A bronchoscopy-associated pseudo-outbreak of Mycobacterium chelonae and Mycobacterium mucogenicum associated with contaminated ice machine water and ice.

A pseudo-outbreak of bronchoscopy-associated Mycobacterium chelonae and M. mucogenicum was traced to contaminated ice machine water and ice. A nonsterile ice bath was used to cool uncapped, sterile, saline syringes used to slow procedural bleeding. Joining the growing evidence of bronchoscopy pseudo-outbreaks, our investigation describes several lessons for future prevention.

Environmental risk of nontuberculous mycobacterial infection: Strategies for advancing methodology.

The National Institute of Allergy and Infectious Diseases organized a symposium in June 2022, to facilitate discussion of the environmental risks for nontuberculous mycobacteria exposure and disease. The expert researchers presented recent studies and identified numerous research gaps. This report summarizes the discussion and identifies six major areas of future research related to culture-based and culture independent laboratory methods, alternate culture media and culturing conditions, frameworks for standardized laboratory methods, improved environmental sampling strategies, validation of exposure measures, and availability of high-quality spatiotemporal data.

Tenets of a holistic approach to drinking water-associated pathogen research, management, and communication.

In recent years, drinking water-associated pathogens that can cause infections in immunocompromised or otherwise susceptible individuals (henceforth referred to as DWPI), sometimes referred to as opportunistic pathogens or opportunistic premise plumbing pathogens, have received considerable attention. DWPI research has largely been conducted by experts focusing on specific microorganisms or within silos of expertise. The resulting mitigation approaches optimized for a single microorganism may have unintended consequences and trade-offs for other DWPI or other interests (e.g., energy costs and conservation). For example, the ecological and epidemiological issues characteristic of Legionella pneumophila diverge from those relevant for Mycobacterium avium and other nontuberculous mycobacteria. Recent advances in understanding DWPI as part of a complex microbial ecosystem inhabiting drinking water systems continues to reveal additional challenges: namely, how can all microorganisms of concern be managed simultaneously? In order to protect public health, we must take a more holistic approach in all aspects of the field, including basic research, monitoring methods, risk-based mitigation techniques, and policy. A holistic approach will (i) target multiple microorganisms simultaneously, (ii) involve experts across several disciplines, and (iii) communicate results across disciplines and more broadly, proactively addressing source water-to-customer system management.

Evaluation of a new culture medium for isolation of nontuberculous mycobacteria from environmental water samples.

Nontuberculous mycobacteria (NTM) are waterborne pathogens commonly found in building water systems where they are a primary concern to vulnerable patient populations and can cause severe disease. The recovery of NTM from environmental samples can be a laborious undertaking and current pre-treatment methods and selective media lack sensitivity. We explored the use of the highly selective Rapidly Growing Mycobacteria (RGM) medium for culturing NTM from environmental water samples compared to existing methods. In total, 223 environmental water samples, including potable and non-potable water, were cultured for NTM using three culture media. In addition to direct culture on RGM medium, each sample was cultured on Middlebrook 7H10 medium and Mitchison 7H11 medium after pre-treatment with 0.2M KCl-HCl. Additionally, 33 distinct species of NTM were inoculated onto RGM medium and 7H10 medium in parallel to directly compare their growth. The use of RGM medium alone without pre-treatment provided a sensitivity (91%) comparable to that offered by culture on both 7H10 and 7H11 with acid pretreatment (combined sensitivity; 86%) with significantly less overgrowth and interference from other organisms on RGM medium. The average concentration of NTM observed on RGM medium alone was comparable to or greater than the NTM concentration on either medium alone or combined. Thirty-three species were examined in parallel and all tested strains of 27 of these species successfully grew on RGM medium, including 19 of 21 from the CDC's healthcare-associated infections species list. RGM medium was successful at recovering environmental NTM without a pre-treatment, greatly reducing labor and materials required to process samples. Simplification of culture processing for environmental NTM will allow for a better assessment of their presence in building water systems and the potential for reduced exposure of susceptible populations.

Evaluation of Recommended Water Sample Collection Methods and the Impact of Holding Time on Legionella Recovery and Variability from Healthcare Building Water Systems.

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

Point-of-use filters for prevention of health care-acquired Legionnaires' disease: Field evaluation of a new filter product and literature review.

BACKGROUND: The Centers for Medicare & Medicaid Services requires that health care facilities assess their building water systems and minimize the risk of growth and spread of Legionella and other waterborne pathogens. Increasingly, point-of-use (POU) filters are being used to prevent exposure to these pathogens. This study provides efficacy and performance specifications (membrane size, pore size, and use restrictions), which will aid in selecting POU filters. METHODS: New faucet and shower filters rated for 62 days of use were evaluated at an acute care facility in Southwestern Ontario, Canada. Five faucets and 5 showers served as controls or were equipped with filters. Hot water samples were collected weekly for 12 weeks and cultured for Legionella, heterotrophic plate count, and Pseudomonas. Literature searches for articles on POU filters used in health care settings were performed using PubMed and Google Scholar. Filter specifications from 5 manufacturers were also compared. RESULTS: The 62-day POU filters installed on both faucets and showers eliminated Legionella and reduced heterotrophic plate count concentrations for 12 weeks. No Pseudomonas was recovered during this study. Twenty peer-reviewed studies are summarized, and 21 features of 53 POU filters have been compiled. CONCLUSIONS: The information provides infection preventionists and facility engineers with information to verify claims from manufacturers and compare differences among POU products, including validated efficacy, filter design, and operational specifications.

Water Quality as a Predictor of Legionella Positivity of Building Water Systems.

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

Environmental sources of community-acquired legionnaires' disease: A review.

BACKGROUND: Most Legionnaires' disease in the US and abroad is community-acquired and believed to be sporadic, or non-outbreak associated. Most patients are exposed to numerous water sources, thus making it difficult to focus environmental investigations. Identifying known sources of sporadic community-acquired Legionnaires' disease will inform future sporadic Legionnaires' disease investigations as well as highlight directions for research. The objective is to summarize and rank sporadic Legionnaires' disease sources based on the level of linkage between the environmental source and cases. METHODS: A PubMed search was conducted using the search terms legion* and (origins or source or transmission) and (sporadic or community-acquired). Studies of nosocomial and/or outbreak-associated disease were excluded from this review. Definite, probable, possible and suspect ranks were assigned to sources based on evidence of linkage to sporadic Legionnaires' disease. RESULTS: The search yielded 196 articles and 47 articles were included in the final review after application of exclusion criteria. A total of 28 sources were identified. Of these, eight were assigned definite rank including residential potable water and car air-conditioner water leakage. Probable rank was assigned to five sources including solar-heated potable water and soil. Possible rank was assigned to nine sources including residential potable water and cooling towers. Suspect rank was assigned to 20 sources including large building water systems and cooling towers. CONCLUSION: Residential potable water, large building water systems and car travel appear to contribute to a substantial proportion of sporadic Legionnaires' disease. Cooling towers are also a potentially significant source; however, definitive linkage to sporadic cases proves difficult. The sources of sporadic Legionnaires' disease cannot be definitively identified for most cases.

Diagnostic testing for Legionnaires' disease.

Legionnaires' disease is commonly diagnosed clinically using a urinary antigen test. The urinary antigen test is highly accurate for L. pneumophila serogroup 1, however other diagnostic tests should also be utilized in conjunction with the urinary antigen as many other Legionella species and serogroups are pathogenic. Culturing of patient specimens remains the gold standard for diagnosis of Legionnaires' disease. Selective media, BYCE with the addition of antibiotics, allows for a high sensitivity and specificity. Culturing can identify all species and serogroups of Legionella. A major benefit of culturing is that it provides the recovery of a patient isolate, which can be used to find an environmental match. Other diagnostic tests, including DFA and molecular tests such as PCR and LAMP, are useful tests to supplement culturing. Molecular tests provide much more rapid results in comparison to culture, however these tests should not be a primary diagnostic tool given their lower sensitivity and specificity in comparison to culturing. It is recommended that all laboratories develop the ability to culture patient specimens in-house with the selective media.

Lack of correlation between Legionella colonization and microbial population quantification using heterotrophic plate count and adenosine triphosphate bioluminescence measurement.

This investigation compared biological quantification of potable and non-potable (cooling) water samples using pour plate heterotrophic plate count (HPC) methods and adenosine triphosphate (ATP) concentration measurement using bioluminescence. The relationship between these measurements and the presence of Legionella spp. was also examined. HPC for potable and non-potable water were cultured on R2A and PCA, respectively. Results indicated a strong correlation between HPC and ATP measurements in potable water (R = 0.90, p < 0.001). In the make-up water and two cooling towers, the correlations between ATP and HPC were much weaker but statistically significant (make-up water: R = 0.37, p = 0.005; cooling tower 1: R = 0.52, p < 0.001; cooling tower 2: R = 0.54, p < 0.001). For potable and non-potable samples, HPC exhibited higher measurement variability than ATP. However, ATP measurements showed higher microbial concentrations than HPC measurements. Following chlorination of the cooling towers, ATP measurements indicated very low bacterial concentrations (<10 colony-forming units (CFU)/mL) despite high HPC concentrations (>1000 CFU/mL) which consisted primarily of non-tuberculous mycobacteria. HPC concentrations have been suggested to be predictive of Legionella presence, although this has not been proven. Our evaluation showed that HPC or ATP demonstrated a fair predictive capacity for Legionella positivity in potable water (HPC: receiver operating characteristic (ROC) = 0.70; ATP: ROC = 0.78; p = 0.003). However, HPC or ATP correctly classified sites as positive only 64 and 62% of the time, respectively. No correlation between HPC or ATP and Legionella colonization in non-potable water samples was found (HPC: ROC = 0.28; ATP: ROC = 0.44; p = 0.193).

Effect of monochloramine treatment on the microbial ecology of Legionella and associated bacterial populations in a hospital hot water system.

Opportunistic pathogens, including Legionella spp. and non-tuberculous mycobacteria, can thrive in building hot water systems despite municipal and traditional on-site chlorine disinfection. Monochloramine is a relatively new approach to on-site disinfection, but the microbiological impact of on-site chloramine use has not been well studied. We hypothesized that comparison of the microbial ecology associated with monochloramine treatment versus no on-site treatment would yield highly dissimilar bacterial communities. Hot water samples were collected monthly from 7 locations for three months from two buildings in a Pennsylvania hospital complex supplied with common municipal water: (1) a hospital administrative building (no on-site treatment) and (2) an adjacent acute-care hospital treated on-site with monochloramine to control Legionella spp. Water samples were subjected to DNA extraction, rRNA PCR, and 454 pyrosequencing. Stark differences in the microbiome of the chloraminated water and the control were observed. Bacteria in the treated samples were primarily Sphingomonadales and Limnohabitans, whereas Flexibacter and Planctomycetaceae predominated in untreated control samples. Serendipitously, one sampling month coincided with dysfunction of the on-site disinfection system that resulted in a Legionella bloom detected by sequencing and culture. This study also demonstrates the potential utility of high-throughput DNA sequencing to monitor microbial ecology in water systems.

Fungal diversity and presence of potentially pathogenic fungi in a hospital hot water system treated with on-site monochloramine.

Currently, our knowledge of fungal ecology in engineered drinking water systems is limited, despite the potential for these systems to serve as a reservoir for opportunistic pathogens. In this study, hot water samples were collected both prior to and following the addition of monochloramine as an on-site disinfectant in a hospital hot water system. Fungal ecology was then analyzed by high throughput sequencing of the fungal ITS1 region. The results demonstrate that the genera Penicillium, Aspergillus, Peniophora, Cladosporium and Rhodosporidium comprised the core fungal biome of the hospital hot water system. Penicillium dominated the fungal community with an average relative abundance of 88.89% (±6.37%). ITS1 sequences of fungal genera containing potential pathogens such as Aspergillus, Candida, and Fusarium were also detected in this study. No significant change in fungal community structure was observed before and after the initiation of on-site monochloramine water treatment. This work represents the first report of the effects of on-site secondary water disinfection on fungal ecology in premise plumbing system, and demonstrates the necessity of considering opportunistic fungal pathogens during the evaluation of secondary premise plumbing disinfection systems.

Evaluation of a new monochloramine generation system for controlling Legionella in building hot water systems.

OBJECTIVE: To evaluate the efficacy of a new monochloramine generation system for control of Legionella in a hospital hot water distribution system. SETTING: A 495-bed tertiary care hospital in Pittsburgh, Pennsylvania. The hospital has 12 floors covering approximately 78,000 m(2). METHODS: The hospital hot water system was monitored for a total of 29 months, including a 5-month baseline sampling period prior to installation of the monochloramine system and 24 months of surveillance after system installation (postdisinfection period). Water samples were collected for microbiological analysis (Legionella species, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Acinetobacter species, nitrifying bacteria, heterotrophic plate count [HPC] bacteria, and nontuberculous mycobacteria). Chemical parameters monitored during the investigation included monochloramine, chlorine (free and total), nitrate, nitrite, total ammonia, copper, silver, lead, and pH. RESULTS: A significant reduction in Legionella distal site positivity was observed between the pre- and postdisinfection periods, with positivity decreasing from an average of 53% (baseline) to an average of 9% after monochloramine application (P<0.5]). Although geometric mean HPC concentrations decreased by approximately 2 log colony-forming units per milliliter during monochloramine treatment, we did not observe significant changes in other microbial populations. CONCLUSIONS: This is the first evaluation in the United States of a commercially available monochloramine system installed on a hospital hot water system for Legionella disinfection, and it demonstrated a significant reduction in Legionella colonization. Significant increases in microbial populations or other negative effects previously associated with monochloramine use in large municipal cold water systems were not observed.

Field evaluation of a new point-of-use faucet filter for preventing exposure to Legionella and other waterborne pathogens in health care facilities.

BACKGROUND: Opportunistic waterborne pathogens (eg, Legionella, Pseudomonas) may persist in water distribution systems despite municipal chlorination and secondary disinfection and can cause health care-acquired infections. Point-of-use (POU) filtration can limit exposure to pathogens; however, their short maximum lifetime and membrane clogging have limited their use. METHODS: A new faucet filter rated at 62 days was evaluated at a cancer center in Northwestern Pennsylvania. Five sinks were equipped with filters, and 5 sinks served as controls. Hot water was collected weekly for 17 weeks and cultured for Legionella, Pseudomonas, and total bacteria. RESULTS: Legionella was removed from all filtered samples for 12 weeks. One colony was recovered from 1 site at 13 weeks; however, subsequent tests were negative through 17 weeks of testing. Total bacteria were excluded for the first 2 weeks, followed by an average of 1.86 log reduction in total bacteria compared with controls. No Pseudomonas was recovered from filtered or control faucets. CONCLUSION: This next generation faucet filter eliminated Legionella beyond the 62 day manufacturers' recommended maximum duration of use. These new POU filters will require fewer change-outs than standard filters and could be a cost-effective method for preventing exposure to Legionella and other opportunistic waterborne pathogens in hospitals with high-risk patients.

Shift in the microbial ecology of a hospital hot water system following the introduction of an on-site monochloramine disinfection system.

Drinking water distribution systems, including premise plumbing, contain a diverse microbiological community that may include opportunistic pathogens. On-site supplemental disinfection systems have been proposed as a control method for opportunistic pathogens in premise plumbing. The majority of on-site disinfection systems to date have been installed in hospitals due to the high concentration of opportunistic pathogen susceptible occupants. The installation of on-site supplemental disinfection systems in hospitals allows for evaluation of the impact of on-site disinfection systems on drinking water system microbial ecology prior to widespread application. This study evaluated the impact of supplemental monochloramine on the microbial ecology of a hospital's hot water system. Samples were taken three months and immediately prior to monochloramine treatment and monthly for the first six months of treatment, and all samples were subjected to high throughput Illumina 16S rRNA region sequencing. The microbial community composition of monochloramine treated samples was dramatically different than the baseline months. There was an immediate shift towards decreased relative abundance of Betaproteobacteria, and increased relative abundance of Firmicutes, Alphaproteobacteria, Gammaproteobacteria, Cyanobacteria and Actinobacteria. Following treatment, microbial populations grouped by sampling location rather than sampling time. Over the course of treatment the relative abundance of certain genera containing opportunistic pathogens and genera containing denitrifying bacteria increased. The results demonstrate the driving influence of supplemental disinfection on premise plumbing microbial ecology and suggest the value of further investigation into the overall effects of premise plumbing disinfection strategies on microbial ecology and not solely specific target microorganisms.

Monochloramine and chlorine dioxide for controlling Legionella pneumophila contamination: biocide levels and disinfection by-product formation in hospital water networks.

Legionella colonization in hospital hot water distribution networks was evaluated following 36 months of continuous treatment with monochloramine and compared with chlorine dioxide. Nitrite, nitrate, chlorite, chlorate, bromide, trihalomethanes and haloacetic acids as well as the biocide concentration at sampled points were measured. Only 8/84 samples treated with monochloramine were found contaminated and after the first 8 months of treatment no Legionella was isolated. Chlorine dioxide was associated with a strong reduction in Legionella contamination compared to pre-treatment, but differences according to the device were observed. Monochloramine between 2 and 3 mg l(-1) and chlorine dioxide between 0.50 and 0.70 mg l(-1) were needed to control Legionella colonization. Comparing no- and post-flush samples, a higher frequency of no-flush positive samples was noted using chlorine dioxide, suggesting an increased risk for patients when they open the tap. No increase in chlorite levels and no water nitrification occurred by using monochloramine. Chlorite at levels exceeding the limit requested for drinking water was measured when chlorine dioxide was applied. In conclusion, we highlight that continuous injection of monochloramine should be considered as an effective alternative to chlorine dioxide in controlling legionellae contamination inside hospital water distribution systems.

Monochloramine use for prevention of Legionella in hospital water systems.

Eradication of Legionella species from water distribution systems especially in hospital settings has proven to be challenging. Legionella species causes Legionnaire's disease that is a potentially fatal respiratory disease often acquired through the aerosolization of contaminated water. Monochloramine has been used successfully in the municipal water systems to eradicate Legionella and there is currently limited data to support its use in the hospital setting. This technology appears to be affordable, safe and effective at penetrating biofilm in water distribution systems.

Computer keyboard covers impregnated with a novel antimicrobial polymer significantly reduce microbial contamination.

BACKGROUND: Contaminated computer keyboards have been acknowledged as a potential source for bacterial transmission between health care providers and patients. Biosafe HM 4100 is an antimicrobial polymer that can be incorporated into the polyurethane material used to make keyboard covers. This study aimed to determine whether plastic keyboard covers containing HM 4100 effectively minimize the survival of bacterial species commonly present on health care environmental surfaces. METHODS: Polyurethane material that contained 0.5% HM 4100, 1% HM 4100, and 1% HM 4100 with spray coating of 1% HM 4100 were tested. In 2 separate experiments, the surfaces of test materials were inoculated with suspensions of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VREF), Escherichia coli, or Pseudomonas aeruginosa. Viability was assessed on the materials at 0, 10, 30, 60, 120, and 240 minutes after inoculation. RESULTS: Maximum reductions in viability were observed for all 4 organisms at the longest tested time period on each test material. Mean reductions on the 0.5% HM 4100 material at 240 minutes were 99.99% for E coli, 97.8% for MRSA, 95.0% for VREF, and 92.1% for P aeruginosa. Mean reductions on the 1% HM 4100 at 120 minutes were 99.9% for VREF, 99.9% for MRSA, 99.9% for P aeruginosa, and 99.5% for E coli. Mean reductions on the 1% HM 4100 plus spray coating at 30 minutes were 99.9% for E coli, 99.8% for VREF, 98.8% for P aeruginosa, and 97.2% for MRSA. CONCLUSIONS: Incorporation of the HM 4100 antimicrobial polymer into polyurethane keyboard material may reduce the hand carriage of bacteria between health care providers and patients.