Genomic characterization of Legionella pneumophila serogroup 1 ST901 isolates responsible for recurrent travel-associated Legionnaires’ disease cases and clusters

Several Travel-associated Legionnaires’ disease (TALD) cases occur annually in Europe. Except from the most obvious sites (cooling towers and hot water systems), infections can also be associated with recreational, water feature, and garden areas of hotels. This argument is of great interest to better comprehend the colonization and to calculate the risk to human health of these sites. From July 2000–November 2017, the public health authorities of the Island of Crete (Greece) inspected 119 hotels associated with TALD, as reported through the European Legionnaires’ Disease Surveillance Network. Five hundred and eighteen samples were collected from decorative fountain ponds, showers near pools and spas, swimming pools, spa pools, garden sprinklers, drip irrigation systems (reclaimed water) and soil. Of those, 67 (12.93%), originating from 43 (35.83%) hotels, tested positive for Legionella (Legionella pneumophila serogroups 1, 2, 3, 6, 7, 8, 13, 14, 15 and non-pneumophila species (L. anisa, L. erythra, L. taurinensis, L. birminghamensis, L. rubrilucens). A Relative Risk (R.R.) > 1 (p < 0.0001) was calculated for chlorine concentrations of less than 0.2 mg/L (R.R.: 54.78), star classification (<4) (R.R.: 4.75) and absence of Water Safety Plan implementation (R.R.: 3.96). High risk (≥104 CFU/L) was estimated for pool showers (16.42%), garden sprinklers (7.46%) and pool water (5.97%).

According to European Guidelines for Legionnaires' Disease prevention and control, travel-associated Legionnaires' disease (TALD) cases are managed differently if classified as sporadic or as part of a cluster and more stringent control measures are deployed after clusters are identified. In this study, we propose to modify the current cluster definition: 'two or more cases of Legionnaires' disease (LD) who stayed at, or visited, the same commercial accommodation site 2-10 days before onset of illness and whose onset is within the same 2-year period' with a new cluster definition, i.e. accommodation sites associated with multiple cases regardless of the time elapsed between them. TALD cases occurred in Italy and in the Balearic Islands between 2005 and 2015 were analysed applying the current European Legionnaires' Disease Surveillance Network (ELDSNet) cluster definition. In a sample of selected accommodation sites with multiple cases, a microbiological study was also conducted. Using the new definition, 63 additional sites (16.4% increase) and 225 additional linked cases (19.5% increase) were identified. Legionella pneumophila sg1 was isolated from 90.7% of the selected accommodation sites. The use of the here proposed TALD cluster definition would warrant a full investigation for each new identified case. This approach should therefore increase the number of sites that will require a risk assessment and, in the presence of an increased risk, the adoption of LD control measures to hopefully prevent additional cases.

In 2008, the European Surveillance Scheme for Travel Associated Legionnaires Disease (EWGLINET) received reports of 866 cases of travel-associated Legionnaires disease, 42 of whom were reported to have died. 824 of the cases were classified as confirmed and 42 were presumptive. As in previous years, a very low proportion of clinical isolates were obtained (63 cases, 7.3%). Males outnumbered females by 2.8:1 in the 2008 dataset and had a median age of 60 years compared with women, whose median age was 63 years. Travel outside Europe was reported for 12% of the cases. The scheme identified 108 new clusters in 2008. Sixteen were located in countries outside EWGLINET and 38 (35.2%) involved only one case from each reporting country, and would not ordinarily have been detected by national surveillance schemes alone. The largest cluster (six cases) was associated with travel to Spain. The 108 clusters were associated with 144 environmental investigations, 35 of which were at re-offending sites, (sites which had previously been investigated and where additional cases had subsequently occurred). At 61 (42.1%) of the sites Legionella species were detected. The names of 12 sites were published on the EWGLINET website.

Between April and May 2025, an outbreak of travel-associated Legionnaires’ disease (TALD) occurred, involving six cases at a hotel in Crete, Greece. Including two cases reported in 2023 and two additional cases from 2016 to 2017, ten cases were associated with this accommodation site. All TALD cases were reported by the European Legionnaires’ Disease Surveillance Network (ELDSNet). In compliance with the European Centre for Disease Prevention and Control (ECDC) surveillance and investigation protocols for hotels associated with the patient’s stay, local public health authorities conducted on-site inspections at the hotel by collecting water samples and performing risk assessments, while simultaneously recording the required epidemiological, environmental, and physicochemical data. A total of 181 statistically analyzed water samples showed positive rates for L. pneumophila of 12.71% (95% CI: 7.86–17.56) for (≥50 CFU/L) and 6.08% (95% CI: 2.60–9.56) for (≥1000 CFU/L). Risk assessments identified 18 stagnation points, systemic maintenance deficiencies, and high cumulative structural (30/52) and water (36/71) system risk scores. Low microbiological positivity of water samples does not necessarily equate to low risk, thus necessitating continuous risk assessment, implementation of Water Safety Plans (WSPs), and integrated monitoring by accommodation facilities to prevent LD cases.

Since its first appearance as a human pathogen in 1976, Legionella pneumophila has been identified as a causative agent of community-acquired pneumonia (CAP). It survives in rivers, bays, lakes, and water reservoirs, and it is categorized as the fourth most common causative agent of CAP leading to hospitalization. We aimed to investigate patterns in which environmental, seasonal and regional factors may affect the prevalence of Legionnaires’ disease in Crete during the last two decades (2000–2022).The data used originated from the national surveillance database and included any person reported with travel-associated Legionnaires’ disease (TALD) between January 2000 and December 2022. Meteorological data were collected from the National Weather Service. The meteorological variables included (max) temperature (in °C), cloudiness (in octas), wind speed (in knots), and relative humidity (RH) (%). The statistical analysis was based on a case-crossover design with 1:1 matching characteristic. We revealed both seasonal and regional effects on the incidence of Legionnaires’ disease. Cases are significantly more frequent in autumn, in comparison to the other three seasons, while Rethymnon is the prefecture with fewer cases in comparison to Chania or Heraklion. In addition, our research showed that the majority of cases occurred during the years 2017–2018. TALD in Crete is significantly associated with temperature in °C and wind speed in knots. Our research suggests that temporal and spatial factors significantly influence disease cases. These results are in line with studies from foreign countries. The study results aspire to expand our knowledge regarding the epidemiological characteristics of Legionnaires’ disease in relation to local, geographical and meteorological factors on the island of Crete.

Travelling and staying in hotels are a known risk factor of infections by Legionella. In Europe, it is known that more than 20% of all declared cases of legionellosis are associated with travelling. In Spain, a country that receives 50 million tourists every year and which has one of the most important tourism industries in the world, the travel-associated Legionnaires' disease (TALD) cases have followed a parallel evolution. The Hotel Inspection Programme, that was in operation for 1 year (June 1999 to July 2000), adequately accomplished the task of providing hygiene-health training of the management and maintenance staff of the hotels involved. Collaterally, it acted as a means of detecting certain generalized hygienic deficiencies in the water and airconditioning systems of the hotels, and is likely to have had a positive influence on their being remedied. The data derived from the evaluations that were made show that the programme was very well received by those taking part and that, consequently, this experience may be used to advantage in other tourism areas in the Mediterranean. Although it is early to venture conclusions on the epidemiological impact of the programme, the data available to us from the first year of its application are very encouraging and suggest that the educational factor may play a key role in the prevention of legionellosis in the tourist sector.

COSMID: A Web-based Tool for Identifying and Validating CRISPR/Cas Off-target Sites.

Travel-associated Legionnaires’ disease (TALD) cluster definition

BackgroundThe detection of a cluster of travel-associated Legionnaires’ disease (TALD) cases in any European Union/European Economic Area (EU/EEA) country prompts action at the accommodation, follow-up by health authorities and reporting of measures taken. Some accommodations incur further cases despite presumed implementation of adequate control measures.AimTo identify factors associated with the occurrence of a further TALD case after the implementation of control measures.MethodsWe conducted a retrospective cohort study of hotel and holiday rental accommodations in the EU/EEA associated with two or more TALD cases with onset dates less than 2 years apart (a ‘cluster’) and notification between 1 June 2011−31 December 2016. We fitted Cox regression models to estimate the association between accommodation characteristics and the occurrence of a further case, defined as any case with onset date after the report on measures taken.ResultsOf the 357 accommodations in the analysis, 90 (25%) were associated with at least one further case after the report on measures taken (12.4/100 accommodation-years). Accommodations associated with two or more cases before the cluster notification were more likely to be associated with a further case, compared with those not previously associated with any case (adjusted hazard ratio 1.85; 95% confidence interval: 1.14–3.02). Neither the detection of Legionella in the water system nor the type of disinfection were found to be associated with the risk of a further case.ConclusionAccommodation size and previous TALD cases were predictive of further Legionnaires’ disease cases after implementation of control measures.

CRISPR-based adaptive and heritable immunity in prokaryotes

Legionnaires' disease and Pontiac fever are both diseases with important public health implications and require prompt and thorough responses to outbreaks for future prevention. Effective methods for defining, diagnosing, reporting and responding to legionellosis outbreaks ideally should be standardized across countries. Therefore, the European Union surveillance methods for countering Legionnaires' disease is a useful model especially for travel-associated Legionnaires' disease (TALD) cases which are on the rise. Multi-country surveillance in the EU and European Economic Area (EEA) has evolved since the first organizational efforts in the 1980's to the currently responsible, European Legionnaires' Disease Surveillance Network (ELDSNet). This chapter outlines the practices of the EU surveillance of Legionnaires' disease including their schemata, definitions, responsibilities of participating members, methods and the results of the data collected since the program's inception. Lastly, improvements must still be made as the incidence of Legionnaires' disease in the EU is likely underestimated due to underreporting and/or underdiagnosis. Nearly 70% of cases are reported from only four countries, France, Germany, Italy and Spain, which represent 50% of the EU population.

Much data has been gathered by the EWGLINET scheme on the distribution of cases of travel-associated Legionnaires' disease (TALD) by country of infection, but less analysis has been carried out on the distribution of these cases within countries. Travel-associated cases with onset in 2002 linked to France, Italy, Spain and Turkey were mapped. Rates of Legionnaires' disease per 100,000 tourists were calculated for internal and foreign visitors for the regions of each country, and mapped. Rates of 1.5 cases/100,000 and 2 cases/100,000 tourists were classified as 'high' and 'very high' respectively. Cases of TALD were concentrated in certain regions, but when rates were calculated using tourist data, the results were relatively constant throughout each country. Rates were higher among foreign visitors than internal visitors; three of the countries had at least one region with 'high' rates, whilst Turkey additionally had three regions with 'very high' rates.

For several years, over 50% of the cases of travel-associated Legionnaires' disease (TALD) reported to the European Working Group for Legionella Infections (EWGLINET) have been among travellers to France, Italy, and Spain. We describe clusters of TALD cases reported in these countries during a four-year period. We analysed data from EWGLINET and from the individual countries. In all three countries, upon notification of a cluster, local health authorities are alerted by the national collaborator and immediately begin an environmental investigation at the accommodation site, which includes risk assessments and analysis of water samples. From July 1, 2002 to June 30, 2006, 2,101 accommodation sites were associated with TALD cases and reported by EWGLINET to Italian, Spanish and French collaborators. Of these, 252 sites (12%) were associated with clusters: 13.8% (96/697) in Italy, 13.2% (81/615) in Spain and 9.5% (75/789) in France. Overall, 641 cases were reported. Hotels, camping sites and ships and other sites represented respectively 83%, 10% and 7% of the total accommodation sites, with similar proportions in the three countries. In 99% of the sites, samples were collected; 62% of them were found to be positive for Legionella. The findings of this study highlight that disinfection and long-term preventive measures were correctly implemented by the large majority of sites. However, additional efforts must be made to further reduce the percentage of re-offending sites so as to reduce the number of accommodations that are contaminated by Legionella.

We determined whether the incidence rates of travel-associated Legionnaires' disease (TALD) in hotels in Germany increased after a previous occurrence and whether control measures required by the European Legionnaires' Disease Surveillance Network after a cluster (>2 cases within 2 years) restored the rate to baseline. We analyzed TALD surveillance data from Germany during 2015-2019; a total of 307 TALD cases (163 domestic, 144 nondomestic) in hotels were reported. The incidence rate ratio was 5.5 (95% CI 3.6-7.9) for a second case and 25 (95% CI 11-50) for a third case after a cluster had occurred, suggesting that control measures initiated after the occurrence of TALD clusters might be inadequate to restore the incidence rate to baseline. Our findings indicate that substantial LD preventive measures should be explored by hotels or other accommodations after the first TALD case occurs to reduce the risk for future infections.

Vital Signs: Deficiencies in Environmental Control Identified in Outbreaks of Legionnaires’ Disease—North America, 2000–2014