Quantication and identication of aerobic bacteria in holy water samples from a German environment Christoph König, Stephanie Tauchnitz, Heike Kunzelmann, Christian Horn, Frithjof Blessing, Matthias Kohl and Markus Egert ABSTRACT Despite its frequent use in many religious institutions, the microbiological quality of holy water is clearly underinvestigated. We analyzed the microbial load of 54 holy water samples, repeatedly taken in ve Roman Catholic churches in the greater area of Villingen-Schwenningen, Germany, by means of aerobic colony counting and Matrix-Assisted Laser Desorption/Ionization (MALDI) Biotyping of representative isolates. Over all samples, colony counting indicated an average aerobic microbial load of 5.85 ± 3.98 × 10 3 colony forming units (CFU) ml 1 (average ± standard error of the mean (SEM)). Urban churches showed signicantly higher contaminations than rural churches, probably owing to a greater number of visitors. Out of 145 bacterial isolates, 63 (43%) were identied to genus level and 39 (27%) to species level. The majority of the identied bacteria were typical human skin commensals, mainly afliated with the genus Staphylococcus. Ten out of 20 (50%) of the identied species were classied as potential pathogens. Appropriate hygiene measures should be taken to control microbial contamination of holy water, e.g., regular water exchange, particularly in highly frequented churches. Christoph König Stephanie Tauchnitz Heike Kunzelmann Frithjof Blessing Matthias Kohl Markus Egert (corresponding author) Faculty of Medical and Life Sciences, Institute of Precision Medicine, Furtwangen University, Jakob-Kienzle-Str. 17, 78054 Villingen-Schwenningen, Germany E-mail: markus.egert@hs-furtwangen.de Christian Horn Frithjof Blessing Institute for Laboratory Medicine, Virchowstr. 10c, 78224 Singen, Germany Key words | aerobic bacteria, bacterial load, holy water, hygiene, MALDI Biotyping INTRODUCTION Holy water is regarded as a symbol of blessing and puri- cation and plays an important role in many ceremonies and customs in Christianity and other religions. For instance, Roman Catholic believers wet their ngers with holy water when entering a church and make the sign of the cross; occasionally they also cross themselves on the forehead, lips and chest. In addition, holy water is used to bless items, food or persons by sprinkling them. Maybe also owing to its religious importance, only a few studies have addressed the microbiological and hygienic properties of holy water so far. Holy water was rst recognized as a source for potentially pathogenic microorganisms in 1992, when Pseudomonas aeruginosa was isolated from infected wounds of a hospital patient after sprinkling with holy water (Greaves & Porter ). In two other cases, nosocomial infections with Acineto- bacter baumanii were traced back to contact with holy water (Rees & Allen ; Michel et al. ). Daschner () reported massive contamination with Pseudomonas spp. in holy water samples from churches and hospitals in Freiburg, Germany. In several Buddhist temples in Thailand, holy water was examined for microbiological contamination, and the presence of Escherichia coli was demonstrated (Phatthararangrong et al. ). High contaminations with coliform bacteria and members of the Enterobacteriaceae family, including a considerable diversity of potentially pathogenic bacteria, were also reported from Spain ( Jurado et al. ). Recently, a landmark study by Kirschner et al. () reported contaminations of holy water fonts and holy springs in Vienna, Austria, with cell counts of up to 10 7 colony forming units (CFU) ml 1 , and the presence of fecal indicator bacteria such as enterococci and E. coli. Aiming at a deeper understanding of the hygienic rel- evance of holy water, the goal of our study was to 823 © IWA Publishing 2017 Journal of Water and Health | 15.5 | 2017 doi: 10.2166/wh.2017.026 Downloaded from https://iwaponline.com/jwh/article-pdf/15/5/823/393495/jwh0150823.pdf by guest on 24 May 2020