Introduction. Microbiological quality of recreational environ- ments included restrooms, is generally assessed by water and surface monitoring. In this study, an environmental monitoring, conducted in spring, of swimming pool restrooms of a recreation center located in the Marche region has been carried out. Seven water samples and seven surface swabs were collected. Moreover, six air samples have been included. The aim of this study was to evaluate if air microbiological monitoring, along with molecular detection in real-time PCR, could give additional useful information about the hygienic conditions of the facility.
Methods. Heterotrophic Plate Count (HPC) both at 22°C (psy- chrophilic) and 37°C (mesophilic) was determined by separate cultures in all samples. The presence of Legionella pneumophila and Pseudomonas aeruginosa was evaluated by both culture and real-time PCR.
Results. The analysis of shower water recorded a HPC load of mesophilic bacteria (37°C) more than 10-fold higher in men restroom, respect to women’s one (> 100 vs < 10 CFU/ml), while in air samples was between < 100 and > 500. Concerning pathogen presence, both species Legionella pneumophila and Pseudomonas aeruginosa were detected only in men restroom, but in different sample types by using different methods (culture and real-time PCR).
Conclusions. Air sampling may offer the advantage of giving more representative data about microbial presence in restrooms, including bacterial species transmitted through aerosol, like Legionella. Moreover, the concurrent use of molecular and micro- biological detection in an integrated approach could offer the advantage of greater sensitivity.
Amagliani G, Parlani ML, Brandi G, Sebastianelli G, Stocchi V, Schiavano GF. Molecular detection of Pseudomonas aeruginosa in recreational water. Int J Environ Health Res 2012; 22(1): 60-70. https:// doi: 10.1080/09603123.2011.588325.
Amagliani G, Schiavano GF, Stocchi V, Bucci G, Brandi G. Application of real-time PCR to Pseudomonas aeruginosa monitoring in a public swimming pool. Microchemical Journal 2013; 110: 656-659. https://doi.org/10.1016/j.microc.2013.08.001.
Blais-Lecours P, Perrott P, Duchaine C. Non-culturable bioaerosols in indoor settings: Impact on health and molecular approaches for detection. Atmospheric Environment 2015; 110: 45-53. https://doi: 10.1016/j.atmosenv.2015.03.039.
Bonadonna L and Ottaviani M. Metodi analitici di riferimento per le acque destinate al consumo umano ai sensi del DL.vo 31/2001. 2007.Metodi microbiologici. [Reference analytical methods for water intended for human consumption according to the Italian Legislative Decree 31/2001. Microbiological methods]. https://doi:10.13140/RG.2.1.1419.6088.
Bouillard L, Michel O, Dramaix M, Devleeschouwer M. Bacterial contamination of indoor air, surfaces, and settled dust, and related dust endotoxin concentrations in healthy office buildings. Ann Agric Environ Med 2005; 12(2): 187-192.
Bouwknegt M, Schijven JF, Schalk JA, de Roda Husman AM. Quantitative risk estimation for a Legionella pneumophila infection due to whirlpool use. Risk Anal 2013; 33(7): 1228-1236. https://doi: 10.1111/j.1539-6924.2012.01909.x
Commission of European Communities. Indoor Air Quality & Its Impact on Man; Report n.12 1993. Commission of European Communities: Luxembourg.
De Filippis P, Mozzetti C, Amicosante M, D'Alò GL, Messina A, Varrenti D, Giammattei R, Di Giorgio F, Corradi S, D'Auria A, Fraietta R, Gabrieli R. Occurrence of Legionella in showers at recreational facilities. Journal of water and health 2017; 15(3): 402-409. https:// doi: 10.2166/wh.2017.296.
Dutil S, Meriaux A, de Latremoille MC, Lazure L, Barbeau J, Duchaine C. Measurement of Airborne Bacteria and Endotoxin Generated During Dental Cleaning. Journal of Occupational and Environmental Hygiene 2009; 6(2): 121-130. https://doi:10.1080/15459620802633957.
Garcia MT, Jones S, Pelaz C, Millar RD, Abu Kwaik Y. Acanthamoeba polyphaga resuscitates viable non-culturable Legionella pneumophila after disinfection. Environ Microbiol 2007; 9(5): 1267-1277.
Grisoli P, Albertoni M, Rodolfi M. Application of Airborne Microorganism Indexes in Offices, Gyms, and Libraries. Applied Sciences 2019; 9(6). https://doi.org/10.3390/app9061101
Heo KJ, Lim CE, Kim HB, Lee BU. Effects of human activities on concentrations of culturable bioaerosols in indoor air environments. Journal of Aerosol Science 2017; 104:58–65. https:// doi: 10.1016/j.jaerosci.2016.11.008.
ISO11731:2017. Water Quality – Enumeration of Legionella. International Organization for Standardization, Geneva, Switzerland, 2017.
Kirschner AKT. Determination of viable legionellae in engineered water systems: Do we find what we are looking for? Water Res 2016, 93:276-288. https://doi:10.1016/j.watres.2016.02.016.
Lee BU, Hong IG, Lee DH, Chong E-S, Jung JH, Lee JH, Kim HJ, Lee IS. Bacterial Bioaerosol Concentrations in Public Restroom Environments. Aerosol and Air Quality Research 2012; 12(2):251-255. https:// doi: 10.4209/aaqr.2011.08.0134.
Lutz JK, Lee J. Prevalence and antimicrobial-resistance of Pseudomonas aeruginosa in swimming pools and hot tubs. Int J Environ Res Public Health 2011; 8(2):554-564. htpps://doi: 10.3390/ijerph8020554.
Mbareche H, Brisebois E, Veillette M, Duchaine C. Bioaerosol sampling and detection methods based on molecular approaches: No pain no gain. Science of the Total Environment 2017; 599:2095-2104. https:// doi: 10.1016/j.scitotenv.2017.05.076.
Omiccioli E, Schiavano GF, Ceppetelli V, Amagliani G, Magnani M, Brandi G. Validation according to ISO/TS 12869:2012 of a molecular method for the isolation and quantification of Legionella spp. in water. Molecular and Cellular Probes 2015; 29(2):86-91. https:// doi: 10.1016/j.mcp.2014.12.004.
Peccia J and Hernandez M. Incorporating polymerase chain reaction-based identification, population characterization, and quantification of microorganisms into aerosol science: A review. Atmospheric Environment 2006; 40(21):3941-3961. https://doi:10.1016/j.atmosenv.2006.02.029
[Proceedings of Italian Guidelines for the Prevention and Control of Legionellosis 2015, approved in State-Regions Conference on 05.07.2015] 2015 Italian.
Schiavano GF, Carloni E, Andreoni F, Magi S, Chironna M, Brandi G, Amagliani G. Prevalence and antibiotic resistance of Pseudomonas aeruginosa in water samples in central Italy and molecular characterization of oprD in imipenem resistant isolates. Plos One 2017; 12(12). https://doi: 10.1371/journal.pone.0189172.
UNI EN ISO 16266:2008 Water quality. Detection and enumeration of Pseudomonas aeruginosa. Method by membrane filtration. International Organization for Standardization, Technical committee ISO/TC 147, Subcommittee SC 4, 2008.
UNI EN ISO 6222, [Water quality - Quantitative assessment of viable microorganisms - Counting of colonies by inoculation on agarized medium] UNI - Ente Nazionale Italiano di Uniﬁcazione. Italian 2001.
WHO Guidelines for Indoor Air Quality: Dampness and Mould, 2009.