Search Results (29)

In hospital environments, pathogenic bacteria spread easily and acquire virulence and antibiotic resistance genes. The aim of the study was an evaluation of the genetic diversity of 109 K. pneumoniae isolates recovered from patients of a district hospital in central Poland. The frequencies of genes coding for β-lactamases, efflux pumps, and virulence factors were determined. Genotyping of the isolates was performed with ERIC (Enterobacterial Repetitive Intergenic Consensus) and REP (Repetitive Element Sequence Based) PCR techniques, with 21 and 19 genotypes being identified, respectively. The bla_SHV-1 (92.7%), bla_CTX-M group 1 (83.5%), bla_TEM-1 (28.4%), bla_NDM-1 (16.5%), bla_VEB-1 (11.0%), bla_CTX-M group 9 (3.7%), bla_KPC (1.8%), bla_IMP, bla_OXA-48, bla_CTX-M group 2, bla_CTX-M groups 8, and 25/26 (0% each) and efflux pumps: AcrAB (100%), tolC (93.6%), and mdtk (60.5%), and virulence genes coding: urease subunit ureA (94.5%) endotoxins wabG (92.7%) and uge (64.2%), and siderophore iucB (3.7%) were detected. The bla_SHV-1, bla_CTX-M group 1, mdtk, tolC, AcrAB (16.5%); bla_SHV-1, bla_CTX-M group 1, tolC, AcrAB (15.6%), and bla_SHV-1, bla_CTX-M group 1, bla_NDM-1, mdtk, tolC, AcrAB (11.9%) were the most common resistance patterns. The distribution of resistance and virulence genes varied more between hospital wards than between different clinical materials. Hospital’s antibiotic-resistant and virulent K. pneumoniae, able to spread among humans, animals, and in the environment, pose a significant threat to public health. Full article

Background/Objectives: Klebsiella pneumoniae ST307 is emerging as a significant global high-risk antimicrobial-resistant (AMR) clone with a notable capacity to acquire and disseminate resistance genes. However, there is limited research on the pathogenicity, virulence, and adaptation of ST307 strains and on the clinical characteristics of infected patients. Methods: In this study, a carbapenem-resistant K. pneumoniae (CRKP) ST307 strain named U989 was isolated from a urine culture of a hospitalized patient in Volos, Greece, in July 2024. Whole-genome sequencing was performed to identify resistance genes to β-lactams bla_KPC-2, bla_CTX-M-15, bla_TEM-1B, bla_OXA-1, bla_OXA-10, bla_SHV-106, and bla_VEB-1 and resistance genes to other antibiotics. Results: A genomic analysis also revealed the presence of virulence factors such as iutA, clpK1, fyuA, fimH, mrkA, Irp2, and TraT and an IncFiB(pQil)/IncFII(K) replicon, which harbors the bla_KPC-2 gene. Additionally, the transposable element Tn4401 was identified as a key vehicle for the mobilization of the bla_KPC-2 resistance gene. Finally, this is the report of a high-risk CRKP ST307 clone expressing KPC-2, SHV-106, CTX-M-15, and VEB-1 bla genes in Greece. Conclusions: The coexistence of these resistance genes in addition to aminoglycoside, quinolone, and other resistance genes results in difficult-to-treat infections caused by respective carrier strains, often requiring the use of last-resort antibiotics and contributing to the global challenge of antimicrobial resistance. Full article

Background/Objectives: The World Health Organization has recognized Pseudomonas aeruginosa as a multidrug-resistant bacterium that presents public health concerns. This study aimed to evaluate the prevalence of MDR P. aeruginosa isolates along with their susceptibility profiles and determine the genetic basis of resistance. Methods: A total of 100 P. aeruginosa isolates were cultured on MacConkey agar with various specimens from patients admitted to ICUs and wards. Species identification was performed for each isolate using the VITEK^® 2 system. Each isolate was tested for susceptibility to specific antibiotics by the broth microdilution method. The resistance genes were detected by molecular methods, i.e., PCR and Sangar sequencing. Results: Among the 100 P. aeruginosa isolates tested phenotypically, 33 MDR P. aeruginosa isolates were detected. The aminoglycoside group of antibiotics showed the least resistance against P. aeruginosa, with increasing resistance to carbapenems and ciprofloxacin. The most prevalent detected genes responsible for resistance were blaVEB, blaVIM, aac (6′)-Ib, and qnr S. DNA sequencing results for the MDR isolates showed that 14 isolates had Thr-83> Ile mutation in gyrA, and 12 isolates had Ser-87>Leu mutation in parC genes. Conclusions: We conclude that the low rates of resistance to certain antibiotics, such as amikacin and piperacillin-tazobactam, seem encouraging to be effective for the treatment of Pseudomonas infections. Furthermore, the prominent mechanisms of resistance to fluoroquinolones in clinical strains of P. aeruginosa include mutations in gyrA and parC genes. These findings highlight the necessity of molecular diagnostics in guiding therapy and the potential need for broader surveillance. Full article

Proteus mirabilis (P. mirabilis) is an opportunistic pathogen involved in urinary tract infections as well as various nosocomial infections. Emerging resistances to beta-lactams in this species complicates potential treatment since it is intrinsically resistant to colistin. Eleven isolates of carbapenem-non-susceptible P. mirabilis were identified in Sousse Hospital, Tunisia, from January 2018 to December 2022. MICs were determined and isolates were sequenced to determine their resistomes, sequence types, virulence factors, and their clonal relationships. Susceptibility testing showed that all isolates were resistant to carbapenems, aminoglycosides, fluoroquinolones, chloramphenicol, and the trimethoprim/sulfamethoxazole combination. They remained susceptible to the aztreonam/avibactam combination. All isolates produced NDM-1 carbapenemase and ArmA 16S rRNA methylase. In addition, one isolate co-produced the bla_VEB-6 gene. All isolates belonged to ST135, and phylogenetic analysis revealed that they were closely related. This study described the first outbreak of NDM-1-producing P. mirabilis in Tunisia. Full article

Background/Objectives: The incidence of Ceftazidime/Avibactam (CZA)-resistant Klebsiella pneumoniae isolate co-producing Klebsiella pneumoniae carbapenemase 2 (KPC-2) and Vietnamese extended-spectrum β-lactamase 25 (VEB-25) has been on the rise in Greece over the past five years. This study investigates the isolation of ST323 K. pneumoniae isolates co-resistant to CZA and cefiderocol (FDC) from colonized and infected patients in a single hospital in Athens. Methods: CZA-resistant K. pneumoniae strains were isolated from 5 ICU patients from 27 December 2023 to 22 January 2024. Antimicrobial susceptibility was tested against a panel of agents. Whole-genome sequencing of the isolates was carried out to identify the acquired resistance genes and mutations that were associated with CZA and FDC resistance. Results: The K. pneumoniae isolates belonged to ST323 and harbored bla_KPC-2 and bla_VEB-25. The isolates had a minimum inhibitory concentration (MIC) of >256 mg/L for CZA and 32 mg/L for FDC, due to the disrupted catecholate siderophore receptor Fiu. bla_VEB-25 was located on an IncC non-conjugative plasmid and on a ~14 kb multidrug resistance (MDR) region comprising 15 further acquired resistance genes. Transformation studies showed that the bla_VEB-25-carrying plasmid provided resistance to most of the β-lactams tested, including CZA. The isolates remained susceptible to carbapenems, imipenem/relebactam, and meropenem/vaborbactam. The plasmid harbored the citrate-dependent iron (III) uptake system (fecIRABCDE), which increased the MIC of FDC from ≤0.08 mg/L to 2 mg/L. Conclusions: The bla_VEB-25 gene was associated with IncC plasmids which are important contributors to the spread of key antibiotic resistance genes. Strict infection control measures must be elaborated upon to prevent the spread of extensively drug-resistant organisms such as those described here. Full article

Extensively drug-resistant P. aeruginosa (XDR-PA) has been highlighted as a serious public health threat. The present study aimed to explore the genomic characteristics of two Vietnamese extended-spectrum β-lactamase-9 (VEB-9)-producing XDR-PA isolates from Bulgaria in comparison to all bla_VEB-9-positive strains with available genomes. The isolates designated Pae51 and Pae52 were obtained from tracheobronchial aspirates of intensive care unit (ICU) patients. Antimicrobial susceptibility testing, whole-genome sequencing, RT-qPCR, and phylogenomic analysis were performed. Pae51 and Pae52 were resistant to most antipseudomonal β-lactams including carbapenems, aminoglycosides, and fluoroquinolones but remained susceptible to colistin and cefiderocol. Numerous resistance determinants were detected: bla_VEB-9, bla_PDC-3, bla_OXA-10, bla_OXA-50, aac(6′)-II, ant(2″)-Ia, ant(3″)-IIa, aph(3′)-IIb, cprP, catB7, dfrB2, sul1, fosA, and tet(A). Both isolates carried complex integrons with bla_VEB-9 and tet(A) embedded next to the conservative 3′ end sequences. A variety of virulence factors were also identified, including the type III secretion system exotoxin U. Pae51 and Pae52 differed by only four SNPs and belonged to the high-risk clone ST357. To our knowledge, this is the first report of bla_VEB-9-positive XDR-PA isolates in Bulgaria presenting a detailed genomic analysis. The development of novel antimicrobial strategies for such pathogens should be an essential part of infection control stewardship practices in ICU wards. Full article

The extended-spectrum β-lactamases (ESβLs) are bacterial enzymes capable of hydrolyzing penicillins, cephalosporins, and aztreonam. The prevalence of ESβL is increasing among clinically significant microorganisms worldwide, drastically reducing the therapeutic management of infectious diseases. The study aimed to determine the drug susceptibility of ESβL-positive clinical isolates acquired from patients hospitalized in Lodz, central Poland, and analyze the prevalence of specific genes, determining acquired resistance in these bacteria. The samples of ESβL-positive clinical isolates were gathered in 2022 from medical microbiological laboratories in the city of Lodz, central Poland. The strains were subjected to biochemical identification and antimicrobial susceptibility testing following EUCAST guidelines. The presence of studied genes (bla_CTX-M, bla_SHV, bla_TEM, bla_PER, bla_VEB) was confirmed by PCR. Over 50% of studied isolates were resistant to gentamicin, cefepime, ceftazidime and ciprofloxacin. The most common ESβL gene was bla_CTX-M. In most isolates, the resistance genes occurred simultaneously. The bla_PER was not detected in any of the tested strains. ESβL-producing strains are largely susceptible to the currently available antibiotics. The observation of the coexistence of different genes in most clinical isolates is alarming. Full article

The metagenomic surveillance of antimicrobial resistance in wastewater has been suggested as a methodological tool to characterize the distribution, status, and trends of antibiotic-resistant bacteria. In this study, a cross-sectional collection of samples of hospital-associated raw and treated wastewater were obtained from February to March 2020. Shotgun metagenomic sequencing and bioinformatic analysis were performed to characterize bacterial abundance and antimicrobial resistance gene analysis. The main bacterial phyla found in all the samples were as follows: Proteobacteria, Bacteroides, Firmicutes, and Actinobacteria. At the species level, ESKAPEE bacteria such as E. coli relative abundance decreased between raw and treated wastewater, but S. aureus, A. baumannii, and P. aeruginosa increased, as did the persistence of K. pneumoniae in both raw and treated wastewater. A total of 172 different ARGs were detected; bla_OXA, bla_VEB, bla_KPC, bla_GES, mphE, mef, erm, msrE, AAC(6′), ant(3″), aadS, lnu, PBP-2, dfrA, vanA-G, tet, and sul were found at the highest abundance and persistence. This study demonstrates the ability of ESKAPEE bacteria to survive tertiary treatment processes of hospital wastewater, as well as the persistence of clinically important antimicrobial resistance genes that are spreading in the environment. Full article

The heterogenicity of antimicrobial resistance genes described in clinically significant bacterial isolates and their potential role in reducing the efficacy of classically effective antibiotics pose a major challenge for global healthcare, especially in infections caused by Gram-negative bacteria. We analyzed 112 multidrug-resistant (MDR) isolates from clinical samples in order to detect high resistance profiles, both phenotypically and genotypically, among four Gram-negative genera (Acinetobacter, Escherichia, Klebsiella, and Pseudomonas). We found that 9.8% of the total selected isolates were classified as extensively drug-resistant (XDR) (six isolates identified as A. baumannii and five among P. pneumoniae isolates). All other isolates were classified as MDR. Almost 100% of the isolates showed positive results for bla_OXA-23 and bla_NDM-1 genes among the A. baumannii samples, one resistance gene (bla_CTX-M) among E. coli, and two genetic determinants (bla_CTX-M and aac(6′)-Ib) among Klebsiella. In contrast, P. aeruginosa showed just one high-frequency antibiotic resistance gene (dfrA), which was present in 68.42% of the isolates studied. We also describe positive associations between ampicillin and cefotaxime resistance in A. baumannii and the presence of bla_VEB and bla_GES genes, as well as between the aztreonam resistance phenotype and the presence of bla_GES gene in E. coli. These data may be useful in achieving a better control of infection strategies and antibiotic management in clinical scenarios where these multidrug-resistant Gram-negative pathogens cause higher morbidity and mortality. Full article

Background: Although ceftazidime/avibactam (CAZ/AVI) has become an important option for treating adults and children, no data or recommendations exist for neonates. We report a neonatal sepsis case due to CAZ/AVI-resistant bla_KPC-2-harboring Klebsiella pneumoniae carrying bla_VEB-25 and the use of a customized active surveillance program in conjunction with enhanced infection control measures. Methods: The index case was an extremely premature neonate hospitalized for 110 days that had been previously treated with multiple antibiotics. Customized molecular surveillance was implemented at hospital level and enhanced infection control measures were taken for early recognition and prevention of outbreak. Detection and identification of bla_VEB-25 was performed using next-generation sequencing. Results: This was the first case of a bloodstream infection caused by KPC-producing K. pneumoniae that was resistant to CAZ/AVI without the presence of a metalo-β-lactamase in the multiplex PCR platform in a neonate. All 36 additional patients tested (12 in the same NICU and 24 from other hospital departments) carried wild-type bla_VEB-1 but they did not harbor bla_VEB-25. Conclusion: The emergence of bla_VEB-25 is signal for the horizontal transfer of plasmids at hospital facilities and it is of greatest concern for maintaining a sharp vigilance for the surveillance of novel resistance mechanisms. Molecular diagnostics can guide appropriate antimicrobial therapy and the early implementation of infection control measures against antimicrobial resistance. Full article

Background: Catheter ablation (CA) has become safe and efficient for the treatment of patients with ventricular extrasystolic beats (VEBs). The three-dimensional electroanatomic mapping (EAM) system allows the elimination of fluoroscopy time during CA procedures. Non-fluoroscopy CA is a challenging procedure requiring intimate knowledge of cardiac anatomy in patients with VEBs. The study aimed to evaluate the efficacy and safety of the non-fluoroscopy CA using the EAM system in patients with VEBs. Methods: Completely fluoroless CA of VEBs guided by EAM was performed in 86% (94 out of 109) of consecutive patients with VEBs. The remaining 15 patients underwent conventional fluoroscopy-guided CA. Demographic and clinical baseline characteristics, procedure parameters, and following complications were obtained from the medical records. Primary outcomes were the acute procedural success rate, the permanent success rate (6-month follow-up), complications, and procedure time. Results: There were no significant differences between groups regarding baseline characteristics. Acute procedural success was achieved in 85 patients (90%) in the non-fluoroscopy group and in 14 patients (93%) in the fluoroscopy group (ns). A long-term success rate was achieved in 82 patients (87%) in the non-fluoroscopy group and in 14 (82%) patients in the fluoroscopy group (ns). The median procedure time was 85 min in the non-fluoroscopy group and 120 min in the fluoroscopy group (p = 0.029). There was only one major complication in the non-fluoroscopy group (ns). Conclusions: Completely fluoroless CA of VEBs guided by EAM is a feasible, safe, and efficient procedure. Full article

The objective of the present study was to genetically characterize ten NDM-1 producing Escherichia coli isolates, recovered from patients in a hospital in Central Greece during the period 2017 to 2021.The isolates were studied by whole genome sequencing to obtain multi-locus sequencing typing (MLST), identification of bla_NDM1-environment, resistome and plasmid content. MLST analysis showed the presence of eight sequence types: ST46* (two isolates), ST46, ST744, ST998, ST410, ST224, ST4380, ST683 and ST12 (one isolate each). Apart of the presence of bla_NDM-1, the isolates carried a combination of various to β-lactams encoding resistance genes: bla_TEM-1B, bla_CTX-15, bla_OXA-1, bla_VIM-1, bla_SHV-5, bla_OXA-16, bla_OXA-10 and bla_VEB-1. Additionally, plurality of resistance genes to aminoglycosides, macrolides, rifamycin, phenicols, sulfonamides and tetracycline was detected. The presence of multiple replicons was observed, with predominance of IncFII and IncFIB. Analysis of bla_NDM-1 genetic environment of the isolates showed that seven had 100% identity with the pS-3002cz plasmid (Accession Number KJ 958927), two with the pB-3002cz plasmid (Accession Number KJ958926) and one with the pEc19397-131 plasmid (Accession Number MG878866). Τhis latter plasmid was derived by the fusion of two, previously identified, plasmids, pAMPD2 and pLK75 (Accession Numbers CP078058 and KJ440076, respectively). The diversity of clones and plasmids of NDM-1 producing E. coli isolated from patients in Greece indicates a continuous horizontal gene transfer. Full article

The COVID-19 pandemic has had a significant impact on numerous industries, including the event industry, resulting in widespread disruptions. The widespread cancellations of festivals have been a direct consequence of the pandemic, and, following the reopening, those that have taken place have had to implement changes in response to the new guidelines and regulations created as a result of the pandemic. In this study, we examine the experiences of festivals held in 2021 in the Veszprém–Balaton 2023 European Capital of Culture region (VEB 2023 region). The study aims to adapt the PwC Risk Management Model and identify its four pillars (1: detect; 2: protect; 3: react; 4: restore) in the case of festivals. Our study outlines how festival organizers have faced unprecedented challenges and risks (detect), how they survived in complete uncertainty (protect), what lessons they learned, and what risk management decisions they made as a challenge of the pandemic period from a consumer perspective (react). The present study utilized a qualitative research methodology and involved conducting structured interviews with a total of 19 event organizers from five different events. In addition, to gain a comprehensive understanding of the sustainable consumption habits of festival visitors, a questionnaire survey was administered yielding 1133 responses. The biggest challenges for the organizers during the times of the pandemic were uncertainty and unpredictability, with increased financial, human, and mental risks and burdens. However, the positive benefits of the pandemic period and the future developmental directions of sustainable festivals were also highlighted (e.g., small-scale, family-friendly events). Regarding the future, the aspects of greening aspirations, a need to reflect on the social-environmental criteria of sustainability, and more flexible management decisions to deal with uncertainty have emerged among the festival organizers (restore). The research has revealed that festival visitors themselves will be important allies in the introduction of sustainable measures. The research result helps festival organizers to adapt more effectively to the new socio-economic circumstances caused by the pandemic. Full article

Automated electrocardiogram (ECG) classification using machine learning (ML) is extensively utilized for arrhythmia detection. Contemporary ML algorithms are typically deployed on the cloud, which may not always meet the availability and privacy requirements of ECG monitoring. Edge inference is an emerging alternative that overcomes the concerns of cloud inference; however, it poses new challenges due to the demanding computational requirements of modern ML algorithms and the tight constraints of edge devices. In this work, we propose a tiny convolutional neural network (CNN) classifier for real-time monitoring of ECG at the edge with the aid of the matched filter (MF) theory. The MIT-BIH dataset with inter-patient division is used for model training and testing. The model generalization capability is validated on the INCART, QT, and PTB diagnostic databases, and the model performance in the presence of noise is experimentally analyzed. The proposed classifier can achieve average accuracy, sensitivity, and F1 scores of 98.18%, 91.90%, and 92.17%, respectively. The sensitivity of detecting supraventricular and ventricular ectopic beats (SVEB and VEB) is 85.3% and 96.34%, respectively. The model is 15 KB in size, with an average inference time of less than 1 ms. The proposed model achieves superior classification and real-time performance results compared to the state-of-the-art ECG classifiers while minimizing the model complexity. The proposed classifier can be readily deployed on a wide range of resource-constrained edge devices for arrhythmia monitoring, which can save millions of cardiovascular disease patients. Full article

This nationwide study aimed to investigate the molecular characteristics of metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa in Serbia, underlying resistance mechanisms, the genetic context of detected MBL genes, and the clonal relationship between isolates harboring genes-encoding MBL. Overall, 320/5334 isolates collected from 2018 to 2021 were identified as P. aeruginosa. Carbapenem-resistant P. aeruginosa (CRPA) were screened for the presence of bla_VIM, bla_IMP, and bla_NDM, genes whereas MBL-positive isolates were tested for the presence of the bla_CTX-M-2, bla_PER, bla_TEM, bla_SHV, bla_VEB, and bla_GES. Multilocus sequence typing and phylogenomic analysis were performed for P. aeruginosa-producing MBL. The majority of the P. aeruginosa isolates were recovered from the lower respiratory tract (n = 120; 37.5%) and wound specimens (n = 108; 33.75%). CRPA isolates accounted for 43.1% (n = 138) of the tested isolates, 31 out of them being bla_NDM-1-positive (22.5%). The colistin resistance rate was 0.3%. MLST analysis revealed the occurrence of ST235 (n = 25) and ST654 (n = 6), mostly confined to Serbia. The distribution of beta-lactamase-encoding genes in these isolates suggested clonal dissemination and possible recombination: ST235/bla_NDM-1, ST235/bla_NDM-1/bla_PER-1, ST654/bla_NDM-1, ST654/bla_NDM-1/bla_PER-1, and ST654/bla_NDM-1/bla_GES-5. High-risk clones ST235 and ST654 identified for the first time in Serbia, are important vectors of acquired MBL and ESBL and their associated multidrug resistance phenotypes represent a cause for considerable concern. Full article