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Persistence of Viable Bacteria in Solar Pasteurised Harvested Rainwater

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Abstract

The aim of the current study was to provide insight into the viable bacterial community that persists in rainwater after solar pasteurisation (SOPAS). Unpasteurised rainwater (n = 8) was collected directly from the rainwater tank, while SOPAS-treated rainwater (n = 8) was collected from the Phungamanzi™ system (connected to the tank) at the temperature ranges of 70 to 79 °C, 80 to 89 °C, and 90 °C and above. Culturable bacteria were isolated and identified using universal 16S ribosmonal ribonucleic acid (rRNA) PCR. Results indicated that Klebsiella spp., Pseudomonas spp., Enterococcus spp. and Bacillus spp. were present in untreated rainwater and sporadically persisted in the SOPAS samples collected at the different temperature ranges. Pseudomonas spp. and Bacillus spp. were the only genera found to persist up to a temperature of 98 °C. Genus-specific polymerase chain reaction (PCR) of total genomic deoxyribonucleic acid (DNA) extracted from the rainwater samples indicated that the most frequently detected genera present after SOPAS were Klebsiella (87.5%) and Pseudomonas (62.5%). The quantification of viable Klebsiella was performed using ethidium monoazide bromide quantitative PCR (EMA-qPCR), which revealed that intact Klebsiella cells persisted up to 98 °C with 9.3 × 104 gene copies mL−1 recorded. While intact Pseudomonas and Legionella cells have been detected in SOPAS-treated rainwater (> 90 °C), this study confirmed that Klebsiella and Bacillus are also viable at temperatures above 90 °C and may potentially result in health risks associated with the utilisation of SOPAS rainwater. Secondary or combination treatment procedures are thus required to effectively reduce the level of these persisting viable opportunistic pathogens.

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References

  • Ahmed, W., Huygens, F., Goonetilleke, A., & Gardner, T. (2008). Real-time PCR detection of pathogenic microorganisms in roof-harvested rainwater in Southeast Queensland, Australia. Applied and Environmental Microbiology, 74, 5490–5496.

    Article  CAS  Google Scholar 

  • Ahmed, W., Goonetilleke, A., & Gardner, T. (2010). Implications of faecal indicator bacteria for the microbiological assessment of roof-harvested rainwater quality in Southeast Queensland, Australia. Canadian Journal of Microbiology, 56(6), 471–479.

    Article  CAS  Google Scholar 

  • Ahmed, W., Hodgers, L., Sidhu, J. P. S., & Toze, S. (2011). Faecal indicators and zoonotic pathogens in household drinking taps fed from rainwater tanks in Southeast Queensland, Australia. Applied and Environmental Microbiology, 78, 219–226.

    Article  CAS  Google Scholar 

  • Ahmed, W., Brandes, H., Gyawali, P., Sidhu, J., & Toze, S. (2014a). Opportunistic pathogens in roof-captured rainwater samples, determined using quantitative PCR. Water Research, 53, 361–369.

    Article  CAS  Google Scholar 

  • Ahmed, W., Richardson, K., Sidhu, J. P. S., Jagals, P., & Toze, S. (2014b). Inactivation of faecal indicator bacteria in a roof-captured rainwater system under ambient meteorological conditions. Journal of Applied Microbiology, 116(1), 199–207.

    Article  CAS  Google Scholar 

  • Allan, B. R., Linseman, M. I., MacDonald, L. A., Lam, J. S., & Kropinski, A. M. (1988). Heat shock response of Pseudomonas aeruginosa. Journal of Bacteriology, 170(8), 3668–3674.

    Article  CAS  Google Scholar 

  • Bédard, E., Prévost, M., & Déziel, E. (2016). Pseudomonas aeruginosa in premise plumbing of large buildings. Microbiology Open, 5(6), 937–956.

    Article  Google Scholar 

  • Bojer, M. S., Struve, C., Ingmer, H., Hansen, D. S., & Krogfelt, K. A. (2010). Heat resistance mediated by a new plasmid encoded Clp ATPase, ClpK, as a possible novel mechanism for nosocomial persistence of Klebsiella pneumoniae. PLoS One, 5(11), e15467.

    Article  CAS  Google Scholar 

  • Bojer, M. S., Krogfelt, K. A., & Struve, C. (2011). The newly discovered ClpK protein strongly promotes survival of Klebsiella pneumoniae biofilm subjected to heat shock. Journal of Medical Microbiology, 60(10), 559–1561.

    Article  Google Scholar 

  • Brisse, S., & Verhoef, J. (2001). Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping. International Journal of Systematic and Evolutionary Microbiology, 51, 915–924.

    Article  CAS  Google Scholar 

  • Chun, J., & Bae, K. S. (2000). Phylogenetic analysis of Bacillus subtilis and related taxa based on partial gyrA sequences. Antonie Van Leeuwenhoek, 78, 123–127.

    Article  CAS  Google Scholar 

  • Delgado-Viscogliosi, P., Solignac, L., & Delattre, J. M. (2009). Viability PCR, a culture-independent method for rapid and selective quantification of viable Legionella pneumophila cells in environmental water samples. Applied and Environmental Microbiology, 75, 3502–3512.

    Article  CAS  Google Scholar 

  • Dineen, S. M., Aranda, R. T., Anders, D. L., & Robertson, J. M. (2010). An evaluation of commercial DNA extraction kits for the isolation of bacterial spore DNA from soil. Journal of Applied Microbiology, 109(6), 1886–1896.

    Article  CAS  Google Scholar 

  • Dobrowsky, P. H., De Kwaadsteniet, M., Cloete, T. E., & Khan, W. (2014a). Distribution of indigenous bacterial pathogens and potential pathogens associated with roof-harvested rainwater. Applied and Environmental Microbiology, 80(7), 2307–2316.

    Article  CAS  Google Scholar 

  • Dobrowsky, P. H., Van Deventer, A., Lombard, M., De Kwaadsteniet, M., Khan, W., & Cloete, T. E. (2014b). Quality of harvested rainwater and the application of point-of-use treatment systems. WRC Report No. TT 603/14, Water Research Commission (WRC), Pretoria.

  • Dobrowsky, P. H., Carstens, M., De Villiers, J., Cloete, T. E., & Khan, W. (2015). Efficiency of a closed-coupled solar pasteurization system in treating roof-harvested rainwater. Science of the Total Environment, 536, 206–214.

    Article  CAS  Google Scholar 

  • Dobrowsky, P. H., Khan, S., Cloete, T. E., & Khan, W. (2016). Molecular detection of Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis as vectors for Legionella spp. in untreated and solar pasteurized harvested rainwater. Parasites & Vectors, 9(1), 539.

    Article  Google Scholar 

  • Dunn, O. J., & Clark, V. A. (1974). Applied statistics: Analysis of variance and regression. London: Wiley.

    Google Scholar 

  • Gikas, G. D., & Tsihrintzis, V. A. (2012). Assessment of water quality of first-flush roof runoff and harvested rainwater. Journal of Hydrology, 466, 115–126.

    Article  CAS  Google Scholar 

  • Hamilton, K., Reyneke, B., Waso, M., Clements, T., Ndlovu, T., Khan, W., DiGiovanni, K., Rakestraw, E., Montalto, F., Haas, C. N., & Ahmed, W. (2019). A global review of the microbiological quality and potential health risks associated with roof-harvested rainwater tanks. npj Clean Water, 2(7).

  • Ito, F., Tamiya, T., Ohtsu, I., Fujimura, M., & Fukumori, F. (2014). Genetic and phenotypic characterization of the heat shock response in Pseudomonas putida. Microbiology Open, 3(6), 922–936.

    Article  CAS  Google Scholar 

  • Jonas, D., Rosenbaum, A., Weyrich, S., & Bhakdi, S. (1995). Enzyme-linked immunoassay for detection of PCR-amplified DNA of Legionellae in bronchoalveolar fluid. Journal of Clinical Microbiology, 33(5), 1247–1252.

    Article  CAS  Google Scholar 

  • King, C. H., Shotts, E. B., Wooley, R. E., & Porter, K. G. (1988). Survival of coliforms and bacterial pathogens within protozoa during chlorination. Applied and Environmental Microbiology, 54(12), 3023–3033.

    Article  CAS  Google Scholar 

  • Kong, R. Y. C., So, C. L., Law, W. F., & Wu, R. S. S. (1999). A sensitive and versatile multiplex PCR system for the rapid detection of enterotoxigenic (ETEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) strains of Escherichia coli. Marine Pollution Bulletin, 38(12), 1207–1215.

    Article  CAS  Google Scholar 

  • Kong, R. Y. C., Lee, S. K. Y., Law, T. W. F., Law, S. H. W., & Wu, R. S. S. (2002). Rapid detection of six types of bacterial pathogens in marine waters by multiplex PCR. Water Research, 36, 2802–2812.

    Article  CAS  Google Scholar 

  • Lücking, G., Stoeckel, M., Atamer, Z., Hinrichs, J., & Ehling-Schulz, M. (2013). Characterization of aerobic spore-forming bacteria associated with industrial dairy processing environments and product spoilage. International Journal of Food Microbiology, 166(2), 270–279.

    Article  Google Scholar 

  • Ludwig, W., & Schleifer, K. H. (2000). How quantitative is quantitative PCR with respect to cell counts? Systematic and Applied Microbiology, 23(4), 556–562.

    Article  CAS  Google Scholar 

  • Mena, K. D., & Gerba, C. P. (2009). Risk assessment of Pseudomonas aeruginosa in water. In Reviews of environmental contamination and toxicology Vol 201 (pp. 71–115). Boston: Springer.

    Chapter  Google Scholar 

  • Michel, R., Burghardt, H., & Bergmann, H. (1995). Acanthamoeba, naturally intracellularly infected with Pseudomonas aeruginosa, after their isolation from a microbiologically contaminated drinking water system in a hospital. International Journal of Hygiene and Environmental Health, 196(6), 532–544.

    CAS  Google Scholar 

  • Moreira Neto, R. F., Calijuri, M. L., De Castro Carvalho, I., & Da Fonseca Santiago, A. (2012). Rainwater treatment in airports using slow sand filtration followed by chlorination: Efficiency and costs. Resources, Conservation and Recycling, 65, 124–129.

    Article  Google Scholar 

  • Ndlovu, T., Le Roux, M., Khan, W., & Khan, S. (2015). Co-detection of virulent Escherichia coli genes in surface water sources. PLoS One, 10(2), 1–12.

    Article  CAS  Google Scholar 

  • Oliver, J. D. (2005). The viable but nonculturable state in bacteria. Journal of Microbiology, 43(1), 93–100.

    Google Scholar 

  • Park, S. H., Oh, K. H., & Kim, C. K. (2001). Adaptive and cross-protective responses of Pseudomonas sp. DJ-12 to several aromatics and other stress shocks. Current Microbiology, 43(3), 176–181.

    Article  CAS  Google Scholar 

  • Rawlings, D. E. (1995). Restriction enzyme analysis of 16S rRNA genes for the rapid identification of Thiobacillus ferooxidans, Thiobacillus thiooxidans, Leptospirillum ferooxidans strains in leaching environments. In C. A. Jerez, T. Vargas, H. Toledo, & J. V. Wiertz (Eds.), International biohydrometallurgical symposium (pp. 9–17). Santiago: University of Chile Press.

    Google Scholar 

  • Reyneke, B., Dobrowsky, P. H., Ndlovu, T., Khan, S., & Khan, W. (2016). EMA-qPCR to monitor the efficiency of a closed-coupled solar pasteurization system in reducing Legionella contamination of roof-harvested rainwater. Science of the Total Environment, 553, 662–670.

    Article  CAS  Google Scholar 

  • Ribeiro, N. F., Heath, C. H., Kierath, J., Rea, S., Duncan-Smith, M., & Wood, F. M. (2010). Burn wounds infected by contaminated water: Case reports, review of the literature and recommendations for treatment. Burns, 36(1), 9–22.

    Article  Google Scholar 

  • Roussel, C., Galia, W., Leriche, F., Chalancon, S., Denis, S., Van de Wiele, T., & Blanquet-Diot, S. (2018). Comparison of conventional plating, PMA-qPCR, and flow cytometry for the determination of viable enterotoxigenic Escherichia coli along a gastrointestinal in vitro model. Applied Microbiology and Biotechnology, 102(22), 9793–9802.

    Article  CAS  Google Scholar 

  • Sakai, H., Procop, G. W., Kobayashi, N., Togawa, D., Wilson, D. A., Borden, L., Krebs, V., & Bauer, T. W. (2004). Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets. Journal of Clinical Microbiology, 42(12), 5739–5744.

    Article  CAS  Google Scholar 

  • Simmons, G., Hope, V., Lewis, G., Whitmore, J., & Gao, W. (2001). Contamination of potable roof-collected rainwater in Auckland, New Zealand. Water Research, 35(6), 1518–1524.

    Article  CAS  Google Scholar 

  • Sommer, B., Marino, A., Solarte, Y., Salas, M. L., Dierolf, C., Valiente, C., Mora, D., Rechsteiner, R., et al. (1997). SODIS—An emerging water treatment process. Journal of Water Supply: Research and Technology – Aqua, 46(3), 127–137.

    Google Scholar 

  • Spilker, T., Coenye, T., Vandamme, P., & LiPuma, J. J. (2004). PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. Journal of Clinical Microbiology, 42, 2074–2079.

    Article  CAS  Google Scholar 

  • Spinks, A. T., Coombes, P., Dunstan, R. H., & Kuczera, G. (2003). Water quality treatment processes in domestic rainwater harvesting systems. Proceedings of the 28th International Hydrology and Water Resources Symposium. 10-13 November 2003. Wollongong, Australia.

  • Spinks, A., Dunstan, R., Harrison, T., Coombes, P., & Kuczera, G. (2006). Thermal inactivation of water-borne pathogenic and indicator bacteria at sub-boiling temperatures. Water Research, 40, 1326–1332.

    Article  CAS  Google Scholar 

  • Srivastava, S., Yadav, A., Seem, K., Mishra, S., Chaudhary, V., & Nautiyal, C. S. (2008). Effect of high temperature on Pseudomonas putida NBRI0987 biofilm formation and expression of stress sigma factor RpoS. Current Microbiology, 56(5), 453–457.

    Article  CAS  Google Scholar 

  • Stenkova, A. M., Isaeva, M. P., & Rasskazov, V. A. (2008). Development of a multiplex PCR procedure for detection of Yersinia genus with identification of pathogenic species (Y. pestis, Y. pseudotuberculosis and Y. enterocolitica). Molecular Genetics, Microbiology and Virology, 23, 119–125.

    Article  Google Scholar 

  • Stewart, P. S., Camper, A. K., Handran, S. D., Huang, C. T., & Warnecke, M. (1997). Spatial distribution and coexistence of Klebsiella pneumoniae and Pseudomonas aeruginosa in biofilms. Microbial Ecology, 33(1), 2–10.

    Article  CAS  Google Scholar 

  • Strauss, A., Dobrowsky, P. H., Ndlovu, T., Reyneke, B., & Khan, W. (2016). Comparative analysis of solar pasteurization versus solar disinfection for the treatment of harvested rainwater. BMC Microbiology, 16(289), 1–16.

    Google Scholar 

  • United Nations. (2015). The millennium development goals report. New York: United Nations Publications Available at: http://www.un.org/millenniumgoals/2015_MDG_Report/pdf/MDG%202015%20rev%20(July%201).pdf.

    Google Scholar 

  • World Health Organisation (WHO). (2008). Guidelines for drinking-water quality. Geneva: World Health Organisation.

    Google Scholar 

  • Zhu, H., Sun, S. J., & Dang, H. Y. (2008). PCR detection of Serratia spp. using primers targeting pfs and luxS genes involved in AI-2-dependent quorum sensing. Current Microbiology, 57(4), 326–330.

    Article  CAS  Google Scholar 

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Acknowledgements

The authors would like to thank Jacques de Villiers from the Crest Organisation (Somerset West, Western Cape) for providing the Phungamanzi™ system.

Funding

The Water Research Commission (WRC K5/2368//3) and the National Research Foundation of South Africa (grant number 90320) funded this project. Opinions expressed and conclusions arrived at are those of the authors and are not necessarily to be attributed to the National Research Foundation.

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Correspondence to Wesaal Khan.

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Clements, T., Reyneke, B., Strauss, A. et al. Persistence of Viable Bacteria in Solar Pasteurised Harvested Rainwater. Water Air Soil Pollut 230, 130 (2019). https://doi.org/10.1007/s11270-019-4184-z

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