A community outbreak of Legionnaires' disease caused by outdoor hot tubs for private use in a hotel.

During the period October-November 2017, an outbreak of Legionnaires' disease involving 27 cases occurred in the tourist area of Palmanova (Mallorca, Spain). The majority of cases were reported by the European Centre of Disease Prevention and Control (ECDC) as travel associated cases of Legionnaires' disease (TALD). Most cases belonged to different hotel cluster alerts. No cases were reported among the local population residing in the area. All tourist establishments associated with one or more TALD cases were inspected and sampled by public health inspectors. All relevant sources of aerosol emission detected were investigated and sampled. The absence of active cooling towers in the affected area was verified, by documents and on-site. Samples from hot tubs for private use located on the terraces of the penthouse rooms of a hotel in the area were included in the study. Extremely high concentrations (> 106 CFU/l) of Legionella pneumophila, including the outbreak strain, were found in the hot tubs of vacant rooms of this hotel thus identifying the probable source of infection. Meteorological situation may have contributed to the geographical distribution pattern of this outbreak. In conclusion, hot tubs for private use located outdoors should be considered when investigating community outbreaks of Legionnaires' disease of unclear origin.

Impact of prolonged hotel closures during the COVID-19 pandemic on Legionella infection risks.

In general, it is accepted that water stagnation and lack or poor maintenance in buildings are risk factors for Legionella growth. Then, in theory, the prolonged hotel closures due to the COVID-19 pandemic may have increased the risk of Legionella infections. However, there are very few field studies comparing the level of Legionella colonization in buildings before the pandemic and the new situation created after the lockdown. The objective of this study was to analyze these differences in a group of hotels that experienced prolonged closures in 2020 due to the COVID-19 pandemic. We have studied the Legionella spp. results, analyzed by standard culture, from the domestic water distribution systems of 73 hotels that experienced closures (from 1 to >4 months) during 2020, immediately after the reopening. The results were compared with those obtained in similar samplings of 2019. For the comparative analysis, we divided the hotels in two groups: Group A that have suffered closures for ≤3 months and Group B that remained closed for more than 3 months, both in relation to the opening period of 2019. In the Group B (36 sites), the frequency of positive samples in the hot water system increased from 6.7% in 2019 to 14.0% in 2020 (p < 0.05). In the Group A (37 sites), no significant differences were observed. No statistically significant differences were observed in terms of positive sites (defined as hotels with at least 1 positive sample), Legionella spp. concentrations and prevalence of Legionella pneumophila sg1 between the samplings of the two periods studied. The results suggest that hotels that suffered the longest prolonged closures (> 3 months) could have carried a higher risk of exposure to Legionella in the domestic hot water system. These findings highlight the importance of adequate preopening cleaning and disinfection procedures for hotel water systems, and the convenience of considering the most effective disinfection methods especially for hot water systems and after prolonged closure periods.

Legionella maioricensis sp. nov., a new species isolated from the hot water distribution systems of a hospital and a shopping center during routine sampling.

Two Legionella-like strains isolated from hot water distribution systems in 2012 have been characterized phenotypically, biochemically and genomically in terms of DNA relatedness. Both strains, HCPI-6T and EUR-108, exhibited biochemical phenotypic profiles typical of Legionella species. Cells were Gram-negative motile rods which grew on BCYEα agar but not on blood agar and displayed phenotypic characteristics typical of the family Legionellaceae, including a requirement for l-cysteine and testing catalase positive. Both strains were negative for oxidase, urease, nitrate reduction and hippurate negative, and non-fermentative. The major ubiquinone was Q12 (59.4 % HCPI-6T) and the dominant fatty acids were C16 : 1 ω7c (28.4 % HCPI-6T, ≈16 % EUR-108), C16 : 0 iso (≈22.5 % and ≈13 %) and C15 : 0 anteiso (19.5 % and ≈23.5 %, respectively). The percent G+C content of genomic DNA was determined to be 39.3 mol %. The 16S rRNA gene, mip sequence and comparative genome sequence-based analyses (average nucleotide identity, ANI; digital DNA-DNA hybridization, dDDH; and phylogenomic treeing) demonstrated that the strains represent a new species of the genus Legionella. The analysis based on the 16S rRNA gene sequences showed that the sequence similarities for both strains ranged from 98.8-90.1 % to other members of the genus. The core genome-based phylogenomic tree (protein-concatemer tree based on concatenation of 418 proteins present in single copy) revealed that these two strains clearly form a separate cluster within the genus Legionella. ANI and dDDH values confirmed the distinctiveness of the strains. Based on the genomic, genotypic and phenotypic findings from a polyphasic study, the isolates are considered to represent a single novel species, for which the name Legionella maioricensis sp. nov. is proposed. The type strain is HCPI-6T (=CCUG 75071T=CECT 30569T).

Low Efficacy of Periodical Thermal Shock for Long-Term Control of Legionella spp. in Hot Water System of Hotels.

Different guidelines and regulations for the prevention of legionellosis in public facilities include the recommendation of a periodical thermal shock in the hot water system. The purpose of this study was to assess the efficacy of periodical thermal shocks along a 1-year period on the presence of Legionella spp. in the domestic hot water system of hotels. The Legionella testing results from the period January-December 2019 coming from a group of 77 hotel facilities in the Balearic Islands (Spain) conducting periodical thermal shocks were analyzed. A second group of 44 hotels operating without periodical thermal shocks was used for a comparative analysis. In the facilities where the periodical thermal shock was performed, 16.0% of the results (429 hot water samples collected) were positive for Legionella spp., compared to 21.1% (298 samples), where periodical thermal shock was not performed. Overall, in the thermal shock group, 32.5% of the sites presented at least 1 positive sample along the period of study versus 45.5% in the control group. None of these differences was statistically significant (p-value > 0.05). These findings suggest that the efficacy of regular thermal shock for long-term control of Legionella spp. in domestic hot water systems of hotels is low.

Growth of Legionella anisa in a model drinking water system to evaluate different shower outlets and the impact of cast iron rust.

Legionella continues to be a problem in water systems. This study investigated the influence of different shower mixer faucets, and the influence of the presence of cast iron rust from a drinking water system on the growth of Legionella. The research is conducted using a model of a household containing four drinking water systems. All four systems, which contained standard plumbing components including copper pipes and a water heater, were filled with unchlorinated drinking water. Furthermore, all systems had three different shower faucets: (A) a stainless-steel faucet, (B) a brass-ceramic faucet, and (C) a brass thermostatic faucet. System 1 was solely filled with drinking water. System 2 was filled with drinking water, and cast iron rust. System 3 was contaminated with Legionella, and system 4 was contaminated with a Legionella, and cast iron rust. During a period of 34 months, 450 cold water samples were taken from 15 sample points of the four drinking water systems, and tested for Legionella according to the Dutch Standard (NEN 6265). In system 4, with added cast iron rust, the stainless-steel mixer faucet (A) had the highest concentration of Legionella at >4.3log10CFU/l (>20,000CFU/l) and was positive in 46.4% of samples. In contrast, the stainless-steel mixer faucet (A) of system 3 without cast iron rust showed 14.3% positive samples with a maximum concentration of 3.9log10CFU/l (7600CFU/l) Legionella. Additionally, both contaminated systems (3 and 4), with the brass thermostatic faucet (C), tested positive for Legionella. System 3 in 85.7% of the samples, with a maximum concentration of 4.38log10CFU/l (24,200CFU/l), and system 4 in 64.3% of the samples with a maximum concentration of 4.13log10CFU/l (13.400CFU/l). These results suggest that both the type of faucet used in a drinking water system and the presence or absence of cast iron rust influence the growth of Legionella.

Genomic Investigation of a Legionellosis Outbreak in a Persistently Colonized Hotel.

OBJECTIVES: A long-lasting legionellosis outbreak was reported between November 2011 and July 2012 in a hotel in Calpe (Spain) affecting 44 patients including six deaths. Intensive epidemiological and microbiological investigations were performed in order to detect the reservoirs. METHODS: Clinical and environmental samples were tested for the presence and genetic characterization of Legionella pneumophila. Six of the isolates were subjected to whole-genome sequencing. RESULTS: Sequencing of 14 clinical and 260 environmental samples revealed sequence type (ST) 23 as the main responsible strain for the infections. This ST was found in the spa pool, from where it spread to other hotel public spaces, explaining the ST23 clinical cases, including guests who had not visited the spa. Uncultured clinical specimens showed profiles compatible with ST23, ST578, and mixed patterns. Profiles compatible with ST578 were obtained by direct sequencing from biofilm samples collected from the domestic water system, which provided evidence for the source of infection for non ST23 patients. Whole genome data from five ST23 strains and the identification of different STs and Legionella species showed that different hotel premises were likely colonized since the hotel opening thus explaining how different patients had been infected by distinct STs. CONCLUSIONS: Both epidemiological and molecular data are essential in the investigation of legionellosis outbreaks. Whole-genome sequencing data revealed significant intra-ST variability and allowed to make further inference on the short-term evolution of a local colonization of L. pneumophila.