbiology Unit, Department of Public Health, Medical School, University of Patras, Patras, Greece.
authors.forename: Apostolos
authors.initials: A
authors.lastname: Vantarakis
authors.affiliation: Institute of Geography and Earth Sciences (IGES), Aberystwyth University, Durham City, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Wyn-Jones
authors.affiliation: Laboratory of Virus Contaminants of Water and Food, Department of Microbiology, University of Barcelona, Barcelona, Spain.
authors.forename: Sílvia
authors.initials: S
authors.lastname: Bofill-Mas
authors.affiliation: Laboratory of Virus Contaminants of Water and Food, Department of Microbiology, University of Barcelona, Barcelona, Spain. Electronic address: rgirones@ub.edu.
authors.forename: Rosina
authors.initials: R
authors.lastname: Girones
mesh_terms: D000818:Animals; D001938:Brazil; D004784:Environmental Monitoring / Q000379:methods*; D005060:Europe; D005618:Fresh Water / Q000821:virology*; D006801:Humans; D012623:Seawater / Q000821:virology*; D014773:Virology / Q000379:methods*; D014873:Water Pollutants
chemical_list: D014873:Water Pollutants
keywords: Adenovirus; Microbial Source Tracking (MST); Polyomavirus; River water; Seawater