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A better understanding of risk factors and the predictive capability of water management program (WMP) data in detecting Legionella are needed to inform the efforts aimed at reducing Legionella growth and preventing outbreaks of Legionnaires' disease. Using WMPs and Legionella testing data from a national lodging organization in the United States, we aimed to (1) identify factors associated with Legionella detection and (2) assess the ability of WMP disinfectant and temperature metrics to predict Legionella detection. We conducted a logistic regression analysis to identify WMP metrics associated with Legionella serogroup 1 (SG1) detection. We also estimated the predictive values for each of the WMP metrics and SG1 detection. Of 5435 testing observations from 2018 to 2020, 411 (7.6%) had SG1 detection, and 1606 (29.5%) had either SG1 or non-SG1 detection. We found failures in commonly collected WMP metrics, particularly at the primary test point for total disinfectant levels in hot water, to be associated with SG1 detection. These findings highlight that establishing and regularly monitoring water quality parameters for WMPs may be important for preventing Legionella growth and subsequent disease. However, while unsuitable water quality parameter results are associated with Legionella detection, this study found that they had poor predictive value, due in part to the low prevalence of SG1 detection in this dataset. These findings suggest that Legionella testing provides critical information to validate if a WMP is working, which cannot be obtained through water quality parameter measurements alone.

Legionellosis is a disease caused by the bacterium Legionella that most commonly presents as Legionnaires' disease (LD), a severe form of pneumonia. From 2015 to 2019, an average of 438 LD cases per year were reported in Canada. However, it is believed that the actual number of cases is much higher, since LD may be underdiagnosed and underreported. The purpose of this study was to develop an estimate of the true incidence of illnesses, hospitalizations, and deaths associated with LD in Canada. Values were derived using a stochastic model, based on Canadian surveillance data from 2015 to 2019, which were scaled up to account for underdiagnosis and underreporting. Overall, there were an estimated 1,113 (90% CrI: 737-1,730) illnesses, 1,008 (90% CrI: 271-2,244) hospitalizations, and 34 (90% CrI: 4-86) deaths due to domestically acquired waterborne LD annually in Canada from 2015 to 2019. It was further estimated that only 36% of illnesses and 39% of hospitalizations and deaths were captured in surveillance, and that 22% of illnesses were caused by Legionella serogroups and species other than Legionella pneumophila serogroup 1 (non-Lp1). This study highlights the true burden and areas for improvement in Canada's surveillance and detection of LD.

Objective Recommendations on the timing of Legionella urinary antigen tests for community-acquired pneumonia patients differ among guidelines in Japan, the United States, and European nations. We therefore evaluated the association between the timing of urinary antigen tests and in-hospital mortality in patients with Legionella pneumonia. Methods We conducted a retrospective cohort study using the Diagnosis Procedure Combination database, a nationwide database of acute care inpatients in Japan. Patients who underwent Legionella urinary antigen tests on the day of admission formed the tested group. Patients who were tested on day 2 of admission or later or were unexamined formed the control group. We performed a propensity score matching analysis to compare in-hospital mortality, length of hospital stay and duration of antibiotics use between the two groups. Results Of the 9,254 eligible patients, 6,933 were included in the tested group. One-to-one propensity score matching generated 1,945 pairs. The tested group had a significantly lower 30-day in-hospital mortality than the control group (5.7 vs. 7.7%; odds ratio, 0.72; 95% confidence intervals, 0.55-0.95; p=0.020). The tested group also showed a significantly shorter length of stay and duration of antibiotics use than the control group. Conclusion Urine antigen testing upon admission was associated with better outcomes in patients with Legionella pneumonia. Urine antigen tests upon admission may be recommended for all patients with severe community-acquired pneumonia.

Legionnaires disease is a serious infection acquired by inhalation of water droplets from human-made building water systems that contain Legionella bacteria. On July 11 and 12, 2022, Napa County Public Health (NCPH) in California received reports of three positive urinary antigen tests for Legionella pneumophila serogroup 1 in the town of Napa. By July 21, six Legionnaires disease cases had been confirmed among Napa County residents, compared with a baseline of one or two cases per year. NCPH requested assistance from the California Department of Public Health (CDPH) and CDC to aid in the investigations. Close temporal and geospatial clustering permitted a focused environmental sampling strategy of high-risk facilities which, coupled with whole genome sequencing results from samples and investigation of water system maintenance, facilitated potential linking of the outbreak with an environmental source. NCPH, with technical support from CDC and CDPH, instructed and monitored remediation practices for all environmental locations that tested positive for Legionella. The investigation response to this community outbreak illustrates the importance of interdisciplinary collaboration by public health agencies, laboratory support, timely communication with the public, and cooperation of managers of potentially implicated water systems. Timely identification of possible sources, sampling, and remediation of any facility testing positive for Legionella is crucial to interrupting further transmission., Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. Karen Relucio reports support from Napa County Health & Human Services Agency for travel to and attendance at the Health Officers’ Association of California Fall Semiannual Meeting in October 2022; serving on the Board of Directors for the Napa Solano Medical Society and the Health Officers Association of California, serving as the current past president and executive committee member of the California Conference of Local Health Officers, and serving as co-chair of the Napa Opioid Safety Coalition; stock option payments made through Perlita’s Daughters LLC from Johnson & Johnson, Medtronic, Pfizer, and GE Healthcare; and managing partnership in Perlita’s Daughters LLC equities, cash and real estate holdings in Illinois unrelated to this outbreak and none related to hotels or health care entities. No other potential conflicts of interest were disclosed.

Background: Legionnaires' disease is caused by inhalation or aspiration of small water droplets contaminated with Legionella, commonly found in natural and man-made water systems and in moist soil. Over the past 5 years, notification rates of this disease have almost doubled in the European Union (EU) / European Environmental Agency (EEA), from 1.4 in 2015 to 2.2 cases per 100,000 population in 2019. Some studies show that the greater presence of the microorganism in the water network and the increase in cases of legionellosis could be related to the variations in some environmental factors, such as air temperature, which may influence the water temperature., Study Design: Climate change is currently a prominent topic worldwide because of its significant impact on the natural environment. It is responsible for the increase in numerous waterborne pathologies. The purpose of this study was to correlate the air temperature recorded in Apulia region from January 2018 to April 2023 with the presence of Legionella in the water networks of public and private facilities and the incidence rates of legionellosis during the same period., Methods: During the period from January 2018 to April 2023, water samples were collected from facilities involved in legionellosis cases and analyzed for Legionella. During the same period, all the cases notified to the regional epidemiological observatory (OER-Apulia) were included in this study. Statistical analyses were conducted using the Shapiro-Wilk test to determine whether the Legionella load was distributed normally, the Wilcoxon rank sum test to compare the air temperatures (average and range) of the negative and positive samples for Legionella detection, and the multivariate analysis (Poisson regression) to compare the Legionella load with the water sample temperature, average air temperature, and temperature range on the day of sampling. The Wilcoxon test for paired samples was used to compare legionellosis cases between the warmer and colder months., Results: Overall, 13,044 water samples were analyzed for Legionella and 460 cases of legionellosis were notified. Legionella was isolated in 20.1% of the samples examined. The difference in the air temperature between negative samples and positive samples was statistically significant (p-value < 0.0001): on days when water samples tested positive for Legionella a higher temperature range was observed than on days when water samples tested negative (p-value = 0.004). Poisson regression showed a direct correlation between Legionella load, water temperature, and average air temperature. The incidence of legionellosis cases in warmer months was higher than in colder months (p-value = 0.03)., Conclusions: Our study highlights a significant increase in the load of Legionella in the Apulian water network, and an association between warmer temperatures and legionellosis incidence. In our opinion, further investigations are needed in different contexts and territories to characterize the epidemiology of legionellosis, and to explain its extreme variability in different geographical areas and how these data may be influenced by different risk factors.

Legionella , the bacterium that causes Legionnaires' disease, can grow and spread in building water systems and devices. The COVID-19 pandemic impacted building water systems through reductions in water usage. Legionella growth risk factors can be mitigated through control measures, such as flushing, to address stagnation, as part of a water management program (WMP). A national lodging organization (NLO) provided WMP data, including Legionella environmental testing results for periods before and during the pandemic. The statistical analysis revealed an increased risk of water samples testing positive for Legionella during the pandemic, with the greatest increase in risk observed at the building's cold-water entry test point. Sample positivity did not vary by season, highlighting the importance of year-round Legionella control activities. The NLO's flushing requirements may have prevented an increased risk of Legionella growth during the pandemic. However, additional control measures may be needed for some facilities that experience Legionella detections. This analysis provides needed evidence for the use of flushing to mitigate the impacts of building water stagnation, as well as the value of routine Legionella testing for WMP validation. Furthermore, this report reinforces the idea that WMPs remain the optimal tool to reduce the risk of Legionella growth and spread in building water systems.

Legionnaires' disease (LD) is a severe pneumonia mainly caused by the bacterium Legionella pneumophila. Although many environmental sources of LD have been described, the sources of the majority of non-outbreak LD cases have not been identified. In several outbreaks in the Netherlands, wastewater treatment plants (WWTPs) were identified as the most likely source of infection. In this study, four criteria for Legionella growth and emission to air and surface waters were selected based on the literature and a risk matrix was drafted. An inventory was made of all WWTPs and their characteristics in the Netherlands. The risk matrix was applied to identify WWTPs at risk for Legionella growth and emission. Wastewater was collected at WWTPs with moderate to high risk for Legionella growth and emission. In 18% of the sampled WWTPs, Legionella spp. was detected using culture methods. The presented risk matrix can be used to assess the risks of Legionella growth and emission for WWTPs and support surveillance by prioritizing WWTPs. When Legionella is detected in the wastewater, it is recommended to take action to prevent emission to air or discharge on surface waters and, if possible, reduce the Legionella concentration.

Legionella pneumonia is a fatal disease caused by Legionella pneumophila, a bacterium belonging to the genus Legionella. The incidence of this disease has been increasing since 2005 and has continued to increase following the COVID-19 pandemic in Japan. Furthermore, Legionella pneumonia mortality rates have increased slightly since the pandemic due to some plausible reasons. The increased proportion of older patients with legionellosis might affect it because advanced age is a major risk factor for disease mortality. Additionally, physicians were focused on COVID-19 while examining febrile patients; therefore, they might have missed the early diagnosis of other respiratory infections, including Legionella pneumonia., Competing Interests: Conflict of Interest The authors declare no conflict of interest., (Copyright © 2023 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.)

Only 4 % of reported Legionnaires' disease (LD) cases are outbreak-associated and the remaining 96 % are sporadic, for which no known source of Legionella is identified. Although outbreaks of LD are linked to cooling towers, decorative fountains, spas and hot tubs, and other sources, the drivers of sporadic LD are less known. Residential premise plumbing is likely an important source of aerosol exposure and there are unique features of premise plumbing which could lead to proliferation of Legionella. A sampling study of Legionella in single-family homes was undertaken in NJ from 2020 to 2021 which included a household characteristic survey and collection of hot water temperature and chlorine residual during sampling. A total of 94 homeowners residing in owner-occupied, single-family units with individual hot water systems were recruited to participate through two mechanisms (1) Legionnaire's disease case-patients and (2) non-case volunteers from each NJ county. Among the 94 single-family homes sampled, 15 % had least one sample positive for Legionella by culture and 57 % had at least one sample with detection of Legionella DNA markers by PCR. Chlorine residual, hot water temperature, and season were independently associated with increased detection of Legionella in home water samples. There was limited or inconsistent evidence of the role of household characteristic factors in Legionella detection. This study identified season, insufficient chlorine residual and hot water temperature as risk factors for Legionella detection in single-family homes. Findings from this work can promote additional partnership between public health and water utilities in improving chlorine residuals in residential communities and educating homeowners on best practices for home water management., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jessie Gleason reports financial support was provided by Centers for Disease Control and Prevention., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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.)

Background: Inhalation of Legionella-containing aerosols generated by cooling towers (CT) and evaporative condensers (EC) where water risk management is not performed correctly has been linked to a high percentage of community outbreaks of Legionnaires' disease (LD). Likewise, microbiological and physicochemical characteristics of the water in these facilities have been associated with this bacterium. The main aim of this study was to assess the risk of Legionella colonization in CT and EC based on the data for microbiological and physicochemical water quality provided by the Environmental Health Department and Laboratory of the City Council of L'Hospitalet de Llobregat (Barcelona, Spain)., Methods: Legionella was analysed in 789 samples collected from 127 CT and EC in 46 companies in Catalonia from 2002 to 2019. A two-step logistic regression analysis was carried out to assess the risk of colonization by Legionella in the studied facilities according to the microbiological (aerobic heterotrophic bacteria) and physicochemical (pH, alkalinity, hardness, turbidity, conductivity, total iron and Langelier Index) water parameters. The optimal cut-off points for the water parameters predictive of Legionella contamination were defined as the values on the receiver operating characteristic (ROC) curve where sensitivity and specificity were jointly maximized., Results: Legionella was isolated in 8.49% of the 789 analysed samples, 22.39% of which were heavily contaminated (with counts higher than 1.0 × 10 4  CFU/l). L. pneumophila was isolated in 82.09% of the samples, with 41.82% belonging to serogroup 1. Logistic regression analysis revealed that aerobic heterotrophic bacteria concentrations ≥6.90 × 10 2  CFU/ml [Odds ratios (OR) (95% CI) = 3.56 (1.39-9.43), p = 0.01], a pH ≥ 8.70 [OR (95% CI) = 3.60 (1.34-10.09), p = 0.01], and water hardness ≥5.72 × 10 2  mg/l [OR (95% CI) = 6.30 (2.34-18.56), p < 0.001] were each independently associated with a higher risk of CT and EC colonization by Legionella., Conclusions: The present study shows the importance of risk assessment for improving the control measures aimed at preventing or reducing Legionella populations in CT and EC, thus minimizing potential dangers for public health., Competing Interests: Declarations of competing interest None., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Cooling towers have been linked to Legionnaires' disease cases and outbreaks. Legionella pneumophila results (from a culture-based method) are presented for 557 cooling towers across the City of Vancouver, Canada for 2021. Results of 10 CFU/mL or greater (defined as exceedances) were reported for 30 cooling towers (5.4%), including six >1,000 CFU/mL, and L. pneumophila serogroup 1 (sg1) was identified in 17 of these cooling towers (out of 28 with serogroup-level analysis). The data indicate highly localised Legionella issues, with exceedances concentrated within 16 facilities, including two hospitals. In the 3 months preceding each cooling tower exceedance, the nearest municipal water sampling station had a free chlorine residual of at least 0.46 mg/L and a temperature of <20 °C. There was not a statistically significant correlation between the L. pneumophila concentration of a cooling tower in exceedance and the municipal water free chlorine residual, temperature, pH, turbidity or conductivity. There was a statistically significant negative correlation between the concentrations of L. pneumophila sg1 and other L. pneumophila serogroups in cooling towers. This unique dataset underscores the pivotal role of building owners and managers in preventing the growth of Legionella bacteria and the value of regulations to verify operations and maintenance practices.

Legionella pneumophila, the primary causative agent of Legionnaires' disease, is classified into at least 15 serogroups (SGs). Before genotyping, serotyping is first performed to limit the sources of L. pneumophila infections that caused an outbreak. In addition to conventional assays using monoclonal or polyclonal antisera, serotyping using multiplex polymerase chain reaction (M-PCR) was recently developed for L. pneumophila. In this study, we applied the M-PCR system to 41 strains that remained to be SGUT (untypable) by slide agglutination tests among the 220 L. pneumophila strains isolated from bath water in Kobe City during 2016-2020, to determine SG-genotypes (SGg) by PCR amplification of the specific target gene of the SGs. Among the 41 SGUT strains, SGg4/10/14 was the most predominant (24/41, 58.5%), followed by SGg1 (7/41, 17.1%). Seven strains, except for the strains determined as SGg1, were identified as belonging to a single SGg by M-PCR serotyping (SGg5 [3/41, 7.3%], SGg8 [3/41, 7.3%], and SGg7 [1/41, 2.4%]). Furthermore, we found that the seven strains identified as SGg1 harbored particular genotypes. In conclusion, the M-PCR serotyping assay will be helpful for investigating the distribution of L. pneumophila in environmental and clinical settings.

Abstract: Legionnaires disease is an important, relatively uncommon, yet well known form of atypical community-acquired pneumonia (CAP). If not appropriately treated in the early stage of infection, it can be fatal. Several factors increase the risk of contracting Legionnaires disease, including age over 50 years, chronic cardiovascular disease, underlying respiratory disease, chronic renal disease, diabetes, smoking, any immunosuppressing condition, travel history, and certain types of employment. Legionnaires disease can present without the usual symptoms associated with pneumonia. The incidence of Legionnaires disease is increasing, and because of its morbidity and mortality, clinicians should consider it in the differential when treating patients with CAP. This article reviews the pathophysiology and treatment of Legionnaires disease and when to refer patients to tertiary centers for higher levels of care, including extracorporeal membrane oxygenation., (Copyright © 2022 American Academy of Physician Associates.)

Currently available methods for the laboratory investigation of Legionella pneumophila outbreaks require organism culture. The ability to sequence L. pneumophila directly from clinical samples would significantly reduce delays. Here, we develop a method for targeted next-generation sequencing (NGS) of selected L. pneumophila genes utilizing a CRISPR/Cas9-based target enrichment system. We determine the method's utility by typing cultured L. pneumophila isolates and subsequently apply the method directly to patient samples. We sequenced 10 L. pneumophila isolates by 2 methods, (i) whole-genome sequencing (WGS) and (ii) targeted (CRISPR/Cas9-based) f inding l ow- a bundance s equences by h ybridization (FLASH)-NGS, sequencing 57 selected genes. The targeted NGS of 57 genes was more efficient than WGS, and phylogenetic analysis of the 57 genes yielded the same classification of the L. pneumophila isolates as that based on analysis of whole-genome data. Furthermore, targeted NGS of L. pneumophila performed directly on patient respiratory samples correctly classified the patients according to their corresponding cultured isolates. This provides proof of concept that targeted NGS can be used to sequence L. pneumophila directly from patient samples. Studies on a larger number of patient samples will further validate this method. Nonetheless, CRISPR/Cas9 targeted NGS methods have the potential to be widely applicable to microbial-outbreak investigations in the future, particularly in the context of difficult and slow-growing organisms. IMPORTANCE The bacterium Legionella pneumophila is responsible for outbreaks of serious and life-threatening pneumonia called Legionnaires' disease. There is a need for new molecular methods that allow investigation of Legionella outbreaks directly from patient samples, without the need for prior microbiological culture, which causes delays. Our study aims to address this problem. We have utilized a CRISPR/Cas9-based targeted next-generation sequencing (NGS) method that can be applied directly on human specimens. Furthermore, we show that analysis of the sequences of a small number of targeted genes offers the same classification of L. pneumophila as that based on data derived from the whole genome. Given the rising interest globally in sequencing pathogens directly from human samples, CRISPR/Cas9 targeted NGS methods have the potential to be widely applicable to microbial-outbreak investigations in the future, particularly in the context of difficult and slow-growing organisms.

Legionella growth in healthcare building water systems can result in legionellosis, making water management programs (WMPs) important for patient safety. However, knowledge is limited on Legionella prevalence in healthcare buildings. A dataset of quarterly water testing in Veterans Health Administration (VHA) healthcare buildings was used to examine national environmental Legionella prevalence from 2015 to 2018. Bayesian hierarchical logistic regression modeling assessed factors influencing Legionella positivity. The master dataset included 201,146 water samples from 814 buildings at 168 VHA campuses. Overall Legionella positivity over the 4 years decreased from 7.2 to 5.1%, with the odds of a Legionella -positive sample being 0.94 (0.90-0.97) times the odds of a positive sample in the previous quarter for the 16 quarters of the 4 year period. Positivity varied considerably more at the medical center campus level compared to regional levels or to the building level where controls are typically applied. We found higher odds of Legionella detection in older buildings (OR 0.92 [0.86-0.98] for each more recent decade of construction), in taller buildings (OR 1.20 [1.13-1.27] for each additional floor), in hot water samples (O.R. 1.21 [1.16-1.27]), and in samples with lower residual biocide concentrations. This comprehensive healthcare building review showed reduced Legionella detection in the VHA healthcare system over time. Insights into factors associated with Legionella positivity provide information for healthcare systems implementing WMPs and for organizations setting standards and regulations.

Background: Guidelines recommend testing hospitalized patients with community-acquired pneumonia (CAP) for Legionella pneumophila only if the infection is severe or risk factors are present. There are no validated models for predicting Legionella., Objective: To derive and externally validate a model to predict a positive Legionella test., Design, Setting and Participants: Diagnostic study of adult inpatients with pneumonia using data from 177 US hospitals in the Premier Healthcare Database (training and hold-out validation sets) and 12 Cleveland Clinic Health System (CCHS) hospitals (external validation set). We used multiple logistic regression to predict positive Legionella tests in the training set, and evaluated performance in both validation sets., Main Outcome and Measures: The outcome was a positive Legionella test. Potential predictors included demographics and co-morbidities, disease severity indicators, season, region, and presence of a local outbreak., Results: Of 166,689 patients hospitalized for pneumonia, 43,070 were tested for Legionella and 642 (1.5%) tested positive. The strongest predictors of a positive test were a local outbreak (odds ratio [OR], 3.4), June-October occurrence (OR, 3.4), hyponatremia (OR, 3.3), smoking (OR, 2.4), and diarrhea (OR, 2.0); prior admission within 6 months (OR, 0.27) and chronic pulmonary disease (OR, 0.49) were associated with a negative test. Model c-statistics were 0.79 in the Premier and 0.77 in the CCHS validation samples. High-risk patients were only slightly more likely to have been tested than lower-risk patients. Compared to actual practice, the model-based testing strategy detected twice as many cases., Conclusions: Although Legionella is an uncommon cause of pneumonia, patient characteristics can identify individuals at high risk, allowing for more efficient testing., (© 2022 The Authors. Journal of Hospital Medicine published by Wiley Periodicals LLC on behalf of Society of Hospital Medicine.)

Exposure models are useful tools for relating environmental monitoring data to expected health outcomes. The objective of this study was to (1) compare two Legionella shower exposure models, and (2) develop a risk calculator tool for relating environmental monitoring data to estimated Legionella infection risks and Legionnaires' Disease (LD) illness risks. Legionella infection risks for a single shower event were compared using two shower Legionella exposure models. These models varied in their description of partitioning of Legionella in aerosols and aerosol deposition in the lung, where Model 1 had larger and fewer aerosol ranges than Model 2. Model 2 described conventional vs. water efficient showers separately, while Model 1 described exposure for an unspecified shower type (did not describe it as conventional or water efficient). A Monte Carlo approach was used to account for variability and uncertainty in these aerosolization and deposition parameters, Legionella concentrations, and the dose-response parameter. Methods for relating infection risks to illness risks accounting for demographic differences were used to inform the risk calculator web application ("app"). Model 2 consistently estimated higher infection risks than Model 1 for the same Legionella concentration in water and estimated deposited doses with less variability. For a 7.8-min shower with a Legionella concentration of 0.1 CFU/mL, the average infection risks estimated using Model 2 were 4.8 × 10 - 6 (SD=3.0 × 10 - 6 ) (conventional shower) and 2.3 × 10 - 6 ). Model 2 was used for app development due to more conservative risk estimates and less variability in estimated dose. While multiple Legionella shower models are available for quantitative microbial risk assessments (QMRAs), they may yield notably different infection risks for the same environmental microbial concentration. Model comparisons will inform decisions regarding their integration with risk assessment tools. The development of risk calculator tools for relating environmental microbiology data to infection risks will increase the impact of exposure models for informing water treatment decisions and achieving risk targets.- 6 ) (water efficient). Average infection risk estimated by Model 1 was 1.1 × 10 -6 (SD=9.7 × 10 - 7 ). Model 2 was used for app development due to more conservative risk estimates and less variability in estimated dose. While multiple Legionella shower models are available for quantitative microbial risk assessments (QMRAs), they may yield notably different infection risks for the same environmental microbial concentration. Model comparisons will inform decisions regarding their integration with risk assessment tools. The development of risk calculator tools for relating environmental microbiology data to infection risks will increase the impact of exposure models for informing water treatment decisions and achieving risk targets., (Copyright © 2022. Published by Elsevier Ltd.)

The distribution of pathogenic Legionella in the environmental soil and water of China has not been documented yet. In this study, Legionella was detected in 129 of 575 water (22.43%) and 41 of 442 soil samples (9.28%) by culture. Twelve Legionella species were identified, of which 11 were disease-associated. Of the Legionella -positive samples, 109 of 129 (84.50%) water and 29 of 41 (70.73%) soil were positive for L. pneumophila, which accounted for about 75% of Legionella isolates in both water and soil, suggesting L. pneumophila was the most frequent species. Soil showed a higher diversity of Legionella spp. as compared with water (0.6279 versus 0.4493). In contrast, serogroup (sg) 1 was more prevalent among L. pneumophila isolates from water than from soil (26.66% versus 12.21%). Moreover, many disease-associated sequence types (STs) of L. pneumophila were found in China. Intragenic recombination was acting on L. pneumophila from both water and soil. Phylogeny, population structure, and molecular evolution analyses revealed a probable existence of L. pneumophila isolates with a special genetic background that is more adaptable to soil or water sources and a small proportion of genetic difference between water and soil isolates. The detection of viable, clinically relevant Legionella demonstrates soil as another source for harboring and dissemination of pathogenic Legionella bacteria in China. Future research should assess the implication in public health with the presence of Legionella in the soil and illustrate the genetic and pathogenicity difference of Legionella between water and soil, particularly the most prevalent L. pneumophila. IMPORTANCE Pathogenic Legionella spp. is the causative agent of Legionnaires' disease (LD), and L. pneumophila is the most common one. Most studies have focused on L. pneumophila from water and clinical samples. However, the soil is another important reservoir for this bacterium, and the distribution of Legionella spp. in water and soil sources has not been compared and documented in China yet. Discovering the distribution of Legionella spp. and L. pneumophila in the two environments may help a deep understanding of the pathogenesis and molecular evolution of the bacterium. Our research systematically uncovered the distributions of Legionella spp. in different regions and sources (e.g., water and soil) of China. Moreover, phylogeny, population structure, and molecular evolution study revealed the possible existence of L. pneumophila with a special genetic background that is more adaptable to soil or water sources, and genetic difference may exist.

Legionella pneumophila (L. pneumophila) is one of the main pathogens, causing pneumonia and respiratory tract infections, especially in patients with ventilator-associated pneumonia (VAP). This study aimed to approve the hypothesis that the serogroup distribution of L. pneumophila isolates from patients is correlated with Legionella strains in the environment. A total of 280 bronchoalveolar lavage (BAL) samples from VAP patients admitted to the intensive care unit (ICU) as well as 116 water samples from different sources in four hospitals in Tehran, Iran, were evaluated for the presence of L. pneumophila infection by culture, nested polymerase chain reaction (PCR), real-time PCR, and sequencing for genetic diversity. The molecular and culture methods found 24 (8.6%) and 5 (1.8%) samples to be positive for L. pneumophila in VAP patients, while they found 23 (19.8%) and 8 (6.9%) positive samples in water resources, respectively. The sequencing results indicated that all positive clinical samples and 14 (60.8%) environmental samples were belonged to L. pneumophila serogroup 1. Smoking, age, length of ICU stay, and duration of ventilator use had strong relationship with L. pneumophila infectivity. In conclusion, this is the first report from Iran to determine minor differences in the serogroup distribution of environmental and clinical strains. However, further studies are needed to confirm this relationship in different regions of Iran., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

BackgroundLegionnaires' disease is a respiratory illness often associated with hotels and travel. Spain is a major tourist destination and one of the European countries with most cases of Legionnaires' disease , both community- and travel-associated. However, the prevalence of Legionella in tourist facilities is unknown.AimThe present investigation aimed to survey the tourist facilities in the Balearic Islands, Spain, for Legionella prevalence.MethodsWe visited tourist facilities in the Balearic Islands in two different periods (2006-2010 and 2015-2018) and took water samples following national and international guidelines. Legionella was investigated by culture methods following international standards (ISO 11731:1998).ResultsWe evaluated 13,472 samples from 465 facilities. Bacteria of the Legionella genus were detected in 65.4% of the surveyed facilities. Contamination of the facilities was significantly higher during the second decade (54.5 vs 78.6%). The most frequent colonisers were L. pneumophila serogroup 2-14. We detected the pathogen in 15.9% and 6.9% of hot and cold water distribution systems samples, respectively. The Legionella contamination rate in cold water systems samples was higher when free chlorine levels were < 0.2 mg/L and at > 25 °C temperatures, while in the hot water systems samples, the contamination rate was higher at < 50 °C. Of the samples from hot tubs, 10.9% were contaminated.Conclusion Legionella prevalence in hotels in the Balearic Islands was high but the contamination rates depended on the installations. Corrective measures are still needed to improve Legionella control.

Legionella is an opportunistic premise plumbing pathogen and causative agent of a severe pneumonia called Legionnaires' Disease (LD). Cases of LD have been on the rise in the U.S. and globally. Although Legionella was first identified 45 years ago, it remains an 'emerging pathogen." Legionella is part of the normal ecology of a public water system and is frequently detected in regulatory-compliant drinking water. Drinking water utilities, regulators and public health alike are increasingly required to have a productive understanding of the evolving issues and complex discussions of the contribution of the public water utility to Legionella exposure and LD risk. This review provides a brief overview of scientific considerations important for understanding this complex topic, a review of findings from investigations of public water and LD, including data gaps, and recommendations for professionals interested in investigating public water utilities. Although the current literature is inconclusive in identifying a public water utility as a sole source of an LD outbreak, the evidence is clear that minimizing growth of Legionella in public water utilities through proper maintenance and sustained disinfectant residuals, throughout all sections of the water utility, will lead to a less conducive environment for growth of the bacteria in the system and the buildings they serve., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Legionnaires' disease (LD) ( Legionella ) is a common cause of community-acquired pneumonia (CAP) in those requiring hospitalization. Geographical variation in the importance of Legionella species as an aetiologic agent of CAP is poorly understood. We performed a systematic review and meta-analysis of population-based observational studies that reported the proportion of Legionella infection in patients with CAP (1 January 1990 to 31 May 2020). Using five electronic databases, articles were identified, appraised and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. The quality of the included studies was assessed using the Newcastle-Ottawa Scale. Univariate and multivariate meta-regression analyses were conducted using study design, WHO region, study quality and healthcare setting as the explanatory variables. We reviewed 2778 studies, of which 219 were included in the meta-analysis. The mean incidence of CAP was 46.7/100,000 population (95% CI: 46.6-46.8). The mean proportion of Legionella as the causative agent for CAP was 4.6% (95% CI: 4.4 to 4.7). Consequently, the mean Legionella incidence rate was 2.8/100,000 population (95% CI: 2.7-2.9). There was significant heterogeneity across all studies I 2 = 99.27% ( p < 0.0001). After outliers were removed, there was a decrease in the heterogeneity ( I 2 = 43.53%). Legionella contribution to CAP has a global distribution. Although the rates appear highest in high income countries in temperate regions, there are insufficient studies from low- and middle-income countries to draw conclusions about the rates in these regions. Nevertheless, this study provides an estimate of the mean incidence of Legionella infection in CAP, which could be used to estimate the regional and global burden of LD to support efforts to reduce the impact of this infection as well as to fill important knowledge gaps.

Legionellae are gram-negative bacteria most commonly found in freshwater ecosystems and purpose-built water systems. In humans, the bacterium causes Legionnaires' disease (LD) or a Pontiac fever. In this study, the different waters (drinking water, pool water, cooling towers) in which Legionella pneumophila has been isolated were studied to assess the possible risk of bacterial spreading in the population. The influence of physical and chemical parameters, and interactions with Acanthamoeba castellanii on L. pneumophila , were analyzed by Heterotrophic Plate Count, the Colony-forming units (CFU) methods, transmission electron microscopy (TEM), and Sequence-Based Typing (SBT) analysis. During the study period (2013-2019), a total of 1932 water samples were analyzed, with the average annual rate of Legionella -positive water samples of 8.9%, showing an increasing trend. The largest proportion of Legionella -positive samples was found in cooling towers and rehabilitation centers (33.9% and 33.3%, respectively). Among the isolates, L. pneumophila SGs 2-14 was the most commonly identified strain (76%). The survival of Legionella was enhanced in the samples with higher pH values, while higher electrical conductivity, nitrate, and free residual chlorine concentration significantly reduced the survival of Legionella. Our results show that growth in amoeba does not affect the allelic profile, phenotype, and morphology of the bacterium but environmental L. pneumophila becomes more resistant to pasteurization treatment.

Legionellosis is a severe pneumonia caused by the inhalation of aerosols containing Legionella, Gram-negative bacteria present in the water systems of touristic-recreational facilities. The purpose of this study was to develop a scoring tool to predict the risk of both environmental contamination and Legionnaires' disease cases in such facilities in the Apulia region of southern Italy. We analyzed 47 structural and management parameters/risk factors related to the buildings, water systems, and air conditioning at the facilities. A Poisson regression model was used to compute an overall risk score for each facility with respect to three outcomes: water samples positive for Legionella (risk score range: 7-54), water samples positive for Legionella with an average load exceeding 1000 colony-forming units per liter (CFU/L) (risk score range: 22-179,871), and clinical cases of Legionnaire's disease (risk score range: 6-31). The cut-off values for three outcomes were determined by receiver operating characteristic curves (first outcome, samples positive for Legionella in a touristic-recreational facility: 19; second outcome, samples positive for Legionella in a touristic-recreational facility with an average load exceeding 1000 CFU/L: 2062; third outcome, clinical cases of Legionnaire's disease in a touristic-recreational facility: 22). Above these values, there was a significant probability of observing the outcome. We constructed this predictive model using 70% of a large dataset (18 years of clinical and environmental surveillance) and tested the model on the remaining 30% of the dataset to demonstrate its reliability. Our model enables the assessment of risk for a touristic facility and the creation of a conceptual framework to link the risk analysis with prevention measures., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: Japan experienced a heavy rainfall event from June 28 to July 8, 2018, and many casualties were caused by both heavy rainfall and flooding. Few studies have investigated patients' characteristics and the causative pathogens of community-acquired pneumonia before and after heavy rainfall events. The aim of the present study was to evaluate the causative pathogens and clinical characteristics of hospitalized patients with community-acquired pneumonia before and after the heavy rainfall event using prospective cohort data., Methods: The study was divided into two periods: July to November 2013-2017 (before heavy rainfall) and July to November 2018 (after heavy rainfall). The patients' clinical characteristics and causative pathogens before and after the heavy rainfall were investigated. Regarding the causative pathogens, adjustments were made for precipitation and seasonal patterns., Results: There were no significant differences in the number and clinical characteristics of patients before and after heavy rainfall. However, the frequency of Legionella pneumonia was significantly higher after than before the heavy rainfall event (8.9% vs 3.0%, P = 0.02) and remained significant after adjusting for precipitation and season. Three of 7 Legionella pneumonia patients engaged in reconstruction work and 2 Legionella pneumonia patients had soil exposure., Conclusions: An increased risk of Legionella pneumonia after not only rainfall and serious flooding, but also following recovery work or soil exposure should be considered., (Copyright © 2021 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Legionnaires Disease incidence has risen in the Netherlands in recent years. For the majority of the cases, the source of infection is never identified. Two Dutch wastewater treatment plants (WWTPs) have previously been identified as source of outbreaks of Legionnaires Disease (LD) among local residents. The objective of this study is to examine if LD patients in the Netherlands are more exposed to aerosols originating from WWTPs than controls., Methods: An atmospheric dispersion model was used to generate nationwide exposure maps of aerosols from 776 WWTPs in the Netherlands. Municipal sewage treatment plants and industrial WWTPs were both included. Exposure of LD cases and controls at the residential address was compared, in a matched case-control design using a conditional logistic regression. Cases were notified LD cases with onset of disease in the period 2013-2018 in the Netherlands (n = 1604)., Results: Aerosols dispersed over a large part of the Netherlands, but modelled concentrations are estimated to be elevated in close proximity to WWTPs. A statistically significant association was found between LD and the calculated annual average aerosol concentrations originating from WWTPs (odds-ratio: 1.32 (1.06-1.63)). This association remained significant when the two outbreak-related WWTPs were removed from the analysis (odds-ratio: 1.28 (1.03-1.58))., Conclusion: LD cases were more exposed to aerosols from WWTPs than controls. This indicates that exposure to aerosols dispersed from WWTPs caused Legionnaires Disease in residents living near WWTPs in the period 2013-2018. In order to investigate which characteristics of WWTPs are associated with an increased LD risk, the WWTP database should be updated and more data is needed on the presence and survival of aerosolized Legionella bacteria to improve the Legionella dispersion modelling. Furthermore, it is recommended to further investigate how aerosol dispersion of WWTPs can effectively be reduced in order to reduce the potential health risk., (Copyright © 2021 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Background: An outbreak of Legionella pneumonia occurred at a university hospital using copper-silver ionization for potable water disinfection. We present the epidemiological and laboratory investigation of the outbreak, and associated case-control study., Methods: Cases were defined by syndrome compatible with Legionella pneumonia with laboratory-confirmed Legionella infection. The water circuit and disinfection system were assessed, and water samples collected for Legionella culture. Whole genome multi-locus sequence typing (wgMLST) was used to compare the genetic similarity of patient and environmental isolates. A case-control study was conducted to identify risk factors for Legionella pneumonia., Results: We identified 13 cases of hospital-acquired Legionella. wgMLST revealed >99.9% shared allele content among strains isolated from clinical and water samples. Smoking (P= .008), steroid use (P= .007), and documented shower during hospitalization (P= .03) were risk factors for Legionella pneumonia on multivariable analysis. Environmental assessment identified modifications to the hospital water system had occurred in the month preceding the outbreak. Multiple mitigation efforts and application of point of use water filters stopped the outbreak., Conclusions: Potable water system Legionella colonization occurs despite existing copper-silver ionization systems, particularly after structural disruptions. Multidisciplinary collaboration and direct monitoring for Legionella are important for outbreak prevention. Showering is a modifiable risk factor for nosocomial Legionella pneumonia. Shower restriction and point-of-use filters merit consideration during an outbreak., (Copyright © 2021 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Since 1978, the New York State Department of Health's public health laboratory, Wadsworth Center (WC), in collaboration with epidemiology and environmental partners, has been committed to providing comprehensive public health testing for Legionella in New York. Statewide, clinical case counts have been increasing over time, with the highest numbers identified in 2017 and 2018 (1,022 and 1,426, respectively). Over the course of more than 40 years, the WC Legionella testing program has continuously implemented improved testing methods. The methods utilized have transitioned from solely culture-based methods for organism recovery to development of a suite of reference testing services, including identification and characterization by PCR and pulsed-field gel electrophoresis (PFGE). In the last decade, whole-genome sequencing (WGS) has further refined the ability to link outbreak strains between clinical specimens and environmental samples. Here, we review Legionnaires' disease outbreak investigations during this time period, including comprehensive testing of both clinical and environmental samples. Between 1978 and 2017, 60 outbreaks involving clinical and environmental isolates with matching PFGE patterns were detected in 49 facilities from the 157 investigations at 146 facilities. However, 97 investigations were not solved due to the lack of clinical or environmental isolates or PFGE matches. We found 69% of patient specimens from New York State (NYS) were outbreak associated, a much higher rate than observed in other published reports. The consistent application of new cutting-edge technologies and environmental regulations has resulted in successful investigations resulting in remediation efforts. IMPORTANCE Legionella, the causative agent of Legionnaires' disease (LD), can cause severe respiratory illness. In 2018, there were nearly 10,000 cases of LD reported in the United States (https://www.cdc.gov/legionella/fastfacts.html; https://wonder.cdc.gov/nndss/static/2018/annual/2018-table2h.html), with actual incidence believed to be much higher. About 10% of patients with LD will die, and as high as 90% of patients diagnosed will be hospitalized. As Legionella is spread predominantly through engineered building water systems, identifying sources of outbreaks by assessing environmental sources is key to preventing further cases LD.

Healthcare-related Legionnaires' disease has a devastating impact on high risk patients, with a case fatality rate of 30-50%. Legionella prevention and control in hospitals is therefore crucial. To control Legionella water colonisation in a hospital setting we evaluated the effect of pipeline improvements and temperature increase, analysing 237 samples over a 2-year period (first year: 129, second year: 108). In the first year, 25.58% of samples were positive for Legionella and 16.67% for amoeba. Assessing the distance of the points analysed from the hot water tank, the most distal points presented higher proportion of Legionella colonisation and lower temperatures (nearest points: 6.4% colonised, and temperature 61.4 °C; most distal points: 50% and temperature 59.1 °C). After the first year, the hot water system was repaired and the temperature stabilised. This led to a dramatic reduction in Legionella colonisation, which was negative in all the samples analysed; however, amoeba colonisation remained stable. This study shows the importance of keeping the temperature stable throughout the circuit, at around 60 °C. Special attention should be paid to the most distal points of the circuit; a fall in temperature at these weak points would favour the colonisation and spread of Legionella, because amoeba (the main Legionella reservoir) are not affected by temperature.

Legionellosis is a serious bacterial infection characterized by atypical pneumonia primarily due to infection with Legionella pneumophila, and bathing can be a potential cause of this infection. Legionellosis was first identified in 1977, and it is caused by Gram-negative bacteria belonging to the genus Legionella. Legionellosis remains an important public health threat, particularly in Japan, where the population is rapidly aging, thereby becoming more at risk of developing severe disease and accompanying life-threatening pneumonia. The bacteria are most commonly transmitted via the inhalation of contaminated aerosols produced and broadcast via water sprays, jets or mists. Infection can also occur via the aspiration of contaminated water or ice, or through inhalation of contaminated dust. Because the signs and symptoms of Legionnaires' disease (LD), as well as radiographic imaging are similar to pneumonia caused by other pathogens, a specific diagnostic test is required, such as a urine antigen detection test. Six clinical and laboratory parameters, a high body temperature, a non-productive cough, low serum sodium and platelet counts, and high lactate dehydrogenase (LDH) and c-reactive protein concentrations can be used to reliably predict the likelihood of LD. The first choices for chemotherapy are fluoroquinolone and macrolide antibiotic drugs. The main goals of LD prevention measures are 1) the prevention of microbial growth and biofilm formation, 2) the removal of all biofilm formed on equipment and in facilities, 3) minimizing aerosol splash and spread, and 4) minimizing bacterial contamination from external sources. It is apparent that, in Japan, where hot spring (onsen) bathing is common among aged people, strict regulations need to be in place - and enforced - to ensure that all Japanese onsens and spas provide a safe environment and undertake regular, effective infection control practices.

We reviewed findings of clinical, epidemiologic, and environmental investigations for 288 confirmed case-patients with Legionnaires' disease reported in Hong Kong, China, during January 2005-December 2015. We found that chronic renal failure/impairment (adjusted odds ratio [aOR] 4.09), chronic pulmonary diseases (aOR 3.22), malignancy (aOR 3.04), and heart diseases (aOR 2.15) were independently associated with a higher risk for severe Legionnaires' disease. However, patients with hyperlipidemia had a lower risk for severe outcome (aOR 0.17). Legionella positivity rate was 22% for 1,904 water samples collected. We found a higher positivity rate in summer months (28%-30%), which corroborated with months of highest rainfalls. Our novel finding that Legionnaires' disease patients with hyperlipidemia had a lower risk for severe outcome deserves further study to confirm the observation and ascertain the underlying reason.

Legionella bacteria can colonise and proliferate in water systems in the built environment and can be spread by aerosol generation. If inhaled by a susceptible individual, this can lead to respiratory infections such as Legionnaires' Disease (LD), or the generally milder Pontiac fever. Evaporative cooling systems (ECS), including cooling towers, used in industrial processes to dissipate excessive heat are prone to contamination by Legionella. From these systems it is possible for contaminated aerosols to be dispersed over a wide area, potentially exposing workers on site, neighbouring workplaces or nearby members of the public. Analysis of reported data on outbreaks of LD in Great Britain, collated for a ten year period, identified 44 separate legionellosis outbreaks of which seven were attributed to ECS and were responsible for 229 infections and 10 fatalities. This prompted an examination of health and safety inspection records which revealed, over a five year period, 321 enforcement actions taken against failings in Legionella control, of which 31% were attributed to cooling towers. Based on this evidence, an intervention programme was undertaken by health and safety inspectors in which 1,906 sites with ECS were inspected. During these inspections, sites were rated against four topics that are used to demonstrate compliance with statutory requirements for Legionella control: Risk Assessment; Written Control Scheme; Implementation of Control Scheme; and Record Keeping. While there was compliance at the majority of sites, breaches of the legislation were found at 625 sites (33% of those inspected), leading to 409 Improvement Notices (compelling dutyholders to make improvements to health and safety breaches of law in a given timeframe) and 12 Prohibition Notices (compelling dutyholders to stop work until they have remedied breaches in health and safety law) being served at 229 sites (12.0% of those inspected). Data from the intervention programme was analysed to identify root causes of these breaches of legislation on Legionella control. The majority of Improvement Notices (53%) were issued for the 'lack of effective implementation of a Written Control Scheme', with 'Risk Assessment' and 'Written Control Scheme' both accounting for 23%. More detailed examination showed major problems to be lack of training; failure to maintain the cleanliness of cooling towers and the water within them; risk assessments either being absent or not up to date, i.e., no longer representing the risks present; and Written Control Schemes being absent or insufficiently detailed. This provides a valuable data resource for dutyholders, so that they can understand where they need to focus to achieve significant improvement in legal compliance and therefore reduce the risk of LD for employees and members of the public affected by their workplace, and valuable data for regulators to target future interventions aimed at improving dutyholder compliance leading to better protection of workers and members of the public., Competing Interests: Declaration of competing interest None., (Crown Copyright © 2019. Published by Elsevier GmbH. All rights reserved.)

Legionella-related disease is caused by an intracellular bacteria mainly living in water. Contamination results from inhalation of Legionella sp containing aerosolized water. Main risk factors are tobacco, immunodeficiency, and advanced age. Antigenuria is the cornerstone of the diagnosis. Immunocompromised patients, more commonly infected with non pneumophilaLegionella, present negative antigenuria, and culture and PCR are essential for the diagnosis. Legionnaires' disease may be severe, especially in elderly and/or immunocompromised patients. Mortality rate varies from 10 % in the general population to 50 % in intensive care. Treatment is based on macrolides or fluoroquinolones. Antibiotic resistance is very rare., (Copyright © 2019. Published by Elsevier Masson SAS.)

Legionnaires' disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large and old buildings, such as hospitals, can be contaminated with legionellae and therefore represent a potential risk for the hospital population. In this study, we demonstrated the constant presence of Legionella in water samples from the water system of a large university hospital in Messina (Sicily, Italy) consisting of 11 separate pavilions during a period of 15 years (2004⁻2018). In total, 1346 hot water samples were collected between January 2004 and December 2018. During this period, to recover Legionella spp. from water samples, the standard procedures reported by the 2000 Italian Guidelines were adopted; from May 2015 to 2018 Italian Guidelines revised in 2015 (ISS, 2015) were used. Most water samples (72%) were positive to L. pneumophila serogroups 2⁻14, whereas L. pneumophila serogroup 1 accounted for 18% and non- Legionella pneumophila spp. Accounted for 15%. Most of the positive samples were found in the buildings where the following critical wards are situated: (Intensive Care Unit) ICU, Neurosurgery, Surgeries, Pneumology, and Neonatal Intensive Unit Care. This study highlights the importance of the continuous monitoring of hospital water samples to prevent the potential risk of nosocomial legionellosis.

Background: Although historically the focus has been placed above all on hospital infections and travel-associated outbreaks, most of the cases of Legionella infection are sporadic and occur in community-dwellers., Objectives: To evaluate the presence and load of Legionella in hot water systems of non-healthcare facilities that host closed communities. Furthermore, we tried to verify the association between Heterotrophic Plate Counts (HPCs) and presence of Legionella., Methods: We collected hot water and biofilm samples from the showerheads of retirement homes and group homes. Samples were tested by culture method for the presence of Legionella. Confirmation and identification were carried out through Latex test and PCR. We determined the HPCs at 22 and 37 °C by the pour plate method. Statistics performed through STATA., Results: We collected 140 hot water and biofilm samples, 95 from 26 retirement homes and 35 from 9 group homes. Legionella was found in 36.8% samples collected from retirement homes and only in 10.3% group homes' samples (p = 0.01). Legionella was identified more frequently in water than in biofilm (29.8% vs 16.9%); just in one case the pathogen was found in the biofilm only. L. pneumophila sg 1 was the pathogen more frequently isolated (65.8%), with an average load of 2720 CFU/L (SD = 8393 CFU/L). We have often noticed a high microbial contamination (67% of HPCs >200 CFU/mL) and identified a higher prevalence of Legionella for intermediate values of HPC 22 °C (p = 0.011). 32% of people hosted in retirement homes were exposed to Legionella., Conclusions: Colonization of water-systems of retirement homes and group homes is anything but occasional, and in our survey it mainly affects the former, moreover often due to L. pneumophila sg 1. The search for the pathogen in the biofilm has proved to be of little use. The relationship between HPC and Legionella deserves further studies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study aimed to assess the colonization of hotel water systems in central Greece and Corfu by Legionella , and to investigate the association between physicochemical parameters and Legionella colonization. Standardized hygiene inspection was conducted in 51 hotels, and 556 water samples were analyzed for Legionella spp . Free chlorine concentration, pH, hardness, conductivity, and trace metals were defined in cold water samples. The results of inspections and chemical analyses were associated with the microbiological results using univariate and logistic regression analysis. According to the score of the checklist used for the inspections, 17.6% of the hotels were classified as satisfactory, 15.7% as adequate, and 66.7% as unsatisfactory. Moreover, 74.5% of the hotels were colonized by Legionella spp. and 31.4% required remedial measures according to the European guidelines. Legionella spp . were isolated in 28% of the samples. Unsatisfactory results of inspections were associated with Legionella presence (relative risk (RR) = 7.67, p -value = 0.043). In hot-water systems, <50 °C temperatures increased the risk of Legionella colonization (RR = 5.36, p -value < 0.001). In cold-water systems, free chlorine concentration <0.375 mg/L (odds ratio (OR) = 9.76, p -value = 0.001), pH ≥ 7.45 (OR = 4.05, p -value = 0.007), and hardness ≥321 mgCaCO₃/L (OR = 5.63, p -value = 0.003) increased the risk, whereas copper pipes demonstrated a protective role (OR = 0.29, p -value = 0.0024). The majority of the hotels inspected were colonized with Legionella . Supplementary monitoring of the risk factors that were identified should be considered., Competing Interests: The authors declare no conflicts of interest.

The incidence of Legionnaires' disease in the United States has been increasing since 2000. Outbreaks and clusters are associated with decorative, recreational, domestic, and industrial water systems, with the largest outbreaks being caused by cooling towers. Since 2006, 6 community-associated Legionnaires' disease outbreaks have occurred in New York City, resulting in 213 cases and 18 deaths. Three outbreaks occurred in 2015, including the largest on record (138 cases). Three outbreaks were linked to cooling towers by molecular comparison of human and environmental Legionella isolates, and the sources for the other 3 outbreaks were undetermined. The evolution of investigation methods and lessons learned from these outbreaks prompted enactment of a new comprehensive law governing the operation and maintenance of New York City cooling towers. Ongoing surveillance and program evaluation will determine if enforcement of the new cooling tower law reduces Legionnaires' disease incidence in New York City.

Between 1 October 2016 and 31 August 2017, 51 Legionnaires' disease (LD) cases from the United Kingdom, Sweden and the Netherlands were identified with associated travel to Dubai. Cases did not all stay in the same accommodation, indicating that no single accommodation could be the source for all these infections. While local investigations continue into other potential sources, clinicians should remain alert to the possibility of LD among travellers returning from Dubai with respiratory illness., (This article is copyright of The Authors, 2017.)

Objectives: Infections caused by Legionella are the leading cause of waterborne disease outbreaks in the United States. We investigated a large outbreak of Legionnaires' disease in New York City in summer 2015 to characterize patients, risk factors for mortality, and environmental exposures., Methods: We defined cases as patients with pneumonia and laboratory evidence of Legionella infection from July 2 through August 3, 2015, and with a history of residing in or visiting 1 of several South Bronx neighborhoods of New York City. We describe the epidemiologic, environmental, and laboratory investigation that identified the source of the outbreak., Results: We identified 138 patients with outbreak-related Legionnaires' disease, 16 of whom died. The median age of patients was 55. A total of 107 patients had a chronic health condition, including 43 with diabetes, 40 with alcoholism, and 24 with HIV infection. We tested 55 cooling towers for Legionella, and 2 had a strain indistinguishable by pulsed-field gel electrophoresis from 26 patient isolates. Whole-genome sequencing and epidemiologic evidence implicated 1 cooling tower as the source of the outbreak., Conclusions: A large outbreak of Legionnaires' disease caused by a cooling tower occurred in a medically vulnerable community. The outbreak prompted enactment of a new city law on the operation and maintenance of cooling towers. Ongoing surveillance and evaluation of cooling tower process controls will determine if the new law reduces the incidence of Legionnaires' disease in New York City.

Legionellae are waterborne bacteria which are capable of causing potentially fatal Legionnaires' disease (LD), as well as Pontiac Fever. Public concern about Legionella exploded following the 1976 outbreak at the American Legion conference in Philadelphia, where 221 attendees contracted pneumonia and 34 died. Since that time, a variety of different control methods and strategies have been developed and implemented in an effort to eradicate Legionella from building water systems. Despite these efforts, the incidence of LD has been steadily increasing in the U.S. for more than a decade. Public health and occupational hygiene professionals have maintained an active debate regarding best practices for management and control of Legionella. Professional opinion remains divided with respect to the relative merits of performing routine sampling for Legionella, vs. the passive, reactive approach that has been largely embraced by public health officials and facility owners. Given the potential risks and ramifications associated with waiting to assess systems for Legionella until after disease has been identified and confirmed, a proactive approach of periodic testing for Legionella, along with proper water treatment, is the best approach to avoiding large-scale disease outbreaks.

Rationale: Legionella testing is not recommended for all patients with pneumonia, but rather for particular patient subgroups. As a result, the overall incidence of Legionella pneumonia may be underestimated., Objectives: To determine the incidence of Legionella pneumonia in a veteran population in an endemic area after introduction of a systematic infectious diseases consultation and testing program., Methods: In response to a 2011-2012 outbreak, the VA Pittsburgh Healthcare System mandated infectious diseases consultations and testing for Legionella by urine antigen and sputum culture in all patients with pneumonia., Measurements and Main Results: Between January 2013 and December 2015, 1,579 cases of pneumonia were identified. The incidence of pneumonia was 788/100,000 veterans per year, including 352/100,000 veterans per year and 436/100,000 veterans per year with community-associated pneumonia (CAP) and health care-associated pneumonia, respectively. Ninety-eight percent of patients with suspected pneumonia were tested for Legionella by at least one method. Legionella accounted for 1% of pneumonia cases (n = 16), including 1.7% (12/706) and 0.6% (4/873) of CAP and health care-associated pneumonia, respectively. The yearly incidences of Legionella pneumonia and Legionella CAP were 7.99 and 5.99/100,000 veterans, respectively. The sensitivities of urine antigen and sputum culture were 81% and 60%, respectively; the specificity of urine antigen was >99.97%. Urine antigen testing and Legionella cultures increased by 65% and 330%, respectively, after introduction of our program., Conclusions: Systematic testing of veterans in an endemic area revealed a higher incidence of Legionella pneumonia and CAP than previously reported. Widespread urine antigen testing was not limited by false positivity.

Legionnaires' disease (LD) is a globally distributed systemic infectious disease. The burden of LD in many regions is still unclear, especially in Asian countries including China. A survey of Legionella infection using real-time PCR and nested sequence-based typing (SBT) was performed in two hospitals in Shanghai, China. A total of 265 bronchoalveolar lavage fluid (BALF) specimens were collected from hospital A between January 2012 and December 2013, and 359 sputum specimens were collected from hospital B throughout 2012. A total of 71 specimens were positive for Legionella according to real-time PCR focusing on the 5S rRNA gene. Seventy of these specimens were identified as Legionella pneumophila as a result of real-time PCR amplification of the dotA gene. Results of nested SBT revealed high genetic polymorphism in these L. pneumophila and ST1 was the predominant sequence type. These data revealed that the burden of LD in China is much greater than that recognized previously, and real-time PCR may be a suitable monitoring technology for LD in large sample surveys in regions lacking the economic and technical resources to perform other methods, such as urinary antigen tests and culture methods.

Aim: To compare the discriminatory power of pulsed-field gel electrophoresis (PFGE) and sequence-based typing (SBT) in Legionella outbreaks for determining the infection source., Materials & Methods: Twenty-five investigations of Legionnaires' disease were analyzed by PFGE, SBT and Dresden monoclonal antibody., Results: The results suggested that monoclonal antibody could reduce the number of Legionella isolates to be characterized by molecular methods. The epidemiological concordance PFGE-SBT was 100%, while the molecular concordance was 64%. Adjusted Wallace index (AW) showed that PFGE has better discriminatory power than SBT (AWSBT→PFGE = 0.767; AWPFGE→SBT = 1). The discrepancies appeared mostly in sequence type (ST) 1, a worldwide distributed ST for which PFGE discriminated different profiles., Conclusion: SBT discriminatory power was not sufficient verifying the infection source, especially in worldwide distributed STs, which were classified into different PFGE patterns.

In 2002, the National Legionella Outbreak Detection Program was implemented in the Netherlands to detect and eliminate potential sources of organisms that cause Legionnaires' disease (LD). During 2002-2012, a total of 1,991 patients with LD were reported, and 1,484 source investigations were performed. Of those sources investigated, 24.7% were positive for Legionella spp. For 266 patients with LD, 105 cluster locations were identified. A genotype match was made between a strain detected in 41 patients and a strain from a source location. Despite the systematic approach used by the program, most sources of LD infections during 2002-2012 remained undiscovered. Explorative studies are needed to identify yet undiscovered reservoirs and transmission routes for Legionella bacteria, and improved laboratory techniques are needed to detect Legionella spp. in clinical samples with a high background of microbial flora (such as soil).

Legionella bacteria are ubiquitous in natural matrices and man-made systems. However, it is not always clear if these reservoirs can act as source of infection resulting in cases of Legionnaires' disease. This review provides an overview of reservoirs of Legionella reported in the literature, other than drinking water distribution systems. Levels of evidence were developed to discriminate between potential and confirmed sources of Legionella. A total of 17 systems and matrices could be classified as confirmed sources of Legionella. Many other man-made systems or natural matrices were not classified as a confirmed source, since either no patients were linked to these reservoirs or the supporting evidence was weak. However, these systems or matrices could play an important role in the transmission of infectious Legionella bacteria; they might not yet be considered in source investigations, resulting in an underestimation of their importance. To optimize source investigations it is important to have knowledge about all the (potential) sources of Legionella. Further research is needed to unravel what the contribution is of each confirmed source, and possibly also potential sources, to the LD disease burden.