The objective of the present study is to report the detection and the molecular characterization of nine bla_NDM-1-positive Pseudomonas aeruginosa isolates, all of which belonged to the epidemic high-risk international clone ST308, and all were isolated from patients in a tertiary care hospital in Central Greece from May to July 2023.The isolates were characterized by whole genome sequencing to obtain multi-locus sequencing typing (MLST) and identify the bla_NDM1-environment and resistome and virulence genes content. In silico MLST analysis showed that all isolates belonged to the high-risk ST308 international clone. All strains possessed 22 different genes, encoding resistance to various antimicrobial agents. Whole genome sequencing revealed that the bla_NDM-1 was chromosomally located within the integrative and conjugative element ICE_Tn43716385 and that it was part of one cassette along with two other resistance genes, floR and msrE. Two additional resistance cassettes were also found in the genome, which included the arrays of aph(6)-Id, aph(3″)-Ib, floR, sul2 and aadA10, qnrVC1, aac(3)-Id, dfrB5, aac(6′)-II. Additionally, the strains possessed various virulence genes, e.g., aprA, exoU, lasA, lasB, toxA, and estA. All of the isolates shared identical genomes, which showed 98% similarity with the P. aeruginosa ST308 genome (acc. no CP020703), previously reported from Singapore. To our knowledge, this is the first report of ST308 bla_NDM-1-positive P. aeruginosa isolation in Europe, which indicates the transmission dynamics of this high-risk clone.

To date, three carbapenem resistance mechanisms have been identified: carbapenemase released from the pathogen, changes in the expression of the outer membrane OprD porin, and overexpression of the efflux pump MexAB-OprM. Twelve carbapenemase-negative carbapenem-resistant Pseudomonas aeruginosa strains, isolated from patients hospitalized at the University Hospital of Larissa, Central Greece, during 2023, which belonged to various sequence types (STs), were selected and were studied focusing on the characterization of their β-lactamases, on changes to OprD and its regulator MexT proteins, and on alterations to the MexAB-OprM regulator proteins encoded by the mexR, nalC, and nalD genes. Whole genome sequencing analysis revealed the presence of β-lactamase encoding genes, with bla_PAO present in all isolates. Additionally, seven different genes of the oxacillinase family (bla_OXA-35, bla_OXA-50, bla_OXA-395, bla_OXA-396, bla_OXA-486, bla_OXA-488, bla_OXA-494) were identified, with each strain harboring one to three of these. Regarding the OprD, five strains had truncated structures, at Loop 2, Loop 3, Loop 4, and Loop 9, while the remaining strains carried previously reported amino acid changes. Further, an additional strain had a truncated MexR; whereas, two other strains had totally modified NalC sequences. The active form of MexT, responsible for the downregulation of OprD production, as the intact sequence of the NalD protein, was found in all the strains studied. It is concluded that the truncated OprD, MexR, and NalC proteins, detected in eight strains, probably led to inactive proteins, contributing to carbapenem resistance. However, four strains carried known modifications in OprD, MexR, and NalC, as previously reported in both susceptible and resistant strains, a finding that indicates the complexity of carbapenem resistance in P. aeruginosa.

The objective of the present study is to report the detection and the molecular characterization of nine bla_NDM-1-positive Acinetobacter baumannii isolates, which were isolated from patients in a tertiary care hospital in Central Greece from December 2022 to August 2023. The isolates were characterized by whole genome sequencing to obtain Pasteur multilocus sequencing typing (MLST) and to identify the bla_NDM-1-environment, resistome, and virulence genes content. In silico MLST analysis showed that the isolates belonged to four different clones (STs 160, 2, 85, and 2493). All strains, apart from the bla_NDM-1-gene, possessed at least eight different genes, encoding resistance to various antimicrobial agents. Whole genome sequencing revealed two different structures of the bla_NDM-1 environment. The first, detected in ST160 strain, was identical with the Tn125, whereas the second, found in STs 2, 85, and 2493 was associated with Tn7382. To our knowledge, after a sole strain reported in 2016 and imported by a patient hospitalized in a Libyan hospital, this is the first report of the emergence of polyclonal bla_NDM-1-positive Acinetobacter baumannii in Greece. Our findings re-emphasize the need to apply diligent surveillance protocols in order to limit the horizontal transfer of the bla_NDM-1 gene to other A. baumannii clones or to other recipient strains.

The objectives of this study were (a) to detect zoonotic gastrointestinal pathogens in faecal samples of horses using the FilmArray^® GI Panel and (b) to identify variables potentially associated with their presence. Faecal samples collected from 224 horses obtained during a countrywide study in Greece were tested by means of the BioFire^® FilmArray^® Gastrointestinal (GI) Panel, which uses multiplex-PCR technology for the detection of 22 pathogens. Gastrointestinal pathogens were detected in the faecal samples obtained from 97 horses (43.3%). Zoonotic pathogens were detected more frequently in samples from horses in courtyard housing (56.0%) than in samples from horses in other housing types (39.7%) (p = 0.040). The most frequently detected zoonotic pathogens were enteropathogenic Escherichia coli (19.2% of horses) and Shiga-like toxin-producing E. coli stx1/stx2 (13.8%). During multivariable analysis, two variables emerged as significant predictors for the outcome ‘detection of at least one zoonotic pathogen in the faecal sample from an animal’: (a) the decreasing age of horses (p = 0.0001) and (b) the presence of livestock at the same premises as the horses (p = 0.013). As a significant predictor for the outcome ‘detection of two zoonotic pathogens concurrently in the faecal sample from an animal’, only the season of sampling of animals (autumn) emerged as significant in the multivariable analysis (p = 0.049). The results indicated a diversity of gastrointestinal pathogens with zoonotic potential in horses and provided evidence for predictors for the infections; also, they can serve to inform horse owners and handlers regarding the possible risk of transmission of pathogens with zoonotic potential. In addition, our findings highlight the importance of continuous surveillance for zoonotic pathogens in domestic animals.

This study aimed to investigate the marked increase in bla_NDM-1-positive Acinetobacter baumannii isolates (20% of all CRAB isolates) observed in a tertiary-care hospital in Central Greece during 2025. Thirty bla_NDM-1-producing carbapenem-resistant A. baumannii (CRAB) isolates were selected and subjected to whole-genome sequencing for molecular typing and characterization of antimicrobial resistance determinants and virulence-associated genes. The majority of the isolates (22/30) belonged to ST2^Pasteur (18 strains), ST85^Pasteur (2 strains) and ST2493^Pasteur (2 strains), all previously identified in 2023. The remaining eight strains belonged to newly detected STs, such as ST1^Pasteur (six strains) and ST2806^Pasteur (two strains). All strains carried the bla_NDM-1 gene on the Tn7382. The high prevalence of bla_NDM-1-positive CRAB represents not only a local threat but also a national concern, as patients from our hospital are frequently transferred to other healthcare facilities or long-term care centers across the country, creating a substantial risk for further dissemination of these strains. Routine molecular screening for bla_NDM-1-positive CRAB may represent a valuable strategy for limiting further dissemination.