Carbapenem Resistance Research Articles

High level non-carbapenemase carbapenem resistance by overlaying mutations of mexR , oprD, and ftsI in Pseudomonas aeruginosa

ABSTRACT Carbapenem-resistant Pseudomonas aeruginosa (CRPA) is a global threat, but the mechanism of non-carbapenemase carbapenem resistance is still unclear. In the current study, we investigated the contributions of point mutations in mexR , oprD , and ftsI to carbapenem resistance in P. aeruginosa during in vivo evolution studies with consecutive clinical isolates. Real-time qPCR and Electrophoretic Mobility Shift Assay demonstrated that MexR (Gln55Pro) mutation increased MexAB efflux pump genes expression by altering MexR’s binding capacity, leading to a four- to eight-fold increase in meropenem MIC in the Pae d1 Green ∆ mexR and PAO1∆ mexR mutants. The OprD (Trp415*) truncation affected porin structure, and the constructed mutant Pae d1 Green oprD Trp415* increased meropenem MIC by 16-fold (from 0.25 to 4 µg/mL). The contribution of ftsI mutation to meropenem resistance was confirmed by clinical linkage analysis and was estimated to cause a two-fold increase in meropenem MIC by comparing the resistant clinical isolate with the Pae d1 Green oprD Trp415*∆ mexR double mutant. The study found that the oprD Trp415* allele alone accounts for the imipenem MIC in clinical isolates, while the ∆ mexR and ftsI Arg504Cys alleles do not contribute to imipenem resistance. In conclusion, we identified and explored the contributions of mexR , oprD, and ftsI mutations to high level non-carbapenemase carbapenem resistance in P. aeruginosa . These findings highlight the interplay of different mutations in causing non-carbapenemase carbapenem-resistance in P. aeruginosa . IMPORTANCE The emergence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) poses a significant global health threat, complicating treatment options for infections caused by this pathogen. Understanding the mechanisms behind non-carbapenemase carbapenem resistance is critical for developing effective therapeutic strategies. This study provides crucial insights into how specific point mutations in key genes- mexR , oprD , and ftsI -contribute to carbapenem resistance, particularly the MexR (Gln55Pro) mutation’s effect on efflux pump expression and the OprD (Trp415*) truncation’s impact on porin structure. The findings elucidate the complex interplay of these mutations, highlighting their roles in conferring high-level resistance, and underscore the imperative for continued research to inform therapeutic strategies against CRPA infections.

Clonal and horizontal transmission of carbapenem-resistant Enterobacterales strains and genes via flies.

Antimicrobial resistance (AMR) is one of the most pressing global public health challenges; in particular, the rapid dissemination of carbapenem-resistant Enterobacterales (CRE) is emerging as a significant concern worldwide. Flies, serving as carriers of pathogens, pose a potential threat in the transmission of antibiotic-resistant bacteria (ARB) between animals and humans. The aim of this study was to evaluate and reveal the potential risk of AMR spread by flies. A total of 450 flies were collected from four farms, four rural areas, and four urban areas in Dengfeng, Henan, China. To select CRE strains on the surface of flies, three flies sampled from the same geographical location were arbitrarily selected and placed into one tube of brain heart infusion broth (BHI), and the supernatant was screened using CHROMagar™ mSuperCARBA culture medium. Different colors and shapes of colonies were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA sequencing. Antimicrobial susceptibility testing for CRE strains was performed using broth microdilution. All CRE strains were whole-genome sequenced. Short-read sequencing was performed using MGISEQ-2000 and long-read sequencing was conducted using GridION. Totally, 150 BHI tubes were screened for CRE strains, and 33 strains were identified as CRE positive. In 24 mSuperCARBA plates, only one species of CRE strain was isolated from each plate. In three plates, two different species of CRE strains were identified in each plate. In one plate, three different species of CRE strains were simultaneously isolated. Carbapenem resistance genes were detected in 81.8% of CRE strains, and blaNDM-1 was predominant (66.7%). No significant correlations between carbapenem-resistant phenotypes and carbapenem resistance genes were observed. The complete genomes of all 33 strains were obtained. Genome analysis revealed that clonal transmission events may have occurred among different farms and rural areas. Phylogenetic analysis revealed that blaNDM-1 IncFII plasmids could break bacterial species barrier for cross-host transmission in diverse areas. To understand and control the transmission of AMR from the perspective of One Health, it is imperative to enhance surveillance of ARB, antibiotic resistance genes, and antibiotic-resistant plasmids in flies.

Laboratory detection of carbapenemases among Gram-negative organisms.

SUMMARYThe carbapenems remain some of the most effective options available for treating patients with serious infections due to Gram-negative bacteria. Carbapenemases are enzymes that hydrolyze carbapenems and are the primary method driving carbapenem resistance globally. Detection of carbapenemases is required for patient management, the rapid implementation of infection prevention and control (IP&C) protocols, and for epidemiologic purposes. Therefore, clinical and public health microbiology laboratories must be able to detect and report carbapenemases among predominant Gram-negative organisms from both cultured isolates and direct from clinical specimens for treatment and surveillance purposes. There is not a "one size fits all" laboratory approach for the detection of bacteria with carbapenemases, and institutions need to determine what fits best with the goals of their antimicrobial stewardship and IP&C programs. Luckily, there are several options and approaches available for clinical laboratories to choose methods that best suits their individual needs. A laboratory approach to detect carbapenemases among bacterial isolates consists of two steps, namely a screening process (e.g., not susceptible to ertapenem, meropenem, and/or imipenem), followed by a confirmation test (i.e., phenotypic, genotypic or proteomic methods) for the presence of a carbapenemase. Direct from specimen testing for the most common carbapenemases generally involves detection via rapid, molecular approaches. The aim of this article is to provide brief overviews on Gram-negative bacteria carbapenem-resistant definitions, types of carbapenemases, global epidemiology, and then describe in detail the laboratory methods for the detection of carbapenemases among Gram-negative bacteria. We will specifically focus on the Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter baumannii complex.

Mutant prevention concentrations, in vitro resistance evolution dynamics, and mechanisms of resistance to imipenem and imipenem/relebactam in carbapenem-susceptible Klebsiella pneumoniae isolates showing ceftazidime/avibactam resistance.

Klebsiella pneumoniae carbapenemase (KPC) variants selected during ceftazidime/avibactam treatment usually develop susceptibility to carbapenems and carbapenem/β-lactamase inhibitors, such as imipenem and imipenem/relebactam. We analyzed imipenem and imipenem/relebactam single-step mutant frequencies, resistance development trajectories and differentially selected resistance mechanisms using two representative K. pneumoniae isolates that had developed ceftazidime/avibactam resistance during therapy (ST512/KPC-31 and ST258/KPC-35). Mutant frequencies and mutant prevention concentrations were measured in Mueller-Hinton agar plates containing incremental concentrations of imipenem or imipenem/relebactam. Resistance dynamics were determined after incubation for 7 days in 10 mL MH tubes containing incremental concentrations of each antibiotic or combination, up to 64 times their baseline MIC. Two colonies per strain from each experiment were characterized by antimicrobial susceptibility testing and whole genome sequencing. The impact of KPC variants identified in resistant mutants on β-lactam resistance was investigated by cloning experiments. Imipenem/relebactam suppressed the emergence of resistant mutants at lower concentrations than imipenem, slowed down resistance development for both strains, and the resulting mutants yielded lower MICs of carbapenems and carbapenem/β-lactamase inhibitors than those selected with imipenem alone. Characterization of resistant mutants revealed that imipenem resistance was mainly caused by inactivation of OmpK36 and mutations in the KPC β-lactamase. Imipenem/relebactam-resistant mutants also maintained OmpK36 alterations, but mutations in KPC were much less frequent compared with those selected with imipenem alone. Genetic and biochemical characterization of the KPC derivatives identified in the resistant mutants confirmed their role in carbapenem resistance. Our data positions imipenem/relebactam as an attractive therapeutic option for combating ceftazidime/avibactam-resistant KPC-producing K. pneumoniae infections.

Rise of Carbapenem-Resistant Escherichia coli and the Concerning Existence of blaNDM-1 Gene in Gut Microbiota of Rural Populations

In rural communities, the blaNDM-1 gene in the gut microbiota may be contributing to the growing issue of antibiotic resistance. This gene enables bacteria to produce enzymes that break down antibiotics, making treatments less effective. The limited healthcare resources and frequent exposure to resistant bacteria in these areas could further amplify this problem, increasing the risk of difficult-to-treat infections among the population. The objective of this study was to evaluate the antimicrobial susceptibility of Escherichia coli [E. coli] strains isolated from healthy individuals and to assess the prevalence of blaNDM-1 gene carriage. The samples were cultured and identified as gut commensal by conventional method. Then antibiotic susceptibility test using the Kirby Bauer disc diffusion method were carried out. Carbapenem-resistant organisms were isolated, and the blaNDM-1 was identified by polymerase chain reaction. This study includes 187 Healthy participants faecal samples were obtained from people in and around the Chengalpattu area. Out of 187 samples, 110 showed E. coli growth, 63 showed Enterococci species growth, and 14 showed no growth. Among the 110 E. coli isolates, 24 (21.8%) were sensitive to all drug classes, and 86 (78.2%) were multidrug resistant. Only 8(7.27%) of the 110 E. coli tested positive for carbapenem resistance. Out of 110 E. coli isolates, only (2.7%) had the blaNDM-1 gene. This study discovered that rural communities that are unlikely to be subjected to obvious antibiotic pressure had a high incidence of MDR microorganism faecal carriage. Finding resistance genes in commensal bacteria is alarming. This has the ability to spread and emerge in the environment.

Co-existence of antibiotic resistance and virulence factors in carbapenem resistant Klebsiella pneumoniae clinical isolates from Alexandria, Egypt.

The emergence and spread of carbapenem resistance among Enterobacteriaceae, particularly Klebsiella pneumoniae, constitute a serious threat to public health, since carbapenems are the last line of defense in the treatment of life-threatening infections caused by drug-resistant Enterobacteriaceae. The current study investigated the co-existence of different virulence factors and carbapenemases in carbapenem-resistant Klebsiella pneumoniae clinical isolates from Alexandria, Egypt. Phenotypic characterization of virulence factors indicated that 41.5% of the isolates were strong biofilm producers, while hypermucoviscosity was detected in 14.9% of the isolates. All isolates harbored five or more virulence factor encoding genes. entB, ycfM, mrkD and fimH were detected in all isolates, while only one isolate was negative for ybtS. uge, iutA, rmpA and kpn were detected in 61 (64.8%), 55 (58.5%), 41 (43.6%) and 27 (28.7%) isolates, respectively, while all isolates lacked magA and k2A. Phenotypic detection of carbapenemases was explored by performing CarbaNP and mCIM/eCIM. CarbaNP test showed positive results in 98.9% of the isolates and positive mCIM tests were observed in all isolates, while 68 (72.3%) isolates showed positive eCIM tests. blaNDM was the most prevalent carbapenemase encoding gene (92.5%) followed by the blaOXA-48 (51.1%), while blaKPC was detected in only one (1.06%) isolate. blaVIM, blaIMP and blaGES were not detected in any of the tested isolates. The widespread of carbapenem-resistant Klebsiella pneumoniae represents a major problem in health care settings. A significant association between certain virulence factors and carbapenemase-encoding genes was observed. Antibiotic stewardship programs and infection control policies should be effectively implemented especially in hospitals to limit the spread of such highly virulent pathogens.

KleTy: integrated typing scheme for core genome and plasmids reveals repeated emergence of multi-drug resistant epidemic lineages in Klebsiella worldwide.

Clinically important lineages in Klebsiella, especially those expressing multi-drug resistance (MDR), pose severe threats to public health worldwide. They arose from the co-evolution of the vertically inherited core genome and horizontal gene transfers by plasmids, which has not been systematically explored. We designed KleTy, which consists of dedicated typing schemes for both the core genome and plasmids in Klebsiella. We compared the performance of KleTy with many state-of-the-art pipelines using both simulated and real data. Employing KleTy, we genotyped 33,272 Klebsiella genomes, categorizing them into 1773 distinct populations and predicting the presence of 87,410 plasmids from 837 clusters (PCs). Notably, Klebsiella is the center of the plasmid-exchange network within Enterobacteriaceae. Our results associated the international emergence of prevalent Klebsiella populations with only four carbapenem-resistance (CR) PCs, two hypervirulent PCs, and two hvCR-PCs encoding both carbapenemase and hypervirulence. Furthermore, we observed the ongoing international emergence of blaNDM, accompanied by the replacement of the previously dominant population, blaKPC-encoding HC1360_8 (CC258), during 2003-2018, with the emerging blaNDM-encoding HC1360_3 (CC147) thereafter. Additionally, expansions of hypervirulent carbapenem-resistant Klebsiella pneumoniae (hvCRKP) were evidenced in both populations, driven by plasmids of MDR-hypervirulence convergences. The study illuminates how the global genetic landscape of Klebsiella has been shaped by the co-evolution of both the core genome and the plasmids, underscoring the importance of surveillance and control of the dissemination of plasmids for curtailing the emergence of hvCRKPs.

Extended-Spectrum Beta-Lactamase-Producing and Multidrug-Resistant Escherichia coli and Klebsiella spp. from the Human–Animal–Environment Interface on Cattle Farms in Burkina Faso

Extended-spectrum beta-lactamase (ESBL)-producing and multidrug-resistant Enterobacterales pose a major threat to both human and animal health. This study assessed the prevalence of ESBL-producing Escherichia coli (ESBL-Ec) and Klebsiella spp. (ESBL-K) on cattle farms in Ouagadougou, Burkina Faso, using a One Health approach. From May 2021 to September 2022, cattle faeces, farmers’ stools, their drinking water and farm soil samples were collected from semi-intensive and traditional farms. An ESBL-selective medium was used to obtain resistant isolates, which were further characterised using biochemical tests. Antimicrobial susceptibility testing was performed using the Kirby–Bauer disc diffusion method. ESBL-Ec and/or ESBL-K were detected in 188 of 322 samples (58.0%). The prevalence of ESBL-Ec isolates was 42.2% (136/322) and that of ESBL-K isolates was 24.5% (79/322). Notably, 156 of the 188 ESBL isolates (83.0%) exhibited multidrug resistance. The highest resistance rates were observed against tetracycline and cotrimoxazole. Importantly, no isolates showed resistance to meropenem, which was used to test for carbapenem resistance. This study highlights the presence of ESBL-Ec and ESBL-K among the humans, animals and environment of the cattle farms. Good hygiene and biosafety practices are essential to limit the potential spread of multidrug-resistant bacteria between different interfaces on farms.

Imipenem exposure influence the expression of quorum sensing receptor sdiA in Escherichia coli.

The increasing trend of carbapenem resistance amongst Escherichia coli poses a major public health crisis and requires active surveillance of resistance mechanisms to control the threat. Quorum Sensing system plays a role in bacterial resistance to antibiotics. Quorum Sensing is a cell-cell communication system where bacteria alter their gene expression in response to specific stimuli. Here, in this study we investigated the transcriptional response of quorum sensing receptor, sdiA in E.coli under sub-inhibitory concentration of carbapenem in presence of quorum sensing signal molecules. Two E.coli isolates harbouring blaNDM were subjected to treatment with 10% SDS for 20 consecutive days of which blaNDM encoding plasmid was successfully eliminated from one isolate. Both the wild type and the cured mutant were then allowed to grow under eight different inducing conditions and the transcriptional response of sdiA gene was studied by quantitative real time PCR method. We found different response levels of sdiA in wild type and cured mutant under exogenous AHL and imipenem and when co-cultured with P.aeruginosa under imipenem stress. This study highlighted that sub-inhibitory concentration of imipenem in combination with AHL is acting as signal to SdiA, a quorum sensing receptor in E.coli.

A comparison of two MALDI-TOF MS based assays for the detection of carbapenemases in Enterobacterales

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.

Antimicrobial Resistance Trends and Epidemiological Characteristics of Isolates from Intra-Abdominal Infections in China: A 6-Year Retrospective Study (2017-2022).

Background: Antimicrobial resistance represents a continuing threat to the health of patients with intra-abdominal infections (IAIs). This study aimed to provide clinicians with guidance to optimize antibiotic therapy. Methods: The clinical data and antibiotic susceptibility results of pathogens from patients with IAIs from 2017 to 2022 were retrospectively collected. The 6-year period was segmented into two stages, namely, the early (2017-2020) and recent stages (2021-2022). The distribution and antibiotic resistance of pathogens were compared between the stages. Results: In total, 5,795 pathogens were isolated from 2,283 patients diagnosed with IAIs. Gram-negative bacteria, Gram-positive bacteria, and fungi accounted for 71.0%, 21.4%, and 7.5% of the isolates, respectively. Klebsiella pneumoniae (1,037, 17.9%) was the primary isolate. The proportion of extended-spectrum β-lactamase-producing Enterobacteriaceae was 89.8% (2,028/2,259), with extended-spectrum β-lactamase-producing Escherichia coli and K. pneumoniae accounting for 27.4% and 43.2%, respectively, of all such isolates. The carbapenem resistance rates of E. coli and K. pneumoniae were 17.1% and 75.9%, respectively. Compared with that in the early stage, the imipenem resistance rate of E. coli was significantly higher in the recent stage (13.8% vs. 25.1%, p < 0.001). Among Gram-positive bacteria, 88 strains of vancomycin-resistant Enterococcus were detected, giving a resistance rate of 10.3%, and the detection rate of methicillin-resistant Staphylococcus aureus was 65.7%. Conclusions: Enterobacteriales and non-fermentative bacteria from IAIs remain highly resistant to carbapenems. The epidemiological characteristics and antibiotic resistance profiles of pathogens in various regions should be closely monitored to mitigate the appearance of drug-resistant bacteria in clinical settings.

Molecular Characterization of Multidrug-Resistant and Hypervirulent New Delhi Metallo-Beta-Lactamase Klebsiella pneumoniae in Lazio, Italy: A Five-Year Retrospective Study

Background/Objectives: Antimicrobial resistance represents a challenge to public health systems because of the array of resistance and virulence mechanisms that lead to treatment failure and increased mortality rates. Although for years the main driver of carbapenem resistance in Italy has been the Klebsiella pneumoniae KPC carbapenemase, recent years have seen an increase in VIM and NDM metallo-beta-lactamases (MBLs). We conducted a five-year survey of New Delhi Metallo-beta-Lactamase (NDM)-producing Klebsiella pneumoniae (NDM-Kpn) clinical isolates from the Lazio region, Italy; the study aimed to elucidate the molecular mechanisms underpinning their resistant and virulent phenotype. Methods: Antimicrobial susceptibility was evaluated by automated systems and broth microdilution. In silico analysis of acquired resistance and virulence genes was performed using whole-genome sequencing (WGS), molecular typing through MLST, and core genome multi-locus sequence typing (cgMLST). Conclusions: A total of 126 clinical NDM-Kpn isolates were collected from 19 distinct hospitals in the Lazio region. Molecular analysis highlighted the existence of NDM-1 (108/126) and NDM-5 (18/126) variants, 18 Sequence Types (STs), and 15 Cluster Types (CTs). Notably, 31/126 isolates displayed a virulence score of 4, carrying ybt, ICEKp, iuc, and rmp genes. This study identified a variety of NDM-Kpn STs, mainly carrying the blaNDM-1 gene, with a significant number linked to high-risk clones. Of these isolates, 24.6% showed high-level resistance and virulence, emphasizing the risk of the spread of strains that combine multi-drug-resistance (MDR) and virulence. Proactive surveillance and international collaborations are needed to prevent the spread of high-risk clones, as well as further research into new antimicrobial agents to fight antibiotic resistance.

Longitudinal genomics reveals carbapenem-resistant Acinetobacter baumannii population changes with emergence of highly resistant ST164 clone

Carbapenem-resistant Acinetobacter baumannii (CRAB) is a persistent nosocomial pathogen that poses a significant threat to global public health, particularly in intensive care units (ICUs). Here we report a three-month longitudinal genomic surveillance study conducted in a Hangzhou ICU in 2021. This followed a three-month study conducted in the same ICU in 2019, and infection prevention and control (IPC) interventions targeting patients, staff and the ICU environment. Most A. baumannii isolated in this ICU in 2021 were CRAB (80.9%; 419/518) with higher-level resistance to carbapenems. This was accompanied by the proportion of global clone 2 (GC2) isolates falling from 99.5% in 2019 to 50.8% (213/419) in 2021. The phylogenetic diversity of GC2 increased, apparently driven by regular introductions of distinct clusters in association with patients. The remaining CRAB (40.2%; 206/419) were a highly clonal population of ST164. Isolates of ST164 carried blaNDM-1 and blaOXA-23 carbapenemase genes, and exhibited higher carbapenem MIC50/MIC90 values than GC2. Comparative analysis of publicly available genomes from 26 countries (five continents) revealed that ST164 has evolved towards carbapenem resistance on multiple independent occasions. Its success in this ICU and global capacity for acquiring resistance determinants indicate that ST164 CRAB is an emerging high-risk lineage of global concern.

Evaluation of biofilm formation and carbapenem resistance in Klebsiella pneumoniae isolated from clinical samples at a rural hospital in western Uttar Pradesh

ABSTRACT Introduction: Klebsiella pneumoniae commonly causes healthcare-associated infections and shows multidrug resistance. K. pneumoniae can produce biofilm. Carbapenem resistance in K. pneumoniae is due to the production of carbapenemases mainly. This study was done to evaluate the formation of biofilm and carbapenemase resistance in K. pneumoniae isolates. Material and Methods: A total of 110 K. pneumoniae isolated from various clinical samples were taken, the antibiotic susceptibility test was done by the Kirby disk diffusion method, and biofilm detection was done by the tissue culture plate method. All the carbapenem-resistant isolates were confirmed by multiplex real-time PCR (mPCR). Those found positive for any of the carbapenemase genes were tested by the modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), and ethylenediamine tetraacetic acid (EDTA)-modified carbapenem inactivation method (eCIM). Results: Out of 110 isolates, 66% (72/110) were carbapenem-resistant (suggestive of carbapenemase producers) by Kirby-Bauer disk diffusion but 58% (42/72) of Klebsiella isolates were confirmed for carbapenemase production by mPCR. Maximum number of carbapenemase gene were New Delhi metallo-β-lactamase (NDM) 52% (N = 22), 29% (N = 12) coproducers (NDM+OXA-48), and lowest in oxacillinase (OXA-48), 19% (N = 8). The overall sensitivity of MHT and mCIM+eCIM was 62% and 93%, and specificity was 88% and 97%, respectively. Our study showed that moderate biofilm producers were 51% (N = 56) K. pneumoniae isolates, strong biofilm producers 27% (N = 30), and 22% (N = 30) were weak/non-biofilm producers. We also found the correlation between biofilm formation and carbapenem-resistant K. pneumoniae (CR-KP) genes was statistically significant with a P value of 0.01*&lt;0.05. Conclusion: Most isolates of K. pneumoniae demonstrated a wide range of antibiotic resistance and were biofilm producers. Our results indicated that the combination of mCIM with eCIM showed high sensitivity and specificity to detect CR-KP compared with MHT.

Prevalence of carbapenem-resistant Enterobacterales (CRE) in Saudi Arabia: A systematic review and meta-analysis

Antimicrobial resistance is a significant public health issue. In addressing the threat of multidrug resistant bacterial infections, carbapenems have been used. The carbapenem-resistant Enterobacterales (CRE) are, however, rapidly expanding worldwide. Since the issue of CRE is also a problem in Saudi Arabia, the current meta-analysis was performed to comprehensively evaluate the resistance rates to the main carbapenem antibiotics and determine the actual prevalence of CRE in the country. Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines was followed. Different web databases including PubMed, Scopus, Web of Science, and ScienceDirect were searched for relevant records. Data were extracted, and summary estimates for resistance to carbapenems were calculated using DerSimonian-Laird method of meta-analysis and the random-effects model. From a total of 787 retrieved records, 69 studies were found fully eligible and were included in the final analyses. More than 50 % of all the studies were conducted after 2010, and the most frequently examined members of the Enterobacterales were Escherichia coli and Klebsiella pneumoniae. The pooled prevalence estimate for imipenem resistance was 6.6 % (95 % CI: 4.7–9.2), 9.1 % (95 % CI: 6.7–12.3) for meropenem, and 18.6 % (95 % CI: 11.9–27.9) for ertapenem. High heterogeneity (I2 > 97 %, p < 0.001) was observed for all the estimates. Compared to other regions of the country, there was higher resistance rates in the Al-Qassim and Al-Jouf provinces. Additionally, resistance to ertapenem was as high as 34.2 % in the most recent study period (2021–2024). Proteus spp was the most prevalent CRE (26.2 %). This review highlights an increasing rate of carbapenem resistance among Enterobacterales, emphasizing the need for collaborative efforts to implement strict infection control and prevention measures. Consistent surveillance is indispensable for safeguarding public health, guiding clinical decisions, and strengthening efforts to tackle the challenges of antibiotic resistance.

Prevalence of some Metallo β-lactamase enzymes genes in P. aeruginosa isolated from different clinical sample in Baghdad, Iraq

The increasing challenge of carbapenem antibiotics resistance caused by Pseudomonas aeruginosa is one of the global healthcare problems. P. aeruginosa is a significant opportunistic infection. For epidemiological reasons, identifying resistance genes is essential, and it is also desirable to quickly identify the techniques for producing carbapenemase enzymes. So, the study aims to determine the prevalence of beta-lactamase encoding genes (blaIMP, blaVIM, bla GIM, and bla SPM) phenotypically and genotypically in P. aeruginosa isolates to address the epidemiological spread of these genes in Baghdad city. The study identified P. aeruginosa isolates from various clinical sources by chemical characterization and VITEK 2 system, the antibiogram test, phenotypic screening carbapenem resistance by Combined disk synergy test (CDST), and conventional PCR were used to detect presence of VIM, IMP, SPM, and GIM genes. Bacterial susceptibility testing revealed (40%) of isolates were resistant to Imipenem and (85%) of them positive to CDST. Genotypic screening on phenotypic Metallo-β-lactamase isolates showed that (100%) isolates contained blaVIM genes, blaGIM genes (88%), whereas (65%) isolates carried blaSPM genes. In this investigation, there was no evidence of blaIMP among carbapenem-resistant isolates. The study appeared high prevalence of multi-drug resistance P. aeruginosa isolates that produce carbapenemase enzymes and having β- lactamase genes in local hospitalized patients compared to global ratio. Expanding the sample size and types of enzymes screening in MDR P. aeruginosa should be the main focus in the future research.

Surveillance of carbapenem resistance in Asian countries: a systematic review and meta-analysis

IntroductionCarbapenems are used to treat multidrug-resistant organisms as last-resort antibiotics. Resistance to carbapenems is an emerging public health threat worldwide, with alarming reported rates in Asian countries. To minimise further spread of resistant strains and guide interventions, it is important to understand the prevalence rates and causative molecular determinants. Therefore, the systematic review and meta-analysis arising from this protocol will aim to comprehensively describe the available surveillance data on carbapenem resistance in Asian countries to provide an in-depth overview of the carbapenem resistance status in this region.Methodology and analysisA systematic review and meta-analysis will be conducted via PubMed, ScienceDirect, Cochrane Library and Web of Science databases. Additional articles and grey literature will be searched in Google Scholar, OpenGrey and Google Chrome by manually searching the reference lists of selected studies. The review question was designed according to components in the ECLIPSE (E-expectations, C-client, L-location, I-impact, P-profile and S-service) framework. Studies conducted with samples other than clinical samples will be excluded. Only original articles published in the English language will be included. The Joanna Briggs Institute critical appraisal tool will be used to assess the quality of the included studies. A random-effects meta-analysis will be performed if the data are sufficiently homogenous. Heterogeneity between studies will be assessed via the I² statistic. The subgroup analysis will be performed considering the type of sample, pathogen type, region/country where the studies were conducted and genetic determinants.Ethics and disseminationAs this study is conducted on the basis of published data, ethical approval is exempt. The findings of this study will be disseminated in a peer-reviewed journal with the intention of providing summarised data on the globally emerging threat of carbapenem resistance and emphasising the need to introduce alternative, more effective treatment options.PROSPERO registration numberCRD42024515806.

Zn(II) enhances the antimicrobial effect of chloroxine and structural analogues against drug-resistant ESKAPE pathogens in vitro

The emerging antibiotic-resistant bacteria, especially the “ESKAPE” pathogens, pose a continuous threat to global health. In this study, we explored metalloantibiotics as promising therapeutics and innovative antimicrobial agents. The role of metal in the antimicrobial activity of chloroxine (5,7-dichloro-8-hydroxyquinoline), as a metalloantibiotic, was investigated by minimal inhibit concentration (MIC) assay and a series of assays, including growth curve, time-killing, and UV–visible spectroscopy and PAR (4-(2-pyridylazo)-resorcinol) competition assays. Both chloroxine and its structural analogues exhibited increased antibacterial potency against Gram-positive bacteria compared to Gram-negative bacteria. The introduction of exogenous manganese or zinc ions significantly boosted chloroxine’s antibacterial efficacy against Gram-negative bacteria, including the notorious ESKAPE pathogens. However, the enhanced antibacterial activity induced by zinc ions could be negated in the presence of copper or ferrous iron ions, as well as changes in oxygen availability, highlighting the involvement of proton motive force, oxidative and antioxidative systems. Notably, chloroxine effectively inhibited the enzymatic activity of superoxide dismutase (SOD). In addition, chloroxine could reverse polymyxin and carbapenem resistance in E. coli in vitro. Therefore, these results suggested that chloroxine with zinc ions are promising therapeutics and antibiotics potentiator to combat multidrug-resistant ESKAPE pathogens

Investigation of Carbapenemase-Producing Pseudomonas aeruginosa at Secondary Care Hospital in Bolu, Turkey.

The global increase in carbapenem resistance poses a significant public health threat due to the potential emergence of multidrug-resistant pathogens and limited treatment options. To learn more about this issue and offer potential solutions, we conducted a study of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infections in a secondary care hospital setting. The study utilized the carbapenem inactivation method (CIM), a leading phenotypic analysis, to determine carbapenemase activity in 63 CRPA isolates. Additionally, polymerase chain reaction (PCR) analysis was conducted to test for the presence of carbapenemase genes associated with the production or expression of various carbapenemase enzymes, including blaKPC, blaNDM, blaVIM, blaOXA-48, blaIMP, and blaGES. Arbitrary primed PCR (AP-PCR) was performed to assess the clonal relationship between different isolates. The isolates were also classified as either health care-associated infections or community-acquired infections, and their clonal relationship and gene positivity were evaluated. A total of 63 CRPA samples underwent evaluation, with 14 isolates determined to be carbapenemase producers via CIM tests. PCR assays revealed that 14 isolates carried carbapenemase genes, with 9 carrying blaNDM, 2 carrying blaGES, 2 carrying blaVIM, and 1 carrying blaIMP. CRPA exhibited a 22% prevalence of carbapenemase genes, of which 64% were attributed to the NDM gene responsible for multidrug resistance. AP-PCR revealed high clonal diversity among the isolates. Molecular epidemiological evaluation also showed no dominant outbreak strain among PA isolates. This study presents significant data on the prevalence and distribution of carbapenemase-producing CRPA strains isolated from secondary health care facilities. Typically, the literature focuses on resistance rates in tertiary care public hospitals. These findings may aid in understanding resistance and its mechanisms, as well as in developing effective treatment strategies and infection control measures.

Carbapenem resistance in gram-negative pathogens in an Iranian hospital: high prevalence of OXA-type carbapenemase genes.

The widespread dissemination of carbapenem-resistant Gram-negative bacteria poses a significant threat to global public health. This study aimed to investigate the prevalence of carbapenem resistance in Gram-negative bacteria isolated from patients at the Children's Medical Center Hospital, Tehran, Iran, to understand the molecular mechanisms underlying this resistance. During the period spanning from June 2019 to June 2020, 777 Gram-negative bacterial strains were isolated. Antibiotic susceptibility testing was performed according to Clinical and Laboratory Standards Institute. Polymerase chain reaction was used to detect carbapenem resistance genes including bla OXA23, bla OXA24, bla OXA48, bla OXA51, bla OXA58, bla OXA143, bla KPC, bla IMP, bla VIM, and bla NDM. Among the total bacterial isolates, 141 (18.1%) exhibited carbapenem resistance. Escherichia coli was the most prevalent (57.4%), followed by Klebsiella pneumoniae (11.3%) and Acinetobacter baumannii (10.6%). Other notable contributors included Enterobacter spp. (5.7%), Salmonella spp. (3.5%), and Stenotrophomonas maltophilia (2.8%). Citrobacter spp., Proteus mirabilis, and Pseudomonas aeruginosa contributed to the distributions of two, one, and three isolates, respectively. Notably, bla OXA48 showed the highest prevalence (33%), followed by bla OXA143 and bla OXA58 (27% and 24%, respectively). In addition, bla OXA24 was present in 11% of the total isolates, bla OXA23 in 10%, and bla NDM in 10%, whereas bla KPC, bla VIM, and bla IMP were not detected. Our study highlights the prevalence of carbapenemase-producing Gram-negative isolates among pediatric patients. Notable resistance patterns, especially in K. pneumoniae and E. coli, underline the urgent need for proactive interventions, including appropriate antibiotic prescription practices and strengthening of antibiotic stewardship programs.