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Purpose: Water-bearing instruments and treatments in dental units produce aerosols originating from the dental unit waterlines (DUWLs), which are often microbially contaminated. Particularly, the presence of Legionella mainly realized as aerosols leads to a risk of infection in patients and dental staff., Methods: Here, we record the general bacteriological status of DUWLs in Germany and investigated the prevalence of Legionella spp., with a focus on identification and occurrence of distinct species considering the various aspects of dental practice such as dental chair equipment, disinfection methods, and temperatures., Results: Out of 3789 water samples of 459 dental practices, collected in the years 2019 and 2020, 36.4% were Legionella positive with predominance of L. anisa (97.89%) identified by MALDI-TOF biotyping. L. pneumophila was detected very rarely. Risk factor analysis revealed that temperatures >20°C are a significant factor for increased Legionella colonization., Conclusion: In order to minimize the risk of infection, routine monitoring of the water quality in dental chair units is recommended with regard to general microbiological loads and to the presence of Legionella as opportunistic pathogen as well as the regular application of routine disinfection procedures., (© 2023. The Author(s).)

Legionella, the causative agent of Legionnaires' disease, is an emerging concern for water utilities. Passaic Valley Water Commission (PVWC) is a public drinking water supplier, which provides treated surface water to approximately 800,000 customers in New Jersey. To evaluate the occurrence of Legionella in the PVWC distribution system, swab, first draw, and flushed cold water samples were collected from total coliform sites (n = 58) during a summer and winter sampling event. Endpoint PCR detection methods were combined with culture for Legionella detection. Among 58 total coliform sites during the summer, 17.2% (10/58) of first draw samples were positive for 16S and mip Legionella DNA markers and 15.5% (9/58) in flushed samples. Across both summer and winter sampling, a total of four out of 58 sites had low-level culture detection of Legionella spp. (0.5-1.6 CFU/mL) among first draw samples. Only one site had both a first and flush draw detection (8.5 CFU/mL and 1.1 CFU/mL) for an estimated culture detection frequency of 0% in the summer and 1.7% in the winter among flushed draw samples. No L. pneumophila was detected by culture. Legionella DNA detection was significantly greater in the summer than in the winter, and detection was greater in samples collected from areas treated with phosphate. No statistical difference was found between first draw and flush sample detection. Total organic carbon, copper, and nitrate were significantly associated with Legionella DNA detection., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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.

Aims: An evaluation was made of the prevalence of Legionella species in hot water distribution systems in the city of Bologna (Italy) and their possible association with bacterial contamination (total counts and Pseudomonadaceae) and the chemical characteristics of the water (pH, Ca, Mg, Fe, Mn, Cu, Zn and Total Organic Carbon, TOC)., Methods and Results: A total of 137 hot water samples were analysed: 59 from the same number of private apartments, 46 from 11 hotels and 32 from five hospitals, all using the same water supply. Legionella species were detected in 40.0% of the distribution systems, L. pneumophila in 33.3%. The highest colonization was found in the hot water systems of hospitals (93.7% of samples positive for L. pneumophila, geometric mean: 2.4 x 10(3) CFU l(-1)), followed by the hotels (60.9%, geometric mean: 127.3 CFU l(-1)) and the apartments with centralized heating (41.9%, geometric mean: 30.5 CFU l(-1)). The apartments with independent heating systems showed a lower level of colonization (3.6% for Legionella species), with no evidence of L. pneumophila. Correlation analysis suggests that copper exerts an inhibiting action, while the TOC tends to favour the development of L. pneumophila. No statistically significant association was seen with Pseudomonadaceae, which were found at lower water temperatures than legionellae and in individual distribution points rather than in the whole network., Conclusions: The water recirculation system used by centralized boilers enhances the spreading of legionellae throughout the whole network, both in terms of the number of colonized sites and in terms of CFU count., Significance and Impact of the Study: Differences in Legionella colonization between types of buildings are not due to a variation in water supply but to other factors. Besides the importance of water recirculation, the study demonstrates the inhibiting action of copper and the favourable action of TOC on the development of L. pneumophila.

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.

A multicentric Italian investigation on legionnaires' disease is in course to clarify host factors as well as pathogen associated characteristics involved in the infection/disease. The main goal of the research plan is to account for some critical aspects concerning identification and prevention of legionellosis. To improve knowledge on factors associated with Legionella spp colonisation in hot waters, to detect cases and to characterize risk factors in subjects which develop pneumonia are specific objectives of the research programme. Preliminary results show that hot waters of houses and hotels are frequently contaminated (22.6% and 54.6%, respectively), mainly by L. pneumophila. Microbial concentrations were low in domestic waters (<1.000 ufc/l), but higher in samples from the hotels (geom. mean 1.85 x 10(3) ufc/l). Warming system, age of the plant, type of building were risk factors significantly associated with Legionella spp positivity. The active surveillance on patients affected by pneumonia with search for Legionella urinary antigen allowed the identification of 34 cases, 3 of which of nosocomial origin, corresponding to 4.2% of the screened pneumonia. After informed consent, 26 subjects were recruited for a case-control-study to clarify risk factors for the disease.

The relationship between Legionella and Pseudomonas spp contamination and mineral content of domestic hot waters was investigated in a cross-sectional Italian survey. Pseudomonas spp contamination was associated with elevated Ca, but lower Fe and free chlorine content in the water. Waters with Cu concentrations > 50 microg/L prevented Legionella spp colonisation (OR = 0.14, 95% CI = 0.02-1.13), whereas the risk of legionellae occurrence increased in waters with Mn levels > 3 microg/L (OR = 2.37, 95% CI = 1.06-5.30). Furthermore, Mn was positively associated with the risk of colonisation by eterotrophic bacteria growing at either 22 and 36 degrees C. Legionella species and serogroups were differently distributed according to Cu and Mn concentrations, suggesting that Legionella strains may have a different sensibility/resistance to trace elements. A specific action of Cu as decontamination factor is suggested and the consideration of Mn as a risk indicator for bacterial colonisation and biofilm presence is proposed.

Background: It's pivotal to control the presence of legionella in sanitary structures. So, it's important to determine the risk factors associated with Legionella colonization in health care centres. In recent years that is why new diagnostic techniques have been developed., Objective: To evaluate risks factors for Legionella colonization using a novel and more sensitive Legionella positivity index., Methods: A total of 204 one-litre water samples (102 cold water samples and 102 hot water samples), were collected from 68 different sampling sites of the hospital water system and tested for Legionella spp. by two laboratories using culture, polymerase chain reaction and a method based on immunomagnetic separation (IMS). A Legionella positivity index was defined to evaluate Legionella colonization and associated risk factors in the 68 water samples sites. We performed bivariate analyses and then logistic regression analysis with adjustment of potentially confounding variables. We compared the performance of culture and IMS methods using this index as a new gold standard to determine if rapid IMS method is an acceptable alternative to the use of slower culture method., Results: Based on the new Legionella positivity index, no statistically significant differences were found neither between laboratories nor between methods (culture, IMS). Positivity was significantly correlated with ambulatory health assistance (p = 0.05) and frequency of use of the terminal points. The logistic regression model revealed that chlorine (p = 0.009) and the frequency of use of the terminal points (p = 0.001) are predictors of Legionella colonization. Regarding this index, the IMS method proved more sensitive (69%) than culture method (65.4%) in hot water samples., Significance: We showed that the frequency of use of terminal points should be considered when examining environmental Legionella colonization, which can be better evaluated using the provided Legionella positivity index. This study has implications for the prevention of Legionnaires' disease in hospital settings., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

e. leoni, g. de luca, p.p. legnani, r. sacchetti, s. stampi and f. zanetti. 2004.An evaluation was made of the prevalence ofLegionellaspecies in hot water distribution systems in the city of Bologna (Italy) and their possible association with bacterial contamination (total counts and Pseudomonadaceae) and the chemical characteristics of the water (pH, Ca, Mg, Fe, Mn, Cu, Zn and Total Organic Carbon, TOC).A total of 137 hot water samples were analysed: 59 from the same number of private apartments, 46 from 11 hotels and 32 from five hospitals, all using the same water supply.Legionellaspecies were detected in 40·0% of the distribution systems,L. pneumophilain 33·3%. The highest colonization was found in the hot water systems of hospitals (93·7% of samples positive forL. pneumophila, geometric mean: 2·4 × 103 CFU l−1), followed by the hotels (60·9%, geometric mean: 127·3 CFU l−1) and the apartments with centralized heating (41·9%, geometric mean: 30·5 CFU l−1). The apartments with independent heating systems showed a lower level of colonization (3·6% forLegionellaspecies), with no evidence ofL. pneumophila. Correlation analysis suggests that copper exerts an inhibiting action, while the TOC tends to favour the development ofL. pneumophila. No statistically significant association was seen with Pseudomonadaceae, which were found at lower water temperatures than legionellae and in individual distribution points rather than in the whole network.The water recirculation system used by centralized boilers enhances the spreading of legionellae throughout the whole network, both in terms of the number of colonized sites and in terms of CFU count.Differences inLegionellacolonization between types of buildings are not due to a variation in water supply but to other factors. Besides the importance of water recirculation, the study demonstrates the inhibiting action of copper and the favourable action of TOC on the development ofL. pneumophila. [ABSTRACT FROM AUTHOR]

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 and Objectives Legionella is a genus of gram-negative bacteria common in soil and aquatic systems and is associated with protists. They have emerged as a pathogenic group due to the increased use and poor maintenance of artificial water environments. This study aims at phenotypic and molecular identification of Legionella in water and swab samples collected from water-based recreational sites. The biofilm-forming ability of Legionella on exposure to various temperatures and iron concentrations was also studied. Methods A total of 60 samples including 30 swab samples and 30 water samples (decorative fountain ponds, swimming pools, garden sprinklers, drip irrigation system) were collected from in and around Mangalore, Karnataka, India. From each source, swab and water samples were collected as per the Indian standard IS: 1622. The collected samples were processed within 4 hours of collection. The samples were subjected to microbiological and chemical estimation followed by filtration through a 0.2 µm membrane filter. Isolation of Legionella from collected samples was performed as per US Centers for Disease Control and Prevention guidelines 2005. The positive isolates were then checked for biofilm-forming ability at various temperatures (25°C, 35°C, and 45°C) and iron concentration (3 mg/L, 30 mg/L, and 300 mg/L) using crystal violet assay. Results Out of 30, water and swab samples tested, one water sample from a garden sprinkler, swimming pool, and one from both water and swab samples from a decorative fountain showed the presence of Legionella. A biofilm study of Legionella at various temperatures and iron concentrations categorized the bacteria as a moderate biofilm former. Conclusion This study revealed that most of the water and swab samples were found to be negative for Legionella that is quite encouraging and the contamination of water systems in recreational facilities can be reduced by decontamination techniques and proper hygienic practices. [ABSTRACT FROM AUTHOR]

Copyright of Epidemiological Review / Przegląd Epidemiologiczny is the property of National Institute of Public Health - National Institute of Hygiene and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

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]

Copyright of SKEI - International Interdisciplinary Journal / Medunarodni Interdisciplinarni Casopis is the property of University Vitez, Bosnia & Herzegovina and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Legionella longbeachae is an important cause of Legionnaires' disease in Australasia and is associated with exposure to potting soils. Our aim was to identify ways to reduce the load of L. longbeachae in potting soils. Inductively-coupled plasma optical emission spectrometry (ICP-OES) of an all-purpose potting mix showed copper (Cu) concentrations (mg/kg) range from 15.8 to 23.6. Zinc (Zn) and manganese (Mn) were significantly higher than Cu ranging from 88.6–106 to 171–203, respectively. Minimal inhibitory and bactericidal concentrations of 10 salts used in the horticultural industry were determined for Legionella species in buffered yeast extract (BYE) broth. For L. longbeachae (n  = 9) the median (range) minimum inhibitory concentration (MIC) (mg/L) of copper sulfate was 31.25 (15.6–31.25), zinc sulfate 31.25 (7.81–31.25), and manganese sulfate 31.25 (7.81–62.5). The MIC and minimum bactericidal concentration (MBC) were within one dilution of each other. Susceptibility to Cu and Zn salts increased as the concentration of pyrophosphate iron in the media decreased. The MIC values for these three metals against Legionella pneumophila (n  = 3) and Legionella micdadei (n  = 4) were similar. Combinations of Cu, Zn, and Mn were additive. Legionella longbeachae has similar susceptibility to Cu and other metal ions in comparison to L. pneumophila. [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]

Copyright of Annali di Igiene, Medicina Preventiva e di Comunità is the property of Societa Editrice Universo s.r.l. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The Federation of Bosnia and Herzegovina is becoming a more attractive tourist destination with a significant increase in the number of tourists and overnight stays. The purpose of this study was to evaluate the presence of Legionella spp. in tourist facilities of FB&H. The aim is to point out the growing trend of the presence of Legionella spp. and to investigate the connection of environmental factors, temperature, and chlorine as disinfectants, with the occurrence of these bacteria in the water systems of tourist and public facilities. The research included the period from 2014 to 2018. Of the total number of analyzed samples (n = 237), the presence of legionella was confirmed in 61 samples (25.74 %), while in 176 (74.26 %) samples it was not confirmed. Among the confirmed cases of legionella (n = 61), the part of those in which the absence of chlorine was determined was 40 (65.58 %). There is a significant statistical difference of positive samples about to with concerning to the presence of chlorine in the water. Of the total number of positive samples (n = 61), 29 were associated with hot water. The mean temperature in the positive samples was 51.79 °C (s.d. = 10.94, min.31.9, max.69). The results of the analysis indicate seasonal trends in the presence of legionella with a peak in months during the preparation for the winter tourist season (fourth quarter). [ABSTRACT FROM AUTHOR]

Legionella pneumophila is an opportunistic bacterial respiratory pathogen that is one of the leading causes of drinking water outbreaks in the United States. Dental offices pose a potential risk for inhalation or aspiration of L. pneumophila due to the high surface area to volume ratio of dental unit water lines—a feature that is conducive to biofilm growth. This is coupled with the use of high-pressure water devices (e.g., ultrasonic scalers) that produce fine aerosols within the breathing zone. Prior research confirms that L. pneumophila occurs in dental unit water lines, but the associated human health risks have not been assessed. We aimed to: (1) synthesize the evidence for transmission and management of Legionnaires' disease in dental offices; (2) create a quantitative modeling framework for predicting associated L. pneumophila infection risk; and (3) highlight influential parameters and research gaps requiring further study. We reviewed outbreaks, management guidance, and exposure studies and used these data to parameterize a quantitative microbial risk assessment (QMRA) model for L. pneumophila in dental applications. Probabilities of infection for dental hygienists and patients were assessed on a per-exposure and annual basis. We also assessed the impact of varying ventilation rates and the use of personal protective equipment (PPE). Following an instrument purge (i.e., flush) and with a ventilation rate of 1.2 air changes per hour, the median per-exposure probability of infection for dental hygienists and patients exceeded a 1-in-10,000 infection risk benchmark. Per-exposure risks for workers during a purge and annual risks for workers wearing N95 masks did not exceed the benchmark. Increasing air change rates in the treatment room from 1.2 to 10 would achieve an ∼85% risk reduction, while utilization of N95 respirators would reduce risks by ∼95%. The concentration of L. pneumophila in dental unit water lines was a dominant parameter in the model and driver of risk. Future risk assessment efforts and refinement of microbiological control protocols would benefit from expanded occurrence datasets for L. pneumophila in dental applications. [ABSTRACT FROM AUTHOR]

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]

Unregulated private wells are understudied potential sources of community‐acquired Legionnaires' disease. Here we conducted a comprehensive survey of 44 homes supplied by private wells in Wake County, North Carolina, quantifying Legionella spp. DNA, Legionella pneumophila DNA, and total bacterial 16S rRNA genes via real‐time polymerase chain reaction in hot and cold drinking water samples, along with culturable L. pneumophila via IDEXX Legiolert in cold drinking water samples. Legionella spp. DNA, L. pneumophila DNA and culturable L. pneumophila were detected in 100, 65·5 and 15·9% of the 44 homes, respectively, and culturable levels were comparable to some municipal surveys applying the same methods. Total coliforms and Escherichia coli were monitored as representative faecal indicators and were found in 20·4 and 0·0% of homes. Within certain sample types, Legionella spp. and L. pneumophila gene copy numbers were positively associated with total bacteria (i.e. total 16S rRNA genes) and water softener use, but were not associated with faecal indicator bacteria, inorganic water parameters or other well characteristics. These findings confirm that occurrence of Legionella and L. pneumophila is highly variable in private wells. Significance and Impact of the Study: Legionella is the leading identified cause of waterborne disease outbreaks associated with US municipal water systems. While Legionella is known to occur naturally in groundwater, prior efforts to characterize its occurrence in unregulated private wells are limited to sampling at the wellhead and not in the home plumbing where Legionella can thrive. This work documents much higher levels of Legionella in home plumbing versus water directly from private wells and examines factors associated with higher Legionella occurrence. [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]

A novel method was used to incorporate in vivo host-pathogen dynamics into a new robust outbreak model for legionellosis. Dose-response and time-dose-response (TDR) models were generated for Legionella longbeachae exposure to mice via the intratracheal route using a maximum likelihood estimation approach. The best-fit TDR model was then incorporated into two L. pneumophila outbreak models: an outbreak that occurred at a spa in Japan, and one that occurred in a Melbourne aquarium. The best-fit TDR from the murine dosing study was the beta-Poisson with exponential-reciprocal dependency model, which had a minimized deviance of 32.9. This model was tested against other incubation distributions in the Japan outbreak, and performed consistently well, with reported deviances ranging from 32 to 35. In the case of the Melbourne outbreak, the exponential model with exponential dependency was tested against non-time-dependent distributions to explore the performance of the time-dependent model with the lowest number of parameters. This model reported low minimized deviances around 8 for the Weibull, gamma, and lognormal exposure distribution cases. This work shows that the incorporation of a time factor into outbreak distributions provides models with acceptable fits that can provide insight into the in vivo dynamics of the host-pathogen system. [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]

Hot water recirculation systems (HWRS) in hotels and nursing homes, which are common in countries such as Spain, have been related to outbreaks of legionellosis. To establish the relationships of microbial and physicochemical parameters, especially protozoa, with the occurrence of Legionella in HWRS, 231 samples from hotels and nursing homes were analysed for Legionella, protozoa, heterotrophic plate counts (HPC) at 22 and 37 °C, Pseudomonas, metals, temperature and others. Legionella pneumophila was the dominant species isolated, and 22 % were sg. 1. The sampling method became particularly important in order to define which factors were involved on the occurrence of Legionella. Results showed that the bacteria and the accompanying microbiota were more abundant in the first flush water whose temperature was lower. The bacteria occurred in those samples with high HPC and were inversely correlated with high temperatures. Multivariate regression showed that a concentration above 1 × 10 CFU/100 mL of HPC at 37 °C, Fe above 0.095 ppm and the presence of protozoa increased significantly the risk of Legionella colonization, while univariant regression showed that the presence of Cu above 0.76 ppm and temperature above 55 °C diminished it. Therefore, to reduce the risk associated with Legionella occurrence in HWRS these parameters should be taken into consideration. [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]

The aim of the present study was to determine the prevalence of Legionella spp. in water systems of hospitals and hotels located in South Western Greece, to study the molecular epidemiology of the isolated strains and their possible association with bacterial contamination (total count and Pseudomonas aeruginosa), the water pH, and temperature. A prevalence survey for Legionella spp. by culturing techniques in water distribution systems of eight hospitals and nine hotels occurred in South Western Greece. Water sampling and microbiological analysis were carried out following the ISO methods. Legionella pneumophila was detected in 33% and 36% of the distribution systems of hospitals and hotels, respectively. Our survey results suggest a frequent prevalence of elevated concentrations of Legionella spp. in water systems of hospitals and hotels. Our investigation has confirmed the need to regularly monitor the microbiological condition of water systems in hospitals and hotels. [ABSTRACT FROM AUTHOR]

Legionella were detected with the direct DNA extraction method, Legionella culture method, and free-living amoebae (FLA) culture method from weak alkaline carbonate spring water in Taiwan. Moreover, we also investigated the existence of Acanthamoeba, Hartmannella, and Naegleria, ubiquitous FLA in aquatic environments, to identify the correlations between existing Legionella. This study reports detecting Legionella in 15 of the 51 weak alkaline carbonate spring water samples (29.4%). This work also found five of the 51 samples (9.8%) analyzed by the direct DNA extraction method, three of the 51 (5.9%) samples analyzed by the Legionella culture method, and 11 of the 51 samples (21.6%) evaluated using the FLA culture method to be positive for Legionella. The most frequently identified Legionella species was the Legionella-like amoebal pathogen ( n = 5), followed by unidentified Legionella spp. ( n = 4), and Legionella pneumophila ( n = 4), Legionella fairfieldensis ( n = 3), and then Legionella rubrilucens ( n = 2). Legionella waltersii was detected once. The occurrence of Acanthamoeba, Hartmannella, and Naegleria were 5.9% (3/51), 52.9% (27/51), and 5.9% (3/51), respectively. All Hartmannella isolates were identified as Hartmannella vermiformis, and Naegleria isolates were all identified as Naegleria australiensis. The three Acanthamoeba isolates were identified as one Acanthamoeba polyphaga and two Acanthamoeba jacobsi. H. vermiformis (40.7%) were Legionella hosts, including all of the amoebae-resistant Legionella detected in the present study. Therefore, the important correlations between Legionella and H. vermiformis require further clarification. The combined results of this survey confirm that Legionella and FLA are ubiquitous in weak alkaline carbonate spring water in Taiwan. [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]

Aims: To compare the standard culture method with a new, rapid test (ScanVIT-Legionella™) using fluorescently labelled gene probes for the detection and enumeration of Legionella spp. The new technique was validated through experiments conducted on both artificially and naturally contaminated water and through an inter-laboratory comparison. Methods and Results: All samples were processed by the ScanVIT test according to the manufacturer's instructions and by a culture method (ISO 11731). ScanVIT detected significantly more positive samples, although concentrations were similar and a strong positive correlation between the two methods was observed ( r = 0·888, P < 0·001). The new test was more accurate in identifying the co-presence of Legionella pneumophila and Leg. non- pneumophila. ScanVIT showed a slightly higher Legionella recovery from water samples artificially contaminated with Leg. pneumophila alone or together with Pseudomonas aeruginosa. Lastly, the inter-laboratory comparison revealed that the ScanVIT test exhibits a lower variability than the traditional culture test (mean coefficient of variation 8·7 vs 16·1%). Conclusions: The results confirmed that the ScanVIT largely overlaps the reference method and offers advantages in terms of sensitivity, quantitative reliability and reduced assay time. Significance and Impact of the Study: The proposed method may represent a useful validated alternative to traditional culture for the rapid detection and quantification of Legionella spp. in water. [ABSTRACT FROM AUTHOR]