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Vierheilig J.,Vienna University of Technology | Farnleitner A.H.,Vienna University of Technology | Farnleitner A.H.,InterUniversity Cooperation Center Water and Health Water and Health | Kollanur D.,InterUniversity Cooperation Center Water and Health Water and Health | And 2 more authors.
Journal of Microbiological Methods | Year: 2012

Two frequently applied genetic Bacteroidetes markers for total fecal pollution (AllBac and BacUni) were found in high numbers in pristine soil samples of two alpine catchment areas casting doubt on their value as fecal indicators. This finding underlines the necessity to evaluate assays locally and against non-intestinal samples before application. © 2012 Elsevier B.V. Source

Bliem R.,Medical University of Vienna | Bliem R.,Armament and Defence Technology Agency | Schauer S.,Medical University of Vienna | Plicka H.,Armament and Defence Technology Agency | And 10 more authors.
Applied and Environmental Microbiology | Year: 2015

Vibrio cholerae is a severe human pathogen and a frequent member of aquatic ecosystems. Quantification of V. cholerae in environmental water samples is therefore fundamental for ecological studies and health risk assessment. Beside time-consuming cultivation techniques, quantitative PCR (qPCR) has the potential to provide reliable quantitative data and offers the opportunity to quantify multiple targets simultaneously. A novel triplex qPCR strategy was developed in order to simultaneously quantify toxigenic and nontoxigenic V. cholerae in environmental water samples. To obtain quality-controlled PCR results, an internal amplification control was included. The qPCR assay was specific, highly sensitive, and quantitative across the tested 5-log dynamic range down to a method detection limit of 5 copies per reaction. Repeatability and reproducibility were high for all three tested target genes. For environmental application, global DNA recovery (GR) rates were assessed for drinking water, river water, and water from different lakes. GR rates ranged from 1.6% to 76.4% and were dependent on the environmental background. Uncorrected and GR-corrected V. cholerae abundances were determined in two lakes with extremely high turbidity. Uncorrected abundances ranged from 4.6 × 102 to 2.3 × 104 cell equivalents liter-1, whereas GR-corrected abundances ranged from 4.7 × 103 to 1.6 × 106 cell equivalents liter-1. GR-corrected qPCR results were in good agreement with an independent cellbased direct detection method but were up to 1.6 log higher than cultivation-based abundances. We recommend the newly developed triplex qPCR strategy as a powerful tool to simultaneously quantify toxigenic and nontoxigenic V. cholerae in various aquatic environments for ecological studies as well as for risk assessment programs. © 2015, American Society for Microbiology. Source

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