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Carbapenem resistant (CRE) and carbapenemase producing Enterobacterales (CPE) in particular, represent a major threat for healthcare systems worldwide. Rapid, reliable, and easy to perform assays are required to enable targeted and effective therapy. MALDI-TOF MS based carbapenemase diagnostics has potential advantages over molecular and phenotypic sensitivity tests, especially in terms of time to result. So far, only one mass spectrometry (MS)-based carbapenemase test system is commercially available for routine use. The aim of this study was to compare the performance of the established system to a novel MS-based test to identify CPE isolates. Forty consecutive CRE isolates (70% CPEs) were pre-screened for carbapenemase activity by routine laboratory methods. Isolates then were tested using the to date only IVD CE certified MALDI-TOF MS carbapenemase detection assay (MBT STAR-Carba IVD Kit; Bruker Daltonics) and a novel test designed for the recently launched EXS2600 MALDI-TOF MS system (Carbapenemase Activity Kit; Zybio). Valid results were obtained for 93% and 85% isolates by the Bruker and the Zybio assay, respectively. Sensitivities, specificities, positive and negative predictive values were 92%, 91%, 96%, and 83% for the Bruker assay and 96%, 64%, 85%, and 88% for the Zybio assay. There are notable differences concerning the handling of the assays. In summary, both systems featured high sensitivities for the detection of carbapenemases, but the Bruker assay yielded less false-positive results. There are advantages and disadvantages concerning the handling for each system, but both proved to be suitable for the use in a routine laboratory. [ABSTRACT FROM AUTHOR]

The increasing prevalence of carbapenem-resistant Enterobacter cloacae complex (CREC) poses great challenges to infection treatment in the clinical setting. In this study, we reported the emergence of carbapenemase in a rare species, Enterobacter chuandaensis, belonging to the Enterobacter cloacae complex (ECC). We elucidated the genetic characteristics of carbapenem-resistant isolate FAHZZU5885, co-harboring blaNDM−1 and blaKPC−2. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and average nucleotide identity (ANI) analysis were used to identify E. chuandaensis. S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) and Southern blotting were used to clarify the number and size of the plasmids in FAHZZU5885. Antimicrobial phenotypes were identified by antimicrobial susceptibility testing (AST), and the characteristics of the strain were examined with whole-genome sequencing (WGS). The conjugation experiment and stability assay were conducted to verify the transferability and stability of the plasmid carrying carbapenemase-encoding genes. E. chuandaensis FAHZZU5885 was isolated from a perianal swab of a patient admitted to the ICU. This strain simultaneously carried blaNDM−1 and two blaKPC−2 genes. FAHZZU5885 was resistant to most of the tested antibiotics except for amikacin, tigecycline, and colistin. Two blaKPC−2 were located separately on two different plasmids, the ~ 120 kb IncFIA-IncFII plasmid and the ~ 80 kb IncR plasmid. Both plasmids shared the conserved sequence klcA-korC-ISkpn6-blaKPC−2-ISkpn27-tnpR-tnpA. The blaNDM−1-bearing plasmid had the potential to transfer and can remain stable after successive passages. In addition, the blaNDM−1 was carried on a ~ 80 kb IncFII plasmid with the conserved sequence ISAba125-blaNDM−1-ble-trpF-dsbD-cutA-groS-groL. In summary, this study marks the first report of the multidrug-resistant E. chuandaensis strain FAHZZU5885 harboring two blaKPC−2-bearing plasmids, indicating the potential for the further dissemination of carbapenemase-encoding genes in novel species. The findings contribute to enhancing our understanding of CREC strains, emphasizing the need for continued and comprehensive surveillance of this species. [ABSTRACT FROM AUTHOR]

Colistin HiMIC Plate Kit (HiMedia Laboratories), a new commercial broth microdilution (BMD) test for colistin susceptibility testing was evaluated. BMD according to ISO standard 20776–1 (2019) with two-fold dilutions from 128 to 0.125 mg/L was used as a reference method. The colistin reference MICs (minimal inhibitory concentration) ranged from 0,25 to 128 mg/L with 15 (20.5%; 15/73) isolates having colistin reference MICs close to the current EUCAST breakpoint (MICs of 2, 4, and 8 mg/L). The study assessed the compliance of a commercial kit with the CLSI criteria, including categorical agreement (CA) and essential agreement (EA ≥90%), very major error (VME rate) <3%, and major error (ME) rate <3%. On 73 carbapenemase-producing Klebsiella pneumoniae isolates Colistin HiMICTM Plate Kit showed CA and EA of 100% (73/73; 95% CI: 0.97–1.00) and 82.2% (60/73; 95% CI: 0.72–0.90), respectively. No ME (false-resistant results) and VME (false-susceptible results) were detected. Kit showed acceptable CA, ME, and VME error parameters, whereas the EA did not meet the ≥90% threshold. Laboratories must check for possible limitations of commercial kits before they can be used for colistin susceptibility testing. [ABSTRACT FROM AUTHOR]

The World Health Organization (WHO) has declared antimicrobial resistance (AMR) as one of the top ten public health threats faced by humanity. Carbapenems are currently the last option β-lactam antibiotics for treatment of both hospital and community acquired infections. The rapid rise and spread of carbapenem resistant organisms (CRO) is a global concern highlighting the need for early detection and isolation of patients. A total of 226 non- repeat, carbapenem resistant Gram-negative isolates were subjected to carbapenemase detection. Three phenotypic methods like Modified Hodge Test (MHT), RAPIDEC® CARBA NP kit (bioMérieux, France) and Chromogenic agar were used and a cartridge based molecular method Xpert® Carba-R (Cepheid) was used on all the study samples for detection of carbapenemases. The results of MHT, RAPIDEC® CARBA NP and CHROMagar were same for both Enterobacteriaceae and Non Fermenters (except for CHROMagar, which is applicable only for Enterobacteriaceae). Xpert® Carba R could not detect genes in all the phenotypically positive organisms for carbapenemase production. However, there was no statistically significant difference between the three phenotypic tests and Xpert® Carba R used for the detection of carbapenem resistance. Preserving the last resort antimicrobials is the need of the hour, as there are few therapeutic options for treating infections caused by carbapenem-resistant bacteria. Early detection, followed by strict implementation of infection prevention and control strategies to stall the dissemination of such organisms is the best way and needs to be implemented on a war footing. [ABSTRACT FROM AUTHOR]

Background: Carbapenem-resistant Enterobacterales (CRE) have been identified as a public health problem. Treatment options for CRE are limited as they are mostly resistant to βeta-lactams, aminoglycosides, and fluoroquinolones as well as carbapenems. The present study aims to evaluate the performance of three phenotypic methods compared to a molecular-based technique for carbapenemase detection in Enterobacterales and determining their applicability in clinical and epidemiological settings. Methods: A total of 1,158 Enterobacterales were isolated from the urine samples in the microbiology laboratory of Alexandria main university hospital during the period from April 2020 to April 2021. Fifty randomly selected (39 Klebsiella and 11 E. coli) Enterobacterales were screened for carbapenem resistance by disc diffusion method. They were subjected to 3 phenotypic tests which are: Carba NP method, Modified carbapenem inactivation method (m CIM) and EDTA- modified carbapenem inactivation method (e CIM). Detection of 5 carbapenemase genes (blaKPC, blaIMP, blaVIM, blaOXA-48 and blaNDM-1) was performed using real time PCR. Results: CRE represented 33% of Enterobacterales isolates. Twenty-six cases (52%) were males and 94% of the cases were above 40 years old. Carba NP test was positive in 43/50 (86%) of the selected isolates, m CIM was positive in 35/50 (70%) and e CIM was positive in 30/50 (60%). The most common carbapenemase gene detected was blaNDM-1 (94%), followed by blaOXA-48 gene (72%) and blaVIM gene (24%). The blaKPC gene and blaIMP gene were not detected. Coexistence of the blaOXA-48 and the blaNDM-1 genes was detected in 48% isolates, while the blaNDM-1, the blaOXA-48 and the blaVIM genes were found in 22% isolates. The sensitivity of Carba NP, m CIM and e CIM was 87.5%, 72.9%, and 85.7% respectively. Conclusion: The study highlights the necessity of early detection of CRE. Carba NP test assists in the rapid identification of carbapenemase production. However, the genotypic test remains the gold standard for detection of CRE. [ABSTRACT FROM AUTHOR]

Background: Continuous monitoring is crucial to detect infection by Carbapenemase-producing Klebsiella pneumoniae (CPKP) in intensive care COVID-19 patients. Aim: to evaluate the CR-KP colonization and co- infection and evaluate the antibiotic resistance pattern. Methods: A cross -sectional study was done on COVID-19 patients with CPKP infection in Zagazig University Hospitals where rectal swab samples were detected using XpertCarba-R Assay(Cepheid, Sunnyvale), while isolates from sputum, urine, and blood culture were identified phenotypically by VITEK® 2 (bioMérieux SA) and modified Hodge test while genotypically by XpertCarba-R Assay(Cepheid, Sunnyvale). Results: CPKP was found in 60 patients out of 140 patients (42.8%), with 34 CPKP identified from rectal swabs, 12 patients infected from diverse locations (blood, respiratory secretions, and urine), and 14 COVID-19 patients developing CPKP co-infection with colonization. A widespread resistance to all antibiotic classes was found on the antibiograms of CPKP isolates and the blaNDM was the most common genotype detected by preliminary PCR testing. Conclusion: Regarding CPKP infection, a continues focus should be taken when dealing with it, particularly among COVID-19 patients. The incidence of multi-drug resistance due to nosocomial infection can be decreased by the rigorous monitoring of colonized patients and expedite the infection control practices. [ABSTRACT FROM AUTHOR]

Introduction: Infections caused by Klebsiella pneumoniae carbapenemase (KPC) pose a significant clinical challenge due to increasing antibiotic resistance. This study analyzes the effectiveness of ceftazidime/avibactam treatment for KPC infections, focusing on 30-day mortality and comparing results with control groups treated with different antibiotics. Material and methods: We conducted a retrospective analysis of six cohort studies, comparing thirty-day mortality in patients treated with ceftazidime/avibactam with control groups using other antibiotics. The studies included patients infected with KPC, and the analysis focused on therapeutic effectiveness. Aim of the study: The aim of the study is to compare the results in terms of 30-day mortality in groups treated with ceftazidime/avibactam compared to control groups using different antibiotics. Conclusions: Data analysis from various studies revealed varied 30-day mortality outcomes in groups treated with ceftazidime/avibactam compared to control groups using different antibiotics. Ceftazidime/avibactam proved to be more effective in all studies, reducing mortality rates compared to other treatment regimens. We emphasize that emerging antibiotic resistance, especially in the case of KPC, requires a comprehensive therapeutic approach. Despite promising ceftazidime/avibactam results, factors such as overall patient health and treatment delays may influence final therapy outcomes. We also highlight controversies regarding combination therapy vs. monotherapy, necessitating further research. Our work underscores the importance of monitoring the effectiveness of KPC infection treatment and exploring new therapeutic strategies. Further clinical studies are essential to develop a fuller understanding and optimal therapeutic protocols in the face of the growing antibiotic resistance problem. [ABSTRACT FROM AUTHOR]

This research aimed to analyze the volatile compounds emitted during the proliferation of Klebsiella pneumoniae (K. pneumoniae) in the laboratory setting using gas chromatography-ion mobility spectrometry (GC-IMS) and to investigate the potential of volatile metabolomics for detecting carbapenemase-producing strains of K. pneumoniae. The volatile metabolomics of K. pneumoniae were comprehensively analyzed using GC-IMS in tryptic soy broth (TSB) as the culture medium. Afterward, the growth stabilization period (T2) served as the primary time point for analysis, with the introduction of imipenem and carbapenemase inhibitors (avibactam sodium or EDTA) during the exponential growth phase (T0) to further investigate alterations in volatile molecules associated with K. pneumoniae. Standard strains were utilized as references, while clinical strains were employed for validation purposes. At T2, a total of 22 volatile organic compounds (VOCs) associated with K. pneumoniae were identified (3 VOCs found in both monomer and dimer forms). Significant differences in VOCs were observed between carbapenemase-negative and carbapenemase-positive strains, both standard and clinical, following the introduction of imipenem. Furthermore, the addition of avibactam sodium led to distinct changes in the VOC content of strains producing class A carbapenemase, while the addition of EDTA resulted in specific alterations in the volatile metabolic profiles of strains producing class B carbapenemase. GC-IMS demonstrated significant promise for analyzing bacterial volatile metabolomics, and its application in evaluating the volatolomics of K. pneumoniae may facilitate the timely detection of carbapenemase-producing strains. [ABSTRACT FROM AUTHOR]

Background: The hospital wastewater system has been reported as a source of nosocomial acquisition of carbapenemase producing Enterobacteriaceae (CPE) in various settings. Cleaning and disinfection protocols or replacement of contaminated equipment often fail to eradicate these environmental reservoirs, which can lead to long-term transmission of CPE. We report a successful multimodal approach to control a New Delhi metallo-beta-lactamase positive Klebsiella pneumoniae (NDM-KP) nosocomial outbreak implicating contamination of sink traps in a low-incidence setting. Methods: Following the incidental identification of NDM-KP in a urine culture of an inpatient, we performed an epidemiological investigation, including patient and environmental CPE screening, and whole genome sequencing (WGS) of strains. We also implemented multimodal infection prevention and control (IPC) measures, namely the isolation of cases, waterless patient care, replacement of contaminated P-traps and connecting pieces, and bleach and steam disinfection of sinks for 6 months, followed by patient and environmental screenings for eradication. Results: Between February and May 2022, five NDM-KP cases were identified in an eight-bed neurosurgical intermediate care unit. Among the eight sink traps of the unit, three were positive for NDM-KP. Patient and environmental isolates belonged to multilocus sequence typing ST-268. All isolate genomes were genetically very similar suggesting cross-transmission and a potential role of the environment as the source of transmissions. Following the introduction of combined IPC measures, no new case was subsequently detected and sink traps remained negative for NDM-KP within 6 months after the intervention. Conclusion: The implementation of multimodal IPC measures, including waterless patient care combined with the replacement and disinfection of P-traps and connecting pieces, was successful in the control of NDM-KP after eight months. In a low-incidence setting, this approach has made it possible to pursue the objective of zero transmission of carbapenemase-producing Enterobacteriaceae (CPE). [ABSTRACT FROM AUTHOR]

Background: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) is one of the main Gram-negative bacterium causes of infections in hospital settings, and the spread of them is a significant challenge to public health. Methods: A total of 30 non-duplicate isolates of CRPA were collected. Antibacterial susceptibility of isolates to antibiotic agents, AmpC β-lactamase production, and biofilm formation were determined. Minimum biofilm inhibitory concentrations (MBIC) of isolates to cefepime (FEP), imipenem (IPM), ceftazidime (CAZ), and meropenem (MEM) were evaluated with/without cloxacillin (CLX). The carbapenemase and 16 S rRNA methylase genes were identified by PCR, and the transcription levels of oprD, ampC, and mexA genes were determined by quantitative real-time PCR (qPCR). ERIC-PCR was used to detect genetic relationships among isolates. Results: All isolates were multidrug resistant (MDR) and strong biofilm producers. The resistance genes including blaNDM, blaIMP, blaVIM, blaSIM, blaGES, and armA were detected in 21 (70%), 6 (20%), 3 (10%), 2 (6.6%), 1 (3.3%), and 17 (56.6%) of the isolates, respectively. CLX at 500 and 1000 µg/mL significantly reduced the level of MIC to MEM, IPM, CAZ, and FEP, also at 2000 µg/mL significantly reduced the level of MBIC to MEM, IPM, CAZ, and FEP. In all isolates, the transcription levels of oprD were significantly downregulated as well as significantly increased for ampC and mexA. ERIC-PCR typing results divided 30 isolates into four clusters A to D. Conclusion: In this study, we reported the spread of different clones of CRPA harboring co-existence of various carbapenemase genes with armA 16 S rRNA methylase for the first time in Kerman, Iran. Also, our isolates had several mechanisms of resistance to carbapenems as well as ability biofilm formation along with resistance to aminoglycosides, the further spread of which could cause serious challenges in our hospital settings. Therefore, serious monitoring is necessary to reduce their prevalence. [ABSTRACT FROM AUTHOR]

Background: This study aims to investigate the frequency of cas1 and cas3 and CRISPR1,2,3 genes in Klebsiella pneumoniae isolates, as well as their connection with antibiotic resistance. Materials and methods: 106 K. pneumoniae isolates were identified by biochemical assays and PCR. The susceptibility to antibiotics was determined by Kirby–Bauer disk diffusion method. Screening of ESBLs was undertaken by using double disk diffusion and standard disk diffusion methods. The E-test and mCIM techniques was used to confirm the disc diffusion-based carbapenem resistance profiles. CRISPR-Cas system genes were identified using PCR. Results: ESBL production was found in 19% of isolates. Carbapenemase production was found in 46% of the isolates. Furthermore, the bacteria were classified as multidrug (76%), extensively drug-resistant (4%), or pan-drug-resistant (2%). When CRISPR/Cas systems were present, antibiotic resistance was lower; conversely, when they were absent, resistance was higher. Conclusions: If the CRISPR/Cas modules aren't present, the bacteria can still acquire foreign DNA, including antibiotic resistance genes. K. pneumoniae isolates with a CRISPR-Cas system were less likely to carry antibiotic-resistance genes than those lacking this defense system. [ABSTRACT FROM AUTHOR]

With the escalation of hospital-acquired infections by multidrug resistant bacteria, understanding antibiotic resistance is of paramount importance. This study focuses on the β-lactamase gene, bla OXA-58, an important resistance determinant identified in a patient-facing hospital wastewater system. This study aimed to characterize the behaviour of the OXA-58 enzyme when expressed using a non-native plasmid and expression host. bla OXA-58 was cloned using a pET28a(+)/ Escherichia coli BL21(DE3) expression system. Nitrocefin hydrolysis and antimicrobial susceptibility of OXA-58-producing cells were assessed against penicillin G, ampicillin, meropenem, and amoxicillin. bla OXA-58 conferred resistance to amoxicillin, penicillin G, and ampicillin, but not to meropenem. This was unexpected given OXA-58's annotation as a carbapenemase. The presence of meropenem also reduced nitrocefin hydrolysis, suggesting it acts as a competitive inhibitor of the OXA-58 enzyme. This study elucidates the phenotypic resistance conferred by an antimicrobial resistance gene (ARG) obtained from a clinically relevant setting and reveals that successful functional expression of ARGs is multifaceted. This study challenges the reliability of predicting antimicrobial resistance based solely on gene sequence alone, and serves as a reminder of the intricate interplay between genetics and structural factors in understanding resistance profiles across different host environments. [ABSTRACT FROM AUTHOR]

Genes conferring antibiotic resistance phenotype, particularly to last resort antibiotics, pose a significant concern globally. Wastewater treatment plant (WWTP) effluent substantially contributes to antibiotic resistance in receiving rivers, threatening human health. Globally, colistin- and carbapenem-resistant Klebsiella pneumoniae infections cause high morbidity and mortality. We investigated colistin-resistant carbapenemase-producing K. pneumoniae (Co-CRKP) isolates in Kathajodi river receiving WWTP effluent, their resistance genes, and pathogenic potential. Four isolates (Co-CRKP-7, Co-CRKP-8, Co-CRKP-10, and Co-CRKP-15) exhibited extensively drug-resistant (XDR) phenotype, harbouring bla TEM-1, bla CTX-M-15, bla NDM-5, and bla OXA-48 genes. Colistin resistance was attributed to mutations in the pmrA and pmrB genes. Virulence genes (fimH, mrkD, entB, iucA, iutA , and irp1), capsular serotypes (K1, K2) and biofilm formation in the isolates explicated their pathogenicity. Furthermore, Inc plasmid replicons (Y, FrepB, P, K/B, L/M, N, FIA, A/C, and FIB) indicated the dissemination potential of the resistance genes in Co-CRKP isolates. The multi-locus sequence typing showed that Co-CRKP-7 and Co-CRKP-8 belonged to ST42, while Co-CRKP-10 and Co-CRKP-15 were ST16 and ST231, respectively. These high-risk clones carrying multidrug resistance and virulence genes, implicated in numerous outbreaks, have spread worldwide. Our findings emphasize the necessity for effective treatment of hospital wastes to restrict the spread of clinical isolates into aquatic environments. [ABSTRACT FROM AUTHOR]

Background: Clinical significance of Eschericia coli is due to its capacity to develop resistance against many antibiotics specially carbapenem by producing carbapenemase which limits treatment options. blaNDM and blaVIM genes are responsible for this multi-drug-resistance. This is an alarming condition for society as well as clinician because they are very difficult to treat due to limited options of antibiotics and its high virulence. Aims and Objectives: The study was conducted to detect antibiogram pattern and prevalence of blaNDM and blaVIM genes among carbapenem resistant E. coli isolates in a tertiary care hospital. Our study will also help to identify molecular basis of Carbapenem resistance including treatment guideline. Materials and Methods: Cross-sectional study was performed during January'2021 to June'2022 time period. Identification of E. coli was followed by standard protocol and antibiotic sensitivity was done by Kirby-Bauer disk-diffusion method following CLSI guideline, using VITEK-2D-SYSTEM. Detection of genes was done by polymerase chain reaction (PCR). Results: Out of 1867 samples, 161 isolates were identified as E. coli, among which 90 were resistant to carbapenem. Among those, 50 (31%) were resistant to all three carbapenems. Among those, 100% were resistant to ampicillin, amoxycillin-clavulanic acid, cefuroxime, cefuroxime-axetil, cefoperazone-sulbactam, ceftriaxone, cefepime, piperacillin-tazobactam, nalidixic acid, ciprofloxacin; 52% were resistant to amikacin; 54% were resistant to gentamicin; 86% were resistant to cotrimoxazole; 22% were resistant to nitrofurantoin and 96% were sensitive to tigecycline; 100% were intermediate sensitive to colistin. blaNDM gene is detected in 17 (34%) isolates and blaVIM gene is detected in 2 (4%) isolates by PCR. Conclusion: E. coli leads to common clinical presentations such as urinary tract infection, blood stream infection, pneumonia, and wound infection. High burden of carbapenem resistant E. coli was associated with urinary indoor samples. Rising incidence of multi-drug-resistance occurs due to lack of infection control measures, irrational use of antibiotics. Continuous surveillance will prevent development of multi-drug-resistance and help to improve treatment guideline. [ABSTRACT FROM AUTHOR]

Carbapenem resistance in Acinetobacter baumannii is a critical global health threat attributed to transferrable carbapenemase genes. Carbapenemase genotyping using polymerase chain reaction (PCR) presents a challenge in resource-limited settings because of its technical requirements. This study designed new loop-mediated isothermal amplification (LAMP) primers using multiple sequence alignment-based workflows, validated the primer performance against multiple target variants in silico, and developed novel LAMP assays (LAntRN-OXA23 and LAntRN-ISAba1) to detect the transferable blaOXA-23-like carbapenemase genes and ISAba1 elements in pure cultures and A. baumannii-spiked serum samples. The designed LAMP primers bind to the conserved regions of their highly polymorphic targets, with their in silico performance comparable with other published primers. The in vitro LAMP assays (using 30 PCR-profiled A. baumannii and 10 standard multidrug-resistant gram-negative isolates) have 100% concordance with the PCR-positive clinical samples, limits of detection as low as 1 pg/µL (200 copies/µL), and specificities of 57.89–100%. Both assays produced positive results when testing DNA samples (extracted using a commercial kit) from blaOXA-23-like and ISAba1-blaOXA-51-like PCR-positive A. baumannii-spiked normal human sera (five set-ups per target). In summary, the LAMP assays accurately detected the target genes and have applications in infection management, control, and point-of-care testing in resource-limited healthcare settings. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Escherichia coli pose a significant threat to global public health due to the dearth of available treatment options, resulting in infections with high mortality and morbidity. The study aimed to investigate the mechanism of carbapenem resistance in a carbapenem non-susceptible E. coli isolate recovered from an urinary tract infection patient admitted to a tertiary referral hospital, through whole-genome sequencing using Illumina NovaSeq 6000 platform. Carbapenemase production followed by antibiotic susceptibility testing were performed following Clinical Laboratory Standard Institute guidelines. Polymerase chain reaction targeting carbapenemase genes was performed followed by an investigation of horizontal transferability. The Center for Genomic Epidemiology database was used to analyze the sequenced data. ST2519 E. coli BJD_EC1808 with a genome size of 5.8 Mb harbored Col440I plasmid and a chromosomally located blaOXA-116 gene with an IS18 element upstream, along with multiple antibiotic resistance genes conferring clinical resistance toward beta-lactams, aminoglycosides, amphenicols, sulfonamides, tetracyclines, trimethoprim, rifampin, macrolide, and streptogramin antibiotics and antiseptics. E. coli ST2519 harboring blaOXA-116 associated with a mobile genetic element exhibiting carbapenem resistance is a public health threat due to its limiting effect on the therapeutic usage of carbapenem and their dissemination into carbapenem non-susceptible phenotypes will contribute to carbapenem resistance burden and, therefore, warrants urgent monitoring and clinical intervention. [ABSTRACT FROM AUTHOR]

Objective: The global emergence of carbapenem‐resistant Klebsiella pneumoniae is considered a significant contemporary concern., as carbapenems are the last resort for treating infections caused by multidrug‐resistant Gram‐negative bacteria. This study aimed to investigate the carbapenem‐resistance genes in extended‐spectrum β‐lactamase producing K. pneumoniae isolates. Methods: Seventy‐five non‐duplicate clinical K. pneumoniae strains were isolated from urine, blood, sputum, and wound samples. Antimicrobial susceptibility tests for 12 different antibiotics were performed using the disk diffusion method, followed by determining minimum inhibitory concentrations of imipenem and meropenem. Phenotypic detection of extended‐spectrum β‐lactamase and carbapenemase enzymes was performed by double‐disc synergy test and modified Hodge test, respectively. PCR assay further investigated resistant isolates for extended‐spectrum β‐lactamase and carbapenemase‐encoding genes. Results: The highest and lowest resistance rates were observed against ampicillin (93.3%) and tigecycline (9.3%). According to phenotypic tests, 46.7% of isolates were positive for extended‐spectrum β‐lactamase enzymes and 52.8% for carbapenemase. A total of 11 isolates contained carbapenemase genes, with blaOXA‐48 (19.4%; 7/36) being the predominant gene, followed by blaNDM (8.3%; 3/36). Conclusion: The study's findings reveal the alarming prevalence of beta‐lactamase enzymes in K. pneumoniae strains. Early detection of carbapenem‐resistant isolates and effective infection control measures are necessary to minimise further spread, as carbapenem resistance has become a public health concern. [ABSTRACT FROM AUTHOR]

Carbapenem-resistance in Enterobacter spp due to acquisition of mobile carbapenemases is of concern. An Enterobacter spp grew on ChromID CARBA medium and was positive for the mCIM carbapenemase detection assay. Susceptibility testing showed resistance to aztreonam and reduced susceptibility to imipenem. Conventional PCR using FRI primers detected a blaFRI gene. Whole genome sequencing reveled a new variant; blaFRI−12 was closest in sequence to blaFRI−5 differing by 13 amino acids and was found on a unique 110Kb IncR plasmid. Given the intrinsic nature of Enterobacter spp. to be carbapenem non-susceptible, blaFRI-types may be under reported globally. [ABSTRACT FROM AUTHOR]

Background: Standard 24-h antibiotic prophylaxis (AP) is widely employed to minimize the risk of infection complications (ICs) within 30 days following a radical cystectomy (RC). However, a considerable variety of prophylaxis protocols do not prevent a high ICs rate after surgery (37–67%). Therefore, antibiotic's type and its duration are still controversial for AP.(Objective: To compare standard 24-h AP with a prolonged 120-h regimen in a multicenter randomized clinical trial. Methods: Patients were randomized in a 1:1 ratio to standard 24-h AP regimen (Group A) versus the prolonged meropenem AP 120-h (Group B). The primary endpoint was an event rate defined as the frequency of ICs within 30 days. The secondary endpoint were biomarker's analysis and antibiotic re-administration rate (ArAR). Results: A total of 92 patients were enrolled. The Clavien-Dindo complications rate did not differ between the groups (p = 0.065), however the overall complication rate was higher in Group A (63.0% vs. 34.8%, p = 0.007). The infection complication rate was 2.75 times higher in the standard antibiotic prophylaxis group: 47.8% compared to 17.4% cases in Group B (p = 0.002). The new prolonged antibiotic regimen decreased the risk of ICs (OR 0.23; 95% CI 0.08–.598; p = 0.003). The event-free survival for ICs of clinical interest in group A was 7.00 days and in group B was 9.00 days (HR = 0.447; 0.191–1.050, p = 0.065). The ArAR was higher in Group A -47.8%, while in Group B it was only in 17.4% of the cases. The incidence of bacteriuria before RC was the same between groups (p = 0.666), however, after stent removal the risk of a positive culture was lower in group B (RR = 0.64; 95% CI 0.37–1.08; p = 0.05). Conclusions: The administration AP over 120-h appears to be safe and feasible, demonstrating a reduction in the total number of complications and ArAR. Trial registration in Clinical Trials: NCT05392634. Trial registration in Clinical Trials: NCT05392634. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant strains of Pseudomonas aeruginosa (CRPA) have become a major health care concern in many countries, against which anti-infective strategies are limited and which require adequate infection control interventions. Knowing the different modes of transmission of CRPA in intensive care units (ICUs) would be helpful to adapt the means of prevention. The aim of this retrospective case-control study was conducted between January 1, 2017 and February 28, 2022 to identify the risk factors for the acquisition of CRPA in ICUs. During the study period, 147 patients were included (49 cases and 98 controls). Among the 49 patients, 31 (63%) acquired CRPA in clusters and 18 (37%) sporadically. A univariate analysis showed that 4 variables were associated with CRPA acquisition, including (1) prior antibiotic prescriptions, (2) admission to rooms 203 and 207, (3) severity of illness at admission, and (4) use of mechanical ventilation. Multivariate analysis identified 3 factors of CRPA acquisition, including admission to room 203 (odds ratio [OR] = 29.5 [3.52-247.09]), previous antibiotic therapy (OR = 3.44 [1.02-11.76]), and severity of condition at admission (OR = 1.02 [1-1.04]). Our study suggests the role of a contaminated environment in the acquisition of CRPA in the ICU, along with antibiotic use. • Inert surfaces and water points are commonly contaminated by Pseudomona s aeruginosa. • There is a relationship between sporadic acquisition and the environment. • Antibiotic prescription is associated with CRPA acquisition. [ABSTRACT FROM AUTHOR]

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Background: Acinetobacter baumannii continued to be an important Gram-negative pathogen of concern in the clinical context. The resistance of this pathogen to carbapenems due to the production of carbapenemases is considered a global threat. Despite the efforts to track carbapenemase synthesis among A. baumannii in the Philippines, local data on its molecular features are very scarce. This study aims to characterize A. baumannii clinical isolates from a Philippine tertiary hospital through genotyping of the pathogen's carbapenemase genes. Methods: Antibiotic susceptibility profiling, phenotypic testing of carbapenemase production, and polymerase chain reaction assays to detect the different classes of carbapenemase genes (class A blaKPC, class B blaNDM, blaIMP, blaVIM, and class D blaOXA-23-like, blaOXA-24/40-like, blaOXA-48-like, blaOXA-51-like, ISAba1-blaOXA-51-like, blaOXA-58-like) were performed in all collected A. baumannii, both carbapenem resistant and susceptible (n = 52). Results: Results showed that the majority of the carbapenem-resistant strains phenotypically produced carbapenemases (up to 84% in carbapenem inactivation methods) and possessed the ISAba1-blaOXA-51-like gene complex (80%). Meanwhile, both carbapenem-resistant and carbapenem-susceptible isolates possessed multi-class carbapenemase genes including blaNDM (1.9%), blaVIM (3.9%), blaOXA-24/40-like (5.8%), blaOXA-58-like (5.8%), blaKPC (11.5%), and blaOXA-23-like (94.2%), which coexist with each other in some strains (17.3%). In terms of the intrinsic blaOXA-51-like (oxaAb) genes, 23 unique alleles were reported (blaOXA-1058 to blaOXA-1080), the majority of which are closely related to blaOXA-66. Isolates possessing these alleles showed varying carbapenem resistance profiles. Conclusions: In summary, this study highlighted the importance of molecular genotyping in the characterization of A. baumannii by revealing the carbapenemase profiles of the pathogen (which may not be captured accurately in phenotypic tests), in identifying potent carriers of transferrable carbapenemase genes (which may not be expressed straightforwardly in antimicrobial susceptibility testing), and in monitoring unique pathogen epidemiology in the local clinical setting. [ABSTRACT FROM AUTHOR]

Background: Wastewaters are considered as important players in the spread of antimicrobial resistance, thus affecting the health of humans and animals. Here, we focused on wastewaters as a possible source of carbapenemase-producing Enterobacterales for the environment. Methods: A total of 180 presumptive coliforms from hospital and municipal wastewaters, and a river in the Czech Republic were obtained by selective cultivation on meropenem-supplemented media and tested for presence of carbapenemase-encoding genes by PCR. Strains carrying genes of interest were characterized by testing antimicrobial susceptibility, carbapenemase production and combination of short- and long- read whole-genome sequencing. The phylogenetic tree including publicly available genomes of Enterobacter asburiae was conducted using Prokka, Roary and RAxML. Results: Three VIM-producing Enterobacter asburiae isolates, members of the Enterobacter cloacae complex, were detected from hospital and municipal wastewaters, and the river. The blaVIM−1 gene was located within a class 1 integron that was carried by different F-type plasmids and one non-typeable plasmid. Furthermore, one of the isolates carried plasmid-borne colistin-resistance gene mcr-10, while in another isolate chromosomally located mcr-9 without colistin resistance phenotype was detected. In addition, the analysis of 685 publicly available E. asburiae genomes showed they frequently carry carbapenemase genes, highlighting the importance of this species in the emergence of resistance to last-line antibiotics. Conclusion: Our findings pointed out the important contribution of hospital and community wastewaters in transmission of multi-drug resistant pathogens. [ABSTRACT FROM AUTHOR]

The worldwide spread of Pseudomonas aeruginosa, especially carbapenem-resistant P. aeruginosa (CRPA), poses a serious threat to global public health. In this research, we collected and studied the clinical prevalence, molecular epidemiology, and resistance mechanisms of CRPA in Fujian, China. Among 167 non-duplicated P. aeruginosa isolates collected during 2019-2021, strains from respiratory specimens and wound secretions of older males in the intensive care unit dominated. Ninety-eight isolates (58.7%) were resistant to at least one tested antibiotic, among which 70 strains were carbapenem-resistant. Moleclar typing of the CRPA isolates revealed they were highly divergent, belonging to 46 different sequence types. It is noteworthy that two previously reported high risk clones, ST1971 specific to China and the globally prevalent ST357, were found. Several carbapenemresistance-related characteristics were also explored in 70 CRPA isolates. Firstly, carbapenemase was phenotypically positive in 22.9% of CRPA, genetically predominant by metallo-b-lactamase (MBL) and co-carrige of different carbapenemase genes. Then, mutations of the carbapenem-specific porins oprD and opdP were commonly observed, with frequencies of 97.1% and 100.0%, respectively. Furthermore, the biofilm formation and relative transcription levels of 8 multidrug efflux pump genes were also found to be increased in 48.6% and 72.9% of CRPA isolates compared to the reference strain PAO1. These findings will help fill the data gaps in molecular characteristics of CRPA on the southeastern coast of China and emphasize the urgent need for data-based specific stewardship for antipseudomonal practices to prevent the dissemination of CRPA. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa (P. aeruginosa) is one of the most common bacteria isolated from clinical samples, with a rising incidence in hospital infections. This pathogen is inherently resistant to many antibacterial agents. This study aimed to investigate the frequency of carbapenemase genes in P. aeruginosa strains isolated from patients admitted to hospitals in northwestern Iran. A total of 500 P. aeruginosa samples were collected from different clinical samples. Antibiotic susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines, and the frequency of the target genes was assessed using polymerase chain reaction (PCR). The antibiotic resistance results of the samples by disc diffusion method showed that imipenem 98.4%, gentamicin 98%, meropenem 91.8%, amikacin 91.6% and cefepime 91% had the highest resistance; also, out of 500 P. aeruginosa isolates, 309 (61.8%) samples were carbapenemase producers. Using the PCR method, it was determined that the blaOXA-48 (39.16%), blaGES (31.72%), and blaIMP (22.01%) genes were the dominant genes. Our results showed that the prevalence of carbapenemase genes in P. aeruginosa strains isolated from patients admitted to hospitals in northwestern Iran is very high; indicating a need for effective infection control measures to prevent the spread of P. aeruginosa in hospitals. [ABSTRACT FROM AUTHOR]

Aim This study reports the presence of carbapenem-resistant Escherichia coli hybrid pathovars and its prevalence in 200 fresh-cut fruits from Accra. Methods and results Standard culture methods were used to quantify microbial indicators and E. coli on fresh-cut fruits retailed in formal and informal outlets in Accra. The Kirby–Bauer disc diffusion method was used to determine the antibiotic resistance profile of E. coli , while multiplex PCR was employed to identify the virulence and carbapenem-resistance genes. Escherichia coli prevalence in cut fruits was 17%, with pawpaw, watermelon, and mixed fruit having higher prevalence than pineapple. Of the 34 E. coli isolates from fresh-cut fruits, 44% showed broad resistance to beta-lactam antibiotics, while 5.9% showed carbapenem resistance. The study identified virulence genes associated with all E. coli isolates, including stx1, stx2, escV , and ipaH , of which 97% were hybrid pathovars bearing genes for Shiga toxin-producing E. coli /enteropathogenic E. coli /enteroinvasive E. coli. The carbapenemase gene, bla IMP, was associated with both carbapenem-resistant E. coli phenotypes identified. Conclusion Despite a low-carbapenem-resistance prevalence observed among E. coli isolates, hypervirulent hybrid strains of E. coli is present in fresh-cut fruits in the sampling area, posing a potential public health risk to fresh-cut fruit consumers. [ABSTRACT FROM AUTHOR]

Escherichia coli is an important opportunistic pathogen, causing several infections in dogs. The antimicrobial resistance of E. coli occurring in companion animals becomes an emerging problem. This study aimed to estimate the prevalence of ESBL-producing E. coli in diarrheic dogs, investigate the occurrence and molecular characterization of carbapenem-resistant isolates, and determine their virulence genes. Fecal samples were collected from 150 diarrheic dogs in Tunisia. E. coli isolates were screened for antimicrobial resistance against 21 antibiotics by the disk diffusion method. The characterization of β-lactamase genes, associated resistance genes, and virulence genes was studied using PCR. Among 95 E. coli strains, 25 were ESBL-producing, and most of them were multidrug-resistant. The most prevalent β-lactamase genes were blaCTX-M1 (n = 14), blaTEM (n = 3), and blaCMY (n = 2). The blaIMP carbapenemase gene was found in two carbapenem-resistant isolates, which showed that carbapenemase-producing E. coli spread to companion animals in Tunisia. Different virulence genes associated with extraintestinal pathogenic E. coli were detected. This is the first report of the characterization of carbapenem resistance and virulence genes in dogs in North Africa. Our study showed that diarrheic dogs in Tunisia can be a potential reservoir of ESBL- or carbapenemase-producing E. coli with a possible risk of transmission to humans. [ABSTRACT FROM AUTHOR]

Multidrug-resistant (MDR) Gram-negative bacterial infections have emerged as a major public health concern. The aim of the present study was to detect the rate of infections due to MDR Gram-negative bacteria (GNB) in a tertiary care hospital, the rate of Carbapenemases and AmpC-b-lactamases production and the Antimicrobial susceptibility test pattern (AST) among MDR GNB. The rate of MDR GNB during the study period was 25.70%. Urine samples showed the highest contribution to the total MDR GNB. Among the total MDR GNB isolates, 166 were randomly selected and included in the present study. A higher rate of MDR GNB was reported among male patients (61.5%) compared to the females (38.5%) and most of them were from the patients aged between 61-70 years (30.7%). The most prevalent MDR GNB was Klebsiella pneumoniae 80 (48.12%), followed by Escherichia coli 43 (25.9%). AST of MDR GNB revealed their significant resistance to b-lactamases/b-lactamases inhibitors, cephalosporins, fluoroquinolones and carbapenem drugs (98%). Of 123 MDR Enterobacterales, 83% of them were found to be Metallo b-lactamase (MBLs) producers by mCIM and eCIM methods. Of 43 MDR non-fermenters, 29 (67.4%) of them were found to be carbapenemase producers by MHT. About 29.51% of MDR GNB isolates were found to be AmpC producers by AmpC disk test. A reliable and rapid phenotypic method to detect carbapenemases and AmpC b-lactamases among MDR GNB in a routine microbiology laboratory method is clinically important to guide antibiotic therapy and implementation of effective infection control practices. [ABSTRACT FROM AUTHOR]

Objective To identify early Klebsiella pneumoniae (KP) infection after liver transplantation and its impact on prognosis. Methods Clinical data of 171 liver transplant recipients were retrospectively analyzed, and they were divided into the non-infection (n=52) and infection groups (n=119) according to the bacterial culture results at postoperative 2 weeks. In the infection group, KP was not detected in 86 cases (non-KP infection group), and KP was cultured in 33 cases (KP infection group). Preoperative, intraoperative and postoperative data were statistically compared between the non-infection and infection groups, and between the non-KP infection and KP infection groups. The risk factors of early KP infection after liver transplantation and the influencing factors of long-term survival of the recipients were analyzed.Results Compared with the non-infection group, model for end-stage liver disease (MELD) score and total bilirubin level were higher, the operation time was longer, the length of postoperative intensive care unit (ICU) stay and the length of hospital stay were longer, the amount of intraoperative red blood cell transfusion was higher, the hospitalization expense was higher, the incidence of severe complications was higher, white blood cell count, absolute neutrophil cell count and neutrophil-to-lymphocyte ratio at postoperative 14 and 30 d were higher, absolute lymphocyte count at postoperative 14 d was lower and hemoglobin level at postoperative 30 d was lower in the infection group. The differences were statistically significant (all P<0.05). Compared with the non-KP infection group, MELD score, total bilirubin level and aspartate aminotransferase (AST) level were higher, the operation time and the length of postoperative ICU stay were longer, the hospitalization expense was higher, the 90-d fatality was higher, the albumin level at postoperative 14 d was lower, and total bilirubin level at postoperative 30 d was higher in the KP infection group. The differences were statistically significant (all P<0.05). Among 33 recipients with KP infection, 16 cases were resistant to carbapenem antibiotics, and 7 of them died within postoperative 90 d. Seventeen cases were intermediate or sensitive to carbapenem antibiotics, and 4 of them died within postoperative 90 d. Preoperative MELD score ≥17 and operation time≥415 min were the independent risk factors for KP infection after liver transplantation (both P<0.05). The length of postoperative ICU stay ≥44 h and KP infection were the independent risk factors for long-term prognosis of liver transplantation (both P<0.05). Conclusions KP infection is an independent risk factor for death after liver transplantation. High preoperative MELD score and long operation time are the independent risk factors for early KP infection after liver transplantation. [ABSTRACT FROM AUTHOR]

Purpose: Carbapenem resistant Pseudomonas aeruginosa (CR-PA) is escalating worldwide and leaves clinicians few therapeutic options in recent years, β-lactam/β-lactamase inhibitor combinations (ceftolozane-tazobactam, ceftazidime-avibactam) and a new siderophore cephalosporin (cefiderocol) have been approved for the treatment of P. aeruginosa infection and have shown potent activity against isolates defined as carbapenem resistant. The aim of this study was to determine the phenotypic profile of these agents against CR-PA in the emerging setting of carbapenemases. Methods: CR-PA clinical isolates were collected from three teaching hospitals in different geographical regions between January 2017-December 2021. All isolates were subjected to phenotypic carbapenemase testing using modified carbapenem inactivation method. MICs were determined by reference broth microdilution and evaluated according to EUCAST standards, while genotypic profiling was determined using PCR methods. Results: 244 CR-PA sourced most frequently from the respiratory tract (32.2%), blood (20.4%) and urine (17.5%) were evaluated. Of all isolates, 32 (13.1%) were phenotypically and 38 (15.6%) were genotypically defined as carbapenemase-positive. The most common carbapenemase was GES (63.1%), followed by VIM (15.8%). The MIC50/90(S%) of ceftazidime/avibactam, ceftolozane/tazobactam and cefiderocol in all CR-PA isolates were 4 and 32 (80%), 1 and > 64 (69%) and 0.25 and 1 mg/L (96%), respectively. Cefiderocol was also the most active agent in carbapenemase-positive isolates (90%). Conslusion: While ceftolozane/tazobactam and ceftazidime/avibactam remained highly active against CR-PA devoid of carbapenemases, cefiderocol provided potent in vitro activity irrespective of carbapenemase production. When considering the potential clinical utility of newer agents against CR-PA, regional variations in carbapenemase prevalence must be considered. [ABSTRACT FROM AUTHOR]

Background: Klebsiella pneumoniae species complex (KpSC) is an important disseminator of carbapenemase‐encoding genes, mainly blaKPC‐2 and blaNDM‐1, from hospitals to the environment. Consequently, carbapenem‐resistant strains can be spread through the agrifood system, raising concerns about food safety. This study therefore aimed to isolate carbapenem‐resistant KpSC strains from the agricultural and environmental sectors and characterize them using phenotypic, molecular, and genomic analyses. Results: Klebsiella pneumoniae and Klebsiella quasipneumoniae strains isolated from soils used for lemon, guava, and fig cultivation, and from surface waters, displayed an extensive drug‐resistance profile and carried blaKPC‐2, blaNDM‐1, or both. In addition to carbapenemase‐encoding genes, KpSC strains harbor a broad resistome (antimicrobial resistance and metal tolerance) and present putative hypervirulence. Soil‐derived K. pneumoniae strains were assigned as high‐risk clones (ST11 and ST307) and harbored the blaKPC‐2 gene associated with Tn4401b and Tn3‐like elements on IncN‐pST15 and IncX5 plasmids. In surface waters, the coexistence of blaKPC‐2 and blaNDM‐1 genes was identified in K. pneumoniae ST6326, a new carbapenem‐resistant regional Brazilian clone. In this case, blaKPC‐2 with Tn4401a isoform and blaNDM‐1 associated with a Tn125‐like transposon were located on different plasmids. Klebsiella quasipneumoniae ST526 also presented the blaNDM‐1 gene associated with a Tn3000 transposon on an IncX3 plasmid. Conclusion: These findings provide a warning regarding the transmission of carbapenemase‐positive KpSC across the agricultural and environmental sectors, raising critical food safety and environmental issues. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The healthcare water environment is a potential reservoir of carbapenem-resistant organisms (CROs). To report the role of the water environment as a reservoir and the infection control measures applied to suppress a prolonged outbreak of Klebsiella pneumoniae carbapenemase-producing Serratia marcescens (KPC-SM) in two intensive care units (ICUs). The outbreak occurred in the ICUs of a tertiary hospital from October 2020 to July 2021. Comprehensive patient contact tracing and environmental assessments were conducted, and a case–control study was performed to identify factors associated with the acquisition of KPC-SM. Associations among isolates were assessed via pulsed-field gel electrophoresis (PFGE). Antibiotic usage was analysed. The outbreak consisted of two waves involving a total of 30 patients with KPC-SM. Multiple environmental cultures identified KPC-SM in a sink, a dirty utility room, and a communal bathroom shared by the ICUs, together with the waste bucket of a continuous renal replacement therapy (CRRT) system. The genetic similarity of the KPC-SM isolates from patients and the environment was confirmed by PFGE. A retrospective review of 30 cases identified that the use of CRRT and antibiotics was associated with acquisition of KPC-SM (P < 0.05). There was a continuous increase in the use of carbapenems; notably, the use of colistin has increased since 2019. Our study demonstrates that CRRT systems, along with other hospital water environments, are significant potential sources of resistant micro-organisms, underscoring the necessity of enhancing infection control practices in these areas. [ABSTRACT FROM AUTHOR]

We report a nosocomial outbreak caused by a multidrug-resistant carbapenemase-producing Klebsiella pneumoniae (MDRCP Kp), that was detected in six patients admitted to the medical intensive care unit between 20th of December 2023 and 15th of January 2024 in Ankara, Turkey. The investigation of this outbreak was started on 29th of December 2023. During the outbreak 11 samples were collected from the six patients with MDRCP Kp. Pulsed-field gel electrophoresis (PFGE) was performed to determine the genetic relatedness and clonality of MDRCP Kp strains. MDRCP Kp was isolated in the tracheal aspiration culture, blood, urine, and screening samples. Five patients with MDRCP Kp colonization developed healthcare-associated infection. In one patient MDRCP Kp was isolated from tracheal aspirate and the screening cultures were considered as colonization not infection. PFGE analysis revealed that all isolates belonged to the same K. pneumoniae clone. MDRCP Kp strain of this outbreak exhibited multidrug resistance and co-produced OXA-48 and NDM-1. This outbreak ended after application of strict infection control measures. An outbreak of MDRCP Kp can occur in hospitals, especially in the intensive care units; thus, it should be detected early by infection control teams. A strong collaboration between infection control team and microbiology laboratory is essential to cope with MDR bacterial outbreaks in hospitals. [ABSTRACT FROM AUTHOR]

VIM-type-producing Gram-negative bacteria (GNB) infections are difficult to treat. This is a retrospective single-center study of 34 patients who received cefiderocol for the treatment of VIM-type-producing GNB infections, including 25 Pseudomonas spp., 7 Enterobacterales, and 5 Achromobacter sp. Primary outcomes were clinical failure (defined as death, lack of clinical improvement, or a switch to another drug) at day 14 and 30-day all-cause mortality. The median age was 59 years (IQR 53.7–73.4), and the median Charlson comorbidity index was 3.5 (IQR 2–5). The main infections were respiratory tract infections (n = 9, 27%) and skin and soft tissue infections (n = 9, 27%). Eight patients exhibited bacteremia. In 9/17 patients with a drainable focus, drainage was performed. The median cefiderocol treatment duration was 13 days (IQR 8–24). Five patients (15%) experienced clinical failure on day 14, and the thirty-day mortality rate was 9/34 (27%); two cases occurred because of an uncontrolled infection source, and one was due to a new infection caused by the same bacteria. The other six deaths were unrelated to the index infection. Five patients experienced microbiological recurrence within three months. Susceptibility testing revealed the development of cefiderocol resistance in 1/7 cases with persistent or recurrent positive cultures. Cefiderocol, even in monotherapy, could be considered for the treatment of VIM-type-producing GNB infections. [ABSTRACT FROM AUTHOR]

Due to the increasing resistance of aerobic and facultative anaerobic Gram-negative rods, ceftazidime-avibactam and ceftolozane-tazobactam have been launched in the market in the last few years. In this study, we analyzed the susceptibility pattern of the major aerobic and facultative anaerobic Gram-negative rods in Hong Kong for ceftazidime-avibactam, ceftolozane-tazobactam, four other broad-spectrum antibiotics commonly used in Hong Kong and colistin. For 300 isolates collected from January to December 2021, non-ESBL-producing Enterobacterales, ESBL-producing Enterobacterales and Pseudomonas aeruginosa were highly susceptible to ceftazidime-avibactam (all 100%) and ceftolozane-tazobactam (98.7%, 99.7% and 94.3%). For 32 archived ESBL-producing Klebsiella pneumoniae isolates collected between January 2014 and March 2023, all were susceptible to ceftazidime-avibactam and ceftolozane-tazobactam. For 101 archived carbapenemase-producing Enterobacterales, their susceptibilities to ceftazidime-avibactam and ceftolozane-tazobactam varied depending on the type of carbapenemase produced. Both had high activities against OXA-producing strains (97.1% and 76.5%, respectively) but were 100% resistant for NDM-producing and NDM+OXA-producing strains. All KPC-producing strains were susceptible to ceftazidime-avibactam but resistant to ceftolozane-tazobactam. Ceftazidime-avibactam and ceftolozane-tazobactam are good alternatives for the management of infections caused by ESBL-producing Enterobacterales and selective strains of carbapenemase-producing Enterobacterales in Hong Kong. [ABSTRACT FROM AUTHOR]

Our aim was to elucidate the resistance mechanisms and assess the combined synergistic and bactericidal activities of aztreonam in combination with ceftazidime/avibactam (CZA), meropenem/vaborbactam (MEV), and imipenem/relebactam (IMR) against Enterobacterales strains producing dual carbapenemases. Methods: Species identification, antimicrobial susceptibility testing and determination of carbapenemase type were performed for these strains. Plasmid sizes, plasmid conjugation abilities and the localization of carbapenemase genes were investigated. Whole-genome sequencing was performed for all strains and their molecular characteristics were analyzed. In vitro synergistic and bactericidal activities of the combination of aztreonam with CZA, MEV and IMR against these strains were determined using checkerboard assay and time-kill curve assay. Results: A total of 12 Enterobacterales strains producing dual-carbapenemases were collected, including nine K. pneumoniae, two P. rettgeri, and one E. hormaechei. The most common dual-carbapenemase gene pattern observed was bla(KPC-2+NDM-5) (n=4), followed by blaKPC-2+IMP-26 (n=3), bla(KPC-2+NDM-1) (n=2), bla(KPC-2+IMP-4) (n=1), bla(NDM-1+IMP-4) (n=1) and bla(KPC-2+KPC-2) (n=1). In each strain, the carbapenemase genes were found to be located on two distinct plasmids which were capable of conjugating from the original strain to the receipt strain E. coli J53. The results of the checkerboard synergy analysis consistently revealed good synergistic effects of the combination of ATM with CZA, MEV and IMR. Except for one strain, all strains exhibited significant synergistic activity and bactericidal activity between 2 and 8 hours. Conclusion: Dual-carbapenemase-producing Enterobacterales posed a significant threat to clinical anti-infection treatment. However, the combination of ATM with innovative β-lactam/β-lactamase inhibitor compounds had proven to be an effective treatment option. [ABSTRACT FROM AUTHOR]

Carbapenemase-producing organisms (CPOs) present a major threat to public health, demanding precise diagnostic techniques for their detection. Discrepancies among the CPO tests have raised concerns, partly due to limitations in detecting bacterial diversity within host specimens. We explored the impact of an unbiased colony selection on carbapenemase testing and assessed its relevance to various tests. Using the FirstAll method for unbiased colony selection to reduce bias, we compared the results from different methods, namely the modified carbapenem inactivation method/EDTA-modified carbapenem inactivation method (mCIM/eCIM), the Carba5, the CPO panel, and the multiplex PCR (MPCR). We compared the FirstAll method to the conventional colony selection for MPCR with seven CPO species. In addition, we evaluated the test performance on seven CPO species using MPCR as a reference and the FirstAll method as the colony-selection method. The results revealed that the selections from the FirstAll method have improved rates of carbapenemase detection, in comparison to approximately 11.2% of the CPO isolates that were noted to be false negatives in the conventional colony-selection methods. Both the Carba5 test and the CPO panel showed suboptimal performance (sensitivity/specificity: Carba5 74.6%/89.5%, CPO panel 77.2%/74.4%) in comparison to the FirstAll method. The Carba5 test provided specific carbapenemase class assignments, but the CPO panel failed in 18.7% of the cases. The Carba5 test and the CPO panel results correlated well with ceftazidime–avibactam minimal inhibitory concentrations (MICs). The concordance for Class A/D with MICs was 94.7% for Carba5 and 92.7% for the CPO panel; whereas for Class B, it was 86.5% for Carba5 and 75.9% for the CPO panel. In conclusion, FirstAll, as the unbiased colony-selection method, was shown to impact carbapenemase testing. With FirstAll, the diagnostic performance of both the Carba5 and the CPO panel was found to be lower. Furthermore, the utilization of ceftazidime–avibactam guided by either the CPO panel or Carba5 was appropriate. [ABSTRACT FROM AUTHOR]

Background. In the context of increasing antimicrobial resistance (AMR), whole-genome sequencing (WGS) of bacteria is considered a highly accurate and comprehensive surveillance method for detecting and tracking the spread of resistant pathogens. Two primary sequencing technologies exist: short-read sequencing (50–300 base pairs) and long-read sequencing (thousands of base pairs). The former, based on Illumina sequencing platforms (ISPs), provides extensive coverage and high accuracy for detecting single nucleotide polymorphisms (SNPs) and small insertions/deletions, but is limited by its read length. The latter, based on platforms such as Oxford Nanopore Technologies (ONT), enables the assembly of genomes, particularly those with repetitive regions and structural variants, although its accuracy has historically been lower. Results. We performed a head-to-head comparison of these techniques to sequence the K. pneumoniae VS17 isolate, focusing on blaNDM resistance gene alleles in the context of a surveillance program. Discrepancies between the ISP (blaNDM-4 allele identified) and ONT (blaNDM-1 and blaNDM-5 alleles identified) were observed. Conjugation assays and Sanger sequencing, used as the gold standard, confirmed the validity of ONT results. This study demonstrates the importance of long-read or hybrid assemblies for accurate carbapenemase resistance gene identification and highlights the limitations of short reads in the context of gene duplications or multiple alleles. Conclusions. In this proof-of-concept study, we conclude that recent long-read sequencing technology may outperform standard short-read sequencing for the accurate identification of carbapenemase alleles. Such information is crucial given the rising prevalence of strains producing multiple carbapenemases, especially as WGS is increasingly used for epidemiological surveillance and infection control. [ABSTRACT FROM AUTHOR]

As bloodstream infections and associated septic shock are common causes of mortality in hospitals, rapid antibiotic susceptibility testing (AST) performed directly on positive blood cultures is needed to implement an efficient therapy in clinical settings. We evaluated the Reveal® rapid AST system on a collection of 197 fully characterized carbapenem-resistant Enterobacterales, including 177 carbapenemase producers (CPE) spiked in blood culture bottles. The clinical categorization based on the Minimal Inhibitory Concentration (MIC) determination of eighteen antimicrobial molecules was compared to the clinical categorization based on the disk diffusion assay as a reference. The Reveal AST system provided results within a mean time to result of 5 h. Overall, the categorical agreement (CA) between the two techniques was 94.1%. The rates of very major errors (VMEs), major errors (MEs) and minor errors (mEs) were 3.8%, 3.7% and 5.6%, respectively. Imipenem was the antimicrobial with the lowest CA rate (78.7%), with rates of 15% VMEs and 10.7% MEs, but the performances were better when considering only the non-CPE category (CA of 89%). On this resistant collection of Enterobacterales with numerous acquired β-lactamases, the Specific Reveal assay proved to be useful for a rapid determination of AST compatible with a quick adaptation of the patient's antimicrobial treatment. [ABSTRACT FROM AUTHOR]

Background: Carbapenem resistance in Pseudomonas aeruginosa is a serious global health problem. We investigated the clonal distribution and its association with the carbapenem resistance mechanisms of carbapenem-non-susceptible P. aeruginosa isolates from three Korean hospitals. Methods: A total of 155 carbapenem-non-susceptible P. aeruginosa isolates collected between 2011 and 2019 were analyzed for sequence types (STs), antimicrobial susceptibility, and carbapenem resistance mechanisms, including carbapenemase production, the presence of resistance genes, OprD mutations, and the hyperproduction of AmpC ß-lactamase. Results: Sixty STs were identified in carbapenem-non-susceptible P. aeruginosa isolates. Two high-risk clones, ST235 (N=41) and ST111 (N=20), were predominant; however, sporadic STs were more prevalent than high-risk clones. The resistance rate to amikacin was the lowest (49.7%), whereas that to piperacillin was the highest (92.3%). Of the 155 carbapenem-non-susceptible isolates, 43 (27.7%) produced carbapenemases. Three metallo-β-lactamase (MBL) genes, blaIMP-6 (N=38), blaVIM-2(N=3), and blaNDM-1(N=2), were detected. blaIMP-6 was detected in clonal complex 235 isolates. Two ST773 isolates carried blaNDM-1 and rmtB. Frameshift mutations in oprD were identified in all isolates tested, regardless of the presence of MBL genes. Hyperproduction of AmpC was detected in MBL gene-negative isolates. Conclusions: Frameshift mutations in oprD combined with MBL production or hyperproduction of AmpC are responsible for carbapenem resistance in P. aeruginosa. Further attention is required to curb the emergence and spread of new carbapenem-resistant P. aeruginosa clones. [ABSTRACT FROM AUTHOR]

Caiyun Li,1,&ast; Fa Zhang,1,&ast; Gang Li,2,3 Wen Wang2,3 1Department of Clinical Laboratory, Nanjing Pukou People’s Hospital, Nanjing, 211800, People’s Republic of China; 2Center of Medical Laboratory, General Hospital of Ningxia Medical University, Yinchuan, 750004, People’s Republic of China; 3Ningxia Key Laboratory of Clinical and Pathogenic Microbiology, General Hospital of Ningxia Medical University, Yinchuan, 750004, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wen Wang, Email 42170662@qq.comObjective: To analyze the prevalence and molecular characteristics of 16S rRNA methylase genes in clinical isolates of carbapenem-resistant Enterobacterales, for clinical doctors provide a reference basis for the rational use of drugs.Methods: The Enterobacterales isolated from our hospital from 2020 to 2022 were selected and identified by VITEK 2 Compact automatic bacterial identification instrument. Resistance genes were detected by polymerase chain reaction.Results: A total of 180 carbapenem-resistant Enterobacterales were isolated, of which 158 (87.8%) were resistant to at least one aminoglycoside. The resistance rates to gentamicin, tobramycin and amikacin were 85.0%,82.8% and 54.4%, respectively. Compared with 16S rRNA methyltransferase negative isolates, the positive isolates were more sensitive to trimethoprim-sulfamethoxazole, tetracycline and minocycline, but had higher resistance rates to aztreonam, tobramycin, gentamicin, amikacin and ciprofloxacin. The resistance rates of 16S rRNA methyltransferase gene positive strains to most commonly used antibiotics were more than 80%. But the rates for colistin and tigecycline were less than 10%. There were 114 strains (63.3%) positive for 16S rRNA methyltransferase genes, mainly rmtB, accounting for 70.2%. The positive rates of other armA, rmtA and armA+rmtB genes were 22.8%, 4.4% and 2.6%, respectively. No rmtC, rmtD, rmtE and npmA genes were detected. In addition, 175 of the 180 carbapenem-resistant Enterobacterales carried at least one carbapenemase genes. The blaKPC was the main one (115, 65.7%). There were 111 (61.7%) strains carried both carbapenemase and 16S rRNA methyltransferase genes, simultaneously. Compared with 16S rRNA methyltransferase negative strains, the positive strains carried more blaKPC genes and less blaNDM genes, with P values of 0.034 and 0.003, respectively.Conclusion: blaKPC and rmtB genes are the main resistance mechanisms of Enterobacterales to carbapenems and aminoglycosides in our hospital. It is necessary to strengthen the detection of multi-drug resistant strains to provide scientific basis for clinical rational drug use.Keywords: enterobacterales, carbapenemase, 16S rRNA methylase

Abstract The increasing prevalence of carbapenem-resistant Enterobacter cloacae complex (CREC) poses great challenges to infection treatment in the clinical setting. In this study, we reported the emergence of carbapenemase in a rare species, Enterobacter chuandaensis, belonging to the Enterobacter cloacae complex (ECC). We elucidated the genetic characteristics of carbapenem-resistant isolate FAHZZU5885, co-harboring bla NDM−1 and bla KPC−2. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and average nucleotide identity (ANI) analysis were used to identify E. chuandaensis. S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) and Southern blotting were used to clarify the number and size of the plasmids in FAHZZU5885. Antimicrobial phenotypes were identified by antimicrobial susceptibility testing (AST), and the characteristics of the strain were examined with whole-genome sequencing (WGS). The conjugation experiment and stability assay were conducted to verify the transferability and stability of the plasmid carrying carbapenemase-encoding genes. E. chuandaensis FAHZZU5885 was isolated from a perianal swab of a patient admitted to the ICU. This strain simultaneously carried bla NDM−1 and two bla KPC−2 genes. FAHZZU5885 was resistant to most of the tested antibiotics except for amikacin, tigecycline, and colistin. Two bla KPC−2 were located separately on two different plasmids, the ~ 120 kb IncFIA-IncFII plasmid and the ~ 80 kb IncR plasmid. Both plasmids shared the conserved sequence klcA-korC-ISkpn6-bla KPC−2 -ISkpn27-tnpR-tnpA. The bla NDM−1-bearing plasmid had the potential to transfer and can remain stable after successive passages. In addition, the bla NDM−1 was carried on a ~ 80 kb IncFII plasmid with the conserved sequence ISAba125-bla NDM−1-ble-trpF-dsbD-cutA-groS-groL. In summary, this study marks the first report of the multidrug-resistant E. chuandaensis strain FAHZZU5885 harboring two bla KPC−2-bearing plasmids, indicating the potential for the further dissemination of carbapenemase-encoding genes in novel species. The findings contribute to enhancing our understanding of CREC strains, emphasizing the need for continued and comprehensive surveillance of this species.

Abstract Carbapenem resistant (CRE) and carbapenemase producing Enterobacterales (CPE) in particular, represent a major threat for healthcare systems worldwide. Rapid, reliable, and easy to perform assays are required to enable targeted and effective therapy. MALDI-TOF MS based carbapenemase diagnostics has potential advantages over molecular and phenotypic sensitivity tests, especially in terms of time to result. So far, only one mass spectrometry (MS)-based carbapenemase test system is commercially available for routine use. The aim of this study was to compare the performance of the established system to a novel MS-based test to identify CPE isolates. Forty consecutive CRE isolates (70% CPEs) were pre-screened for carbapenemase activity by routine laboratory methods. Isolates then were tested using the to date only IVD CE certified MALDI-TOF MS carbapenemase detection assay (MBT STAR-Carba IVD Kit; Bruker Daltonics) and a novel test designed for the recently launched EXS2600 MALDI-TOF MS system (Carbapenemase Activity Kit; Zybio). Valid results were obtained for 93% and 85% isolates by the Bruker and the Zybio assay, respectively. Sensitivities, specificities, positive and negative predictive values were 92%, 91%, 96%, and 83% for the Bruker assay and 96%, 64%, 85%, and 88% for the Zybio assay. There are notable differences concerning the handling of the assays. In summary, both systems featured high sensitivities for the detection of carbapenemases, but the Bruker assay yielded less false-positive results. There are advantages and disadvantages concerning the handling for each system, but both proved to be suitable for the use in a routine laboratory.

Abstract This research aimed to analyze the volatile compounds emitted during the proliferation of Klebsiella pneumoniae (K. pneumoniae) in the laboratory setting using gas chromatography-ion mobility spectrometry (GC-IMS) and to investigate the potential of volatile metabolomics for detecting carbapenemase-producing strains of K. pneumoniae. The volatile metabolomics of K. pneumoniae were comprehensively analyzed using GC-IMS in tryptic soy broth (TSB) as the culture medium. Afterward, the growth stabilization period (T2) served as the primary time point for analysis, with the introduction of imipenem and carbapenemase inhibitors (avibactam sodium or EDTA) during the exponential growth phase (T0) to further investigate alterations in volatile molecules associated with K. pneumoniae. Standard strains were utilized as references, while clinical strains were employed for validation purposes. At T2, a total of 22 volatile organic compounds (VOCs) associated with K. pneumoniae were identified (3 VOCs found in both monomer and dimer forms). Significant differences in VOCs were observed between carbapenemase-negative and carbapenemase-positive strains, both standard and clinical, following the introduction of imipenem. Furthermore, the addition of avibactam sodium led to distinct changes in the VOC content of strains producing class A carbapenemase, while the addition of EDTA resulted in specific alterations in the volatile metabolic profiles of strains producing class B carbapenemase. GC-IMS demonstrated significant promise for analyzing bacterial volatile metabolomics, and its application in evaluating the volatolomics of K. pneumoniae may facilitate the timely detection of carbapenemase-producing strains.

Abstract Background The hospital wastewater system has been reported as a source of nosocomial acquisition of carbapenemase producing Enterobacteriaceae (CPE) in various settings. Cleaning and disinfection protocols or replacement of contaminated equipment often fail to eradicate these environmental reservoirs, which can lead to long-term transmission of CPE. We report a successful multimodal approach to control a New Delhi metallo-beta-lactamase positive Klebsiella pneumoniae (NDM-KP) nosocomial outbreak implicating contamination of sink traps in a low-incidence setting. Methods Following the incidental identification of NDM-KP in a urine culture of an inpatient, we performed an epidemiological investigation, including patient and environmental CPE screening, and whole genome sequencing (WGS) of strains. We also implemented multimodal infection prevention and control (IPC) measures, namely the isolation of cases, waterless patient care, replacement of contaminated P-traps and connecting pieces, and bleach and steam disinfection of sinks for 6 months, followed by patient and environmental screenings for eradication. Results Between February and May 2022, five NDM-KP cases were identified in an eight-bed neurosurgical intermediate care unit. Among the eight sink traps of the unit, three were positive for NDM-KP. Patient and environmental isolates belonged to multilocus sequence typing ST-268. All isolate genomes were genetically very similar suggesting cross-transmission and a potential role of the environment as the source of transmissions. Following the introduction of combined IPC measures, no new case was subsequently detected and sink traps remained negative for NDM-KP within 6 months after the intervention. Conclusion The implementation of multimodal IPC measures, including waterless patient care combined with the replacement and disinfection of P-traps and connecting pieces, was successful in the control of NDM-KP after eight months. In a low-incidence setting, this approach has made it possible to pursue the objective of zero transmission of carbapenemase-producing Enterobacteriaceae (CPE).

Abstract Background Carbapenem-resistant Pseudomonas aeruginosa (CRPA) is one of the main Gram-negative bacterium causes of infections in hospital settings, and the spread of them is a significant challenge to public health. Methods A total of 30 non-duplicate isolates of CRPA were collected. Antibacterial susceptibility of isolates to antibiotic agents, AmpC β-lactamase production, and biofilm formation were determined. Minimum biofilm inhibitory concentrations (MBIC) of isolates to cefepime (FEP), imipenem (IPM), ceftazidime (CAZ), and meropenem (MEM) were evaluated with/without cloxacillin (CLX). The carbapenemase and 16 S rRNA methylase genes were identified by PCR, and the transcription levels of oprD, ampC, and mexA genes were determined by quantitative real-time PCR (qPCR). ERIC-PCR was used to detect genetic relationships among isolates. Results All isolates were multidrug resistant (MDR) and strong biofilm producers. The resistance genes including bla NDM, blaIMP, bla VIM, bla SIM, bla GES, and armA were detected in 21 (70%), 6 (20%), 3 (10%), 2 (6.6%), 1 (3.3%), and 17 (56.6%) of the isolates, respectively. CLX at 500 and 1000 µg/mL significantly reduced the level of MIC to MEM, IPM, CAZ, and FEP, also at 2000 µg/mL significantly reduced the level of MBIC to MEM, IPM, CAZ, and FEP. In all isolates, the transcription levels of oprD were significantly downregulated as well as significantly increased for ampC and mexA. ERIC-PCR typing results divided 30 isolates into four clusters A to D. Conclusion In this study, we reported the spread of different clones of CRPA harboring co-existence of various carbapenemase genes with armA 16 S rRNA methylase for the first time in Kerman, Iran. Also, our isolates had several mechanisms of resistance to carbapenems as well as ability biofilm formation along with resistance to aminoglycosides, the further spread of which could cause serious challenges in our hospital settings. Therefore, serious monitoring is necessary to reduce their prevalence.

Abstract Objective We aim to validate and evaluate a new rapid and simplified method, called Blood-rsCDM, for the detection and characterization of carbapenemase using 3-aminophenylboronic acid (APBA) and ethylenediaminetetraacetic acid (EDTA) β-lactamase inhibitors from positive blood cultures. Method We utilized a panel of 172 Enterobacterales strains, including blaKPC (77), blaNDM (48), blaIMP (9), blaVIM (2), blaOXA-181 (2), blaKPC and blaNDM (6), as well as 28 carbapenem-susceptible Enterobacterales isolates, to assess the performance of Blood-rsCDM and the EDTA-carbapenem inactivation method (eCIM). Carbapenemase class was determined using specific inhibitors at 4 h and 6 h by Blood-rsCDM. Results Blood-rsCDM exhibited a sensitivity of 97.9% at both time points, with a specificity of 100%, regardless of the culture duration. The sensitivity of eCIM was 94.4%, with a specificity of 100%. Blood-rsCDM accurately characterized KPC-producing isolates as 77/77, metallo-β-lactamases (MBLs) as 58/59, and KPC and NDM carbapenemases as 6/6 at 4 h. There was no difference in results between the 4 h and 6 h time points. However, Blood-rsCDM could not differentiate OXA-181-producing strains. For eCIM, the characterization numbers for KPC-, OXA-181-, and MBLs-producing strains were 77/77, 2/2, and 57/59, respectively, but it failed to detect the coproduction of KPC and NDM isolates. Conclusion Blood-rsCDM accurately discriminates carbapenemase within 4 h and is capable of directly differentiating multi-enzyme (KPC and NDM) presence from positive blood culture broths. Therefore, Blood-rsCDM represents a rapid, simple, easy-to-read, and accurate tool that can be utilized in resource-limited settings.

Abstract Background This study aims to investigate the frequency of cas1 and cas3 and CRISPR1,2,3 genes in Klebsiella pneumoniae isolates, as well as their connection with antibiotic resistance. Materials and methods 106 K. pneumoniae isolates were identified by biochemical assays and PCR. The susceptibility to antibiotics was determined by Kirby–Bauer disk diffusion method. Screening of ESBLs was undertaken by using double disk diffusion and standard disk diffusion methods. The E-test and mCIM techniques was used to confirm the disc diffusion-based carbapenem resistance profiles. CRISPR-Cas system genes were identified using PCR. Results ESBL production was found in 19% of isolates. Carbapenemase production was found in 46% of the isolates. Furthermore, the bacteria were classified as multidrug (76%), extensively drug-resistant (4%), or pan-drug-resistant (2%). When CRISPR/Cas systems were present, antibiotic resistance was lower; conversely, when they were absent, resistance was higher. Conclusions If the CRISPR/Cas modules aren’t present, the bacteria can still acquire foreign DNA, including antibiotic resistance genes. K. pneumoniae isolates with a CRISPR-Cas system were less likely to carry antibiotic-resistance genes than those lacking this defense system.

We examined the emergence and characteristics of oxacillinase-484–producing Enterobacterales in France during 2012–2023. Genomic analysis identified 2 predominant sequence types in Escherichia coli: ST410 and ST1722. Plasmid analysis revealed that blaOXA-484 genes were carried mostly on an IncX3-type plasmid associated with genetic elements including insertion sequences IS3000 and ISKpn19.

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a global threat, causing serious community- and hospital-acquired infections with limited treatment options. Understanding the local epidemiology of CRKP is crucial for guiding antimicrobial stewardship and infection control measures. This study aimed to investigate the phenotypic characteristics, antimicrobial resistance profiles, and carbapenemase genes among CRKP isolates from the patients in Najaf, Iraq. Methods: This cross-sectional research was performed at diverse hospitals and centers in Najaf, Iraq, from September 2023 to March 2024. The study was approved by the Ethics Committee of Kufa University. CRKP isolates were collected from various health-care facilities and tested for hypermucoviscosity, antimicrobial susceptibility, and carbapenemase production using phenotypic methods. The presence of carbapenemase genes (blaNDM, blaVIM, blaKPC, blaOXA-48, blaOXA-23, and blaOXA-51) was examined by polymerase chain reaction. Results: Of the 27 CRKP isolates, 9 (33.3%) exhibited a hypermucoviscous phenotype. Regarding antimicrobial resistance, 12 (44.4%) were multidrug resistant, 14 (51.9%) were extensively drug resistant, and 1 (3.7%) was pandrug resistant. Phenotypic carbapenemase production was detected in 5 (18.5%) and 11 (40.7%) isolates by the modified Hodge test and E-test metallo-β-lactamase strips, respectively. Molecular analysis revealed that all CRKP harbored the blaOXA-51 gene, whereas blaNDM, blaOXA-23, and blaVIM were detected in 70.4%, 40.7%, and 11.1% of the isolates, respectively. Neither blaKPC nor blaOXA-48 genes were found. In addition, 22 (81.5%) isolates carried multiple carbapenemase genes. Conclusions: The high prevalence of blaOXA-51 and blaNDM carbapenemases, along with the high rates of multidrug resistance among CRKP isolates in Najaf, Iraq, are alarming. This necessitates the immediate implementation of effective antimicrobial stewardship and infection control measures to prevent the further spread of these difficult-to-treat pathogens.