Search Results (383)

The global dissemination of carbapenem resistance is predominantly facilitated by plasmid-mediated carbapenemase genes, notably blaOXA-48-like genes. A comprehensive understanding of their evolutionary relationships and structural conservation is essential for monitoring their spread and informing therapeutic strategies. This study aimed to investigate the phylogenetic relationships and structural conservation of blaOXA-48-like carbapenemase genes in multiple Gram-negative bacterial species. We analysed blaOXA-48-like carbapenemase sequences obtained from a hospital in Pakistan and compared them with globally reported variants retrieved from GenBank. Carbapenemase gene sequences (blaOXA-48-like, blaNDM, and blaVIM) were analyzed using maximum-likelihood phylogenetics (MEGA11, Tamura–Nei model, 1000 bootstrap replicates). Comparative global sequences were retrieved from GenBank. Structural modeling of blaOXA-48-like genes was performed using SWISS-MODEL Workspace with the template PDB 3HBR, followed by validation using GMQE, QMEANDisCo, and Ramachandran plot analyses. Phylogenetic analysis revealed a tight clustering of blaOXA-48-like genes across A. baumannii, K. pneumoniae, and E. meningoseptica, showing high similarity to globally distributed plasmid-associated sequences. Structural modeling demonstrated strong conservation of the enzyme, with preserved catalytic residues (Ser70, Lys73, Ser118, Trp157, and Tyr211) and minimal structural deviation (RMSD < 0.3 Å). blaOXA-48-like carbapenemases exhibit strong phylogenetic conservation and structural stability across species and regions, consistent with the horizontal dissemination of blaOXA-48-like genes across bacterial hosts. These findings indicate that blaOXA-48-like carbapenemases have high evolutionary stability. Full article

To improve clinical decision-making about Carbapenem-resistant Gram-negative bacteria (CR-GNB) infections and halt the spread of resistant microbes, quicker and less expensive diagnostic techniques are required. Thus, the purpose of this study was to thoroughly evaluate the diagnostic efficiency (sensitivity, specificity, and concordance) of direct-from-specimen multiplex lateral flow immunoassay (LFIA) across diverse raw clinical specimens and pathogen types from critically sick patients. A total of 300 non-duplicate samples were tested to detect CR-GNB. Five major Carbapenemase genes were detected directly from the specimen using carbapenem-resistant K.N.I.V.O. detection K-Set and from culture using culture-enhanced multiplex PCR. Turnaround time (TAT) of each method was calculated. The direct LFIA revealed 100% specificity for NDM, KPC, and IMP enzymes in all tested clinical matrices (blood, urine, and respiratory samples). The study demonstrated 100% sensitivity and specificity with perfect categorical agreement (κ = 1.000) for the blaKPC in the Klebsiella pneumoniae and for blaOXA-48 and blaIMP in the Acinetobacter baumannii; however, sensitivity of blaVIM was significantly diminished across all isolates and samples. TAT decreased significantly (p < 0.001) from 30 to 70 h to about 50 min. The tested direct LFIA facilitates the prompt enhancement of lifesaving tailored antibiotic treatment for severe illnesses. Full article

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a critical public health threat because infections caused by this pathogen are associated with high morbidity, mortality, and limited effective therapeutic options. Whilst the majority of studies have concentrated on inter-patient bacterial transmission, within-host genomic analysis offers unprecedented resolution for tracking dynamic clone predominance, plasmid rearrangements, and microevolution under clinical selection pressures. Methods and Results: Whole-genome sequencing (WGS) of nine isolates recovered from oral and rectal swabs revealed an exceptional case of CRKP clonal turnover in an intensive care unit (ICU) patient. Three distinct high-risk clones were identified during the 18 days of surveillance: an initial ST101 (Clonal Group (CG) 101) strain (days 1–7) followed by concurrent colonization with ST395 (carrying bla_NDM-5) and ST512 lineages (both CG258, days 11–18). Conclusions: This study describes a rare instance of within-host heterogeneity of CRKP, involving three distinct STs spanning two CGs. Whole-genome analysis revealed potential structural rearrangements of resistance- and virulence-associated plasmids between coexisting lineages. These genomic shifts likely reflect rapid adaptation under the intense selective pressure of broad-spectrum antibiotic therapy, culminating in the persistence of a less virulent yet multidrug-resistant ST512 clone and a favorable clinical outcome with patient recovery. Full article

The emergence of plasmid-mediated tigecycline resistance in food-producing animals poses a significant threat to global public health by compromising a last-resort antimicrobial. This study investigated the prevalence, antimicrobial susceptibility profiles, and molecular genetic characteristics of tigecycline-resistant Escherichia coli (E. coli) isolated from broiler cecal samples in the Jinan region of Shandong Province, China, between 2020 and 2024. Antimicrobial susceptibility testing showed that high-level resistance to colistin (87.5%, 14/16), florfenicol (81.3%, 13/15), and enrofloxacin (75.0%, 12/16) was observed. Notably, a single isolate was resistant to meropenem (6.3%, 1/16). Whole-genome sequencing and subsequent in silico analysis demonstrated that all 16 tigecycline-resistant isolates harbored the tigecycline resistance gene tet(X4), the primary determinant of resistance. Molecular typing identified a diverse population structure, with sequence type 224 (ST224) being the dominant clone, accounting for 37.5% (6/16) of the isolates. The genetic milieu of resistance was complex, characterized by the co-existence of tet(X4) with multiple other clinically important resistance genes, including the mobile colistin resistance gene mcr-1 (87.5%, 14/16) and various extended-spectrum β-lactamase genes such as blaCTX-M variants and, critically, blaNDM-5. Furthermore, an array of virulence factor genes was identified, with a particularly high prevalence of the toxin gene astA (68.8%, 11/16) and the bacteriocin gene cma (50.0%, 8/16), indicating pathogenic potential. This convergence of resistance and virulence in a foodborne pathogen highlights an urgent need for continuous One Health surveillance to mitigate the risks posed by these potentially untreatable “superbugs” to both animal and human health. Full article

Growing resistance to carbapenem antibiotics is a major public health problem as these antibiotics are considered the last line of therapy for infections caused by multidrug-resistant (MDR) Gram-negative bacteria. The rapid emergence and dissemination of carbapenem-resistant bacterial strains are mainly due to horizontal gene transfer (HGT) within or between bacterial cells via the mobilome. The aim of this article is to discuss the role of mobile genetic elements (MGEs) that capture and disseminate resistance determinants of carbapenem antibiotics, as a comprehensive review integrating the combined role of plasmids, transposons and integrons. It attempts to systematically fill the gap by investigating the role of these MGEs in the acquisition, mobilization and dissemination of genes encoding carbapenemases across clinically important bacteria. Various types of plasmids such as IncF and IncH in Klebsiella pneumoniae, IncL/M in Enterobacter cloacae, IncX3 in Escherichia coli and IncA/C₂ in Salmonella enterica carry important genes encoding carbapenemases. The rapid distribution of transposons among bacterial species is one of the main contributing factors in the dissemination of carbapenem-resistant isolates. Transposons including Tn4401 carrying bla_KPC in K. pneumoniae and Tn1721 carrying bla_KPC in E. coli; Tn2006, Tn2007, Tn2008 and Tn2009 carrying bla_OXA-23 in Acinetobacter baumannii; Tn1696 carrying bla_IMP-4 in Pseudomonas aeruginosa; Tn125 carrying bla_NDM in E. coli; and Tn6306 carrying bla_IMI in Raoultella ornithinolytica encode different types of carbapenemases. Integrons mainly belonging to class 1 capture resistance determinants for metallo-carbapenemases such as NDM-, VIM-, SIM- and IMP-type enzymes in P. aeruginosa, A. baumannii, K. pneumoniae and E. coli and can promote the transcription and expression of these determinants. These findings are useful for understanding the genetics of carbapenem resistance and additional knowledge on MGEs may provide avenues for screening of resistance to these antibiotics in clinical settings. Full article

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a significant pathogen for both hospital-acquired and community-acquired infections, characterized by its strong epidemic potential and high mortality rate, posing a severe threat to global public health. CRKP spreads widely across the globe through the horizontal transfer of plasmid-mediated resistance genes such as *blaKPC*, *blaNDM*, and *blaOXA-48*. The clinical treatment options for this bacterium are limited, and its resistance has been increasing year by year, urgently necessitating the development of new antimicrobial drugs or alternative strategies. In recent years, the CRISPR-Cas system has shown great potential in the diagnosis and treatment of CRKP, including rapid detection and identification, gene editing, antimicrobial strategies, and resistance inhibition. For instance, CRISPR-Cas12a/13a can be used for the rapid detection and identification of CRKP, while CRISPR-Cas9/Cas3 can target resistance genes to reverse the resistance of strains. With the advancement of delivery and biotechnologies, the CRISPR-Cas system is expected to become an important tool against drug-resistant CRKP. This review focuses on the application of the CRISPR-Cas system in the detection and treatment of CRKP, analyzing its technical advantages, limitations, and future development directions. Full article

Background: The emergence and spread of urinary carbapenem-resistant organisms (CROs) are a major public health concern, particularly in Sri Lanka. Therefore, we aimed to detect and genotypically characterize CROs in urinary tract infections (UTIs) and their clinical outcomes. Methods: Urinary CROs were collected from two hospitals in Sri Lanka from January to December, 2023. Among 7640 urine samples, 100 CROs were identified by disk diffusion method, and 99 were detected by BD Pheonix^TM automated system. The presence of eight carbapenemase genes; bla_KPC, bla_NDM, bla_VIM, bla_IMP, bla_OXA-23, bla_OXA-48, bla_OXA-51, and bla_OXA-58, among 97 CROs was detected by a multiplex PCR kit. Results: Out of 99 urinary CROs, K. pneumoniae (33.3%; n = 33/97) was the most common species. Among the 97 isolates tested by PCR, a single carbapenemase gene was detected in 35.05% (34/97), while two or more genes co-occurred in 39.18% (38/97). The most frequently identified gene was bla_OXA-51 (47.4%), followed by bla_OXA-58 (41.2%). Most patients (95.74%; n = 90/97) showed clinical improvement within seven days of treatment. Among the 93 patients discharged and followed for three months, 74.20% (n = 69/93) experienced at least one mild UTI recurrence. A total of 10 patients died during the study period. Of which, four (40%) during hospitalization and six (60%) during follow-up, though none of the deaths were attributed to UTIs. Conclusions: K. pneumoniae, showed the highest carbapenemase gene diversity. Recurrent UTIs were observed during the follow-up period. Continuous surveillance and implementation of targeted infection control programs are needed to minimize further emergence and spread of carbapenemase genes. Full article

Patients undergoing cervical conization for cervical intraepithelial neoplasia grade II (CIN II) may develop postoperative bloodstream infections (BSIs), and multidrug-resistant (MDR) Escherichia coli isolates with distinct resistance profiles can complicate antimicrobial management. In this case-based study, two E. coli strains, HBFY-1 and HBFY-2, were recovered from blood cultures obtained from a 38-year-old CIN II patient with postoperative fever. The isolates were characterized by antimicrobial susceptibility testing, whole-genome sequencing, conjugation assays, and comparative genomics against publicly available genomes matched by sequence type and serotype. Fever occurred on postoperative day 2. HBFY-1 was fluoroquinolone-resistant; belonged to ST744/O101:H9; carried the extended-spectrum β-lactamase (ESBL) gene bla_CTX-M-27; was phenotypically confirmed as an ESBL producer; and grouped within a multi-source near-neighbor clade consistent with a conserved fluoroquinolone-associated resistance backbone in ST744/O101:H9. HBFY-2 was carbapenem-resistant; belonged to ST48/O113:H32; carried bla_NDM-5 on an IncY-associated plasmid bin; was phenotypically confirmed as a metallo-carbapenemase producer; and did not harbor any ESBL gene. Within the matched ST48/O113:H32 dataset, bla_NDM-5 was detected only in HBFY-2, which clustered within an Asia-enriched lineage, including China-derived human and swine genomes. The bla_CTX-M-27-associated and bla_NDM-5-associated elements were transferred to E. coli C600 at frequencies of 5.3 × 10⁻² and 4.6 × 10⁻⁶, respectively, and transfer of the bla_NDM-5-associated element imposed no detectable growth penalty under the tested conditions. As this study is based on a single clinical case, the findings should be interpreted cautiously, yet they still highlight the potential value of integrating susceptibility testing with rapid genomic characterization for identifying mobilizable carbapenem-resistance platforms. Full article

Wastewater surveillance is an effective approach for monitoring populations of antibiotic-resistant bacteria and tracking the spread of antimicrobial resistance (AMR) across different settings. In this study, hospital and municipal wastewater were collected monthly for 12 months from multiple locations in the greater Pittsburgh area to quantify the presence of antibiotic-resistant bacteria and investigate their genomic diversity. After quantitative culturing on six different selective media types, a total of 150 isolates were speciated by 16S rRNA sequencing, which revealed diverse pathogenic and non-pathogenic taxa, including Klebsiella spp. (n = 28), Pseudomonas spp. (n = 20) and Aeromonas spp. (n = 37). A subset of isolates (n = 46) underwent whole genome sequencing, which identified several antibiotic resistance genes of clinical concern, such as bla_KPC (n = 17), bla_NDM (n = 6) and bla_IMP (n = 6), and revealed genetic similarities between wastewater isolates and clinical isolates collected from infected patients at a Pittsburgh-area medical center. In addition, analysis of plasmids carried by wastewater isolates revealed closely related plasmids present in isolates from different species and sampling locations. Overall, these findings suggest that both hospital and municipal wastewater act as interconnected reservoirs of antimicrobial resistance. Integrating wastewater surveillance with clinical and genomic data could enable the early detection of emerging resistance threats and support proactive infection-control strategies. Full article

Objectives: This study evaluated the diagnostic performance of the lateral flow immunochromatographic assay (RESIST-ACINETO, Coris BioConcept) for the rapid detection of the major carbapenemases in Acinetobacter baumannii. Methods: Blood culture isolates collected between 2014 and 2024 with meropenem MIC ≥ 4 mg/L were retrieved, re-identified by MALDI-TOF MS, and susceptibility was confirmed by broth microdilution. Carbapenemase genes (bla_OXA-23, bla_OXA-40, bla_NDM) were detected using multiplex PCR, which served as the reference standard. All isolates were tested using the RESIST ACINETO assay, and diagnostic accuracy parameters were calculated. Results: A total of 114 isolates were recovered and confirmed as A. baumannii. Among 93 carbapenem-non-susceptible isolates, 97.8% (91/93) were correctly identified by the assay. The test showed 99.1% sensitivity and 99.1% specificity, with most positive results appearing within 3–10 min. Two discrepant results were observed (one false positive, one false negative), while all meropenem-susceptible isolates tested negative. Conclusions: The RESIST ACINETO assay provides rapid, accurate detection of carbapenemases in A. baumannii, significantly reducing turnaround time compared with conventional workflows. Its performance supports integration into routine diagnostics to enhance timely resistance confirmation and infection-control interventions. Full article

Carbapenems comprise a class of β-lactam antibiotics with broad-spectrum hydrolytic activity and are often reserved as last-line agents for the treatment of serious multidrug-resistant (MDR) bacterial infections. Clinically important nosocomial MDR Gram-negative bacteria (GNB) include Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Carbapenem resistance among these organisms is predominantly mediated by the production of β-lactamases called carbapenemases, such as K. pneumoniae carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), imipenemase (IMP), Verona integron-encoded metallo-β-lactamase (VIM), and selected oxacillinase (OXA)-type carbapenemases. These enzymes degrade carbapenems, significantly compromising their clinical efficacy. To address escalating antimicrobial resistance, novel next-generation β-lactamase inhibitors (BLIs), partnered with established β-lactams (BLs), have been approved or are currently under development to inhibit carbapenemase activity. The present narrative review aims to synthesize the most current information on the major carbapenemases and discusses recently approved and investigational BL/BLI combination therapies in terms of their mechanisms of action, spectrum of activity, gaps in coverage, and available clinical and in vitro evidence. Development of resistance to novel BL/BLI combinations is also examined. Comparative analysis of inhibitory spectra and microbiological coverage indicates a continued need for metallo-β-lactamase inhibitors with direct pan-inhibitory activity, pathogen-specific BL/BLI regimens for carbapenem-resistant A. baumannii, and carbapenemase-targeted agents effective in the context of non-enzymatic resistance mechanisms. Treatment-emergent resistance to novel BL/BLIs and limitations in activity profiles underscore the critical need for continued innovation in pipeline development, vigilant global and local surveillance of carbapenemase epidemiology, and robust antimicrobial stewardship strategies to aid in preserving the efficacy of the antibacterial drug armamentarium. Full article

In 2023, the European Centre for Disease Prevention and Control surveillance report highlighted an increasing number of carbapenem-resistant Escherichia coli isolates carrying the less common bla_NDM-5 variant in Europe. The aim of this study was to investigate the molecular epidemiology of NDM-producing (New Delhi metallo-β-lactamase) E. coli isolates collected in Croatia over a one-year period. A total of 160 carbapenemase-producing E. coli isolates were reported through national surveillance in Croatia between March 2023 and March 2024. Whole-genome sequencing was performed on 22 NDM-producing E. coli isolates. Phylogenetic analysis identified 17 sequence types, indicating high diversity and polyclonal spread. High variability in resistome profiles and co-occurrence of resistance genes across multiple antimicrobial classes indicate multidrug resistance. The predominant bla_NDM variant was bla_NDM-1 (77.27%), followed by bla_NDM-5 (22.73%). Co-occurrence of bla_NDM with extended-spectrum β-lactamase (ESBL) encoding genes was detected in 12/22 isolates (54.55%). Plasmid analysis identified 22 different replicon types, with IncFII (54.54%) and IncA/C2 (45.45%) being the most frequent. Our findings provide insights into the molecular epidemiology of NDM-producing E. coli at the national level, highlighting the presence of the bla_NDM-5 variant. These results emphasize the need for genomic surveillance and strengthened infection control strategies to better understand and limit its spread. Full article

Background: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a critical pathogen associated with severe hospital infections and high antimicrobial resistance. Despite the global significance of A. baumannii, there are limited data from Costa Rica regarding the resistance rate and genomic characteristics of CRAB. Methods: This study aimed to provide initial and exploratory epidemiological data on infections caused by A. baumannii and CRAB isolated in Costa Rica and to gain insights on the genome of selected strains, focusing on their resistance determinants and phylogenetic relationships. Results: Based on data from five main hospitals in Costa Rica, resistance rate to carbapenems was estimated at 9.8% to imipenem and 6.1% to meropenem. From 190 carbapenem-resistant clinical isolates available in a local collection, seven A. baumannii strains were identified, all showing resistance to carbapenems and carrying the bla_NDM-1 gene. Whole-genome sequencing of two strains yielded two distinct MLST profiles (Pasteur scheme: ST-150 for strain IPAT15 and ST-250 for IPAT72), as well as variations in the number and identity of plasmids, genomic islands, and other elements of the mobilome. Both isolates carried ten antimicrobial resistance genes, which are predicted to be harbored in plasmids for IPAT15, unlike the chromosomal determinants in IPAT72. A pangenome analysis of 878 genomes from a public database identified over 51,000 genes, with only 1338 (2.6%) forming the core genome. Phylogenetic analysis and assignation of international clones (ICs) showed predominance of IC2. Isolates from Costa Rica clustered near IC9 and shared some resistance determinants, but they were not directly assigned to an IC. Conclusions: Overall, this study provides exploratory insights regarding the occurrence of CRAB in Costa Rica using epidemiological and genomic data, with profiles that are comparable to other regions in Latin America and diverse genomic resistance determinants. While this study does not show the whole landscape of CRAB in Costa Rica, these data constitute an initial approach for improving clinical management and public health responses to CRAB infections, to ultimately improve outcomes for patients affected by this pathogen. Full article

Background/Objectives: Multidrug-resistant Salmonella enterica serovar Enteritidis, especially those isolated from humans, remains a public concern. In the present study, S. Enteritidis strain 31404 was obtained clinically from a fecal sample of a fifteen-year-old girl, who was positive for bla_NDM-13. Methods: Antibiotic susceptibility testing and whole genome sequencing were performed. Core genome MLST and hierarchical clustering (HierCC) were performed using EnteroBase. Population structure analysis of 57 S. Enteritidis isolates collected between 2023 and 2025 in Jiaxing city was conducted. A comparative structure analysis of bla_NDM-13-positive plasmids was also performed. Results: S. Enteritidis strain 31404 was resistant to 13 antimicrobial agents. We found that strain 31404 belonged to ST11 and carried resistance genes, such as bla_NDM-13, bla_CTX-M-14, ble_MBL, fosA3, qnrS, and tet (A). bla_NDM-13 was located on an IncI1-I (α) plasmid designated as p31404-NDM13. S. Enteritidis isolate 31404 was closely related to PNUSAS514422, which was isolated from the United States in 2025. Comparative genetic environment related to bla_NDM-13-positive plasmids available in the NCBI database indicates that ΔTn125-mediated contexts were commonly associated with bla_NDM-13. IS1294 (IS91 family), which replaces ISAba125, is likely to mobilize bla_NDM-13. Conclusions: The findings in this study provide insights into the molecular characterization and diversification of bla_NDM-13. The identification of bla_NDM-13-containing transferable plasmids in different serotypes of Salmonella isolates (such as S. Rissen, S. Typhimurium, and S. Enteritidis) in different cities in China highlights the risk of the spread of carbapenem-resistant genes among Salmonella isolates. Full article

Salmonella is a major cause of foodborne illness worldwide, posing significant risks to public health and food safety. This study investigated the prevalence, serovar distribution, genotypic characteristics, and antimicrobial resistance (AMR) profiles of Salmonella. A total of 2515 food samples were collected from retail markets, supermarkets, and food processing facilities, and 13,670 stool samples were obtained from sentinel hospitals across 14 cities in Liaoning. The Kruskal–Wallis test was used to compare genetic features among serovars, followed by Dunn’s post hoc test for pairwise comparisons. A total of 314 Salmonella strains were identified, with raw poultry showing the highest detection rate (28.88%) among food sources and children aged 0–6 years (3.47%) the highest among the clinical age groups. Among food samples, S. Enteritidis was the most prevalent serovar (42.6%), and it was also the most common in clinical samples (35.8%); in contrast, S. 4,[5],12:i:- was dominant in pediatric clinical cases. According to AMR analysis, 90.13% of strains were resistant to at least one antibiotic and 67.83% were multidrug-resistant (MDR), with the highest resistance to ampicillin (68.47%). Analysis revealed that S. 4,[5],12:i:- harbored the ASSuT resistance module (blaTEM-1B, aph(3″)-Ib/aph(6)-Id, sul2, tet(B)). Extensive MDR phenotypes were observed in S. Indiana and S. Kentucky, associated with abundant insertion sequences (IS) and resistance genes (ARGs), including clinically critical determinants (blaNDM-9, mcr-1.1, rmtB). The highest mean virulence factor (VF) count (111.17) was observed in S. Enteritidis, contributing to its epidemiological success. Conversely, S. Indiana and S. Kentucky, predominantly food-associated serovars, exhibited reduced virulence but served as critical AMR reservoirs. These findings highlight the epidemiological characteristics and AMR risks of Salmonella in food and clinical settings, providing critical data for food safety and clinical antimicrobial stewardship. Full article