Growth kinetics of environmental Legionella pneumophila isolated from industrial wastewater

Wastewater treatment plants are environmental niches for Legionella pneumophila, the most commonly identified causative agent of severe pneumonia known as Legionnaire's disease. In the present study, Legionella pneumophila's concentrations were monitored in an industrial wastewater treatment plant and environmental isolates were characterized concerning their growth kinetics with respect to temperature and their inhibition by organic acids and ammonium. The results of the monitoring study showed that Legionella pneumophila occurs in activated sludge tanks operated with very different sludge retention times, 2.5 days in a complete-mix reactor, and 10 days in a membrane bioreactor, indicating that this bacterium can grow at different rates, despite the same wastewater temperature of 35 degrees C. The morphology of Legionella cells is different in both reactors; in the membrane bioreactor, the bacteria grow in clusters, while in the complete-mix reactor, filaments predominate demonstrating a faster growth rate. Legionella pneumophila concentrations in the complete-mix reactor and in the membrane bioreactor were within the range 3 x 10(1) to 4.8 x 10(3) GU/mL and 3 x 10(2) to 4.7 x 10(3) GU/mL, respectively. Environmental Legionella pneumophila SG2-14 isolates showed distinct temperature preferences. The lowest growth rate was observed at 28 degrees C, and the highest 0.34 d(-1) was obtained at 42 degrees C. The presence of high concentrations of organic acids and ammonium found in anaerobically pre-treated wastewater caused growth inhibition. Despite the increasing research efforts, the mechanisms governing the growth of Legionella pneumophila in wastewater treatment plants are still unclear. New innovative strategies to prevent the proliferation of this bacterium in wastewater are in demand

Whole genome sequence analysis reveals the broad distribution of the RtxA type 1 secretion system and four novel putative type 1 secretion systems throughout the Legionella genus.

Type 1 secretion systems (T1SSs) are broadly distributed among bacteria and translocate effectors with diverse function across the bacterial cell membrane. Legionella pneumophila, the species most commonly associated with Legionellosis, encodes a T1SS at the lssXYZABD locus which is responsible for the secretion of the virulence factor RtxA. Many investigations have failed to detect lssD, the gene encoding the membrane fusion protein of the RtxA T1SS, in non-pneumophila Legionella, which has led to the assumption that this system is a virulence factor exclusively possessed by L. pneumophila. Here we discovered RtxA and its associated T1SS in a novel Legionella taurinensis strain, leading us to question whether this system may be more widespread than previously thought. Through a bioinformatic analysis of publicly available data, we classified and determined the distribution of four T1SSs including the RtxA T1SS and four novel T1SSs among diverse Legionella spp. The ABC transporter of the novel Legionella T1SS Legionella repeat protein secretion system shares structural similarity to those of diverse T1SS families, including the alkaline protease T1SS in Pseudomonas aeruginosa. The Legionella bacteriocin (1-3) secretion systems T1SSs are novel putative bacteriocin transporting T1SSs as their ABC transporters include C-39 peptidase domains in their N-terminal regions, with LB2SS and LB3SS likely constituting a nitrile hydratase leader peptide transport T1SSs. The LB1SS is more closely related to the colicin V T1SS in Escherichia coli. Of 45 Legionella spp. whole genomes examined, 19 (42%) were determined to possess lssB and lssD homologs. Of these 19, only 7 (37%) are known pathogens. There was no difference in the proportions of disease associated and non-disease associated species that possessed the RtxA T1SS (p = 0.4), contrary to the current consensus regarding the RtxA T1SS. These results draw into question the nature of RtxA and its T1SS as a singular virulence factor. Future studies should investigate mechanistic explanations for the association of RtxA with virulence

Legionella and legionellosis in touristic-recreational facilities. Influence of climate factors and geostatistical analysis in Southern Italy (2001-2017)

Legionella is the causative agent of Legionnaires' disease, a flu-like illness normally acquired following inhalation or aspiration of contaminated water aerosols. Our recent studies revealed that climatic parameters can increase the number of reported cases of community-acquired Legionnaires' disease. Here, we evaluated the presence of Legionella in water networks and the distribution of Legionnaires' disease cases associated with touristic-recreational facilities in the Apulia region (southern Italy) during the period 2001-2017 using geostatistical and climatic analyses. Geostatistical analysis data revealed that the area with the highest concentration of Legionella in water systems also had the greatest number of cases of Legionnaires' disease associated with touristic-recreational facilities. Climatic analysis showed that higher daily temperature excursion (difference between maximum and minimum temperature) on the day of sampling was more often associated with Legionella-positive samples than Legionella-negative samples. In addition, our data highlighted an increased risk of Legionnaires' disease with increases in precipitation and average temperature and with decreases in daily temperature excursion (difference between maximum and minimum temperature over the course of 24 h in the days of incubation period of disease) and minimum temperature. Healthcare professionals should be aware of this phenomenon and be particularly vigilant for cases of community-acquired pneumonia during such climatic conditions and among the tourist population. The innovative geo-statistical approach used in this study could be applied in other contexts when evaluating the effects of climatic conditions on the incidence of Legionella infections

Optimizing production efficiencies of hot water units using building energy simulations : trade-off between Legionella pneumophila contamination risk and energy efficiency

The energy needed for domestic hot water represents an important share in the total energy use of well-insulated and airtight buildings. One of the main reasons for this high energy demand is that hot water is produced at temperatures above 60°C to mitigate the risk of contaminating the hot water system with Legionella pneumophila. However, this elevated temperature is not necessary for most domestic hot water applications, and has a negative effect on the efficiency of hot water production units. A simulation model has been developed which proposes an alternative to this constant 60°C by predicting the Legionella pneumophila concentration dynamically throughout the hot water system. Based on this knowledge, a hot water controller is added to the simulation model that sets a lower hot water comfort temperature in combination with heat shocks. In this paper, the simulation model is used to estimate the energy saving potential in a case study building, at the level of the heat production system by reaching higher production efficiencies. Three different production units, namely an electric boiler, heat pump and solar collector have been investigated. The controller is expected to become an alternative for the current, energy intensive, high temperature tap water heating systems

Water, water everywhere but not a drop to drink?

Biofilms are emerging as an increasing problem as medical technology advances. Dental practice is no exception and interest in the role of biofilms within dental units as a possible source of cross-infection is intensifying. It is difficult to quantitate the risks associated with aerosolised bacteria for the majority of patients seen in general practice. However, it seems prudent to eliminate this source of infection during treatment of compromised patients. This article attempts to provide a brief overview of current concepts and problems in this area of infection control

Occupational risk for Legionella infection among dental healthcare workers: meta-analysis in occupational epidemiology

Objective The occupational risk for Legionella infection among dental healthcare workers (DHCWs) is conjectured because of the risk of routine inhalation of potentially contaminated aerosols produced by the dental instruments. Nevertheless, occupational epidemiology studies are contrasting. This metaanalysis assessed the level of scientific evidence regarding the relative occupational risk for Legionella infection among DHCWs. Methods Literature search was performed without time and language restrictions, using broad data banks (PubMed, Scopus, Web of Science, GOOGLE Scholar) and generic keywords (‘legionella’ AND ‘dent*’). Analytical cross-sectional studies comparing prevalence of high serum Legionella antibody levels in DHCWs and occupationally unexposed individuals were considered. The relative occupational risk was assessed through prevalence ratio (PR) with 95% CI. Between-study heterogeneity was assessed (Cochran’s Q test) and was used to choose the meta-analytic method. Study quality (modified Newcastle-Ottawa Scale) and publication bias (Begg and Mazumdar’s test, Egger and colleagues’ test, trim and fill R0 method) were assessed formally and considered for the sensitivity analysis. Sensitivity analysis to study inclusion, subgroup analyses (dental staff categories; publication year, before vs after 1998, ie, 5 years after the release by the Centers for Disease Control and Prevention of the infection control guidelines in dental healthcare setting) were performed. Results Seven studies were included (2232 DHCWs, 1172 occupationally unexposed individuals). No evidence of publication bias was detected. The pooled PR estimate was statistically non-significant at 95% level (1.7; 95% CI 0.8 to 3.2), study-quality adjustment did not change the PR considerably (PR, 1.5; 95% CI 0.5 to 4.1). PR was statistically significant before 1998 and no longer significant after 1998. Subgroup analysis according to DHCW categories was inconclusive. Conclusions There is no scientific evidence that DHCWs are at high occupational risk. The differences between former and recent studies could be due to different characteristics of municipal water systems and the infection control guideline dissemination

Septic Arthritis Caused by Legionella dumoffii in a Patient with Systemic Lupus Erythematosus-Like Disease

We describe a patient with systemic lupus erythematosus (SLE)-like disease on immunosuppressive treatment who developed septic arthritis of the knee involving Legionella dumoffii. Cultures initially remained negative. A broad-range 16S PCR using synovial fluid revealed L. dumoffii rRNA genes, a finding that was subsequently confirmed by positive Legionella culture results

Legionella pneumophila strain 130b evades macrophage cell death independent of the effector SidF in the absence of flagellin

International audienceThe human pathogen Legionella pneumophila must evade host cell death signaling to enable replication in lung macrophages and to cause disease. After bacterial growth, however, L. pneumophila is thought to induce apoptosis during egress from macrophages. The bacterial effector protein, SidF, has been shown to control host cell survival and death by inhibiting pro-apoptotic BNIP3 and BCL-RAMBO signaling. Using live-cell imaging to follow the L. pneumophila-macrophage interaction, we now demonstrate that L. pneumophila evades host cell apoptosis independent of SidF. In the absence of SidF, L. pneumophila was able to replicate, cause loss of mitochondria membrane potential, kill macrophages, and establish infections in lungs of mice. Consistent with this, deletion of BNIP3 and BCL-RAMBO did not affect intracellular L. pneumophila replication, macrophage death rates, and in vivo bacterial virulence. Abrogating mitochondrial cell death by genetic deletion of the effectors of intrinsic apoptosis, BAX, and BAK, or the regulator of mitochondrial permeability transition pore formation, cyclophilin-D, did not affect bacterial growth or the initial killing of macrophages. Loss of BAX and BAK only marginally limited the ability of L. pneumophila to efficiently kill all macrophages over extended periods. L. pneumophila induced killing of macrophages was delayed in the absence of capsase-11 mediated pyroptosis. Together, our data demonstrate that L. pneumophila evades host cell death responses independently of SidF during replication and can induce pyroptosis to kill macrophages in a timely manner

Presence of Legionellaceae in warm water supplies and typing of strains by polymerase chain reaction

Outbreaks of Legionnaire's disease present a public health challenge especially because fatal outcomes still remain frequent. The aim of this study was to describe the abundance and epidemiology of Legionellaceae in the human-made environment. Water was sampled from hot-water taps in private and public buildings across the area of Göttingen, Germany, including distant suburbs. Following isolation, we used polymerase chain reaction in order to generate strain specific banding profiles of legionella isolates. In total, 70 buildings were examined. Of these 18 (26%) had the bacterium in at least one water sample. Legionella pneumophila serogroups 1, 4, 5 and 6 could be identified in the water samples. Most of the buildings were colonized solely by one distinct strain, as proven by PCR. In three cases equal patterns were found in separate buildings. There were two buildings in this study where isolates with different serogroups were found at the same time