title: EMA-qPCR to monitor the efficiency of a closed-coupled solar pasteurization system in reducing Legionella contamination of roof-harvested rainwater.
abstract: Solar pasteurization is effective in reducing the level of indicator organisms in stored rainwater to within drinking water standards. However, Legionella spp. were detected at temperatures exceeding the recommended pasteurization temperatures using polymerase chain reaction assays. The aim of the current study was thus to apply EMA quantitative polymerase chain reaction (EMA-qPCR) to determine whether the Legionella spp. detected were intact cells and therefore possibly viable at pasteurization temperatures >70°C. The BacTiter-Glo™ Microbial Cell Viability Assay was also used to detect the presence of ATP in the tested samples, as ATP indicates the presence of metabolically active cells. Chemical analysis also indicated that all anions and cations were within the respective drinking water guidelines, with the exception of iron (mean: 186.76 μg/L) and aluminium (mean: 188.13 μg/L), which were detected in the pasteurized tank water samples at levels exceeding recommended guidelines. The BacTiter-Glo™ Microbial Cell Viability Assay indicated the presence of viable cells for all pasteurized temperatures tested, with the percentage of ATP (in the form of relative light units) decreasing with increasing temperature [70-79°C (96.7%); 80- 89°C (99.2%); 90-95°C (99.7%)]. EMA-qPCR then indicated that while solar pasteurization significantly reduced (p<0.05) the genomic copy numbers of intact Legionella cells in the pasteurized tank water (~99%), no significant difference (p>0.05) in the mean copy numbers was detected with an increase in the pasteurization temperature, with 6 × 10(3) genomic copies/mL DNA sample obtained at 95°C. As intact Legionella cells were detected in the pasteurized tank water samples, quantitative microbial risk assessment studies need to be conducted to determine the potential health risk associated with using the water for domestic purposes.
authors.affiliation: Department of Microbiology, Faculty of Science, University of Stellenbosch, Private Bag X1, Stellenbosch 7602, South Africa.
authors.forename: B
authors.initials: B
authors.lastname: Reyneke
authors.affiliation: Department of Microbiology, Faculty of Science, University of Stellenbosch, Private Bag X1, Stellenbosch 7602, South Africa.
authors.forename: P H
authors.initials: PH
authors.lastname: Dobrowsky
authors.affiliation: Department of Microbiology, Faculty of Science, University of Stellenbosch, Private Bag X1, Stellenbosch 7602, South Africa.
authors.forename: T
authors.initials: T
authors.lastname: Ndlovu
authors.affiliation: Department of Biomedical Sciences, Faculty of Health and Wellness Sciences, Cape Peninsula University of Technology, P.O. Box 1906, Bellville 7535, South Africa.
authors.forename: S
authors.initials: S
authors.lastname: Khan
authors.affiliation: Department of Microbiology, Faculty of Science, University of Stellenbosch, Private Bag X1, Stellenbosch 7602, South Africa. Electronic address: wesaal@sun.ac.za.
authors.forename: W
authors.initials: W
authors.lastname: Khan
mesh_terms: D007875:Legionella / Q000254:growth & development*; D050296:Microbial Viability; D061352:Pasteurization / Q000379:methods*; D011891:Rain; D013472:Sunlight; D013696:Temperature; D014871:Water Microbiology; D018508:Water Purification / Q000379:methods*
keywords: EMA-qPCR; Legionella; Rainwater harvesting; Solar pasteurization