Legionella Growth Research Articles

The importance of clinical surveillance in detecting legionnaires' disease outbreaks: a large outbreak in a hospital with a Legionella disinfection system-Pennsylvania, 2011-2012.

Healthcare-associated Legionnaires' disease (LD) is a preventable pneumonia with a 30% case fatality rate. The Centers for Disease Control and Prevention guidelines recommend a high index of suspicion for the diagnosis of healthcare-associated LD. We characterized an outbreak and evaluated contributing factors in a hospital using copper-silver ionization for prevention of Legionella growth in water. Through medical records review at a large, urban tertiary care hospital in November 2012, we identified patients diagnosed with LD during 2011-2012. Laboratory-confirmed cases were categorized as definite, probable, and not healthcare associated based on time spent in the hospital during the incubation period. We performed an environmental assessment of the hospital, including collection of samples for Legionella culture. Clinical and environmental isolates were compared by genotyping. Copper and silver ion concentrations were measured in 11 water samples. We identified 5 definite and 17 probable healthcare-associated LD cases; 6 case patients died. Of 25 locations (mostly potable water) where environmental samples were obtained for Legionella-specific culture, all but 2 showed Legionella growth; 11 isolates were identical to 3 clinical isolates by sequence-based typing. Mean copper and silver concentrations were at or above the manufacturer's recommended target for Legionella control. Despite this, all samples where copper and silver concentrations were tested showed Legionella growth. This outbreak was linked to the hospital's potable water system and highlights the importance of maintaining a high index of suspicion for healthcare-associated LD, even in the setting of a long-term disinfection program.

New antimicrobial contact catalyst killing antibiotic resistant clinical and waterborne pathogens.

Microbial growth on medical and technical devices is a big health issue, particularly when microorganisms aggregate to form biofilms. Moreover, the occurrence of antibiotic-resistant bacteria in the clinical environment is dramatically growing, making treatment of bacterial infections very challenging. In search of an alternative, we studied a novel antimicrobial surface coating based on micro galvanic elements formed by silver and ruthenium with surface catalytic properties. The antimicrobial coating efficiently inhibited the growth of the nosocomial pathogens Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis and Enterococcus faecium as demonstrated by the growth inhibition on agar surface and in biofilms of antibiotic resistant clinical E. faecalis, E. faecium, and S. aureus isolates. It also strongly reduced the growth of Legionella in a drinking water pipeline and of Escherichia coli in urine. We postulate a mode of action of the antimicrobial material, which is independent of the release of silver ions. Thus, the novel antimicrobial coating could represent an alternative to combat microbial growth avoiding the toxic side effects of high levels of silver ions on eukaryotic cells.

Evaluation of the Effectiveness of a 4-Months Continuous Injection of a Gas Mixture (CO<sub>2</sub> and Inert Gases) on <i>Legionella</i> Contamination of a Hot Water Distribution System

Within the biofilm and scales Legionella is less far susceptible to the effects of the most frequently used biocides. The objective of this study was to evaluate the effect of a 4-months continuous injection of a gas mixture (CO2 and inert gas) in the hot water distribution system of a large hotel colonized by L. pneumophila sg3 on limiting biofilm formation and scales and in turn Legionella growth. Before the continuous injection of the gas mixture, out of the 15 sampling points examined every month 60% were colonized by Legionella (mean concentrations of 102 cfu/L in the boilers and the return loop, and 10 4 cfu/L in taps and showers). One week after the injection of the gas mixture and daily fluxing of the distal outlets, the level of colonization decreased ( 10 4 cfu/L) in all the sampling points. Thus, cleaning of the boilers was performed and distal outlets were again fluxed daily. One week after the level of contamination decreased again ( 10 4 cfu/L. Results indicate that the gas mixture was able to reduce the level of colonization by Legionella only because associated to the fluxing of the distal outlets.

Your garden hose: a potential health risk due to Legionella spp. growth facilitated by free-living amoebae.

Common garden hoses may generate aerosols of inhalable size (≤10 μm) during use. If humans inhale aerosols containing Legionella bacteria, Legionnaires' disease or Pontiac fever may result. Clinical cases of these illnesses have been linked to garden hose use. The hose environment is ideal for the growth and interaction of Legionella and free-living amoebae (FLA) due to biofilm formation, elevated temperatures, and stagnation of water. However, the microbial densities and hose conditions necessary to quantify the human health risks have not been reported. Here we present data on FLA and Legionella spp. detected in water and biofilm from two types of garden hoses over 18 months. By culturing and qPCR, two genera of FLA were introduced via the drinking water supply and reached mean densities of 2.5 log10 amoebae·mL(-1) in garden hose water. Legionella spp. densities (likely including pathogenic L. pneumophila) were significantly higher in one type of hose (3.8 log10 cells·mL(-1), p < 0.0001). A positive correlation existed between Vermamoebae vermiformis densities and Legionella spp. densities (r = 0.83, p < 0.028). The densities of Legionella spp. identified in the hoses were similar to those reported during legionellosis outbreaks in other situations. Therefore, we conclude that there is a health risk to susceptible users from the inhalation of garden hose aerosols.

Artificial intelligence in public health prevention of legionelosis in drinking water systems.

Good quality water supplies and safe sanitation in urban areas are a big challenge for governments throughout the world. Providing adequate water quality is a basic requirement for our lives. The colony forming units of the bacterium Legionella pneumophila in potable water represent a big problem which cannot be overlooked for health protection reasons. We analysed several methods to program a virtual hot water tank with AI (artificial intelligence) tools including neuro-fuzzy systems as a precaution against legionelosis. The main goal of this paper is to present research which simulates the temperature profile in the water tank. This research presents a tool for a water management system to simulate conditions which are able to prevent legionelosis outbreaks in a water system. The challenge is to create a virtual water tank simulator including the water environment which can simulate a situation which is common in building water distribution systems. The key feature of the presented system is its adaptation to any hot water tank. While respecting the basic parameters of hot water, a water supplier and building maintainer are required to ensure the predefined quality and water temperature at each sampling site and avoid the growth of Legionella. The presented system is one small contribution how to overcome a situation when legionelosis could find good conditions to spread and jeopardize human lives.

Solar Heating and Cooling System with Absorption Chiller and Latent Heat Storage – A Research Project Summary

A reliable solar thermal cooling and heating system with high solar fraction and seasonal energy efficiency ratio (SEER) is preferable. By now, bulky sensible buffer tanks are used to improve the solar fraction for heating purposes. During summertime when solar heat is converted into useful cold by means of sorption chillers the waste heat dissipation to the ambient is the critical factor. If a dry cooler is installed the performance of the sorption machine suffers from high cooling water temperatures, especially on hot days. In contrast, a wet cooling tower causes expensive water treatment, formation of fog and the risk of legionella and bacterial growth. To overcome these problems a latent heat storage based on a cheap salt hydrate has been developed to support a dry cooler on hot days, whereby a constant low cooling water temperature for the sorption machine is ensured. Therefore the need of a wet cooling tower is avoided and neither make-up water nor maintenance is needed. The same storage serves as additional low temperature heat storage for heating purposes allowing optimal solar yield due to constant low storage temperatures. Four pilot installations between 7kW and 90kW nominal cooling capacity were equipped with latent heat storages between 80 kWh and 240 kWh energy content. Annual in situ measurement data shows a positive effect on the seasonal energy efficiency ratio (SEER) for cooling up to 11.4. Furthermore simulation results under different climatic conditions indicate raising efficiency up to 64% compared to a system with solely dry re-cooling. Long-term test bench measuring data concerning performance and durability as well as a new approach for a state of charge detection for latent heat storages are presented as well.

Characterization of the tail-specific protease (Tsp) from Legionella.

Bacterial tail-specific proteases (Tsps) have been attributed a wide variety of functions including intracellular virulence, cell wall morphology, proteolytic signal cascades and stress response. This study tested the hypothesis that Tsp has a key function for the transmissive form of Legionella pneumophila. A tsp mutant was generated in Legionella pneumophila 130b and the characteristics of this strain and the isogenic wild-type were examined using a range of growth and proteomic analyses. Recombinant Tsp protein was also produced and analyzed. The L. pneumophila tsp mutant showed no defect in growth on rich media or during thermo-osmotic stress conditions. In addition, no defects in cellular morphology were observed when the cells were examined using transmission electron microscopy. Purified recombinant Tsp was found to be an active protease with a narrow substrate range. Proteome analysis using iTRAQ (5% coverage of the proteome) found that, of those proteins detected, only 5 had different levels in the tsp mutant compared to the wild type. ACP (Acyl Carrier Protein), which has a key role for Legionella differentiation to the infectious form, was reduced in the tsp mutant; however, tsp(-) was able to infect and replicate inside macrophages to the same extent as the wild type. Combined, these data demonstrate that Tsp is a protease but is not essential for Legionella growth or cell infection. Thus, Tsp may have functional redundancy in Legionella.

Secreted Pyomelanin of Legionella pneumophila Promotes Bacterial Iron Uptake and Growth under Iron-Limiting Conditions

Iron acquisition is critical to the growth and virulence of Legionella pneumophila. Previously, we found that L. pneumophila uses both a ferrisiderophore pathway and ferrous iron transport to obtain iron. We now report that two molecules secreted by L. pneumophila, homogentisic acid (HGA) and its polymerized variant (HGA-melanin, a pyomelanin), are able to directly mediate the reduction of various ferric iron salts. Furthermore, HGA, synthetic HGA-melanin, and HGA-melanin derived from bacterial supernatants enhanced the ability of L. pneumophila and other species of Legionella to take up radiolabeled iron. Enhanced iron uptake was not observed with a ferrous iron transport mutant. Thus, HGA and HGA-melanin mediate ferric iron reduction, with the resulting ferrous iron being available to the bacterium for uptake. Upon further testing of L. pneumophila culture supernatants, we found that significant amounts of ferric and ferrous iron were associated with secreted HGA-melanin. Importantly, a pyomelanin-containing fraction obtained from a wild-type culture supernatant was able to stimulate the growth of iron-starved legionellae. That the corresponding supernatant fraction obtained from a nonpigmented mutant culture did not stimulate growth demonstrated that HGA-melanin is able to both promote iron uptake and enhance growth under iron-limiting conditions. Indicative of a complementary role in iron acquisition, HGA-melanin levels were inversely related to the levels of siderophore activity. Compatible with a role in the ecology and pathogenesis of L. pneumophila, HGA and HGA-melanin were effective at reducing and releasing iron from both insoluble ferric hydroxide and the mammalian iron chelates ferritin and transferrin.

The role of water in healthcare-associated infections

The aim is to discuss the epidemiology of infections that arise from contaminated water in healthcare settings, including Legionnaires' disease, other Gram-negative pathogens, nontuberculous mycobacteria, and fungi. Legionella can colonize a hospital water system and infect patients despite use of preventive disinfectants. Evidence-based measures are available for secondary prevention. Vulnerable patients can develop healthcare-associated infections with waterborne organisms that are transmitted by colonization of plumbing systems, including sinks and their fixtures. Room humidifiers and decorative fountains have been implicated in serious outbreaks, and pose unwarranted risks in healthcare settings. Design of hospital plumbing must be purposeful and thoughtful to avoid the features that foster growth and dissemination of Legionella and other pathogens. Exposure of patients who have central venous catheters and other invasive devices to tap water poses a risk for infection with waterborne pathogens. Healthcare facilities must conduct aggressive clinical surveillance for Legionnaires' disease and other waterborne infections in order to detect and remediate an outbreak promptly. Hand hygiene is the most important measure to prevent transmission of other Gram-negative waterborne pathogens in the healthcare setting.

IcmQ in the Type 4b Secretion System Contains an NAD+ Binding Domain

A Type 4b secretion system (T4bSS) is required for Legionella growth in alveolar macrophages. IcmQ associates with IcmR, binds to membranes, and has a critical role in the T4bSS. We have now solved a crystal structure of IcmR-IcmQ to further our understanding of this complex. This structure revealed an amphipathic four-helix bundle, formed by IcmR and the N-terminal domain of IcmQ, which is linked to a novel C-terminal domain of IcmQ (Qc) by a linker helix. The Qc domain has structural homology with ADP ribosyltransferase domains in certain bacterial toxins and binds NAD(+) with a dissociation constant in the physiological range. Structural homology andmolecular dynamics were used to identify an extended NAD(+) binding site on Qc, and the resulting model was tested by mutagenesis and binding assays. Based on the data, we suggest that IcmR-IcmQ binds to membranes, where it may interact with, or perhaps modify, a protein in the T4bSS when NAD(+) is bound.

Iron metabolism and resistance to infection by invasive bacteria in the social amoeba Dictyostelium discoideum

Dictyostelium cells are forest soil amoebae, which feed on bacteria and proliferate as solitary cells until bacteria are consumed. Starvation triggers a change in life style, forcing cells to gather into aggregates to form multicellular organisms capable of cell differentiation and morphogenesis. As a soil amoeba and a phagocyte that grazes on bacteria as the obligate source of food, Dictyostelium could be a natural host of pathogenic bacteria. Indeed, many pathogens that occasionally infect humans are hosted for most of their time in protozoa or free-living amoebae, where evolution of their virulence traits occurs. Due to these features and its amenability to genetic manipulation, Dictyostelium has become a valuable model organism for studying strategies of both the host to resist infection and the pathogen to escape the defense mechanisms. Similarly to higher eukaryotes, iron homeostasis is crucial for Dictyostelium resistance to invasive bacteria. Iron is essential for Dictyostelium, as both iron deficiency or overload inhibit cell growth. The Dictyostelium genome shares with mammals many genes regulating iron homeostasis. Iron transporters of the Nramp (Slc11A) family are represented with two genes, encoding Nramp1 and Nramp2. Like the mammalian ortholog, Nramp1 is recruited to phagosomes and macropinosomes, whereas Nramp2 is a membrane protein of the contractile vacuole network, which regulates osmolarity. Nramp1 and Nramp2 localization in distinct compartments suggests that both proteins synergistically regulate iron homeostasis. Rather than by absorption via membrane transporters, iron is likely gained by degradation of ingested bacteria and efflux via Nramp1 from phagosomes to the cytosol. Nramp gene disruption increases Dictyostelium sensitivity to infection, enhancing intracellular growth of Legionella or Mycobacteria. Generation of mutants in other “iron genes” will help identify genes essential for iron homeostasis and resistance to pathogens.

Predictors of Legionella occurrence in dental unit waterlines of a highly colonized dental hospital

Introduction. Legionella is frequently detected in Dental Unit Waterlines (DUWLs). Although such a high occurrence is not necessarily associated with high risk for Legionnaire's disease among patients and staff, it is prudent to monitor DUWLs for Legionella periodically. Since this procedure is long and expensive, surrogate markers are frequently used. Aim. To investigate whether surrogate markers are predictive of Legionella detection in DUWLs in a highly colonized dental hospital. Material and methods. DUWLs from a dental hospital where legionellae were detected intermittently throughout a period of ten years was considered. The investigated predictors were total viable flora (TVF) at 37°C and at 22°C, Pseudomonas (legionellae competitor) occurrence and season. Multivariate analysis was made and, using the best fitting logistic regression model, the probability to detect legionellae in water from DUWLs was estimated. Results. Legionellae were detected in 52% water samples collected in summertime and never detected in wintertime at levels ranging between 0 and 200 colony forming units(CFU)/L. The odds ratio of legionellae occurrence were 25.0 for Pseudomonas undetected vs. detected, 108.3 for summertime vs. wintertime, 2.2-2.3 for TVF levels at 37°C and 22°C >200 CFU/mL vs. ≤200 CFU/mL. A 29% probability to detect legionellae from DUWLs, where Pseudomonas was undetected, TVF levels were >200 CFU/mL and in summertime, was estimated. Conclusion. Despite legionellae were ubiquitous in the dental hospital during the study period, in the most favourable conditions for Legionella growth (lack of competitor, high biofilm and hot weather), legionellae were detected in almost one third of DUWLs.

Legionella pneumophila found in windscreen washer fluid without added screenwash

A paper by Wallensten et al. [1] identified an epidemiological association between Legionnaires’ disease cases and the absence of screenwash in windscreen washer fluid and a follow up pilot study had identified Legionella species in one vehicle. The research extrapolated that as many as 20 % of sporadic community acquired cases may be attributed to this exposure and prevention by the use of screenwash could be readily achieved and could theoretically prevent 50 United Kingdom (UK) cases each year. However, Legionella pneumophila has never been grown from windscreen washer fluid and it is not known whether or not screenwash inhibits or prevents growth of Legionella. We undertook an open cross-sectional study aiming to investigate whether L. pneumophila can be detected in vehicle windscreen washer fluid reservoirs and to establish whether or not the addition of screenwash can inhibit the growth of L. pneumophila and at what concentration. 30 vehicles were sampled, six of which had no screenwash added and 24 used screenwash. L. pneumophila serogroup 1 was not detected in any of the cars with screenwash but was found in one of the other vehicles. The level of contamination was 6,000 colony forming units per litre (cfu/l). The positive vehicle was unusual in that it had never had screenwash added to it. Subsequently, a laboratory-based experimental study was also undertaken to see if screenwash had a bactericidal effect on Legionella. Standard screenwash was found to be effective against Legionella at very low concentrations (10 ppm). This is the first time that L. pneumophila has been identified in windscreen washer fluid and the first time that screenwash has been shown to be effective against its growth.

Epidemiological investigation of a legionellosis outbreak in a Slovenian nursing home, August 2010

August 2010 marked the beginning of the largest outbreak of legionellosis in a Slovenian nursing home. This article presents our experiences with the outbreak investigation. In order to collect the necessary patient epidemiological data, we used individual epidemiological questionnaires. Samples were available from 15 patients and were subject to laboratory investigation. Urine and sputum samples were difficult to obtain due to the underlying diseases of the patients. Serological diagnostics constituted an important part of the epidemiological investigation. Sixty-four environmental samples were taken to identify the sources of infection. By genotyping, we assessed the affinity of the allelic profile of Legionella pneumophila serogroup 1 in environmental samples and in patient samples. Legionnaires' disease was diagnosed in 10 patients based on a combination of various tests. Legionella pneumophila serogroup 1, Legionella pneumophila serogroups 2-14, and Legionella sp., in concentrations of < 10 to 61,000 CFU/l, were isolated from 51 out of 64 environmental samples. The source of the outbreak was confirmed by genotyping the isolates from patients and the isolates from the water supply system. The 2 isolates had identical allelic profiles corresponding to that of L. pneumophila serogroup 1 allelic profile 2,3,9,10,2,1,6, designated sequence type 23 (according to the European Working Group for Legionella Infections). We describe a Legionella pneumophila serogroup 1 outbreak in a Slovenian nursing home. The source of infection was demonstrated using sequence-based typing. Water flow disturbances were determined as the most probable cause of Legionella growth. Overall, the risk of a Legionella outbreak is underestimated in Slovenia.

Legionellen im Trinkwasserbereich

Although much is known about Legionella and the illness they cause, the relationship between their concentration in water and the risk of infection remains unclear. A comprehensive body of experience shows that the growth of Legionella in heated water distribution systems can be effectively controlled by following the preventive measures described in the generally acknowledged codes of practice. As orientation to trigger action, a technical action level of 100 colony-forming units in 100 ml water has found wide acceptance as the maximally tolerable concentration of Legionella in drinking water hygiene. However, this value as well as the concept for controlling the growth of Legionella in installations has developed historically. In part, the focus differs between drinking water hygiene and hospital hygiene. Also, there is no harmonized European approach for controlling Legionella. In spite of such differences, there are important international parallels in the assessment of the occurrence of Legionella, in experience with controlling them, and in recommendations for prevention. There is a need particularly for adequate studies to clarify the risk of infection as well as for the publication of existing data and experience showing the efficacy of measures for prevention, disinfection and system upgrading. Such data are necessary to support evidence-based prevention of Legionella infections and to create a better epidemiological data base in Germany. One chance for reaching this target would be to improve practices in diagnosis, reporting and central data evaluation—not only of illness, but including also data on Legionella occurrence. More frequent Legionella testing of pneumonia patients is a prerequisite for this, but particularly also for the rapid identification and removal of the source of infection. Further requirements include better training of planers, plumbers, and operators of drinking water installations about the approach to preventing Legionella contamination of drinking water installations described in standards and guidelines. The further development of practicable concepts for effective Legionella prevention requires good collaboration between public authorities responsible for drinking water and hospital hygiene on the federal and state levels and experts for the prevention of infection, hygiene, and sanitary installations.

A rapid detection method using flow cytometry to monitor the risk of Legionella in bath water

Legionella species are the causative agents of human legionellosis, and bathing facilities have been identified as the sources of infection in several outbreaks in Japan. Researchers in Japan have recently reported evidence of significant associations between bacterial counts and the occurrence of Legionella in bathing facilities and in a hot tub model. A convenient and quantitative bacterial enumeration method is therefore required as an indicator of Legionella contamination or disinfection to replace existing methods such as time-consuming Legionella culture and expensive Legionella-DNA amplification. In this study, we developed a rapid detection method (RDM) to monitor the risk of Legionella using an automated microbial analyzing device based on flow cytometry techniques to measure the total number of bacteria in water samples within two minutes, by detecting typical patterns of scattered light and fluorescence. We first compared the results of our RDM with plate counting results for five filtered hot spring water samples spiked with three species of bacteria, including Legionella. Inactivation of these samples by chlorine was also assessed by the RDM, a live/dead bacterial fluorescence assay and plate counting. Using the RDM, the lower limit of quantitative bacterial counts in the spiked samples was determined as 3.0×103(3.48log)countsmL−1. We then used a laboratory model of a hot tub and found that the RDM could monitor the growth curve of naturally occurring heterotrophic bacteria with 1 and 2days' delayed growth of amoeba and Legionella, respectively, and could also determine the killing curve of these bacteria by chlorination. Finally, samples with ≥3.48 or <3.48logtotal bacterial countsmL−1 were tested using the RDM from 149 different hot tubs, and were found to be significantly associated with the positive or negative detection of Legionella with 95% sensitivity and 84% specificity. These findings indicated that the RDM can be used for Legionella control at bathing facilities, especially those where the effectiveness of chlorine is reduced by the presence of Fe2+, Mn2+, NH4+, skin debris, and/or biofilms in the water.

Parameters predictive of Legionella contamination in hot water systems: Association with trace elements and heterotrophic plate counts

The contamination of hot water samples with Legionella spp. was studied in relation to temperature, total hardness, trace element concentrations (iron, zinc, manganese, and copper) and heterotrophic plate counts (HPC) at both 22 and 37 °C. Factor analysis and receiver operating characteristic (ROC) curves were used to establish the cut-off of water parameters as predictors for Legionella contamination. Legionella spp. was isolated in 194 out of 408 samples (47.5%), with Legionella pneumophila being the most common (92.8%). After multiple logistic regression analysis, the risk for legionellae colonisation was positively associated with Mn levels >6 μg l −1, HPC at 22 °C >27 CFU l −1, and negatively with temperature >55 °C and Cu levels >50 μg l −1. Multiple regression analysis revealed that Legionella spp. counts were positively associated with Mn, HPC at 37 °C and Zn and negatively associated with temperature. Only 1 out of the 97 samples (1%) having a Mn concentration, an HPC at 22 °C and an HPC at 37 °C below the respective median values exhibited a Legionella spp. concentration exceeding 10 4 CFU l −1 vs. 41 out of the 89 samples (46.1%) with the three parameters above the medians. Our results show a qualitative and quantitative relationship between Legionella spp., the Mn concentration and heterotrophic plate counts in hot water samples from different buildings, suggesting that these parameters should be included in a water safety plan. The role of manganese in biofilm formation and its possible involvement in the mechanisms favouring Legionella survival and growth in water niches should be investigated further.

Cluster of Legionnaires’ disease in a newly built block of flats, Denmark, December 2008 – January 2009

During December 2008 to January 2009, two persons contracted Legionnaires’ disease in a newly built block of flats in a suburb of Copenhagen in Denmark. Polymerase chain reaction and culture was used to diagnose Legionnaires’ disease in this cluster. Isolates from both patients tested positive for Legionella pneumophila serogroup 1 subgroup Philadelphia sequence type 1 and the same strain was detected in hot water samples taken from the residential area indicating that the hot water supply system was the most likely source of infection. Legionella was not detected in the cold water. Two interventions were conducted to limit the Legionella colonisation of the piping and storage tanks and the effect was monitored by investigating water samples from various sites in the block of flats. Only the second intervention had a sufficient effect on the Legionella colonisation. The cluster described here points to several risk factors regarding growth of Legionella in hot water systems: (i) stagnancy of water from when the building is constructed and piping installed and until residents move in, (ii) stagnancy and low temperature (from room temperature to approximately 38°C) of water in shower hoses and (iii) failure in operation of and control measures for the hot water system.

A New Twist in the Regulation of Legionella Replication through ASC and Caspase-1

A New Twist in the Regulation of Legionella Replication through ASC and Caspase-1

A preliminary assessment of the occupational risk of acquiring Legionnaires' disease for people working in telephone manholes, a new workplace environment for Legionella growth

Telephone manholes (TMs) are underground wells, used in Italy by the telecommunication companies to locate telephone networks. Following a fatal case of Legionnaires' disease (LD), acquired during working activity in a TM, we investigated whether Legionella was present in TMs and could be a risk for manhole workers (MWs). Three hundred fifty-three environmental samples were collected from 100 TMs, and serum antibody titer against Legionella pneumophila and Legionella species non-pneumophila was determined from both MWs and control non-manhole workers. L pneumophila and Legionella species non-pneumophila were detected in 28% of water samples, in 8% of the biofilm, and in 6% of sediment matrices taken in TMs, in a concentration range of 10(2) to 10(4) colony-forming units/L. No Legionella was found in TM air samples. Although there was a statistically significant difference (P = .027) in antibody titer to L pneumophila serogroup 1 (Lp1) between MWs and non-manhole workers, a multivariate logistic regression analysis showed a significant association between antibody against Lp1 and both age group and the practice of aquatic sports. Although further investigations will be performed to quantify the risk of acquiring legionellosis, this preliminary study demonstrates for the first time the presence of Legionella, including human pathogenic species, in a working environment such as TM.