Hospital Water Systems Research Articles

Occurrence of chlorine-resistant Pseudomonas aeruginosa in hospital water systems: threat of waterborne infections for patients

BackgroundSeveral healthcare-associated infection outbreaks have been caused by waterborne Pseudomonas aeruginosa exhibiting its ability to colonize water systems and resist conventional chlorine treatment. This study aims to investigate the occurrence of Pseudomonas aeruginosa in hospital drinking water systems and the antimicrobial resistance profiles (antibiotic and chlorine resistance) of isolated strains.MethodsWe investigated the presence of Pseudomonas aeruginosa in water and biofilms developed in nine hospital water systems (n = 192) using culture-based and molecular methods. We further assessed the survival of isolated strains after exposure to 0.5 and 1.5 ppm concentrations of chlorine. The profile of antibiotic resistance and presence of antibiotic resistance genes in isolated strains were also investigated.ResultsUsing direct PCR method, Pseudomonas aeruginosa was detected in 22% (21/96) of water and 28% (27/96) of biofilm samples. However, culturable Pseudomonas aeruginosa was isolated from 14 samples. Most of P. aeruginosa isolates (86%) were resistant to at least one antibiotic (mainly β-lactams), with 50% demonstrating multidrug resistance. Moreover, three isolates harbored intI1 gene and two isolates contained blaOXA−24,blaOXA−48, and blaOXA−58‌ genes. Experiments with chlorine disinfection revealed that all tested Pseudomonas aeruginosa strains were resistant to a 0.5 ppm concentration. However, when exposed to a 1.5 ppm concentration of chlorine for 30 min, 60% of the strains were eliminated. Interestingly, all chlorine-resistant bacteria that survived at 30-minute exposure to 1.5 ppm chlorine were found to harbor the intI1 gene.ConclusionsThe detection of antimicrobial resistant Pseudomonas aeruginosa in hospital water systems raises concerns about the potential for infections among hospitalized patients. The implementation of advanced mitigation measures and targeted disinfection methods should be considered to tackle the evolving challenges within hospital water systems.

Sanitary Waters: Is It Worth Looking for Mycobacteria?

The freshwater environment is suitable for nontuberculous mycobacteria (NTMs) growth. Their high adaptability represents a considerable risk for sanitary water systems, which are a potential vector for NTMs transmission. This study investigated the occurrence of NTMs, such as Mycobacterium saskatchewanense, in hospital water systems to support the surveillance and control of potentially pathogenic NTMs. We analyzed 722 ultrapure dialysis fluid samples from Emilia Romagna Dialysis Services. Among these, 35 samples were found to be positive for M. saskatchewanense. The strains were characterized using whole-genome sequencing (WGS) and variability analysis was carried out along the whole M. saskatchewanense genome. This investigation revealed the exclusive presence of M. saskatchewanense in these dialysis machines, with low genetic variability among all strains (with a low number of different alleles: <15). The strong similarity among the strain groups was also confirmed in the WGS-based ML tree, with very few significant nodes, and no clusters were identified. This research highlights the necessity of implementing surveillance protocols and investigating any potential link to human infections, as well as stressing the urgency of enhancing surveillance and infection control measures.

Curcumin-mediated photodynamic disinfection strategy with specific spectral range for mucoid Pseudomonas Aeruginosa from hospital water

BackgroundHospital water systems represent critical environments for the transmission of pathogens, including multidrug-resistant strains like mucoid Pseudomonas aeruginosa (M-PA). Conventional disinfection methods often struggle to eradicate these pathogens effectively, highlighting the need for innovative approaches. ObjectiveThis study aimed to develop an enhanced photodynamic disinfection strategy targeting M-PA from hospital water systems, using curcumin-mediated photodynamic inactivation (PDI) with specific spectral range. MethodsAn M-PA strain isolated from hospital water was subjected to photodynamic treatment using curcumin as the photosensitizer. The efficacy of different wavelengths of light and varying concentrations of curcumin, with and without Tris-EDTA adjuvants, was evaluated through bacterial enumeration, ROS level measurements, transcriptome analysis, and assessment of virulence factors and biofilm formation. In vivo experiments utilizing a DSS-induced colitis mouse model assessed the protective effects of the photodynamic treatment against M-PA infection. ResultsOur findings demonstrated that the combination of curcumin-mediated PDI with specific spectral range effectively reduced M-PA counts in water, particularly when supplemented with Tris-EDTA. Transcriptome analysis revealed significant downregulation of virulence-related genes under sublethal photodynamic conditions. Furthermore, photodynamic treatment inhibited pyocyanin production and biofilm formation in M-PA, highlighting its potential to disrupt pathogenicity mechanisms. In vivo experiments showed that PDI attenuated M-PA-induced colitis in mice, indicating its protective efficacy. ConclusionThis study presents a promising photodynamic disinfection strategy for combating M-PA from hospital water. By optimizing curcumin-mediated PDI with specific spectral range and adjuvants, our approach demonstrates substantial efficacy in reducing bacterial counts, inhibiting virulence factors, and preventing M-PA-associated colitis.

Effectiveness of water system chemical disinfection against Pseudomonas aeruginosa infections, despite a not-so-obvious connection

Pseudomonas aeruginosa is a well-recognized opportunistic pathogen frequently responsible for hospital-acquired infections. Acquisition routes of P aeruginosa are both endogenous and exogenous, including transmission from a portion of the hospital water system. The impact of disinfection procedures of the water system and description routes of P aeruginosa transmission in a surgical intensive care unit over a 2-year period were investigated. Two distinct periods A and B were considered, respectively, before and after the disinfection. Fourier transform infrared spectroscopy was used to compare isolates recovered from patients and tap water. Overall, 21.3% of tap water samples were positive but with a significantly lower rate in period B. Concomitantly, the prevalence of patients positive for P aeruginosa decreased from 2.6% to 1%, suggesting a correlation between the presence of environmental sources and patient contaminations. The results revealed that 18% of patients were involved in cross-transmission events not related to any isolate recovered from water, suggesting transmission through care practices. Conversely, only 1 environmental transmission event was suspected in a patient. Although the link between the hospital environment and patients was unclear, HCW-associated care practices could be related to contaminated point-of-use waters and then indirect spreading to patients.

The Contribution of Legionella anisa to Legionella Contamination of Water in the Built Environment.

Legionella bacteria can proliferate in poorly maintained water systems, posing risks to users. All Legionella species are potentially pathogenic, but Legionella pneumophila (L. pneumophila) is usually the primary focus of testing. However, Legionella anisa (L. anisa) also colonizes water distribution systems, is frequently found with L. pneumophila, and could be a good indicator for increased risk of nosocomial infection. Anonymized data from three commercial Legionella testing laboratories afforded an analysis of 565,750 water samples. The data covered July 2019 to August 2021, including the COVID-19 pandemic. The results confirmed that L. anisa commonly colonizes water distribution systems, being the most frequently identified non-L. pneumophila species. The proportions of L. anisa and L. pneumophila generally remained similar, but increases in L. pneumophila during COVID-19 lockdown suggest static water supplies might favor its growth. Disinfection of hospital water systems was effective, but re-colonization did occur, appearing to favor L. pneumophila; however, L. anisa colony numbers also increased as a proportion of the total. While L. pneumophila remains the main species of concern as a risk to human health, L. anisa's role should not be underestimated, either as a potential infection risk or as an indicator of the need to intervene to control Legionella's colonization of water supplies.

Fungal presence and health implications in hospital water systems

ABSTRACT Given the increasing occurrence of invasive fungal infections and the limited efficacy of modern antifungal medications, it is crucial to disseminate information regarding the potential sources of nosocomial mycoses through the One Health approach. This study investigated the presence and antifungal susceptibility of fungi in biofilm and water samples obtained from the drinking water distribution system (DWDS) of hospitals. The positivity rate for fungi in biofilm and water samples was 41% and 9%, respectively, with Aspergillus species, a significant causative agent of nosocomial mycoses, being the predominant fungi identified. Analysis of antifungal susceptibility test revelead a comparable resistance profile between some isolated species from the DWDS and those reported for certain clinical samples. While further research is required to determine the specific contribution of waterborne fungi to nosocomial fungal infections, our results emphasize the importance of controlling biofilm formation within DWDSs, particularly in high-risk hospital wards.

Use of a peracetic acid (PAA) disinfectant to reduce total viable bacteria count in hospital wastewater drains

The periphery of the hospital water system interfaces at multiple points with patients and staff in clinical areas. This comprises mostly sinks and showers and presents a significant infection control risk. Wastewater drains in particular act as a reservoir of pathogens that can be transmitted to patients. Numerous strategies have been investigated as potential methods to reduce biofilm and bacterial load including regular application of biocidal chemicals. Traditional methods of assessing the efficacy of such products relies on culture-based microbiological techniques, usually targeting a limited range of key pathogens. We assessed the efficacy of a peracetic acid containing drain disinfectant product on seven clinical handwash basin drains, taking daily samples over six weeks (before, during and after use of the drain disinfectant product). We used a rapid, culture-independent estimation of total bacterial viable count (TVC) to assess efficacy. We applied long-read metagenomic sequencing to study the entire drain microbiome, which allowed taxonomic changes to be documented following use of the drain disinfectant product. All samples were found to be heavily contaminated, however the drain disinfectant product reduced the TVC from an estimated mean of 4228cfu/mL to 2874cfu/mL. This reduction was sustained in the two weeks following cessation of the product. Long-read metagenomic sequencing showed a microbiome dominated with Gram-negative organisms, with some taxonomic shifts in samples before and after application of the drain disinfectant. The impact on hospital-acquired infections from reducing bioburden in hospital drains by approximately a third, along with any associated changes in bacterial composition, needs evaluation in future studies.

Bactericidal effectors of the Stenotrophomonas maltophilia type IV secretion system: functional definition of the nuclease TfdA and structural determination of TfcB.

Stenotrophomonas maltophilia expresses a type IV protein secretion system (T4SS) that promotes contact-dependent killing of other bacteria and does so partly by secreting the effector TfcB. Here, we report the structure of TfcB, comprising an N-terminal domain similar to the catalytic domain of glycosyl hydrolase (GH-19) chitinases and a C-terminal domain for recognition and translocation by the T4SS. Utilizing a two-hybrid assay to measure effector interactions with the T4SS coupling protein VirD4, we documented the existence of five more T4SS substrates. One of these was protein 20845, an annotated nuclease. A S. maltophilia mutant lacking the gene for 20845 was impaired for killing Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Moreover, the cloned 20845 gene conferred robust toxicity, with the recombinant E. coli being rescued when 20845 was co-expressed with its cognate immunity protein. The 20845 effector was an 899 amino-acid protein, comprised of a GHH-nuclease domain in its N-terminus, a large central region of indeterminant function, and a C-terminus for secretion. Engineered variants of the 20845 gene that had mutations in the predicted catalytic site did not impede E. coli, indicating that the antibacterial effect of 20845 involves its nuclease activity. Using flow cytometry with DNA staining, we determined that 20845, but not its mutant variants, confers a loss in DNA content of target bacteria. Database searches revealed that uncharacterized homologs of 20845 occur within a range of bacteria. These data indicate that the S. maltophilia T4SS promotes interbacterial competition through the action of multiple toxic effectors, including a potent, novel DNase.IMPORTANCEStenotrophomonas maltophilia is a multi-drug-resistant, Gram-negative bacterium that is an emerging pathogen of humans. Patients with cystic fibrosis are particularly susceptible to S. maltophilia infection. In hospital water systems and various types of infections, S. maltophilia co-exists with other bacteria, including other pathogens such as Pseudomonas aeruginosa. We previously demonstrated that S. maltophilia has a functional VirB/D4 type VI protein secretion system (T4SS) that promotes contact-dependent killing of other bacteria. Since most work on antibacterial systems involves the type VI secretion system, this observation remains noteworthy. Moreover, S. maltophilia currently stands alone as a model for a human pathogen expressing an antibacterial T4SS. Using biochemical, genetic, and cell biological approaches, we now report both the discovery of a novel antibacterial nuclease (TfdA) and the first structural determination of a bactericidal T4SS effector (TfcB).

EOC 75 - Routine Maintenance Treatment of Hospital Water Systems with Monochloramine Effectively Controls Legionella Growth

EOC 75 - Routine Maintenance Treatment of Hospital Water Systems with Monochloramine Effectively Controls Legionella Growth

Antibiotic resistance pattern of waterborne causative agents of healthcare-associated infections: A call for biofilm control in hospital water systems

BackgroundIn recent years, the global spread of antimicrobial resistance has become a concerning issue, often referred to as a “silent pandemic”. Healthcare-associated infections (HAIs) caused by antibiotic-resistant bacteria (ARB) are a recurring problem, with some originating from waterborne route. The study aimed to investigate the presence of clinically relevant opportunistic bacteria and antibiotic resistance genes (ARGs) in hospital water distribution systems (WDSs). MethodsWater and biofilm samples (n = 192) were collected from nine hospitals in Isfahan and Kashan, located in central Iran, between May 2022 and June 2023. The samples were analyzed to determine the presence and quantities of opportunistic bacteria and ARGs using cultural and molecular methods. ResultsStaphylococcus spp. were highly detected in WDS samples (90 isolates), with 33 % of them harboring mecA gene. However, the occurrences of E. coli (1 isolate), Acinetobacter baumannii (3 isolates), and Pseudomonas aeruginosa (14 isolates) were low. Moreover, several Gram-negative bacteria containing ARGs were identified in the samples, mainly belonging to Stenotrophomonas, Sphingomonas and Brevundimonas genera. Various ARGs, as well as intI1, were found in hospital WDSs (ranging from 14 % to 60 %), with higher occurrences in the biofilm samples. ConclusionOur results underscore the importance of biofilms in water taps as hotspots for the dissemination of opportunistic bacteria and ARG within hospital environments. The identification of multiple opportunistic bacteria and ARGs raises concerns about the potential exposure and acquisition of HAIs, emphasizing the need for proactive measures, particularly in controlling biofilms, to mitigate infection risks in healthcare settings.

Sources, transmissionand hospital-associated outbreaks of nontuberculous mycobacteria: a review.

Nontuberculous mycobacteria (NTM) are widespread environmental organisms found in both natural and man-made settings, such as building plumbing, water distribution networks and hospital water systems. Their ubiquitous presence increases the risk of transmission, leading to a wide range of human infections, particularly in immunocompromised individuals. NTM primarily spreads through environmental exposures, such as inhaling aerosolized particles, ingesting contaminated food and introducing it into wounds. Hospital-associated outbreaks have been linked to contaminated medical devices and water systems. Furthermore, the rising global incidence, prevalence and isolation rates highlight the urgency of addressing NTM infections. Gaining a thorough insight into the sources and epidemiology of NTM infection is crucial for devising novel strategies to prevent and manage NTM transmission and infections.

Persistence of viable but nonculturable Legionella pneumophila state in hospital water systems: A hidden enemy?

There is little evidence of the long-term consequences of maintaining sanitary hot water at high temperatures on the persistence of Legionella in the plumbing system. The aims of this study were to describe the persistence and genotypic variability of L. pneumophila in a hospital building with two entirely independent hot water distribution systems, and to estimate the thermotolerance of the genotypic variants by studying the quantity of VBNC L. pneumophila. Eighty isolates from 55 water samples obtained between the years 2012–2017 were analyzed. All isolates correspond to L. pneumophila serogroup 6. The isolates were discriminated in four restriction patterns by pulsed-field gel electrophoresis. In one installation, pattern A + Aa predominated, accounting for 75.8 % of samples, while the other installation exhibited pattern B as the most frequent (81.8 % of samples; p < 0.001). The mean temperature of the isolates was: 52.6 °C (pattern A + Aa) and 55.0 °C (pattern B), being significantly different. Nine strains were selected as representative among patterns to study their thermotolerance by flow-cytometry after 24 h of thermic treatment. VBNC bacteria were detected in all samples. After thermic treatment at 50 °C, 52.0 % of bacteria had an intact membrane, and after 55 °C this percentage decreased to 23.1 %. Each pattern exhibited varying levels of thermotolerance. These findings indicate that the same hospital building can be colonized with different predominant types of Legionella if it has independent hot water installations. Maintaining a minimum temperature of 50 °C at distal points of the system would allow the survival of replicative L. pneumophila. However, the presence of Legionella in hospital water networks is underestimated if culture is considered as the standard method for Legionella detection, because VBNC do not grow on culture plates. This phenomenon can carry implications for the Legionella risk management plans in hospitals that adjust their control measures based on the microbiological surveillance of water.

Assessment of macrolide susceptibility of Legionella pneumophila isolated from public buildings' water systems in Turkiye.

Legionella pneumophila (Lp) is aerobic, non-spore forming Gram-negative bacteria, which is ubiquitous in freshwater habitats, such as rivers and hot springs, as well as colonizing artificial aquatic environments. The ability of Lp to grow intracellularly within pulmonary macrophages is a prerequisite for the development of infection. Therefore, macrolides can achieve appropriate therapeutic concentrations in eukaryotic cells, such as azithromycin. This study aimed to investigate the macrolides susceptibility of Lp. Pre-flash water samples (n=143) were collected from the public buildings (hospitals and hotels) water system in Istanbul. Colonies were confirmed as Lp ST1, ST2-14, and non-pneumophila Lp using latex agglutination kit. 30 Lp were detected in hospital (n=23) and hotel (n=7) water systems using latex agglutination. Regardless of serotype and excluding strains without zone formation (≥256 mg/L), the main MIC values of azithromycin, erythromycin and clarithromycin were 0.61 mg/L (range 0.047-1 mg/L), 0.47 mg/L (range 0.047-1 mg/L) and 0.44 mg/L (range 0.047-1 mg/L), respectively. The MIC50 and MIC90 values for macrolides were 0.5 and 3 mg/L for azithromycin, respectively; 0.38 and 1 mg/L for erythromycin, respectively; and 0.5 and 1 mg/L for clarithromycin, respectively. We compared the MIC values of the strains for all antimicrobial agents tested without serotype discrimination. We did not find a significant difference between the MIC values of the antibiotics (p=0.16). Although the data obtained from our study do not fully reflect the breakpoints, further epidemiological studies are needed to standardize MIC values.

Occurrence of Legionella sp. bacteria in hot water systems in hospitals and nursing homes in the Świętokrzyskie Province of south-eastern Poland.

The occurrence of bacteria of the genus Legionella sp. in hot water supply systems in buildings is a real threat to human health, especially for patients in hospitals and residents of nursing homes. The aim of the study was determination of the degree of colonization of hot water systems in hospitals and nursing homes in the Świętokrzyskie Province in south-east Poland. Between 2014 - 2018, samples were collected from hot water systems in 30 hospitals and 32 nursing homes in order to determine the degree of contamination. 631 samples collected of the bacteria Legionella sp were analyzed. Excessive contamination (≥ 100CFU/100 ml) with the bacteria Legionella sp. was detected in 12.12% (n=71) of samples. Contamination was significantly more frequently detected in samples from hot water systems in nursing homes, compared to hospitals (16.48% vs. 10.37%). Above-standard contamination of hot water systems with Legionella sp. bacteria occurred in 34.43% of the facilities, and was more frequent in hospitals (41.38%) than in the nursing homes (28.13%). In 21.3% of the facilities, contamination was detected many times during the study period. Excessive contamination of hot water systems with Legionella sp. bacteria in the examined facilities was a common phenomenon. The presence of the pathogen in the installations of these facilities may constitute a considerable health hazard for patients and residents.

Determination of Some Chemicals Compounds in the water system of the Al-Ramadi teaching hospital for maternity and children

Determination of Some Chemicals Compounds in the water system of the Al-Ramadi teaching hospital for maternity and children

Incidence of co-resistance to antibiotics and chlorine in bacterial biofilm of hospital water systems: Insights into the risk of nosocomial infections

The presence of biofilms in drinking water distribution systems (DWDS) in healthcare settings poses a considerable risk to the biological security of water, particularly when the biofilm bacteria demonstrate antimicrobial resistance characteristics. This study aimed to investigate the occurrence of antibiotic-resistant bacteria (ARB) in biofilms within DWDS of hospitals. The chlorine resistance of the isolated ARB was analyzed, and then chlorine-resistant bacteria (CRB) were identified using molecular methods. Additionally, the presence of several antibiotic resistance genes (ARGs) was monitored in the isolated ARB. Out of the 41 biofilm samples collected from hospitals, ARB were detected in 32 (78%) of the samples. A total of 109 colonies of ARB were isolated from DWDS of hospitals, with β-lactam resistant bacteria, including ceftazidime-resistant and ampicillin-resistant bacteria, being the most frequently isolated ARB. Analyzing of ARGs revealed the highest detection of aac6, followed by sul1 gene. However, the β-lactamase genes blaCTX-M and blaTEM were not identified in the ARB, suggesting the presence of other β-lactamase genes not included in the tested panel. Exposure of ARB to free chlorine at a concentration of 0.5 mg/l showed that 64% of the isolates were CRB. However, increasing the chlorine concentration to 4 mg/l decreased the high fraction of ARB (91%). The domi‌‌nant CRB identified were Sphingomonas, Brevundimonas, Stenotrophomonas, Bacillus and Staphylococcus with Bacillus exhibiting the highest frequency. The results highlight the potential risk of biofilm formation in the DWDS of hospitals, leading to the dissemination of ARB in hospital environments, which is a great concern for the health of hospitalized patients, especially vulnerable individuals. Surveillance of antimicrobial resistance in DWDS of hospitals can provide valuable insights for shaping antimicrobial use policies and practices that ensure their efficacy.

The composition of planktonic prokaryotic communities in a hospital building water system depends on both incoming water and flow dynamics.

In recent years, the frequency of nosocomial infections has increased. Hospital water systems support the growth of microbes, especially opportunistic premise plumbing pathogens. In this study, planktonic prokaryotic communities present in water samples taken from hospital showers and hand basins, collected over three different sampling phases, were characterized by 16S rRNA gene amplicon sequencing. Significant differences in the abundance of various prokaryotic taxa were found through univariate and multivariate analysis. Overall, the prokaryotic communities of hospital water were taxonomically diverse and dominated by biofilm forming, corrosion causing, and potentially pathogenic bacteria. The phyla Proteobacteria, Actinobacteriota, Bacteroidota, Planctomycetota, Firmicutes, and Cyanobacteria made up 96% of the relative abundance. The α-diversity measurements of prokaryotic communities showed no difference in taxa evenness and richness based on sampling sites (shower or hand basins), sampling phases (months), and presence or absence of Vermamoeba vermiformis. However, β-diversity measurements showed significant clustering of prokaryotic communities based on sampling phases, with the greatest difference observed between the samples collected in phase 1vs phase 2/3. Importantly, significant difference was observed in prokaryotic communities based on flow dynamics of the incoming water. The Pielou's evenness diversity index revealed a significant difference (Kruskal Wallis, p < 0.05) and showed higher species richness in low flow regime (< 13 minutes water flushing per week and ≤ 765 flushing events per six months). Similarly, Bray-Curtis dissimilarity index found significant differences (PERMANOVA, p < 0.05) in the prokaryotic communities of low vs medium/high flow regimes. Furthermore, linear discriminant analysis effect size showed that several biofilm forming (e.g., Pseudomonadales), corrosion causing (e.g., Desulfobacterales), extremely environmental stress resistant (e.g., Deinococcales), and potentially pathogenic (e.g., Pseudomonas) bacterial taxa were in higher amounts under low flow regime conditions. This study demonstrated that a hospital building water system consists of a complex microbiome that is shaped by incoming water quality and the building flow dynamics arising through usage.

Stagnation arising through intermittent usage is associated with increased viable but non culturable Legionella and amoeba hosts in a hospital water system.

Hospital water systems are a significant source of Legionella, resulting in the potentially fatal Legionnaires' disease. One of the biggest challenges for Legionella management within these systems is that under unfavorable conditions Legionella transforms itself into a viable but non culturable (VBNC) state that cannot be detected using the standard methods. This study used a novel method (flow cytometry-cell sorting and qPCR [VFC+qPCR] assay) concurrently with the standard detection methods to examine the effect of temporary water stagnation, on Legionella spp. and microbial communities present in a hospital water system. Water samples were also analyzed for amoebae using culture and Vermamoeba vermiformis and Acanthamoeba specific qPCR. The water temperature, number and duration of water flow events for the hand basins and showers sampled was measured using the Enware Smart Flow® monitoring system. qPCR analysis demonstrated that 21.8% samples were positive for Legionella spp., 21% for L. pneumophila, 40.9% for V. vermiformis and 4.2% for Acanthamoeba. All samples that were Legionella spp. positive using qPCR (22%) were also positive for VBNC Legionella spp.; however, only 2.5% of samples were positive for culturable Legionella spp. 18.1% of the samples were positive for free-living amoebae (FLA) using culture. All samples positive for Legionella spp. were also positive for FLA. Samples with a high heterotrophic plate count (HPC ≥ 5 × 103 CFU/L) were also significantly associated with high concentrations of Legionella spp. DNA, VBNC Legionella spp./L. pneumophila (p < 0.01) and V. vermiformis (p < 0.05). Temporary water stagnation arising through intermittent usage (< 2 hours of usage per month) significantly (p < 0.01) increased the amount of Legionella spp. DNA, VBNC Legionella spp./L. pneumophila, and V. vermiformis; however, it did not significantly impact the HPC load. In contrast to stagnation, no relationship was observed between the microbes and water temperature. In conclusion, Legionella spp. (DNA and VBNC) was associated with V. vermiformis, heterotrophic bacteria, and stagnation occurring through intermittent usage. This is the first study to monitor VBNC Legionella spp. within a hospital water system. The high percentage of false negative Legionella spp. results provided by the culture method supports the use of either qPCR or VFC+qPCR to monitor Legionella spp. contamination within hospital water systems.

Tap water as the source of a Legionnaires' disease outbreak spread to several residential buildings and one hospital, Finland, 2020 to 2021.

In Finland, all microbiology laboratories notify Legionella findings and physicians notify Legionnaires' disease (LD) cases to the National Infectious Disease Register. All cases are interviewed, and water samples obtained from potential places of exposure. Legionella isolates from humans and water are compared by whole genome sequencing (WGS). In March 2021, Legionella pneumophila serogroup 1 (Lp 1) pneumonia cases increased in one Finnish city (120,000 inhabitants) where single LD cases are detected annually. We identified 12 LD cases, nine living in different residential buildings and three nosocomial, linked by identical human and/or water isolates. Three of these cases were from January 2020, October 2020 and February 2021 and identified retrospectively. Eleven were diagnosed by urinary antigen test, 10 by PCR and five by culture; age ranged between 52 and 85 years, and 10 had underlying diseases. Nine of 12 homes of LD cases and 15 of 26 water samples from the hospital were positive for Lp 1, with concentrations up to 640,000 cfu/L. Water samples from regional storage tanks were negative. Positivity in homes and the hospital suggested inadequate maintenance measures. Enhanced surveillance combined with WGS was crucial in detecting this unusual LD outbreak related to domestic and hospital water systems.

Challenges in the Hospital Water System and Innovations to Prevent Healthcare-Associated Infections

Challenges in the Hospital Water System and Innovations to Prevent Healthcare-Associated Infections