Search Results (1,052)

Background/Objectives: Bacterial colonization is an under-recognized component of antimicrobial resistance (AMR), particularly among cardiovascular outpatients who frequently interact with healthcare services. While surveillance has traditionally focused on multidrug-resistant (MDR) pathogens causing clinical infection, the ecological characteristics of colonization including multisite carriage, polymicrobial communities, and commensal or low-virulence organisms recovered on antimicrobial-containing media remain poorly defined. This study investigated the burden and ecological patterns of colonization by organisms with reduced antimicrobial susceptibility in cardiovascular outpatients. Methods: In this cross-sectional study, 36 adult cardiovascular outpatients were screened using sputum, nasal, and skin specimens (108 samples) on antimicrobial-supplemented selective media. Isolates were identified by MALDI-TOF mass spectrometry, susceptibility was determined using the VITEK 2 system, and colonization patterns were analyzed using exact statistical methods. Results: Colonization by organisms with reduced antimicrobial susceptibility was detected in 61.1% of patients (95% CI, 44.9–75.2), and at least one validated MDR pathogen in 52.8% (95% CI, 37.0–68.0). Sputum showed the highest colonization prevalence. Multisite colonization occurred in 72.7% of colonized patients, with 40.9% positive at all three anatomical sites. Polymicrobial colonization was common (mean 1.9 species per colonized patient). Resistant commensal bacteria including Neisseria flavescens, Neisseria mucosa, Moraxella catarrhalis, and Lacticaseibacillus casei were recovered alongside recognized pathogens and showed reduced susceptibility across multiple antibiotic classes. Colonization showed significant concordance across anatomical sites, indicating a whole-body colonization state rather than a series of independent site events. Prior hospitalization and concurrent antibiotic therapy were significantly associated with colonization, whereas prior percutaneous coronary intervention and individual comorbidities were not. Conclusions: Cardiovascular outpatients frequently harbour complex bacterial communities characterized by multisite colonization, polymicrobial carriage, commensal and low-virulence organisms recovered on selective media, and MDR pathogens. These findings highlight bacterial colonization as a potential community reservoir of AMR and support targeted pre-procedural screening together with community antimicrobial stewardship. As a single-centre, cross-sectional study of 36 patients, these results are exploratory and hypothesis-generating and require confirmation in larger, multicentre cohorts. Full article

Listeria spp. can persist in livestock-production environments and may be introduced through animal feed, fecal contamination, or unhygienic milk-production practices. This cross-sectional study assessed the occurrence and distribution of Listeria spp. in animal feed, fecal material, and raw milk collected from cattle, sheep, and goat farms across the seven geographical regions of Kosovo. Between December 2021 and April 2022, 189 samples were collected from 60 distinct farms representing 63 farm-species units. One animal-feed sample, one pooled fecal sample, and one pooled raw-milk sample were obtained from each unit. The samples were analyzed according to ISO 11290-1:2017, and presumptive isolates were evaluated by phenotypic and biochemical tests and confirmed to species level using MALDI-TOF mass spectrometry. Water activity was also measured in all 63 feed samples. Listeria spp. was detected in 3/189 matrix-level samples analyzed (1.6%). This value represents a descriptive sample-level detection proportion across the three matrices rather than a conventional prevalence estimate. All isolates were identified as Listeria innocua. Two positive samples were hay samples from separate farms in the Gjilan region, representing 2/63 feed samples (3.2%) and 2/24 hay samples (8.3%). The third positive sample was a fecal sample collected from a goat farm in the Gjakova region, representing 1/63 fecal samples (1.6%). Listeria spp. was not detected in any of the raw-milk samples (0/63; 0%) under the conditions of this cross-sectional study. No farm-level co-occurrence among feed, fecal material, and raw milk was observed. These findings indicate a low number of Listeria spp. detections among the samples analyzed, with positive samples limited to two hay samples and one fecal sample. Given the small number of positive samples, no conclusions can be drawn regarding differences among feed categories, animal species, sample matrices, or geographical regions. Continued attention to feed storage, farm hygiene, and milk production practices remains important for limiting the persistence and dissemination of Listeria within ruminant-production environments. Full article

Surface-assisted laser desorption/ionization mass spectrometry imaging (SALDI-MSI) using steel target coated with silver nanoparticles (AgNPs) by electrodeposition was applied for the direct visualization of metabolites in bulb tissue of Narcissus pseudonarcissus. Fresh bulb cross-sections were transferred onto an AgNP-SALDI target by a simple tissue imprint procedure and analyzed using a MALDI TOF mass spectrometer operating in positive-ion reflectron mode. Ion images were generated after total ion current normalization and metabolite annotation was performed based on accurate mass measurements, characteristic silver adduct formation, database searches and literature data. Twenty-one ion images representing seventeen putatively annotated metabolites were selected for detailed discussion. The putatively annotated compounds included primary metabolites (histidine, malic acid, succinic acid, thiamine, coenzyme A and acetyl-coenzyme A), phytohormones (indole-3-acetic acid, indole-3-butyric acid, 3-indolepropionic acid, 4-chloroindole-3-acetic acid and abscisic acid), flavonoids and characteristic Amaryllidaceae alkaloids, including galanthamine, lycoramine, crinine, assoanine, habranthine and 5,6-dihydrobicolorine. Distinct spatial distributions were observed for individual metabolites, reflecting the metabolic heterogeneity of bulb tissues. The results demonstrate that AgNPs-SALDI-MSI provides a rapid, matrix-free approach for in situ visualization of low-molecular-weight metabolites in plant tissues while preserving their spatial organization, making it a promising tool for plant metabolomics and phytochemical investigations. Full article

Background: Candida auris (C. auris) has emerged as a multidrug-resistant fungal pathogen causing healthcare-associated outbreaks worldwide, particularly in intensive care units (ICUs). Its environmental persistence, multidrug resistance, and high mortality make it a major challenge for infection prevention and clinical management. This study aimed to investigate the clinical characteristics, clinical exposures, treatment outcomes, and mortality of adult ICU patients with C. auris candidemia during the first 30 months after the opening of a tertiary care hospital. Methods: This retrospective observational study included all adult patients (≥18 years) admitted to five ICUs between 1 July 2020 and 31 December 2022 who developed culture-confirmed C. auris candidemia. Isolates were identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) with the Microflex LT/SH Smart MS system (Bruker Daltonics, Bremen, Germany) according to routine microbiological laboratory procedures. Demographic characteristics, underlying diseases, invasive procedures, clinical exposures, microbiological findings, treatment, APACHE II scores, and clinical outcomes were retrospectively analyzed. Results: During the 30-month study period, 3716 adult patients were admitted to the ICUs, and candidemia developed in 432 (11.6%). Of these episodes, 22 (5.1%) were caused by C. auris. The first C. auris candidemia case occurred 184 days after hospital opening. The mean age was 55 ± 17 years, and overall ICU mortality was 77.3%. Sepsis (36.4%), malignancy (22.7%), and cerebrovascular disease (18.2%) were the most common reasons for ICU admission. All patients had received broad-spectrum antibiotics, whereas central venous catheterization and invasive mechanical ventilation were present in 77.3% of cases. The mean interval from ICU admission to candidemia was 49 ± 15 days. Caspofungin was used as first-line antifungal therapy in all patients. Non-survivors had significantly higher APACHE II scores (32.8 ± 4.9 vs. 26.4 ± 6.6, p = 0.029) and predicted mortality rates (80.1% vs. 53.3%, p = 0.012). Antifungal treatment duration was significantly shorter among non-survivors (p < 0.001), whereas no significant differences were observed in age, ICU length of stay, or time to blood culture clearance. Conclusions: C. auris candidemia was observed in critically ill patients with prolonged ICU stays and frequent invasive interventions and was associated with high all-cause ICU mortality. Higher APACHE II scores were associated with mortality in this cohort, reflecting greater baseline severity of illness. Given the small sample size and descriptive, exploratory nature of the study, these findings should be interpreted with caution. The effectiveness of specific infection-control, diagnostic, or therapeutic interventions was not evaluated. Larger multicenter prospective studies are needed to better characterize the epidemiology, clinical outcomes, and factors associated with mortality in C. auris candidemia. Full article

Endophytic bacteria associated with medicinal plants represent a potential source of microorganisms with useful enzymatic and antagonistic properties. In this study, 14 bacterial isolates were recovered from the leaves, roots, flowers, and petioles of Inula helenium and screened for morphological, biochemical, and hydrolytic characteristics. Isolate IH-B3, which showed the most pronounced starch- and casein-hydrolysis zones during preliminary screening, was selected for further characterization. MALDI-TOF mass spectrometry assigned the isolate to Bacillus subtilis with an identification score of 2.26, and the strain was designated B. subtilis UzMU25. Its 1444 bp 16S rRNA gene sequence was deposited in GenBank under accession number PZ593820. The strain was catalase- and lecithinase-positive but gelatinase- and hemolysis-negative, and it grew at pH 6.5 and in the presence of horse bile under the qualitative assay conditions used. Hydrolysis-zone diameters ranged from 46 to 52 mm for amylase, 34 to 42 mm for protease, and 10 to 16 mm for lipase activity. In direct antagonism assays, UzMU25 inhibited all six bacterial and fungal test organisms, producing inhibition zones of 26–36 mm. Biofilm biomass varied with incubation time and reached its highest corrected OD₅₉₀ value at 12 h (0.780 ± 0.025). Antibiotic-disc testing showed a 10 mm inhibition zone for vancomycin, indicating reduced susceptibility under the applied conditions and requiring further investigation of its genetic basis and potential transferability. No mortality was observed during the preliminary acute oral study following administration of the tested preparation at doses up to 10,000 mg kg⁻¹, and no visible dermal or conjunctival irritation was detected. Overall, UzMU25 exhibited preliminary enzymatic, antagonistic, and biofilm-forming characteristics of biotechnological interest. However, whole-genome antimicrobial-resistance screening and more comprehensive phenotypic and toxicological evaluations are required before the strain can be recommended for probiotic or other practical applications. Full article

Antimicrobial resistance (AMR) in aquaculture environments has become an increasing concern due to the dissemination of antibiotic-resistant bacteria in aquatic ecosystems. This study investigated the occurrence of presumptive antimicrobial-resistant bacteria in commercial fish farms in Hajdú-Bihar county, Hungary. Between 2023 and 2024, water samples were collected from fishponds from Hajdúszoboszló, Biharugra, and Debrecen, followed by membrane filtration with 0.45 µm pore size mixed cellulose ester membrane filters, bacterial isolation using selective media including vancomycin-resistant enterococci (VRE) agar, Eosin Methylene Blue (EMB) agar, and Endo’s medium. The phenotypic characterization revealed the presence of diverse bacterial populations that could be associated with potential antimicrobial resistance. Bacterial identification using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) showed several bacterial genera, predominantly Acinetobacter, Pseudomonas, Enterococcus, Bacillus, and Escherichia. A total of 16 presumptive antimicrobial-resistant bacterial isolates recovered from selective media were identified using MALDI-TOF MS. Several detected species, including Acinetobacter baumannii, Pseudomonas aeruginosa, Enterococcus faecium, and Enterococcus faecalis, are recognized as opportunistic bacteria associated with antimicrobial resistance dissemination. The findings demonstrate that aquaculture waters in Hajdú-Bihar county may serve as reservoirs of antimicrobial-resistant bacteria. Full article

Background: Bacteria of the genus Providencia are emerging pathogens associated with a wide range of infections, particularly in healthcare settings. In veterinary medicine, infections caused by these organisms are uncommon, and, to date, there have been no reports of carbapenem-resistant Providencia spp. causing infection in companion animals. Methods and Results: This study describes a comprehensive investigation of bacterial endocarditis caused by carbapenem-resistant Providencia rettgeri in a cat, including clinical, imaging, pathological, microbiological, and genomic findings. A 1.5-year-old mixed-breed female cat was admitted to a referral veterinary hospital with an extensive pelvic limb wound and a recent history of laparotomy. During hospitalization, a heart murmur was detected, and echocardiographic examination revealed vegetation-like lesion on the mitral valve. Despite intensive treatment, the patient’s clinical condition progressively deteriorated over a 15-day period, ultimately leading to euthanasia. Necropsy confirmed bacterial endocarditis with evidence of systemic septic embolization. P. rettgeri was isolated from blood collected from both heart ventricles and from an endocardial swab and its identity was confirmed by MALDI-ToF mass spectrometry. Antimicrobial susceptibility testing demonstrated resistance to meropenem. Whole-genome sequencing confirmed the species identification and revealed a Col3M plasmid carrying multiple antimicrobial resistance genes, including the carbapenemase gene bla_NDM-1. Discussion: To the best of our knowledge, this is the first report of carbapenem-resistant Providencia spp. causing infection in a companion animal. This case highlights the emergence of Providencia spp. as potential veterinary pathogens capable of causing severe, invasive and fatal infections. Full article

Background: Klebsiella pneumoniae is an opportunistic zoonotic pathogen that can cause respiratory diseases in chickens. The aim of this study was to investigate the biofilm formation, resistance, and virulence of K. pneumoniae isolates from chickens in Henan Province, China, between 2023 and 2025, and to analyze the associations among these characteristics. Methods: We identified the isolates through blood agar culture, Gram staining and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Hypermucoviscosity phenotype, biofilm-forming ability, antimicrobial resistance and virulence genes were assessed by the string test, crystal violet assay, disk diffusion and polymerase chain reaction (PCR), respectively. Results: A total of 102 K. pneumoniae isolates were identified. Of these, 81.4% were biofilm-forming strains, and 15.7% exhibited the hypermucoviscosity phenotype. The susceptibility profiles of the K. pneumoniae isolates indicated high resistance to penicillins (>90.0%) and low resistance to carbapenems (<10%), with 88.2% identified as multidrug-resistant. The results of virulence gene detection indicated that luxS (79.4%), mrkD (75.5%), and uge (73.5%) were the most prevalent virulence genes, followed by wabG (49.0%), ybtA (34.3%), and iucA (20.6%). The association analysis indicated that biofilm formation was significant association with multidrug-resistant and the carriage of virulence genes. Compared with non–biofilm–forming isolates, biofilm–forming isolates showed significantly higher resistance to ceftiofur and florfenicol, as well as higher detection rates of luxS, mrkD, and uge (p < 0.05). Conclusions: The data obtained in this study reveal the pathogenic potential and multidrug–resistant characteristics of this pathogen, highlighting the need for surveillance and monitoring. Full article

Background: Ignatzschineria species are rare emerging human pathogens that are typically associated with myiasis. Methods: We report the first case of Ignatzschineria indica bacteraemia in Greece in a male patient presenting with maggot-infested wounds of the thorax, shoulders, and arms. The isolate was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and antimicrobial susceptibility testing was performed according to EUCAST guidance. A review of the published literature on I. indica bacteraemia was also conducted. Results: The patient was diagnosed with bacteraemia caused by a multisusceptible I. indica isolate and was successfully treated with antibiotic therapy, resulting in complete recovery. This represents the first reported case of I. indica infection in Greece. Review of the literature showed that most published I. indica isolates were susceptible to a broad range of antimicrobial agents, and all reported patients survived, although some required limb amputation because of severe underlying soft tissue disease. Conclusions: Ignatzschineria indica infection should be considered in patients with neglected, maggot-infested wounds. Increased awareness of this emerging pathogen, together with the use of modern microbiological identification methods, may improve recognition of these uncommon infections. Socio-economic inequalities and the changing distribution of insect vectors due to climate change may contribute to the occurrence of such infections. Full article

Stray cats may act as reservoirs of antimicrobial-resistant and potentially pathogenic bacteria, representing a potential public health concern within a One Health framework. This study investigated the occurrence of antimicrobial resistance (AMR), multidrug resistance (MDR), and virulence-associated traits in commensal Escherichia coli isolated from free-living cat colonies in southern Spain. A total of 169 rectal swabs were collected from cats belonging to feline colonies and shelters in Benalmádena (Málaga, Spain). Bacterial isolation and identification were performed using selective culture media, conventional biochemical tests, and MALDI-TOF mass spectrometry. Antimicrobial susceptibility was determined by minimum inhibitory concentration (MIC) testing against 15 antimicrobial agents, and isolates were screened for selected virulence-associated genes associated with major diarrheagenic E. coli pathotypes. A total of 68 E. coli isolates (40.2%) were recovered. The highest resistance frequencies were observed for sulfamethoxazole (25.0%) and ampicillin (20.6%), whereas all isolates showed high susceptibility rates to most of the antimicrobials tested, including azithromycin (100%), as well as gentamicin, amikacin, cefotaxime, ceftazidime, meropenem, colistin, chloramphenicol and tigecycline (98.5%). Six isolates (8.8%) were classified as multidrug-resistant. In addition, six isolates (8.8%) were identified as atypical enteropathogenic E. coli (aEPEC), although none exhibited a multidrug resistance phenotype. These findings demonstrate the presence of both AMR and virulence-associated traits among commensal E. coli circulating in free-living cats. The inclusion of feline colonies or shelters in AMR surveillance programmes may contribute valuable information for integrated One Health monitoring strategies. Full article

Dentin derived from extracted human teeth has emerged as a promising autogenous biomaterial for bone regeneration owing to its structural and biological similarity to bone tissue. Growing clinical interest in dentin-derived graft materials has also stimulated the development of tooth-banking concepts, in which extracted teeth are preserved for future regenerative applications. However, microbiological contamination remains one of the principal concerns associated with long-term storage before clinical use. This in vitro study evaluated the microbiological status of dentin biomaterials prepared from human teeth stored under long-term refrigerated conditions (approximately 4 °C) and subsequently processed using three commercially available dentin-processing systems: BonMaker (BM), Tooth Transformer (TT), and Smart Dentin Grinder (SDG). A total of 72 extracted teeth collected between 2018 and 2025 were allocated to experimental groups and processed into 24 pooled dentin samples. Five freshly extracted teeth served as controls. Culture-based microbiological assessment was performed before and after dentin processing using Schaedler Broth K3 culture media under aerobic and anaerobic conditions. Microorganisms isolated from positive cultures were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Baseline microbiological assessment detected bacterial contamination in 2 of 24 pooled dentin samples (8.3%) and in 4 of 5 control teeth (80.0%). The identified microorganisms included Staphylococcus epidermidis, Actinomyces viscosus, Streptococcus anginosus, Streptococcus mutans, and Staphylococcus hominis. No microbiologically detectable bacterial growth by culture-based methods was observed following processing with BM, TT, or SDG, irrespective of storage duration, including teeth preserved under refrigerated conditions for up to seven years. Within the limitations of this culture-based investigation, the evaluated dentin-processing protocols effectively eliminated microbiologically detectable cultivable bacterial contamination from long-term refrigerated dentin samples. These findings provide preliminary culture-based microbiological evidence supporting the feasibility of long-term refrigerated tooth preservation under the investigated storage conditions as one component of future tooth-banking research. Since culture-based microbiological methods do not detect viable but non-culturable microorganisms or other non-cultivable pathogens, the present findings should not be interpreted as evidence of complete sterility, overall biological safety, or the superiority of refrigerated storage over alternative preservation methods. Further studies employing molecular microbiological techniques together with biological and physicochemical analyses are required to comprehensively validate long-term tooth-banking protocols. Full article

Neonatal sepsis remains a leading cause of morbidity and mortality in the first month of life, yet its presentation is nonspecific, and its microbiological confirmation is uniquely difficult. Low-circulating bacterial loads, small obtainable blood volumes, frequent intrapartum antibiotic exposure, and the ambiguity of skin-commensal isolates erode the sensitivity and specificity of blood culture, the long-standing reference standard, so that most evaluated infants are treated empirically despite sterile cultures. This review synthesizes current evidence on the integrated microbiological evaluation of the newborn: the host- and specimen-related diagnostic challenge; the etiologic spectrum and its antimicrobial-resistance burden; conventional culture; rapid and molecular methods, including mass spectrometry and syndromic and broad-range PCR; metagenomic and cell-free-DNA sequencing; host-response biomarkers; and antimicrobial-susceptibility determination. The evidence indicates that newer methods accelerate or supplement culture but do not yet replace it, each addressing one limitation while introducing caveats of its own. Notably, much of the evidence for direct-from-blood molecular testing, microbial cell-free DNA, and metagenomic sequencing is extrapolated from adults, whereas neonatal data are stronger for several biomarkers. The greatest benefit arises from integrating complementary modalities within diagnostic- and antimicrobial-stewardship frameworks. Neonatal-specific validation, standardized thresholds, and outcome-driven trials remain the principal priorities. Full article

MicroRNAs (miRNAs) serve as pivotal post-transcriptional modulators of gene expression that govern animal growth and development processes. The present study aimed to investigate the relationship between a single-nucleotide polymorphism (SNP) located within the pre-miR-1453 precursor and growth traits, carcass traits, body size traits, and serum biochemical parameters in an F₂ chicken resource population derived from Gushi and Anka crosses (n = 860). Genotyping of the rs16159272 polymorphism (+31 bp C > T) was performed using MALDI-TOF mass spectrometry, and population genetic parameters, secondary structure prediction, and association analyses were conducted. The results showed that the SNP was moderately polymorphic (PIC = 0.3174) and significantly associated with hatch weight (p < 0.01; CT > TT > CC), as well as body weight at 2 and 4 weeks of age (p < 0.05; TT > CT > CC), but not with carcass or body size traits (p > 0.05). For serum biochemical parameters, total protein, cholinesterase, and creatine phosphokinase showed extremely significant differences across genotypes (p < 0.01), whereas albumin, globulin, and lactate dehydrogenase showed significant differences (p < 0.05). Structural prediction revealed that the C > T mutation increased the minimum free energy by 3.8 kcal/mol, reducing the stability of the pre-miR-1453 stem-loop. Target gene prediction and enrichment analysis indicated that pre-miR-1453 may participate in the regulation of chicken growth and serum biochemical parameters through pathways related to neuron development, cytoskeletal dynamics, and energy metabolism. These findings suggest that the pre-miR-1453 rs16159272 polymorphism could act as a candidate genetic indicator for marker-aided breeding in poultry. Full article

Lys49 phospholipase A2 (Lys49-PLA₂) homologues are catalytically inactive snake venom toxins with diverse biological activities, but their functional characterization requires recombinant production strategies to overcome the limitations inherent to native venom-derived proteins. Here, we produced recombinant BaMtx (rBaMtx), a Lys49-PLA₂ homologue from Bothrops atrox, in the Pichia pastoris KM71 expression system and evaluated whether its biochemical and biological properties were preserved. rBaMtx was secreted into the culture medium, purified by cation-exchange chromatography, and characterized by SDS-PAGE, Western blotting, RP-HPLC (85.9% purity), and MALDI-TOF mass spectrometry, confirming a molecular mass of 13,821 Da. Despite lacking PLA₂ activity, rBaMtx retained pronounced myotoxicity in vivo, inducing dose-dependent plasma creatine kinase release and skeletal muscle damage comparable to or greater than native BaMtx. In vitro, rBaMtx reduced viability of murine 4T1 breast carcinoma cells (IC₅₀ = 60.94 µg/mL), altered cell morphology, modestly increased IL-1β release, and produced context-dependent effects in paired human tumor and non-tumor cell models, differing from native BaMtx and crude venom. These findings establish rBaMtx as a reproducible model for investigating the mechanisms and biomedical potential of catalytically inactive Lys49-PLA₂ homologues. Full article

Microbial contamination in hospitality settings remains understudied despite the high density of human contact and diverse operational activities that characterize hotel environments. This study aimed to characterize microbial contamination patterns and environmental hygiene risks across multiple functional areas of family-oriented hotels. A cross-sectional environmental microbiological investigation was conducted in family hotels in Bavaria, Germany. A total of 225 environmental surface samples were collected from guest rooms, child areas, food-related areas, service environments, and water-exposed locations. Bacterial isolates were identified using culture-based microbiology and MALDI-TOF mass spectrometry. Microbial prevalence, contamination severity, microbial richness, and pathogen prevalence were assessed using mixed-effects ordinal logistic regression, generalized additive model (GAM), and integrated hygiene profile, all performed in R (version 4.5.1). Microbial occurrence patterns differed markedly across functional areas. Contamination category distributions differed significantly among areas, with food-related environments showing the strongest enrichment in the highest contamination category (72.7%). Food-related areas showed significantly greater odds of severe contamination than guest rooms (OR = 8.37, 95% CI: 1.89–37.00), child areas (OR = 6.16, 95% CI: 1.20–31.46), and water-exposed environments (OR = 12.34, 95% CI: 2.45–62.10). Microbial richness differed across operational zones (p = 0.048) and was positively associated with contamination severity (β = 0.141, p < 0.001). GAM revealed a significant non-linear richness–contamination relationship (p < 0.001). Integrated hygiene profile consistently identified food-related and service areas as the highest risk environments. Environmental hygiene risks in hospitality settings display functional-area heterogeneity, highlighting the need for targeted, area-specific hygiene management strategies. Full article