Search Results (126)

Legionella pneumophila is widely recognized as the principal organism responsible for Legionnaires’ disease. Modifications of surface-exposed lipopolysaccharide in L. pneumophila are key determinants of bacterial adaptation to host cells. Using a mutant strain deficient in N-methylation of legionaminic acid and fluorescence-based biophysical analyses, it was demonstrated that N-methyl groups linked to the acetimidoylamino group of legionaminic acid modulate the physicochemical properties of the bacterial cell surface and influence the interaction of L. pneumophila with macrophages. Loss of N-methyl groups reduced the efficiency of bacterial interaction with THP-1-derived macrophages and impaired intracellular proliferation. In addition, the presence of N-methyl groups in legionaminic acid contributes to increased TNF-α production in THP-1 macrophages stimulated with L. pneumophila sg1 LPS, without affecting IL-6 induction, suggesting that N-methylation of legionaminic acid may skew the early pro-inflammatory response toward TNF-α-dominated signaling during L. pneumophila infection. Full article

Travel-associated Legionnaires’ disease (TALD) investigations in hotels have generated extensive environmental monitoring data. However, the occupational implications for workers who operate, maintain, clean, or inspect the same systems are rarely assessed. We developed a hybrid framework integrating a semi-quantitative environmental hazard model with deterministic Quantitative Microbial Risk Assessment (QMRA). In the first model, culture concentration bands were combined with physicochemical deviation indicators (temperature, free residual chlorine, and pH) to derive point-level hazard ($H_i$) and zone-level hazard (${\overline{H}}_z$). In the second model, a job-based presence matrix was combined with zone-specific serogroup-based severity using a simplified World Health Organization (WHO)-style 3 × 3 likelihood–severity approach. Legionella pneumophila (≥50 CFU/L) was detected in 29.94% of water samples and was significantly associated with low chlorine (<0.2 mg/L; RR 2.90) and hot water temperature < 55 °C (RR 3.07). To support comparative occupational exposure stratification, QMRA was applied to estimate the daily inhaled dose ($\mathrm{d}$) for 15 worker groups, indicating variability in modeled biological exposure across occupational categories. Within this framework, modeled occupational exposure potential was shaped by the combined influence of pathogen concentration and assumed exposure duration. Under the hazard model, the highest zone-level hazard estimate was observed in kitchens and food and beverage (F&B) areas (${\overline{H}}_z$ = 2.607), followed by machinery rooms (${\overline{H}}_z$ = 2.022) and guest rooms (${\overline{H}}_z$ = 1.874). These findings support the integration of worker protection into water safety management, particularly in areas and groups overlooked in routine investigations. Full article

In Europe, travel-associated Legionnaires’ disease (TALD) cases require timely environmental investigations to support risk assessment, rapid control measures, and prompt reporting of investigation findings to the European Legionnaires’ Disease Surveillance Network (ELDSNet). This study evaluated TALD-related environmental investigations conducted during 2025 and early 2026 in Crete, Greece, following notifications through ELDSNet. Overall, 30 notifications corresponded to 24 unique confirmed TALD cases with illness onset in 2025 and 24 implicated hotels, with some cases involving stays in multiple hotels and Regional Units and clusters identified in some implicated hotels. The investigation framework combined microbiological, physicochemical, and operational data, focusing on delays from symptom onset, notification, sampling, and laboratory reporting. Overall, 516 environmental samples were collected, of which 503 yielded valid analytical results. Among the 503 samples analyzed, Legionella spp. were detected at ≥50 colony-forming units per liter (CFU/L) in 127 samples (25.25%). This included 123 samples positive for L. pneumophila (24.45%), of which 31 were serogroup 1 (6.16%). Concentrations exceeding the 1000 CFU/L threshold were recorded in 53 samples (10.54%). Operational indicators varied, with median values of 31.0 days for reporting delay (RD), 14.5 days from notification to first sampling (TTF), 47.5 days from symptom onset to first sampling (TDS), and 67.0 days from symptom onset to first laboratory result (OELR). The findings underscore the necessity to document response delays, enhance inspector capacity and cross-regional coordination, and integrate microbiological results with operational indicators. This integration is crucial for facilitating earlier environmental risk assessments, expediting reporting, and implementing more effective TALD public health interventions. Full article

Legionnaires’ disease is a rare cause of atypical pneumonia associated with a high mortality rate among untreated patients. In Romania, the disease has historically been underreported due to insufficient surveillance and limited diagnostic capacity. The aim of this study was to describe the characteristics of Legionnaires’ disease cases admitted to a specialized infectious disease hospital between 2023 and 2024, with a particular focus on a cluster associated with travel to a spa resort. Most cases included in our study (31/36) were confirmed by urinary antigen testing, while one case was confirmed by a significant increase in the level of specific antibodies against Legionella pneumophila serogroup 1 in paired serum samples. The most frequently reported symptom was fever (28/32), followed by chills (24/32). Among the 32 confirmed cases, 3 patients died. Two cases were identified as part of a family cluster involving a father and son who had undergone physiotherapy at a balneary resort. Both patients presented with fever and gastrointestinal symptoms, and radiological investigations confirmed mixed pneumonia associated with an intense inflammatory syndrome. In the father’s case, hepatic involvement and interstitial nephritis were also identified. Early diagnosis based on epidemiological data, clinical predictive scores, and laboratory investigations would allow timely administration of targeted antibiotic therapy and may contribute to reduced mortality. Full article

Travel-associated Legionnaires’ disease (TALD) events can generate public concern when environmental surveillance findings are communicated without an adequate explanation of the results. This study examined how surveillance data on Legionella spp. were framed and amplified during a TALD-related investigation in Crete, Greece, from June to July 2025. A mixed infodemiology and environmental surveillance approach was applied, including the analysis of 95 online media items across nine languages, Google Trends search-interest data, and hotel water-system surveillance data from epidemiologically linked facilities. Sampling conducted in a limited number of hotels associated with TALD cases indicated that approximately 50% of the water samples exceeded the laboratory reporting limit of ≥50 CFU/L for Legionella spp., a numerically correct but context-specific finding. Numerical misframing occurred in 83.7%, 41.7%, and 18.2% of Greek, German, and English language items, respectively, with significant differences across language markets (χ² (8) = 43.75, p < 0.0001; Cramér’s V = 0.679). Public search-interest signals were transient and geographically limited. Environmental surveillance showed no increase in Legionella pneumophila risk, with similar proportions of samples ≥50 CFU/L in the pre-/peri-infodemic (January–July 2025) and post-infodemic (August–November 2025) periods (23.11% [95% CI: 18.21–28.87] vs. 24.45% [19.34–30.41]) and similar exceedance of ≥1000 CFU/L (13.45% [9.69–18.36] vs. 14.41% [10.45–19.55]). Overall, the loss of contextual interpretation of surveillance results and conflation of laboratory reporting limits with regulatory thresholds were associated with inconsistent public risk perception, without evidence of increased environmental hazard. Full article

Legionella species are ubiquitous bacteria found worldwide in water, moist environments, soils, and compost. Infection occurs through the inhalation of aerosols, leading to either Pontiac fever or Legionnaires’ disease (LD). Current routine diagnostics typically combine culture-based isolation with Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) for species identification and the Latex Agglutination Test (LAT) for serotyping. However, this workflow is fragmented: MALDI-TOF MS lacks serogroup-specific resolution, while LAT relies on subjective visual interpretation. Therefore, this study evaluated Fourier-transform infrared spectroscopy (FT-IR) as a rapid, high-resolution typing method for Legionella isolates to assess its potential as a single-step diagnostic tool. A total of 200 clinical and environmental Legionella isolates were analyzed using FT-IR, including L. pneumophila serogroups (SG) 1–15 and various non-pneumophila species. Spectral data were analyzed using Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA). While MALDI-TOF MS provided accurate species identification, FT-IR spectroscopy demonstrated superior typing capabilities by successfully distinguishing L. pneumophila SG 1 distinct from the SG 2–15 complex and allowing for clear discrimination of most non-pneumophila species. Additionally, FT-IR resolved isolates that showed ambiguous or non-reactive results in LAT. These findings demonstrate that FT-IR overcomes the serotyping limitations of MALDI-TOF MS and offers a more objective, cost-efficient extension to the current multi-step routine, potentially closing the diagnostic gap between simple species identification and deep strain characterization. Full article

Legionella pneumophila (Lp) remains one of the major causes of community- and hospital-acquired pneumonia, yet its diagnosis and source attribution continue to pose significant challenges. Here, we describe the case of an immunocompromised patient who developed Legionnaires’ disease during hospitalization. Following activation of the hospital’s internal surveillance system, Lp and Legionella anisa (L. anisa) were recovered from multiple water distribution points using a simplified culture-based protocol. Whole-genome sequencing (WGS) demonstrated that all environmental isolates belonged to a single clonal strain, whereas the clinical isolate was genetically unrelated, thereby excluding the hospital water system as the source of infection. Although not implicated in the patient’s disease, the detection of both Lp and L. anisa within the plumbing system highlighted underlying structural contamination and the potential masking effect of non-L. pneumophila species during culture-based surveillance. These findings support the integration of conventional microbiological methods with high-resolution genomic tools to enhance surveillance accuracy, support outbreak investigations, and strengthen public health responses. Overall, this case underscores the value of WGS as a decisive tool for source attribution, including the robust exclusion of a suspected nosocomial source, in complex clinical and environmental scenarios. Full article

Legionella pneumophila (L. pneumophila), the primary causative agent of Legionnaires’ disease, is a waterborne bacterial pathogen that poses significant public health concern. This opportunistic pathogen commonly inhabits both natural and man-made water systems, particularly drinking water distribution systems (DWDSs), where it can proliferate and pose a risk to human health. In this study, we evaluated the potential of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) for rapid and accurate subtyping of L. pneumophila. Our analysis included 70 L. pneumophila strains collected from the Middle East, representing one of the largest and most comprehensive MALDI-TOF MS-based subtyping of strains from this geographically underrepresented region. These strains, representing three Multi-Locus Variable Number Tandem Repeat Analysis (MLVA-8) genotypic groups (GT4, GT6, and GT15), have been extensively characterized in previous studies for their virulence traits, cytotoxicity patterns, and antimicrobial susceptibility profiles. Our findings revealed distinct genotype-associated spectral signatures with 30 discriminatory m/z peaks (p ≤ 0.005). These markers enabled accurate genotype-level classification, achieving over 85% classification accuracy with a Random Forest model and over 71% accuracy using a Decision Tree algorithm. Importantly, the m/z peak at 5358 was uniquely present in the GT15 strains, whereas m/z 5353 was consistently detected in both GT4 and GT6 isolates, demonstrating the potential of specific mass peaks to serve as reliable genotype markers. Furthermore, GT15 strains consistently formed a separate cluster in both Principal Component Analysis (PCA) and hierarchical analyses, whereas GT4 and GT6 exhibited partial overlap, reflecting their exceptionally high genomic similarity. Full article

Legionnaires’ disease remains a relevant public health concern, with transmission linked to droplets from diverse aquatic environments, and its burden across Europe, including in Portugal, has been trending up. Vehicle windshield washer reservoirs have been proposed as potential, yet underexplored, habitats for Legionella spp. In this study, we investigated 62 windshield washer fluid samples collected in central Portugal. Cultivation on selective BCYE agar supplemented with GVPC and subsequent molecular identification revealed no evidence of Legionella spp. However, 23 morphologically distinct bacterial isolates were recovered, and sequencing confirmed diverse taxa from the genera Brevundimonas, Sphingomonas, Ralstonia, and Xanthobacter. These findings indicate that washer reservoirs can sustain microbial communities characterized by environmental resilience and biofilm-forming potential, traits that overlap with ecological niches exploited by Legionella. Although no Legionella was detected, this work represents the first systematic survey of windshield washer reservoirs in Portugal, emphasizing their potential role as overlooked microbial ecosystems and highlighting the importance of continued surveillance. Broader characterization of microbial communities in such artificial aquatic systems may yield insights into microbial interactions that shape pathogen persistence and suppression. Full article

Legionella pneumophila, a waterborne pathogen naturally present in freshwater and capable of colonizing artificial water systems, is responsible for Legionnaires’ disease (LD), a severe form of pneumonia transmitted through inhalation of contaminated aerosols. Virulence of Legionella strains is affected by the plasticity of their genome, shaped by horizontal gene transfer and recombination events. Thus, contaminated water systems can host diverse Legionella populations with a distinct virulence potential. Here, we compare the genomic diversity of Legionella pneumophila strains isolated in water systems of academic buildings, together with their cytotoxicity and intracellular replication in THP-1-like macrophages. A six-year environmental surveillance revealed Legionella pneumophila contamination in 20 out of the 50 monitored sites, identifying five serogroups (sg) and 13 Sequence Types (STs). Phylogenetic investigations based on core genome multilocus sequence typing (cgMLST) and comparative genomics of representative isolates of each ST showed a broad diversity and a heterogeneous virulence repertoire, especially within the Dot/Icm and Lvh secretion systems. Following macrophage infection, a strain-dependent cytotoxicity and intracellular replication was observed, underlying significant pathogenic diversity within the same species and stage-dependent infection dynamics. Together, these results showed strain-specific genetic and phenotypic virulence traits to be considered during risk assessment in environmental surveillance. Full article

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. Full article

Urban settings in developing countries present unique challenges such as high population density, inadequate water infrastructure and water supply, all factors that contribute to the growing threat of premise plumbing pathogens such as Legionella. Water droplets from showers and faucets aerosolise Legionella, which, when inhaled, invade the human respiratory tract to manifest as Legionnaires’ disease. Densely populated, high-rise buildings present an ideal case study for investigating the presence of Legionella. The aim of this study was to investigate the occurrence of Legionella pneumophila (L. pneumophila) in water systems of 15 high-rise buildings in Hillbrow, Johannesburg, South Africa. A total of 67 hot- and cold-water samples and 121 swab samples were collected and analysed for the presence of Legionella pneumophila. Samples were analysed using the Legiolert assay, the South African National Standard (SANS) 11731:2017 method, and the amoeba enrichment method for detecting amoeba-associated Legionella. Molecular confirmation of the pathogen was conducted using conventional PCR and quantitative real-time PCR targeting the mip gene. Legionella pneumophila was found in 93% (14/15) of the buildings that were sampled and was more prevalent in cold-water samples (65%) compared to warm-water (35%) samples. All buildings were positive (100%) for the growth of free-living amoeba (FLA) from water and swab samples. Of these samples, three were confirmed positive for L. pneumophila by PCR and the sequencing alignment results confirmed the identity and relatedness of the isolates to L. pneumophila. Full article

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. Full article

In this paper, the authors introduce the transmuted geometric integer-valued autoregressive model with periodicity, designed specifically to analyze epidemiological and public health time series data. The model uses a transmuted geometric distribution as a marginal distribution of the process. It also captures varying tail behaviors seen in disease case counts and health data. Key statistical properties of the process, such as conditional mean, conditional variance, etc., are derived, along with estimation techniques like conditional least squares and conditional maximum likelihood. The ability to provide k-step-ahead forecasts makes this approach valuable for identifying disease trends and planning interventions. Monte Carlo simulation studies confirm the accuracy and reliability of the estimation methods. The effectiveness of the proposed model is analyzed using three real-world public health datasets: weekly reported cases of Legionnaires’ disease, syphilis, and dengue fever. Full article