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      Legionella Colonization of Hotel Water Systems in Touristic Places of Greece: Association with System Characteristics and Physicochemical Parameters

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          Abstract

          This study aimed to assess the colonization of hotel water systems in central Greece and Corfu by Legionella, and to investigate the association between physicochemical parameters and Legionella colonization. Standardized hygiene inspection was conducted in 51 hotels, and 556 water samples were analyzed for Legionella spp. Free chlorine concentration, pH, hardness, conductivity, and trace metals were defined in cold water samples. The results of inspections and chemical analyses were associated with the microbiological results using univariate and logistic regression analysis. According to the score of the checklist used for the inspections, 17.6% of the hotels were classified as satisfactory, 15.7% as adequate, and 66.7% as unsatisfactory. Moreover, 74.5% of the hotels were colonized by Legionella spp. and 31.4% required remedial measures according to the European guidelines. Legionella spp. were isolated in 28% of the samples. Unsatisfactory results of inspections were associated with Legionella presence (relative risk (RR) = 7.67, p-value = 0.043). In hot-water systems, <50 °C temperatures increased the risk of Legionella colonization (RR = 5.36, p-value < 0.001). In cold-water systems, free chlorine concentration <0.375 mg/L (odds ratio (OR) = 9.76, p-value = 0.001), pH ≥ 7.45 (OR = 4.05, p-value = 0.007), and hardness ≥321 mgCaCO 3/L (OR = 5.63, p-value = 0.003) increased the risk, whereas copper pipes demonstrated a protective role (OR = 0.29, p-value = 0.0024). The majority of the hotels inspected were colonized with Legionella. Supplementary monitoring of the risk factors that were identified should be considered.

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          Most cited references37

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          Diagnosis of Legionella infection.

          Legionellae, which are important causes of pneumonia in humans, continue to be incorrectly labeled as exotic pathogens. The ability to diagnose Legionella infection is limited by the nonspecific nature of clinical features and the shortcomings of diagnostic tests. Despite recent improvements, existing diagnostic tests for Legionella infection either lack sensitivity for detecting all clinically important legionellae or are unable to provide results within a clinically useful time frame. Understanding local Legionella epidemiology is important for making decisions about whether to test for Legionella infection and which diagnostic tests to use. In most situations, the use of both the urinary antigen test plus sputum culture is the best diagnostic combination. Polymerase chain reaction (PCR) is a promising tool, but standardized assays are not commercially available. Further work needs to focus on the development of urinary antigen tests assays that detect a wider range of pathogenic legionellae and on the development of standardized PCR assays.
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            Impact of Water Chemistry, Pipe Material and Stagnation on the Building Plumbing Microbiome

            A unique microbiome establishes in the portion of the potable water distribution system within homes and other buildings (i.e., building plumbing). To examine its composition and the factors that shape it, standardized cold water plumbing rigs were deployed at the treatment plant and in the distribution system of five water utilities across the U.S. Three pipe materials (copper with lead solder, CPVC with brass fittings or copper/lead combined pipe) were compared, with 8 hour flush cycles of 10 minutes to simulate typical daily use patterns. High throughput Illumina sequencing of 16S rRNA gene amplicons was employed to profile and compare the resident bulk water bacteria and archaea. The utility, location of the pipe rig, pipe material and stagnation all had a significant influence on the plumbing microbiome composition, but the utility source water and treatment practices were dominant factors. Examination of 21 water chemistry parameters suggested that the total chlorine concentration, pH, P, SO4 2- and Mg were associated with the most of the variation in bulk water microbiome composition. Disinfectant type exerted a notably low-magnitude impact on microbiome composition. At two utilities using the same source water, slight differences in treatment approaches were associated with differences in rare taxa in samples. For genera containing opportunistic pathogens, Utility C samples (highest pH of 9–10) had the highest frequency of detection for Legionella spp. and lowest relative abundance of Mycobacterium spp. Data were examined across utilities to identify a true universal core, special core, and peripheral organisms to deepen insight into the physical and chemical factors that shape the building plumbing microbiome.
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              Impact of water heater temperature setting and water use frequency on the building plumbing microbiome

              Hot water plumbing is an important conduit of microbes into the indoor environment and can increase risk of opportunistic pathogens (for example, Legionella pneumophila). We examined the combined effects of water heater temperature (39, 42, 48, 51 and 58 °C), pipe orientation (upward/downward), and water use frequency (21, 3 and 1 flush per week) on the microbial composition at the tap using a pilot-scale pipe rig. 16S rRNA gene amplicon sequencing indicated that bulk water and corresponding biofilm typically had distinct taxonomic compositions (R 2 Adonis=0.246, P Adonis=0.001), yet similar predicted functions based on PICRUSt analysis (R 2 Adonis=0.087, P Adonis=0.001). Although a prior study had identified 51 °C under low water use frequency to enrich Legionella at the tap, here we reveal that 51 °C is also a threshold above which there are marked effects of the combined influences of temperature, pipe orientation, and use frequency on taxonomic and functional composition. A positive association was noted between relative abundances of Legionella and mitochondrial DNA of Vermamoeba, a genus of amoebae that can enhance virulence and facilitate replication of some pathogens. This study takes a step towards intentional control of the plumbing microbiome and highlights the importance of microbial ecology in governing pathogen proliferation.

                Author and article information

                Journal
                Int J Environ Res Public Health
                Int J Environ Res Public Health
                ijerph
                International Journal of Environmental Research and Public Health
                MDPI
                1661-7827
                1660-4601
                30 November 2018
                December 2018
                : 15
                : 12
                : 2707
                Affiliations
                [1 ]Department of Hygiene and Epidemiology, Medical School, University of Thessaly, 41222 Larissa, Greece; mkiritsi@ 123456med.uth.gr (M.A.K.); mouchtourib@ 123456med.uth.gr (V.A.M.); akatsioul@ 123456med.uth.gr (A.K.); elkost@ 123456med.uth.gr (E.K.); nakulasb@ 123456yahoo.gr (V.N.); marinahatzinikou@ 123456yahoo.gr (M.H.)
                [2 ]Regional Public Health Laboratory of Thessaly, 41222 Larissa, Greece
                Author notes
                [* ]Correspondence: xhatzi@ 123456med.uth.gr
                Article
                ijerph-15-02707
                10.3390/ijerph15122707
                6313630
                30513698
                e5195ac2-2641-4248-9ac2-c2aad7a0b6f9
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 05 November 2018
                : 25 November 2018
                Categories
                Article

                Public health
                hotel-associated,legionnaires’ disease,greece,physicochemical,cold water
                Public health
                hotel-associated, legionnaires’ disease, greece, physicochemical, cold water

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