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A cross-sectional multicenter survey of Italian hotels was conducted to investigate Legionella spp. contamination of hot water. Chemical parameters (hardness, free chlorine concentration, and trace element concentrations), water systems, and building characteristics were evaluated to study risk factors for colonization. The hot water systems of Italian hotels were strongly colonized by Legionella; 75% of the buildings examined and 60% of the water samples were contaminated, mainly at levels of > or =10(3) CFU liter(-1), and Legionella pneumophila was the most frequently isolated species (87%). L. pneumophila serogroup 1 was isolated from 45.8% of the contaminated sites and from 32.5% of the hotels examined. When a multivariate logistic model was used, only hotel age was associated with contamination, but the risk factors differed depending on the contaminating species and serogroup. Soft water with higher chlorine levels and higher temperatures were associated with L. pneumophila serogroup 1 colonization, whereas the opposite was observed for serogroups 2 to 14. In conclusion, Italian hotels, particularly those located in old buildings, represent a major source of risk for Legionnaires' disease due to the high frequency of Legionella contamination, high germ concentration, and major L. pneumophila serogroup 1 colonization. The possible role of chlorine in favoring the survival of Legionella species is discussed.

We investigated Legionella and Pseudomonas contamination of hot water in a cross-sectional multicentric survey in Italy. Chemical parameters (hardness, free chlorine, and trace elements) were determined. Legionella spp. were detected in 33 (22.6%) and Pseudomonas spp. in 56 (38.4%) of 146 samples. Some factors associated with Legionella contamination were heater type, tank distance and capacity, water plant age, and mineral content. Pseudomonas presence was influenced by water source, hardness, free chlorine, and temperature. Legionella contamination was associated with a centralized heater, distance from the heater point >10 m, and a water plant >10 years old. Furthermore, zinc levels of <100 microg/L and copper levels of >50 microg/L appeared to be protective against Legionella colonization [corrected]. Legionella species and serogroups were differently distributed according to heater type, water temperature, and free chlorine, suggesting that Legionella strains may have a different sensibility and resistance to environmental factors and different ecologic niches.

Legionella contamination in public water systems poses significant health risks, particularly in schools where vulnerable populations, including children, regularly use these facilities. This study investigates the presence of Legionella in the hot water systems from 49 primary schools across two municipalities in the Danish capital region. Water samples were collected from taps in each school, and both first-flush and stabile temperature samples were analysed for Legionella contents. The findings revealed that 97% of schools in Municipality 1 and 100% in Municipality 2 had Legionella in their hot water systems. The content of Legionella colonies was significantly higher in schools in Municipality 1, which was probably because of overall lower water temperatures. At stabile temperatures, 76% and 50% of the schools in the two municipalities exceeded the European Union's recommended limit of 1000 CFU/L. Stabile peripheral water temperatures were achieved after 3 min. Tap water temperatures above 54 °C and central tank temperatures above 59 °C were associated with Legionella contents below 1000 CFU/L. This study highlights the need for more stringent Legionella control procedures in schools, including higher water temperatures and refining Legionella reducing interventions with the addition of regular flow and draining procedures. [ABSTRACT FROM AUTHOR]

Legionella is the pathogen that causes Legionnaires' disease, an increasingly prevalent and sometimes fatal disease worldwide. In 2021, 97% of cases in Europe were caused by Legionella pneumophila. We present a mathematical model that can be used by public health officials to assess the effectiveness and efficiency of different Legionella monitoring and control strategies to inform government requirements to prevent community-acquired Legionnaires' disease in non-hospital buildings. This simulation model was built using comprehensive data from multiple scientific and field-based sources. It is a tool for estimating the relative economic and human costs of monitoring and control efforts targeting either L. pneumophila or Legionella species and was designed to analyze the potential application of each approach to specific building classes across Italy. The model results consistently showed that targeting L. pneumophila is not only sufficient but preferable in optimizing total cost (direct and economic) for similar human health benefits, even when stress-tested with extreme inputs. This cost–benefit analytical tool allows the user to run different real-life scenarios with a broad range of epidemiological and prevalence assumptions across different geographies in Italy. With appropriate modifications, this tool can be localized and applied to other countries, states, or provinces. [ABSTRACT FROM AUTHOR]

Background: Legionella are parasites of freshwater protozoa, responsible for Legionellosis. Legionella can be found in a variety of aquatic environments, including rivers, lakes, and springs, as well as in engineered water systems where they can potentially lead to human disease outbarks. Legionella are considered to be predominantly freshwater organisms with a limited ability to proliferate in saline environments. Exposure of Legionella to high sodium concentrations inhibits growth and virulence of laboratory strains, particularly under elevated temperatures. Nonetheless, Legionella have been identified in some saline environments where they likely interact with various protozoan hosts. In this work, we examine how these selection pressures, sodium and grazing, help shape Legionella ecology within natural environments. Utilizing Legionella-specific primers targeting a variable region of the Legionella 16S rRNA gene, we characterized Legionella abundance, diversity, and community composition in natural spring clusters of varying sodium concentrations, focusing on high sodium concentrations and elevated temperatures. Results: We observed the highest abundance of Legionella in spring clusters of high salinity, particularly in combination with elevated temperatures. Legionella abundance was strongly related to sodium concentrations. The Legionella community structure in saline environments was characterized by relatively low diversity, compared to spring clusters of lower salinity. The community composition in high salinity was characterized by few dominant Legionella genotypes, not related to previously described species. Protozoan microbial community structure and composition patterns resembled those of Legionella, suggesting a common response to similar selection pressures. We examined Legionella co-occurrence with potential protozoan hosts and found associations with Ciliophora and Amoebozoa representatives. Conclusions: Our results indicate that selection forces in saline environments favor a small yet dominant group of Legionella species that are not closely related to known species. These novel environmental genotypes interact with various protozoan hosts, under environmental conditions of high salinity. Our findings suggest that alternative survival mechanisms are utilized by these species, representing mechanisms distinct from those of well-studied laboratory strains. Our study demonstrate how salinity can shape communities of opportunistic pathogens and their hosts, in natural environments, shedding light on evolutionary forces acting within these complex environments. Dch3Ld1J5jASmW1xvkYvDB Video Abstract [ABSTRACT FROM AUTHOR]

Legionella pneumophila (Lp) colonizes aquatic environments and is a potential pathogen to humans, causing outbreaks of Legionnaire's disease. It is mainly associated with contaminated cooling towers (CTs). Several regulations, including Spanish legislation (Sl), have introduced the analysis of heterotrophic plate count (HPC) bacteria and Legionella spp. (Lsp) in management plans to prevent and control Legionella outbreaks from CTs. The 2003 Sl for CTs (RD 865/2003) considered that concentrations of HPC bacteria ≤10,000 cfu/mL and of Lsp ≤100 cfu/L are safe; therefore, no action is required, whereas management actions should be implemented above these standards. We have investigated to what extent the proposed standard for HPC bacteria is useful to predict the presence of Lsp in cooling waters. For this, we analyzed Lsp and HPC concentrations, water temperature, and the levels of chlorine in 1376 water samples from 17 CTs. The results showed that in the 1138 water samples negative for Legionella spp. (LN), the HPC geometric mean was significantly lower (83 cfu/mL, p < 0.05) than in the positive Lsp. samples (135 cfu/mL). Of the 238 (17.3%) LP samples, 88.4% (210/238) were associated with values of HPC ≤10,000 cfu/mL and most of them showed HPC concentrations ≤100 (53.7%). In addition, a relatively low percentage of LP (28/238, 11.6%) samples were associated with HPC bacteria concentrations >10,000 cfu/mL, indicating that this standard does not predict the colonization risk for Legionella in the CTs studied. The present study has demonstrated that a threshold concentration ≤100 cfu/mL of HPC bacteria could better predict the higher concentration of Legionella in CTs, which will aid in preventing possible outbreaks. [ABSTRACT FROM AUTHOR]

In general, it is accepted that water stagnation and lack or poor maintenance in buildings are risk factors for Legionella growth. Then, in theory, the prolonged hotel closures due to the COVID-19 pandemic may have increased the risk of Legionella infections. However, there are very few field studies comparing the level of Legionella colonization in buildings before the pandemic and the new situation created after the lockdown. The objective of this study was to analyze these differences in a group of hotels that experienced prolonged closures in 2020 due to the COVID-19 pandemic. We have studied the Legionella spp. results, analyzed by standard culture, from the domestic water distribution systems of 73 hotels that experienced closures (from 1 to >4 months) during 2020, immediately after the reopening. The results were compared with those obtained in similar samplings of 2019. For the comparative analysis, we divided the hotels in two groups: Group A that have suffered closures for ≤3 months and Group B that remained closed for more than 3 months, both in relation to the opening period of 2019. In the Group B (36 sites), the frequency of positive samples in the hot water system increased from 6.7% in 2019 to 14.0% in 2020 (p < 0.05). In the Group A (37 sites), no significant differences were observed. No statistically significant differences were observed in terms of positive sites (defined as hotels with at least 1 positive sample), Legionella spp. concentrations and prevalence of Legionella pneumophila sg1 between the samplings of the two periods studied. The results suggest that hotels that suffered the longest prolonged closures (> 3 months) could have carried a higher risk of exposure to Legionella in the domestic hot water system. These findings highlight the importance of adequate preopening cleaning and disinfection procedures for hotel water systems, and the convenience of considering the most effective disinfection methods especially for hot water systems and after prolonged closure periods. [ABSTRACT FROM AUTHOR]

Thermal facilities comprise a wide spectrum of recreational and therapeutic activities with the number of customers increasing over time despite the break that occurred during the first phases of the COVID-19 pandemic. In order to evaluate the potential role played by this setting in the risk of contracting legionellosis, we conducted a cross-sectional study to evaluate the presence of Legionella spp. in some Italian thermal facilities. Specifically, we retrospectively analyzed the results of a 16-year surveillance carried out before the COVID-19 pandemic. Of 409 samples, 70 (17.1%) were positive with Legionella spp. but there was an overall decrease over time. L. pneumophila 2–14 were by far the most common detected serotypes, while L. pneumophila 1 accounted for only 8.8%. Of all the different kind of samples, swabs and municipal water samples were the most contaminated. Moreover, in the positive samples, bacterial load was often at intermediate values. In only a small percentage of samples was load high and, in this case, L. pneumophila 1 was often the most common strain. Our results show the importance of a continuous monitoring of Legionella risk in these settings, even more so now after the COVID-19 pandemic and the prolonged break in activities. Water is the natural environment of Legionella spp. and environment in general plays a crucial role in the transmission of these bacteria; therefore, it would be useful to frame this infection in a "One Health" key. [ABSTRACT FROM AUTHOR]

Objective: Legionella bacteria are waterborne environmental pathogens that are considered a public health problem because they cause Legionnaires' disease, which is a nationally notifiable disease. Materials and Methods: Legionella analysis was performed in a total of 651 water samples collected during the years 2015 (450) - 2016 (201). Water samples were collected from hospitals (64.66%), hotels (15.05%), the automotive industry (14.43%) and from the buildings (5.83%) in the Central Anatolia Region. After the isolation of Legionella by the filtration and culturing method, serogroup and subtypes were determined via latex agglutination tests and the direct fluorescent antibody method. Results: In 2015, the Legionella positivity rate was 8.6%, where 28.2% of detections were from L. pneumophila serogroup-1. Six isolates were found to be Philadelphia, four were Olda, and one was Bellingham subgroup. Overall, 64.1% were L. pneumophila serogroup 2-14. Moreover, 14 isolates were SG-5, 10 were SG-6, and one was SG-10. 7.7% were unidentified Legionella species. The Legionella species identified were L. micdadei and L. longbeachae. In 2016, the Legionella positivity rate was 10.4%, with 28.6% of them being from L. pneumophila serogroup-1. Four were found to be Olda and two were Philadelphia subgroups. Overall, 66.6% of them were L. pneumophila serogroup 2-14. Moreover, six of them were SG-5, four were SG-6, and four were SG-2. 4.7% were unidentified Legionella species. There was only one species detected as L. micdadei. Conclusion: It has been observed that the distribution of Legionella has exhibited diversity in different water systems throughout the Central Anatolia Region. [ABSTRACT FROM AUTHOR]

Legionella colonization of water supply pipes is a significant public health problem. The objective of this work was to evaluate Legionella colonization in hotel hot water systems and to investigate the relationship between metal concentrations, piping materials (galvanized iron pipes and plastic pipes), and Legionella proliferation. Concentrations of calcium and magnesium ions and the presence of Legionella pneumophila were determined in a total of 108 water samples from the hot water systems of four hotels in Split-Dalmatia County over a 12-month period, and additional data on piping materials were collected. L. pneumophila was isolated in 23.1% of all samples—in 28.8% (15/52) of water samples from galvanized iron pipes and in 17.8% (10/56) of samples from plastic pipes. L. pneumophila serogroups 2–14 were isolated from all samples. This study found higher prevalence of L. pneumophila at higher concentrations of Ca and Mg ions (except for Mg and plastic pipes). The metal parts of the water supply may be important factors in Legionella contamination due to the possibility of lime scale or roughness of the pipes. Higher Ca and Mg ion concentrations increased the risk of Legionella colonization. [ABSTRACT FROM AUTHOR]

Legionella is a ubiquitous bacterium that lives in freshwater environments and colonizes human-made water systems. Legionella pneumophila is the most virulent species, and risk factors for Legionnaires' disease include increasing age, smoking, chronic diseases, and immunodeficiency. For this reason, it is very important to assess and monitor hospital water systems in order to prevent legionellosis. We have monitored a large hospital in Palermo for four years. To determine the presence of microorganisms, according to national guidelines, we used the culture method, which is considered the gold standard for Legionella detection. Sampling was divided into five macro-areas, and a total of 251 samples were collected during the period of investigation, 49% of which were Legionella spp.-positive and 51% were Legionella spp.-negative. Positive samples with L. pneumophila. sgr 2-15 were most frequent in the Underground (55.6%, p = 0.0184), Medicine (42.9%, p = 0.0184) and Other (63.2%, p = 0.002) areas; while positive samples for L. pneumophila sgr 1 were less frequent in the Underground (0.0%, p = 0.0184) and Surgery areas (4.5%, p = 0.033), and for Legionella anisa, were less frequent in the Medicine (4.1%, p = 0.021), Oncohematology (0.0%, p = 0.0282), and Other (0.0%, p = 0.016) areas. Finally, no significant differences were observed among the areas for each isolate considered. The surveillance carried out in these years demonstrates the importance of monitoring, which allows us to analyze the conditions of hospital facilities and, therefore, prevent Legionella spp. infections. [ABSTRACT FROM AUTHOR]

When stay-at-home orders were issued to slow the spread of COVID-19, building occupancy (and water demand) was drastically decreased in many buildings. There was concern that widespread low water demand may cause unprecedented Legionella occurrence and Legionnaires' disease incidence. In lieu of evidenced-based guidance, many people flushed their water systems as a preventative measure, using highly variable practices. Here, we present field-scale research from a building before, during, and after periods of low occupancy, and controlled stagnation experiments. We document no change, a > 4-log increase, and a > 1.5-log decrease of L. pneumophila during 3- to 7-week periods of low water demand. L. pneumophila increased by > 1-log after precautionary flushing prior to reoccupancy, which was repeated in controlled boiler flushing experiments. These results demonstrate that the impact of low water demand (colloquially called stagnation) is not as straight forward as is generally assumed, and that some flushing practices have potential unintended consequences. In particular, stagnation must be considered in context with other Legionella growth factors like temperature and flow profiles. Boiler flushing practices that dramatically increase the flow rate and rapidly deplete boiler temperature may mobilize Legionella present in biofilms and sediment. [ABSTRACT FROM AUTHOR]

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Featured Application: This critical review is highly timely and the need of the hour, given the prolonged and unwanted building closures during the COVID-19 pandemic. Extended water age in premise plumbing greatly increases the risk of opportunistic premise plumbing pathogens. This review critically analyses the chemical and physical parameters that influence the occurrence of opportunistic pathogens in the drinking water distribution system, specifically in premise plumbing. A comprehensive literature review reveals significant impacts of water age, disinfectant residual (type and concentration), temperature, pH, and pipe materials. Evidence suggests that there is substantial interplay between these parameters; however, the dynamics of such relationships is yet to be elucidated. There is a correlation between premise plumbing system characteristics, including those featuring water and energy conservation measures, and increased water quality issues and public health concerns. Other interconnected issues exacerbated by high water age, such as disinfectant decay and reduced corrosion control efficiency, deserve closer attention. Some common features and trends in the occurrence of opportunistic pathogens have been identified through a thorough analysis of the available literature. It is proposed that the efforts to reduce or eliminate their incidence might best focus on these common features. [ABSTRACT FROM AUTHOR]

The aim of the study was to assess the risk posed by Legionella bacteria in a public building in Krakow. An old building with internal installation risers of different ages, as well as draw-off points of different types, was selected for testing. Samples were collected during two campaigns. In one sample of the first series of tests, no bacteria were found. During the second series of tests, no Legionella bacilli were found in just one sample and in one sample only 4 colony-forming units were detected. At the remaining draw-off points (water taps), the bacteria count detected were greater than the maximum threshold allowed by legal regulations (admissible threshold for public utility buildings - 100 cfu/100 mL). No morphological differences were observed with respect to the occurrence of specific serogroups. In 14 samples, Legionella pneumophila serogroups 2-14 were found, while the Legionella pneumophilaserogroup 1 was only found in one sample. The risk assessment was also carried out based on a semi-quantitative risk matrix approach and as a quantitative microbial risk assessment. The risk matrix approach was successfully implemented for the recognition of the potential risk associated with the Legionella occurrence in a water system. The calculated annual cumulative risk is high. The research shows that even if the weekly inhalation exposure dose (and therefore the calculated risk) is high, the number of Legionella pneumophila illness cases found can be equal to zero. This is probably due to the large uncertainty associated with QMRA determination. The size of the room in which the contaminated water is used also affects the possibility of infection. [ABSTRACT FROM AUTHOR]

Background: Legionella have emerged as a pathogenic group due to increased use and poor maintenance of artificial water environments. World Health Organization considers Legionella associated diseases to be underestimated, especially in developing countries due to difficulty in detection. The aim of this study was isolation and culture of Legionella spp. from artificial water systems and to test the importance of iron concentration which can be developed as a reliable chemical marker. Method: Thirty samples each were collected from drinking water coolers, cooling towers and shower heads fitted in different houses of Mangaluru city. Isolation and identification of the bacteria were carried out as per the standard protocols followed by determination of iron content prescribed by IS 3025 (Part 53). Conclusion: Our study shows that concentrations of iron, is an important factor that increases the likelihood of Legionella spp. Statistically, a concentration of 300 mg Fe/L shows a positive correlation with Legionella presence. Therefore, monitoring suspect sites for iron concentration and chelation of available iron, can be developed as respective assays for identifying Legionella spp. Furthermore, we show that PCR based detection of Legionella is a more robust method than the classical 3-plate method, especially for virulent strains. [ABSTRACT FROM AUTHOR]

Legionella spp. are considered an important cause of potentially preventable morbidity and mortality, making environmental surveillance a crucial component of risk assessment plans. In this work, 20,319 water samples were collected in 3,983 environmental surveys during a 16-year period by ARPA, the Regional Agency for Environmental Protection, Friuli Venezia Giulia, and the results were studied to better understand the diffusion mechanisms of Legionella. The data showed a strong seasonal signal, a prevalence of L. pneumophila serogroup 2–15 in most environments (63% of positive samples), a prevalence of L. pneumophila serogroup 1 in swimming pool-associated environments (82% of positive samples), a persistent presence of Legionella in hospitals and a recurrent presence of Legionella in other facilities such as hotels, possibly years after interventions, highlighting the difficulty of eradicating the bacteria. Retrospective spatio-temporal analyses on geocoded historical data were carried out with SaTScan using an ordinal model with risk as a covariate to identify potential clusters with an excess of cases in the higher-risk categories. Although no outbreaks occurred during the period of study, such analyses identified spatially restricted zones with unusual contamination, which sometimes were also areas in which several surveys triggered by notifications of clinical cases were performed. Simulations of periodic prospective analyses permitted the assessment of the efficacy of the method in early detection of such clusters. The proposed method may be a useful tool in environmental surveillance, prevention and control of Legionella. [ABSTRACT FROM AUTHOR]

Background. Nosocomial water systems may be contaminated by Legionella spp; therefore, health care facilities represent a potential health risk for patients and health care staff. Active, well-planned clinical and environmental surveillance in hospitals is the most important instrument of prevention. Aim and Methods. The aim of the present article was to outline the state of the art in legionellosis control and prevention among Italian health care facilities by reporting some experiences in the field. Results. Our results showed that Legionella spp. are largely reported as both hospital water system contaminants and etiological agents in water-related health care-associated infections (HCAI) in Italy. Among the numerous sources of HCAI, water is the most investigated, although it has been demonstrated that air sampling may provide additional information for risk assessment. Conclusions. More appropriate risk assessment is needed, especially in large facilities. In addition, more sensitive diagnostic tests should be used and dedicated training courses should be implemented in health care facilities. [ABSTRACT FROM AUTHOR]

Hot water premise plumbing has emerged as a critical nexus of energy, water, and public health. The composition of hot water microbiomes is of special interest given daily human exposure to resident flora, especially opportunistic pathogens (OPs), which rely on complex microbial ecological interactions for their proliferation. Here, we applied shotgun metagenomic sequencing to characterize taxonomic and functional shifts in microbiomes as a function of water heater temperature setting, stagnation in distal pipes, and associated shifts in water chemistry. A cross-section of samples from controlled, replicated, pilot-scale hot water plumbing rigs representing different temperature settings (39, 42, and 51°C), stagnation periods (8 h vs. 7 days), and time-points, were analyzed. Temperature setting exhibited an overarching impact on taxonomic and functional gene composition. Further, distinct taxa were selectively enriched by specific temperature settings (e.g., Legionella at 39°C vs. Deinococcus at 51°C), while relative abundances of genes encoding corresponding cellular functions were highly consistent with expectations based on the taxa driving these shifts. Stagnation in distal taps diminished taxonomic and functional differences induced by heating the cold influent water to hot water in recirculating line. In distal taps relative to recirculating hot water, reads annotated as being involved in metabolism and growth decreased, while annotations corresponding to stress response (e.g., virulence disease and defense, and specifically antibiotic resistance) increased. Reads corresponding to OPs were readily identified by metagenomic analysis, with L. pneumophila reads in particular correlating remarkably well with gene copy numbers measured by quantitative polymerase chain reaction. Positive correlations between L. pneumophila reads and those of known protozoan hosts were also identified. Elevated proportions of genes encoding metal resistance and hydrogen metabolism were noted, which was consistent with elevated corrosion-induced metal concentrations and hydrogen generation. This study provided new insights into real-world factors influencing taxonomic and functional compositions of hot water microbiomes. Here metagenomics is demonstrated as an effective tool for screening for potential presence, and even quantities, of pathogens, while also providing diagnostic capabilities for assessing functional responses of microbiomes to various operational conditions. These findings can aid in informing future monitoring and intentional control of hot water microbiomes. [ABSTRACT FROM AUTHOR]

Legionella pneumophila, the causative agent of Legionnaires' disease, is widely distributed throughout natural and artificial water systems and can replicate in macrophages and amoebae. Amoebae are the natural hosts of L. pneumophila, whereas macrophages are incidentally infected. The life cycle of L. pneumophila comprises a replicative phase within the Legionella-containing vacuole (LCV) and a transmissive phase during which bacterial cells become motile and are released via killing of the host. Although the host death mechanisms induced by L. pneumophila have been studied, the expression patterns of related L. pneumophila genes have not been reported. The present study compared the expression patterns of host cell death-associated genes in L. pneumophila grown in the human monocytic cell line THP-1 and Acanthamoeba castellanii. Notably, when L. pneumophila was grown in THP-1, expression of the gene flaA, which is involved in the induction of pyroptosis, was downregulated during the course of infection. In contrast, sdhA associated indirectly with host death, was upregulated. Expression of the genes vipD and sidF, which are involved in the induction and suppression of apoptosis, changed by less than 2-fold. Notably, a lower percentage of pyroptotic cells was observed among infected THP-1 cells relative to uninfected cells, and the latter exhibited stronger expression of caspase-1. A different pattern was observed when L. pneumophila was grown in A. castellanii: flaA and vipD were activated, whereas sdhA and sidF were downregulated during the later stage of replication. The percentage of non-viable (annexin-V+ PI+ or annexin-V+PI−) A. castellanii organisms increased with Legionella infection, and the expression of metacaspase-1, which is involved in encystation was up-regulated at late infection time. In summary, L. pneumophila can multiply intracellularly in both amoebae and macrophages to induce cell death and secondary infection, and this characteristic is essential for its survival in water and the lungs. The gene expression profiles observed in this study indicated the increased cytotoxicity of L. pneumophila in A. castellanii, suggesting an increased adaptation of Legionella to this host. [ABSTRACT FROM AUTHOR]

We have monitored the presence of bacteria belonging to the genus Legionella in the plumbing of buildings at the University of Perugia (Italy).More than 300 water samples were collected from 156 control-point taps in 41 buildings comprised in the eight campuses of the University. Legionella was absent in most samples, while it was found in only 12 buildings (29% of the total). Molecular analysis indicated the presence of L. pneumophila (serogroups 1, 8 and 6-10), L. taurinensis and L. anisa. In only three cases contamination levels were above the limit at which remedial actions are required, according to international guidelines. In two buildings, where the water temperature could be raised and maintained above 60 °C, thermal disinfection was effective in eradicating Legionella. Conversely, in buildings where contaminations were caused by heat exchangers that produced hot water at a maximum temperature of 50 °C, a chemical disinfection with silver hydrogen peroxide was carried out but was effective only in the short term. In this case study, Legionella contaminations and remediation effectiveness strongly depended on the network and heating-system characteristics, indicating how a multidisciplinary approach that integrates microbiological analysis with hydraulic surveys is necessary for an effective definition of Legionella prevention and control strategies. [ABSTRACT FROM AUTHOR]

Legionella is an opportunistic waterborne pathogen of increasing public health significance. Pasteurization, otherwise known as super-heat and flush (increasing water temperature to above 70°C and flushing all outlets), has been identified as an important mechanism for the disinfection of Legionella in manufactured water systems. However, several studies have reported that this procedure was ineffective at remediating water distribution systems as Legionella was able to maintain long term persistent contamination. Up to 25% of L. pneumophila cells survived heat treatment of 70°C, but all of these were in a viable but non-culturable state. This demonstrates the limitations of the culture method of Legionella detection currently used to evaluate disinfection protocols. In addition, it has been demonstrated that pasteurization and nutrient starvation can select for thermal tolerant strains, where L. pneumophila was consistently identified as having greater thermal tolerance compared to other Legionella species. This review demonstrates that further research is needed to investigate the effectiveness of pasteurization as a disinfection method. In particular, it focuses on the potential for pasteurization to select for thermal tolerant L. pneumophila strains which, as the primary causative agent of Legionnaires disease, have greater public health significance compared to other Legionella species. [ABSTRACT FROM AUTHOR]

Background and Aims: Several studies were investigated legionella contamination in natural and man- made water resources. The aim of this research was systematically review of legionella water contamination in natural and man-made resources in Iran and foreign countries. Materials and methods: In this systematic review study, the required data was collected using suitable keywords through PubMed, Science direct, Springer link, Google scholar, SID, Iranmedex, Irandoc and Magiran databases. Background and Aim: Several studies were investigated legionella contamination in natural and man- made water resources. The aim of this research was systematically review of legionella water contamination in natural and man-made resources. Materials and Methods: In this systematic review study, the required data was collected using suitable keywords through PubMed, Science direct, Springer link, Google scholar, SID, Iranmedex, Irandoc and Magiran databases. The search was conducted without publication date limitation. Survey and selection of articles was conducted based on PRISMA checklist and Cochrane quality assessment standards. Out of 1386 articles, 56 articles were considered after excluding the remaining articles which were not related to the study objectives. Identification and isolation of legionella with PCR technique and culture in BCYE is one of the article selection criteria. The relevant data were classified in extracted table and analyzed manually. Excel 2007 software was used for to draw diagrams. Due to heterogeneity of data meta-analysis could not be conducted. Results: Out of 56 articles, 9 and 47 articles were published in Iran and foreign countries respectively. In Iran, 5.7-70% of samples were contaminated. In Iran's hospitals 2.85-41.75% of samples were contaminated by Legionella pneumophila. In foreign countries, 0-100% of samples were contaminated and in these countries' hospitals Legionella pneumophila contamination was 17%-98.7%. Conclusion: Although in Iran legionella water contamination is lower than foreign countries but, based on WHO guideline (1CFU/L) for legionella, planning for control of this bacteria and relevance infectious is one of the health priorities. The search was conducted without publication date limitation. Survey and selection of articles was conducted based on PRISMA checklist and Cochrane quality assessment standards. Out of 1386 articles, 56 articles were considered after excluding the remaining articles which were not related to the study objectives. Identification and isolation of legionella with PCR technique and culture in BCYE is one of the article selection criteria. The relevant data were classified in extracted table and analyzed manually. Excel 2007 software was used for to draw diagrams. Due to heterogeneity of data meta-analysis could not be conducted. Results: Out of 56 articles, 9 and 47 articles were published in Iran and foreign countries respectively. In Iran, 5.7 - 70% of samples were contaminated. In Iran's hospitals 2.85-41. 75% of samples were contaminated by Legionella pneumophila. In foreign countries, 0-100% of samples were contaminated and in these countries' hospitals Legionella pneumophila contamination was 17%-98. 7%. Conclusion: The results of this study revealed that the conventional water treatment processes were not effective for legionella elimination and eradication of this agent should be considered in point off-use with effective techniques. [ABSTRACT FROM AUTHOR]

Background: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa are opportunistic premise plumbing pathogens (OPPPs) that persist and grow in household plumbing, habitats they share with humans. Infections caused by these OPPPs involve individuals with preexisting risk factors and frequently require hospitalization. Objectives: The objectives of this report are to alert professionals of the impact of OPPPs, the fact that 30% of the population may be exposed to OPPPs, and the need to develop means to reduce OPPP exposure. We herein present a review of the epidemiology and ecology of these three bacterial OPPPs, specifically to identify common and unique features. Methods: A Water Research Foundation-sponsored workshop gathered experts from across the United States to review the characteristics of OPPPs, identify problems, and develop a list of research priorities to address critical knowledge gaps with respect to increasing OPPP-associated disease. Discussion: OPPPs share the common characteristics of disinfectant resistance and growth in biofilms in water distribution systems or premise plumbing. Thus, they share a number of habitats with humans (e.g., showers) that can lead to exposure and infection. The frequency of OPPPinfected individuals is rising and will likely continue to rise as the number of at-risk individuals is increasing. Improved reporting of OPPP disease and increased understanding of the genetic, physiologic, and structural characteristics governing the persistence and growth of OPPPs in drinking water distribution systems and premise plumbing is needed. Conclusions: Because broadly effective community-level engineering interventions for the control of OPPPs have yet to be identified, and because the number of at-risk individuals will continue to rise, it is likely that OPPP-related infections will continue to increase. However, it is possible that individuals can take measures (e.g., raise hot water heater temperatures and filter water) to reduce home exposures. [ABSTRACT FROM AUTHOR]

Background: Legionellosis is an uncommon form of pneumonia. After a clinical encounter, the necessary antibiotic treatment is available if the diagnosis is made early in the illness. Before the clinical encounter, early detection of the main pathogen involved, Legionella pneumophila, in hazardous environments is important in preventing infectious levels of this bacterium. In this study a qualitative test based on combined magnetic immunocapture and enzyme-immunoassay for the fast detection of Legionella pneumophila in water samples was compared with the standard method, in both comparative and collaborative trials. The test was based on the use of anti-Legionella pneumophila antibodies immobilized on magnetic microspheres. The final protocol included concentration by filtration, resuspension and immunomagnetic capture. The whole assay took less than 1 hour to complete. Results: A comparative trial was performed against the standard culture method (ISO 11731) on both artificially and naturally contaminated water samples, for two matrices: chlorinated tap water and cooling tower water. Performance characteristics of the test used as screening with culture confirmation resulted in sensitivity, specificity, false positive, false negative, and efficiency of 96.6%, 100%, 0%, 3.4%, and 97.8%, respectively. The detection limit at the level under which the false negative rate increases to 50% (LOD50) was 93 colony forming units (CFU) in the volume examined for both tested matrices. The collaborative trial included twelve laboratories. Water samples spiked with certified reference materials were tested. In this study the coincidence level between the two methods was 95.8%. Conclusion: Results demonstrate the applicability of this immunosensing technique to the rapid, simple, and efficient detection of Legionella pneumophila in water samples. This test is not based on microbial growth, so it could be used as a rapid screening technique for the detection of L. pneumophila in waters, maintaining the performance of conventional culture for isolation of the pathogen and related studies [ABSTRACT FROM AUTHOR]

Background: Legionella is the causative agent of Legionnaires' disease, and hot springs are a major source of outbreaks of this disease. It is important from a public health perspective to survey hot spring environments for the presence of Legionella. Methods: Prospective surveillance of the extent of Legionella pollution was conducted at three hot spring recreational areas in Beijing, China in 2011. Pulsed-field gel electrophoresis (PFGE) and sequence-based typing (SBT) were used to describe the genetic polymorphism of isolates. The intracellular growth ability of the isolates was determined by interacting with J774 cells and plating the dilutions onto BCYE agar plates. Results: Overall, 51.9% of spring water samples showed Legionella-positive, and their concentrations ranged from 1 CFU/liter to 2,218 CFU/liter. The positive rates of Legionella were significantly associated with a free chlorine concentration of ≥0.2 mg/L, urea concentration of ≥0.05 mg/L, total microbial counts of ≥400 CFU/ml and total coliform of ≥3 MPN/L (p<0.01). The Legionella concentrations were significantly associated with sample temperature, pH, total microbial counts and total coliform (p<0.01). Legionella pneumophila was the most frequently isolated species (98.9%), and the isolated serogroups included serogroups 3 (25.3%), 6 (23.4%), 5 (19.2%), 1 (18.5%), 2 (10.2%), 8 (0.4%), 10 (0.8%), 9 (1.9%) and 12 (0.4%). Two hundred and twenty-eight isolates were analyzed by PFGE and 62 different patterns were obtained. Fifty-seven L. pneumophila isolates were selected for SBT analysis and divided into 35 different sequence types with 5 main clonal groups. All the 57 isolates had high intracellular growth ability. Conclusions: Our results demonstrated high prevalence and genetic polymorphism of Legionella in springs in Beijing, China, and the SBT and intracellular growth assay results suggested that the Legionella isolates of hot spring environments were pathogenic. Improved control and prevention strategies are urgently needed. [ABSTRACT FROM AUTHOR]

In this study, we investigated the distribution of Legionella in sources of spring water and wastewater throughout Taiwan. Legionella were detected with culture method and PCR method from water samples, combined with molecular methods that allow the taxonomic identification of Legionella species. Factors associated with the distribution of Legionella including the physical-chemical and microbiological water quality characteristics were also measured. Legionella were detected in 18 of the 68 samples. Additionally, 16 of the 68 samples analyzed by polymerase chain reaction (PCR) and eight of the 68 samples evaluated using the culture method were found to be positive for Legionella. The Legionella species identified included Legionella pneumophila, unnamed Legionella genotypes, uncultured Legionella spp., and Legionella lytica. Legionella species were found in water with temperatures ranging from 10.0 to 50.6°C and with pH values ranging from 5.59 to 9.32. Significant differences (Mann-Whitney U test, P < 0.05) were observed in the presence/absence of Legionella in samples with different pH values when Legionella were detected by the culture method. The results of this survey confirmed that Legionella is ubiquitous in the water of spring recreational areas in Taiwan. Long-term investigations of the occurrence of Legionella in hot spring recreational areas throughout Taiwan are needed. [ABSTRACT FROM AUTHOR]

This is a cross-sectional study of the incoming and distal outlet water quality from 41 dental units in Istanbul, carried out to compare the total microbial loads using traditional culture method versus epifluorescence microscopy. The possible presence of Legionella pneumophila using traditional culture method was also analyzed. One hundred and twenty three samples were taken from the high-speed handpiece lines, air–water syringe lines and source (incoming) water supplies from 41 dental units. The samples were assayed for live/dead bacteria, heterotrophic bacterial counts and presence of L. pneumophila bacteria. Thirty nine out of 41 dental units (91%) were not able to meet the standard limit of 200 CFU/ml in dental unit waters. The live bacterial counts were 1–1.5 orders of magnitude higher than aerobic mesophilic heterotrophic bacteria. L. pneumophila (serogroup 2–14) was isolated from five out of 41 units. Some dental units were using commercially bottled (19 l) drinking water as a source. The source water of eight dental unit was heavily contaminated which were fed up by commercially bottled drinking water. [ABSTRACT FROM AUTHOR]

Currently, the investigation of Legionella ecology falls into two distinct areas of research activity: (1) that Legionella multiply within water sources by parasitizing amoebic or ciliate hosts or (2) that Legionella grows extracellularly within biofilms. Less focus has been given to the overlaps that may occur between these two areas or the likelihood that Legionella employs multiple survival strategies to persist in water sources. It is likely that Legionella interacts with protozoa, bacteria, algae, fungi, etc., and biofilm components in a more complex fashion than multiplication or death due to the presence or absence of single components of these complex microbial systems. This paper addresses gaps that exist in the understanding of Legionella ecology and serves to pinpoint areas of future research. To assume that only one other class of organism is important to Legionella ecology may limit our understanding of how this bacterium proliferates in heated water sources and also limit our strategies for its control in the built environment. [ABSTRACT FROM AUTHOR]

Legionella species are increasingly recognized as a cause of both healthcare- and community-acquired pneumonia (so-called “Legionnaire’s disease”). These pathogens are ubiquitous in the environment, but environmental factors in the occurrence of sporadic legionellosis remain poorly understood. We analyzed all legionellosis cases identified in the Greater Toronto Area of Ontario from 1978 to 2006, and evaluated seasonal and environmental patterns in legionellosis case occurrence by using both negative binomial models and case-crossover analysis. A total of 837 cases were reported during the study period. After adjusting for seasonal effects, changes in the local watershed, rather than weather, were the strongest contributors to legionellosis risk. A 3.6-fold increase (95% confidence interval (CI), 2.4–5.3) in odds of disease was identified with decreasing watershed levels approximately 4 weeks before case-occurrence. We also found a 33% increase (95% CI, 8–64%) in odds of disease with decreasing lake temperature during the same period and a 34% increase (95% CI, 14–57%) with increasing humidity 5 weeks before case-occurrence. We conclude that local watershed ecology influences the risk of legionellosis, notwithstanding the availability of advanced water treatment capacity in Toronto. Enhancement of risk might occur through direct contamination of water sources or via introduction of micronutrients or commensal organisms into residential and hospital water supplies. These observations suggest testable hypotheses for future empiric studies. [ABSTRACT FROM AUTHOR]

The water used in dental unit waterlines (DUWLs) acts as a coolant for the high-speed equipments and as an irrigant during dental treatments. There are kind of water tanks. DUWLs provide a favorable environment for microbial biofilm and multiplation primarily due to the high surface in the tubing and the character of fluid dynamics in narrow, smooth-walled waterlines. Biofilms can harbour opportunist pathogens such as Legionella sp., Pseudomonas sp. Several studies have shown that DUWLs have high levels of microbial contamination. Presence of high level of microbial contamination is an important problem for dentists and dental patients who are immunocompromised. We collected water samples from DUWLs of 20 private dental offices. We have determined that only 2 (3.4%) out of 59 dental unit water samples were found to meet the standard (<200 CFU·ml−1) for DUWLs water quality by American Dental Association (ADA). Of the 59 water samples examined, 14 (24%) were positive for Pseudomonas sp. and 18 (30.5%) were positive for fungi. The most common 14 bacterial strains and seven fungi were isolated. Of bacterial strains, 57.1% were identified: Majority of the bacterial species isolated from our samples was identified as Pseudomonas fluorescens, Pasteurella haemolytica, Photobacterium damsela, Ochrobacter anthropi, Moraxella sp., Aspergillus flavus, Penicillium expansum. Legionella sp. were not detected in all water samples. [ABSTRACT FROM AUTHOR]

Different guidelines and regulations for the prevention of legionellosis in public facilities include the recommendation of a periodical thermal shock in the hot water system. The purpose of this study was to assess the efficacy of periodical thermal shocks along a 1-year period on the presence of Legionella spp. in the domestic hot water system of hotels. The Legionella testing results from the period January–December 2019 coming from a group of 77 hotel facilities in the Balearic Islands (Spain) conducting periodical thermal shocks were analyzed. A second group of 44 hotels operating without periodical thermal shocks was used for a comparative analysis. In the facilities where the periodical thermal shock was performed, 16.0% of the results (429 hot water samples collected) were positive for Legionella spp., compared to 21.1% (298 samples), where periodical thermal shock was not performed. Overall, in the thermal shock group, 32.5% of the sites presented at least 1 positive sample along the period of study versus 45.5% in the control group. None of these differences was statistically significant (p-value > 0.05). These findings suggest that the efficacy of regular thermal shock for long-term control of Legionella spp. in domestic hot water systems of hotels is low. [ABSTRACT FROM AUTHOR]

In this study, droplet digital PCRTM (ddPCRTM) was used to characterize total Legionella spp. and five specific Legionella species from source (groundwater) to exposure sites (taps and cooling towers). A total of 42–10 L volume water samples were analyzed during this study: 12 from a reservoir (untreated groundwater and treated water storage tanks), 24 from two buildings (influents and taps), and six from cooling towers, all part of the same water system. The approximate water age (time in the system) for all sample locations are as follows: ~4.5, 3.4, 9.2, 20.8, and 23.2 h (h) for the groundwater to the reservoir influent, reservoir influent to the reservoir effluent, reservoir effluent to building Fa (building names are abbreviated to protect the privacy of site location), building ERC and the cooling towers, respectively. Results demonstrated that gene copies of Legionella spp. (23S rRNA) were significantly higher in the cooling towers and ERC building (p < 0.05) relative to the reservoir and building Fa (closest to reservoir). Legionella spp. (23S rRNA) were found in 100% (42/42) of water samples at concentrations ranging from 2.2 to 4.5 Log10 GC/100 mL. More specifically, L. pneumophila was found in 57% (24/42) of the water samples, followed by L. bozemanii 52% (22/42), L. longbeachae 36% (15/42), L. micdadei 23% (10/42) and L. anisa 21% (9/42) with geometric mean concentrations of 1.7, 1.7, 1.4, 1.6 and 1.7 Log10 GC/100 mL, respectively. Based on this study, it is hypothesized that water age in the distribution system and the premise-plumbing system as well as building management plays a major role in the increase of Legionella spp., (23S rRNA) and the diversity of pathogenic species found as seen in the influent, and at the taps in the ERC building—where the building water quality was most comparable to the industrial cooling towers. Other pathogenic Legionella species besides L.pneumophila are also likely amplifying in the system; thus, it is important to consider other disease relevant species in the whole water supply system—to subsequently control the growth of pathogenic Legionella in the built water environment. [ABSTRACT FROM AUTHOR]

Pathogenic Legionella species grow optimally inside free-living amoebae to concentrations that increase risks to those who are exposed. The aim of this study was to screen a complete drinking water system and cooling towers for the occurrence of Acanthamoeba spp. and Naegleria fowleri and their cooccurrence with Legionella pneumophila, Legionella anisa, Legionella micdadei, Legionella bozemanii, and Legionella longbeachae. A total of 42 large-volume water samples, including 12 from the reservoir (water source), 24 from two buildings (influents to the buildings and exposure sites (taps)), and six cooling towers were collected and analyzed using droplet digital PCR (ddPCR). N. fowleri cooccurred with L. micdadei in 76 (32/42) of the water samples. In the building water system, the concentrations of N. fowleri and L. micdadei ranged from 1.5 to 1.6 Log10 gene copies (GC)/100 mL, but the concentrations of species increased in the cooling towers. The data obtained in this study illustrate the ecology of pathogenic Legionella species in taps and cooling towers. Investigating Legionella's ecology in drinking and industrial waters will hopefully lead to better control of these pathogenic species in drinking water supply systems and cooling towers. [ABSTRACT FROM AUTHOR]

Hotel water systems colonized with Legionella spp. have been the source of travel-associated Legionnaires' disease, and cases, clusters and outbreaks continue to be reported worldwide each year. A total of 132 hotels linked with travel-associated Legionnaires' disease, as reported through the European Legionnaires' Disease Surveillance Network, were inspected and tested for Legionella spp. during 2000–2019 by the public health authorities of the island of Crete (Greece). A total of 3311 samples were collected: 1885 (56.93%) from cold water supply systems, 1387 (41.89%) from hot water supply systems, 37 (1.12%) were swab samples and two (0.06%) were soil. Of those, 685 (20.69%), were collected from 83 (62.89%) hotels, testing positive (≥50 CFU/L) for Legionella pneumophila) serogroups 1–10, 12–14 and non-pneumophila species (L. anisa, L. erythra, L. tusconensis, L. taurinensis, L. birminghamensis, L. rubrilucens, L. londiniesis, L. oakridgensis, L. santicrusis, L. brunensis, L. maceacherii). The most frequently isolated L. pneumophila serogroups were 1 (27.92%) and 3 (17.08%). Significantly higher isolation rates were obtained from hot water supply systems (25.96%) versus cold water systems (16.98%) and swab samples (13.51%). A Relative Risk (R.R.) > 1 (p < 0.0001) was calculated for hot water temperature <55 °C (R.R.: 4.43), chlorine concentrations <0.2 mg/L (R.R.: 2.69), star ratings <4 (R.R.: 1.73) and absence of Water Safety Plan implementation (R.R.: 1.57). [ABSTRACT FROM AUTHOR]

Healthcare facilities are high-risk environments for Legionella disease (LD), including Legionnaires' disease, but transmission in these settings is often overlooked. We used the LD database at the U.S. Department of Veterans Affairs (VA) national healthcare system to assess the type of healthcare exposure for LD cases. Cases were extracted from the database for 1 September 2012 through 31 July 2019, focusing on cases with an overnight stay at a VA facility during the 10-day exposure window prior to symptom onset. Patient medical charts were reviewed for demographics and types of healthcare setting exposure(s). There were 99 LD cases in the cohort: 31.3% were classified as having definite VA exposure, 37.4% were classified as possible VA with inpatient exposure, and 31.3% were classified as possible VA with both inpatient and outpatient exposure. For definite VA LD cases, 67.7% had some type of exposure in the long-term care setting. While 63% of the 99 cases had exposure in the acute care setting only, both the long-term care and acute care settings contributed substantially to the total number of exposure days. A review of patient movement during the exposure period showed the variable and sometimes extensive use of the VA system, and it provides insights useful for epidemiologic investigations and potential preventive actions. [ABSTRACT FROM AUTHOR]

Legionella spp. and other opportunistic premise plumbing pathogens (OPPPs), including Pseudomonas aeruginosa, Mycobacterium avium, Stenotrophomonas maltophilia, and Acinetobacter baumannii, are normal inhabitants of natural waters, drinking water distribution systems and premise plumbing. Thus, humans are regularly exposed to these pathogens. Unfortunately, Legionella spp. and the other OPPPs share a number of features that allow them to grow and persist in premise plumbing. They form biofilms and are also relatively disinfectant-resistant, able to grow at low organic matter concentrations, and able to grow under stagnant conditions. Infections have been traced to exposure to premise plumbing or aerosols generated in showers. A number of measures can lead to reduction in OPPP numbers in premise plumbing, including elevation of water heater temperatures. [ABSTRACT FROM AUTHOR]

The West Bank can be considered as a high-risk area for Legionella prevalence in drinking water due to high ambient temperature, intermittent water supply, frequent pressure loss, and storage of drinking water in roof containers. To assess occurrence of Legionella species, especially L. pneumophila, in the drinking water of the West Bank, the drinking water distribution systems of eight hospitals were sampled over a period of 2.3 years covering the seasonal cycle and the major geographic regions. To gain insight into potential environmental drivers, a set of physico-chemical and microbiological parameters was recorded. Sampling included drinking water and biofilm analyzed by culture and PCR-based methods. Cultivation led to the isolation of 180 strains of L. pneumophila that were genotyped by Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA). Surprisingly, the abundance of culturable L. pneumophila was low in drinking water of the sampling sites, with only three out of eight sites where Legionella was observed at all (range: 30–500 CFU/Liter). By contrast, biofilm and PCR-based analyses showed a higher prevalence. Statistical analyses with physico-chemical parameters revealed a decrease of L. pneumophila abundance for water and biofilm with increasing magnesium concentrations (>30 mg/L). MLVA-genotype analysis of the L. pneumophila isolates and their spatial distribution indicated three niches characterized by distinct physico-chemical parameters and inhabited by specific consortia of genotypes. This study provides novel insights into mechanisms shaping L. pneumophila populations and triggering their abundance leading to an understanding of their genotype-specific niches and ecology in support of improved prevention measures. [ABSTRACT FROM AUTHOR]

Water safety and management programs (WSMP) utilize field measurements to evaluate control limits and monitor water quality parameters including Legionella presence. This monitoring is important to verify that the plan is being implemented properly. However, once it has been determined when and how to sample for Legionella, it is important to choose appropriate collection and processing methods. We sought to compare processing immediate and flushed samples, filtration of different volumes collected, and sample hold times. Hot water samples were collected immediately and after a 2-min flush. These samples were plated directly and after filtration of either 100 mL, 200 mL, or 1 L. Additionally, unflushed samples were collected and processed immediately and after 1, 24, and 48 h of hold time. We found that flushed samples had significant reductions in Legionella counts compared to immediate samples. Processing 100 mL of that immediate sample both directly and after filter concentration yielded the highest concentration and percent sample positivity, respectively. We also show that there was no difference in culture values from time 0 compared to hold times of 1 h and 24 h. At 48 h, there were slightly fewer Legionella recovered than at time 0. However, Legionella counts were so variable based on sampling location and date that this hold time effect was minimal. The interpretation of Legionella culture results depends on the sample collection and processing methods used, as these can have a huge impact on the success of sampling and the validation of control measures. [ABSTRACT FROM AUTHOR]

Growth of Legionella pneumophila and other opportunistic pathogens (OPs) in drinking water premise plumbing poses an increasing public health concern. Premise plumbing is constructed of a variety of materials, creating complex environments that vary chemically, microbiologically, spatially, and temporally in a manner likely to influence survival and growth of OPs. Here we systematically review the literature to critically examine the varied effects of common metallic (copper, iron) and plastic (PVC, cross-linked polyethylene (PEX)) pipe materials on factors influencing OP growth in drinking water, including nutrient availability, disinfectant levels, and the composition of the broader microbiome. Plastic pipes can leach organic carbon, but demonstrate a lower disinfectant demand and fewer water chemistry interactions. Iron pipes may provide OPs with nutrients directly or indirectly, exhibiting a high disinfectant demand and potential to form scales with high surface areas suitable for biofilm colonization. While copper pipes are known for their antimicrobial properties, evidence of their efficacy for OP control is inconsistent. Under some circumstances, copper's interactions with premise plumbing water chemistry and resident microbes can encourage growth of OPs. Plumbing design, configuration, and operation can be manipulated to control such interactions and health outcomes. Influences of pipe materials on OP physiology should also be considered, including the possibility of influencing virulence and antibiotic resistance. In conclusion, all known pipe materials have a potential to either stimulate or inhibit OP growth, depending on the circumstances. This review delineates some of these circumstances and informs future research and guidance towards effective deployment of pipe materials for control of OPs. [ABSTRACT FROM AUTHOR]

Exposure to Legionella spp. contaminated aerosols in hotel settings confers risk for travel-associated Legionnaire's disease (TALD). In this study, we investigated the prevalence of Legionella contamination and its molecular diversity in hotels and resorts across Israel. The study was comprised of a convenience sample of water systems from 168 hotels and resorts countrywide, routinely inspected between March 2015 and February 2017. Isolation and quantitation of Legionella were performed in a water laboratory using the ISO 11731 method. The distribution of Legionella isolates was analyzed according to geography and source. The genetic diversity of a subset of isolates was analyzed by sequence-based typing (SBT) at the National Reference Laboratory for Legionella and compared to the national database. Out of 2830 samples tested, 470 (17%) obtained from 102 different premises (60% of hotels) were positive for Legionella spp. In 230 samples (49% of all positive, 8% of total samples), accounting for 37% of hotels, Legionella spp. counts exceeded the regulatory threshold of 1000 CFU/L. The most frequently contaminated water sources were cooling towers (38%), followed by faucets, hot tubs, water lines, and storage tanks (14–17% each). Furthermore, 32% and 17% of samples obtained from cooling towers and hot tubs, respectively, exceeded the regulatory thresholds. SBT was performed on 78 strains and revealed 27 different sequence types (STs), including two novel STs. The most prevalent STs found were ST1 (26%), ST87 (10%), ST93 (6%), and ST461 and ST1516 (5% each). Several L. pneumophila STs were found to be limited to certain geographical regions. This is the first study to investigate the prevalence and diversity of Legionella in hotels and resorts in Israel during non-outbreak environmental inspections. These findings will inform risk assessment, surveillance, and control measures of TALD. [ABSTRACT FROM AUTHOR]

Testing drinking water systems for the presence of Legionella colonization is a proactive approach to assess and reduce the risk of Legionnaires' disease. Previous studies suggest that there may be a link between Legionella positivity in the hot water return line or certain water quality parameters (temperature, free chlorine residual, etc.) with distal site Legionella positivity. It has been suggested that these measurements could be used as a surrogate for testing for Legionella in building water systems. We evaluated the relationship between hot water return line Legionella positivity and other water quality parameters and Legionella colonization in premise plumbing systems by testing 269 samples from domestic cold and hot water samples in 28 buildings. The hot water return line Legionella positivity and distal site positivity only demonstrated a 77.8% concordance rate. Hot water return line Legionella positivity compared to distal site positivity had a sensitivity of 55% and a specificity of 96%. There was poor correlation and a low positive predictive value between the hot water return line and distal outlet positivity. There was no correlation between Legionella distal site positivity and total bacteria (heterotrophic plate count), pH, free chlorine, calcium, magnesium, zinc, manganese, copper, temperature, total organic carbon, or incoming cold-water chlorine concentration. These findings suggest that hot water return line Legionella positivity and other water quality parameters are not predictive of distal site positivity and should not be used alone to determine the building's Legionella colonization rate and effectiveness of water management programs. [ABSTRACT FROM AUTHOR]

Occurrence of Legionella pneumophila can be relevant to the installation age and the presence of heterotrophic plate counts (HPCs). This research illustrates L. pneumophila contamination of hospital water in accordance with the installation age and the presence of HPCs. One hundred and fifty samples were collected from hot and cold water systems and cultured on R2A and BCYE agar. L. pneumophila identification was done via specific biochemical tests. HPCs and L. pneumophila were detected in 96 and 37.3 % of the samples, respectively. The mean of HPCs density was 947 ± 998 CFU/ml; therefore, 52 % of the samples had higher densities than 500 CFU/ml. High densities of HPCs (>500 CFU/ml) led to colonization of L. pneumophila (≥1000 CFU/ml), mainly observed in cooling systems, gynecological, sonography, and NICU wards. Chi2 test demonstrated that higher densities (>500 CFU/ml) of HPCs and L. pneumophila contamination in cold water were more frequent than warm water (OR: 2.3 and 1.49, respectively). Univariate regressions implied a significant difference between HPCs density and installation age in positive and negative tests of L. pneumophila (OR = 1.1, p < 0.001, OR = 1.2, p < 0.001). Mann-Whitney U test implied the significant effects of HPCs and installation age on L. pneumophila occurrences (p < 0.001). Spearman correlation and multivariate linear regression revealed significant differences between L. pneumophila and HPCs densities (rs = 0.33, p < 0.001 and ß = 0.11, p = 0.02), but nonsignificant difference with installation age (rs = 0.33, p < 0.001 and ß = 0.0, p = 0.91). The occurrence of L. pneumophila, HPCs, and installation age are relevant; so, plumbing system renovation with appropriate materials and promotion of the effective efforts for hospital's water quality assurance is highly recommended. [ABSTRACT FROM AUTHOR]