Quantitative Microbial Source Tracking (QMST)

The increasing demand for the ability to precisely describe the nature and extent of faecal contamination of water resources led to the recent development of an innovative quantitative microbial source tracking (QMST) approach by our research group. The so called QMST BacH/R method represents the first approach for the sensitive and specific DNA-based quantification and discrimination of human versus ruminant faecal contamination in spring water and respective drinking water resources. The QMST BacH/R consists of quantitative TaqMan minor-groove binder real-time PCR assays, quantifying host specific genetic markers in highly abundant faecal members of the bacterial phylum Bacteroidetes. The high sensitivity and specificity offered by the approach promises unique perspectives in the field of water safety evaluation and monitoring. However, basic research and method development have focused on Eastern Austria and the spring environments of the Northern Calcareous Alps, a geographically rather confined area. On the road to an attractive analytical QMST approach with market potential, it is essential that the practical applicability under routine laboratory conditions as well as a broad geographical applicability can be guaranteed. Therefore the aim of the proposed project is the extension of the QMST BacH/R approach in two dimensions: the extension towards routine applicability (modules A1, A2) and the extension towards the wider geographical applicability (modules B1, B2). A defined target number cell standards (DeTaCS), carrying target marker sequences in a controlled single copy, will be developed to make results from QMST comparable (module A1). Compatibility to ISO methods for faecal indicator enumeration will be subsequently evaluated for the standardised approach. Various aquatic resources under human and/or ruminant faecal influence, but strongly differing in environmental characteristics, will be investigated throughout Eastern Austria, the area where specificity of the approach has already been proven (module A2). The applicability of the QMST BacH/R approach will be extended to the Central European Region (CE-region) by testing the specificity and sensitivity on selected faecal sources from this area. Potentially present new and abundant sequence targets will be incorporated into the QMST BacH/R approach (module B1). Finally, the applicability of the extended and standardised QMST BacH/R approach will be demonstrated in the CE-region under field test conditions during the Joint Danube Survey 2007, organised by the International Commission for the Protection of the Danube River (module B2). The realisation of the proposed improved and standardised QMST BacH/R kit prototype will offer a new tool for the field of health related water microbiology. It will probably be the first step towards a tool box of commercially available QMST products for water safety management on global scale.