Detection of carbapenem resistance among third-generation cephalosporin-resistant Enterobacterales from small-scale poultry farms in peri-urban Lusaka, Zambia

Enterobacteriaceae are common pathogens causing bloodstream infection (BSI) in sub-Saharan Africa and frequently express third-generation cephalosporin (3GC) resistance; however, the impact of 3GC resistance on clinical outcomes is rarely studied. We conducted a single-site prospective cohort study at Tygerberg Hospital, Cape Town, South Africa to examine the feasibility of measuring impacts of 3GC resistance in Enterobacteriaceae BSI. We included patients with 3GC-susceptible and 3GC-resistant BSIs and matched each BSI patient to two uninfected patients. We determined the concordance of initial antibiotic treatment with the corresponding isolate's susceptibility profile. We performed exploratory impact analysis using multivariable regression models. Between 1 June 2017 and 31 January 2018, we matched 177 Enterobacteriaceae BSI patients to 347 uninfected patients. Among these BSIs, 35% were phenotypically 3GC resistant. Parameters describing clinical comorbidity showed strong associations with mortality. We found that 18% of 3GC-R and 3% of 3GC-S BSI patient received non-concordant initial therapy. In multivariable Cox regression, we found a mortality impact over their matched patients for both 3GC-R (cause-specific hazard ratio 23.77; 95% CI 5.12-110.3) and 3GC-S (HR 7.49; 95%CI 3.08-18.19) BSI. There was a nonsignificant ratio of these ratios (HR 3.18; 95% CI 0.54-18.70), limited by the small sample size. This form of impact estimation was feasible in one hospital in South Africa where 3GC-R status was associated with non-concordant initial antibiotic treatment. There was a possible increase in mortality among individuals with 3GC-resistant Enterobacteriaceae, but with broad confidence intervals. These analytical approaches could be applied to larger datasets to improve precision of estimates.

Objectives: Although antimicrobial resistance (AMR) disproportionately affects low- and middle-income countries (LMICs), primary clinical data on AMR burden from LMICs are lacking, particularly from the Pacific Islands. We adapted recent World Health Organization methodology to measure the impact of third-generation cephalosporin (3GC) resistance on mortality and excess length of hospital stay among inpatients with Enterobacterales bloodstream infection (BSI) in Fiji. Methods: We conducted a prospective cohort study of inpatients with Enterobacterales BSIs at Colonial War Memorial Hospital in Suva. We collected demographic, clinical, and microbiological data, and we stored bacterial isolates for confirmatory testing and molecular genomics in Melbourne, Australia. We employed cause-specific Cox proportional hazards models to estimate the effect of 3GC-resistance on hazard of in-hospital mortality and discharge alive (competing outcomes), and we used multistate modelling to estimate the excess length of hospital stay associated with 3GCR. Results: From July 2020 to February 2021, we identified 162 consecutive Enterobacterales BSIs, and 66 (40.7%) were 3GC resistant. The crude mortality rates for patients with 3GC-susecptible and 3GC-resistant BSIs were 16.7% (16 of 96) and 30.3% (20 of 66), respectively. Also, 3GC resistance was not associated with either in-hospital mortality (aHR, 1.67; 95% CI, 0.80–3.49) or discharge alive (aHR, 0.75; 95% CI, 0.50–1.12). However, patient comorbidities and acuity of illness were associated with in-hospital mortality. Furthermore, 3GC-resistance was associated with an increased length of stay of 2.6 days (95% CI, 2.5–2.8). Overall, 3GC-resistance was more common among patients with hospital-associated than community-acquired infection, but genomics did not identify clonal transmission. Conclusions: Among patients with Enterobacterales BSIs, mortality was relatively high, and 3GC resistance was common. Also, 3GC resistance was associated with increased hospital length of stay but not with in-hospital mortality after adjusting for potential confounders. Accurate estimates of the burden of AMR are important, especially from LMICs. Such knowledge can inform policy decisions, guide allocation of limited resources, and assist the evaluation of future interventions to address AMR.

BackgroundThis study aimed to identify characteristics of third-generation cephalosporin (3GC) resistance in Escherichia coli bateremia (ECB) and Klebsiella pneumoniae bacteremia (KPB) in patients with liver cirrhosis (LC), and to investigate the effects of appropriateness of empirical antibiotic treatment on outcomes.MethodsWe retrospectively collected demographic, clinical and microbiological information on all ECB and KPB episodes in LC patients ≥ 18 years of age hospitalized to a tertiary-care teaching hospital in South Korea from 2007 to 2018. Clinical characteristics associated with 3GC resistance and treatment failure were analyzed using a multivariate logistic regression model. Treatment failure was defined as persistent bacteremia for ≥ 7 days, or relapsed bacteremia ≤ 30 days, or all-cause mortality ≤ 30 days.Results3GC resistance rates of E. coli were 30.3% overall and increased significantly during the study period (P=0.001), while the rates of K. pneumoniae were not changed (24.3% overall) (P=0.994). Of total 356 ECB and KPB episodes, 112 were caused by 3GC resistant strains. The factor associated with 3GC resistance was isolation of 3GC resistant strain ≤ 1 year in both ECB (OR, 7.754; 95% CI, 2.094~28.716) and KPB (OR, 2.774; 1.318~5.838). In ECB, beta-lactam or fluoroquinolone treatment ≤ 30 days was another factor associated with 3GC resistance (OR, 2.774; 95% CI, 1.318~5.838), but not in KPB. The factor associated with treatment failure was high MELD score in both ECB (OR, 1.193 at 1 increase; 95% CI, 1.118~1.272) and KPB (OR, 1.163; 95% CI 1.083~1.250). Additionally, in ECB, non-alcoholic LC (OR 3.262; 95% CI 1.058~10.063), high Charlson Comorbidity Index (OR, 1.285; 95% CI 1.066~1.548), and inappropriate empirical antibiotic treatment (OR, 3.194; 95% CI 1.207~8.447) were associated with treatment failure.ConclusionDuring the study period, 3GC resistance increased in ECB, but not in KPB. In ECB, the severity of the underlying disease and the appropriateness of empirical antibiotics were associated with treatment failure, but there was no correlation in KPB. In ECB of LC patients, the appropriateness of empirical antibiotics was a factor associated with treatment outcome, and is the only correctable factor in the clinical setting.DisclosuresAll Authors: No reported disclosures

Escherichia coli is a normal inhabitant of the intestinal microbiota of chickens, a small proportion of which may be avian pathogenic E. coli (APEC) or potential extraintestinal pathogenic E. coli (ExPEC), capable of causing disease in humans. These E. coli may also be resistant to antimicrobials of critical importance in human or veterinary health. This study aims to 1) determine the prevalence of antimicrobial resistance (AMR) and resistance genes, multidrug resistance (MDR), chromosomal mechanisms of quinolone-resistance and virulence profiles of E. coli isolated from healthy chicken farms in the region of Dakar, Senegal, 2) investigate the spread of third-generation cephalosporins (3GC) resistance in E. coli isolated from healthy chicken farms with respect to virulence and resistance genes, serogroups, Pulsed-Field Gel Electrophoresis (PFGE), phylogenetic groups, plasmid types and transferability and 3) determine whether nonsusceptibility against 3GC on farms could be linked to risk factors. More than 68% of isolates from environmental faecal and drinking water samples, carcasses and carcass washes collected on 32 healthy chicken farms were multidrug resistant (MDR), resistance to antimicrobials critical in human health (3GC or ciprofloxacin) being found in all types of samples. Ciprofloxacin resistance was due to mutations in the gyrA and parC genes, 95% of tested farms harboring isolates carrying three mutations, in gyrA (Ser83Ile and Asp87Asn) and parC (Ser80Ile). Nine of the 32 farms (28.1%) demonstrated the presence of one or more 3GC-nonsusceptible indicator isolates but none of the potential risk factors were significantly associated with this presence on farms. Following ceftriaxone enrichment, presumptive extended-spectrum beta-lactamase/AmpC-beta-lactamase (ESBL/AmpC)-producer isolates were found in 17 of the 32 farms. 3GC resistance was mediated by blaCMY-2 or blaCTX-M genes, blaCTX-M being of genotypes blaCTX-M-1, blaCTX-M-8 and for the first time in chickens in Senegal, the genotype blaCTX-M-15. Clonally related ESBL/AmpC-producer isolates were found on different farms. In addition, blaCTX-M genes were identified on replicon plasmids I1 and K/B and blaCMY-2 on K/B, I1 and B/O. These plasmids were found in isolates of different clusters. In addition, 18 isolates, some of which were ESBL/AmpC-producers, were defined as potential human ExPEC. In conclusion, E. coli isolates potentially pathogenic for humans and demonstrating MDR, with resistance expressed against antimicrobials of critical importance in human health were found in healthy chickens in Senegal. Our results suggest that both clonal spreading and horizontal gene transfer play a role in the spread of 3GC-resistance and that chickens in Senegal could be a reservoir for AMR and ExPEC for humans. These results highlight the importance of raising awareness about compliance with biosecurity measures and prudent use of antimicrobials.

Third-generation cephalosporin resistance in clinical isolates of Enterobacterales collected between 2016–2018 from USA and Europe: genotypic analysis of β-lactamases and comparative in vitro activity of cefepime/enmetazobactam

In Zambia, 40% of clinical Gram-negative bacteria are either Escherichia coli or Klebsiella pneumoniae, with a high third-generation cephalosporin (3GC) resistance prevalence. Therefore, 3GC resistance surveillance is a crucial indicator for guiding focused intervention policies. However, the lack of genotypic diagnostic tools limits the ability to elucidate trends, especially in peri-urban and rural areas of developing countries. This study aimed to develop a rapid, cost-effective tool for the genotypic surveillance of 3GC resistance. Here, 900 stool samples collected from patients in Kafue (peri-urban, n = 400) and Katete (rural, n = 500) districts of Zambia were used for bacterial isolation on MacConkey agar supplemented with 1 μg/ml cefotaxime. Isolated 3GC-resistant strains were characterized by sequencing the 16S rRNA gene and screening for blaCTX-M and blaTEM genes using single polymerase chain reaction (PCR). Furthermore, selected 3GC-resistant strains were subjected to whole-genome sequencing (WGS) using MiSeq/HiSeq (n = 34) and MinION (n = 1). Using the data from this and other previous studies, we developed a rapid PCR-dipstick DNA chromatography-based tool for detecting blaCTX-M, blaTEM, E. coli-specific yaiO, and K. pneumoniae-specific khe genes. The prevalence of isolated 3GC resistant strains was 15.4% (139/900), dominated by E. coli (102/139, 73.4%). On PCR, the blaCTX-M gene was detected in 72.7% (101/139) of the isolates, while blaTEM was found in 46.8% (65/139) of the strains. The developed tool displayed a high level of agreement with WGS and single PCR/Sanger sequencing, with sensitivity and specificity ≥ 95% and Kappa ≥ 0.95 for each of the four target genes. We envisage that the simplicity and adaptability of this tool will be a significant advantage for the surveillance of 3GC resistance in Zambia and elsewhere.

BackgroundMonitoring pathogens of bloodstream infections (BSI) and their antibiotic susceptibility is important to guide empiric antibiotic treatment strategies and prevention programs.This study assessed the epidemiology of BSI and antibiotic resistance patterns at the German Federal State of Thuringia longitudinally.MethodsA surveillance network consisting of 26 hospitals was established to monitor BSIs from 01/2015 to 12/2019. All blood culture results, without restriction of age of patients, of the participating hospitals were reported by the respective microbiological laboratory. A single detection of obligate pathogens and a repeated detection of coagulase-negative staphylococci, Bacillus spp., Corynebacterium spp., Micrococcus spp. and Propionibacterium spp., within 96 h were regarded as a relevant positive blood culture. If one of the aforementioned non-obligate pathogens has been detected only once within 96 h, contamination has been assumed. Logistic regression models were applied to analyse the relationship between resistance, year of BSI and hospital size. Generalized estimating equations were used to address potential clustering.ResultsA total of 343,284 blood cultures (BC) of 82,527 patients were recorded. Overall, 2.8% (n = 9571) of all BCs were classified as contaminated. At least one relevant pathogen was identified in 13.2% (n = 45,346) of BCs. Escherichia coli (25.4%) was the most commonly detected pathogen, followed by Staphylococcus aureus (15.2%), Staphylococcus epidermidis (8.1%) and Klebsiella pneumoniae (4.6%). In S. aureus, we observed a decline of methicillin resistance (MRSA) from 10.4% in 2015 to 2.5% in 2019 (p < 0.001). The rate of vancomycin resistance in Enterococcus faecium (VRE) has increased from 16.7% in 2015 to 26.9% in 2019 (p < 0.001), with a peak in 2018 (42.5%). In addition, we observed an increase of Cefotaxime (3GC) resistance in E. coli from 10.7% in 2015 to 14.5% in 2019 (p = 0.007) whereas 3GC resistance in K. pneumoniae was stable (2015: 9.9%; 2019: 7.4%, p = 0.35). Carbapenem resistance was less than 1% for both pathogens. These patterns were robustly observed across sensitivity analyses.ConclusionsWe observed evidence for a decline in MRSA, an increase in VRE and a very low rate of carbapenem resistance in gram-negative bacteria. 3GC resistance in E. coli increased constantly over time.

A gradual rise in drug-resistant trends among Gram-negative organisms, especially carbapenem-resistant (CR) Enterobacteriaceae (CRE), CR-Pseudomonas aeruginosa, and extensively-drug-resistant (XDR) Acinetobacter baumannii, poses an enormous threat to healthcare systems worldwide. In the last decade, many pharmaceutical companies have devoted enormous resources to the development of new potent antibiotics against XDR Gram-negative pathogens, particularly CRE. Some of these novel antibiotics against CRE strains are β-lactam/β-lactamase-inhibitor combination agents, while others belong to the non-β-lactam class. Most of these antibiotics display good in vitro activity against the producers of Ambler class A, C, and D β-lactamase, although avibactam and vaborbactam are not active in vitro against metallo-β-lactamase (MβL) enzymes. Nevertheless, in vitro efficacy against the producers of some or all class B enzymes (New Delhi MβL, Verona integron-encoded MβL, etc) has been shown with cefepime-zidebactam, aztreonam-avibactam, VNRX-5133, cefiderocol, plazomicin, and eravacycline. As of Feburary 2019, drugs approved for treatment of some CRE-related infections by the US Food and Drug Administration included ceftazidime-avibactam, meropenem-vaborbactam, plazomicin, and eravacycline. Although active against extended-spectrum and AmpC β-lactamase-producing Enterobacteriaceae, delafloxacin does not show in vitro activity against CRE. Murepavadin is shown to be specifically active against CR- and colistin-resistant P. aeruginosa strains. Despite successful development of novel antibiotics, strict implementation of an antibiotic stewardship policy in combination with theuse of well-established phenotypic tests and novel multiplex PCR methods for detection of the most commonly encountered β-lactamases/carbapenemases in hospitals is important for prescribing effective antibiotics against CRE and decreasing the resistance burden due to CRE.

BackgroundEscherichia coli is the most common cause of bloodstream infections (BSIs) and mortality is an important aspect of burden of disease. Using a multinational population-based cohort of E. coli BSIs, our objectives were to evaluate 30-day case fatality risk and mortality rate, and determine factors associated with each.MethodsDuring 2014–2018, we identified 30-day deaths from all incident E. coli BSIs from surveillance nationally in Finland, and regionally in Sweden (Skaraborg) and Canada (Calgary, Sherbrooke, western interior). We used a multivariable logistic regression model to estimate factors associated with 30-day case fatality risk. The explanatory variables considered for inclusion were year (2014–2018), region (five areas), age (< 70-years-old, ≥70-years-old), sex (female, male), third-generation cephalosporin (3GC) resistance (susceptible, resistant), and location of onset (community-onset, hospital-onset). The European Union 28-country 2018 population was used to directly age and sex standardize mortality rates. We used a multivariable Poisson model to estimate factors associated with mortality rate, and year, region, age and sex were considered for inclusion.ResultsFrom 38.7 million person-years of surveillance, we identified 2961 30-day deaths in 30,923 incident E. coli BSIs. The overall 30-day case fatality risk was 9.6% (2961/30923). Calgary, Skaraborg, and western interior had significantly increased odds of 30-day mortality compared to Finland. Hospital-onset and 3GC-resistant E. coli BSIs had significantly increased odds of mortality compared to community-onset and 3GC-susceptible. The significant association between age and odds of mortality varied with sex, and contrasts were used to interpret this interaction relationship. The overall standardized 30-day mortality rate was 8.5 deaths/100,000 person-years. Sherbrooke had a significantly lower 30-day mortality rate compared to Finland. Patients that were either ≥70-years-old or male both experienced significantly higher mortality rates than those < 70-years-old or female.ConclusionsIn our study populations, region, age, and sex were significantly associated with both 30-day case fatality risk and mortality rate. Additionally, 3GC resistance and location of onset were significantly associated with 30-day case fatality risk. Escherichia coli BSIs caused a considerable burden of disease from 30-day mortality. When analyzing population-based mortality data, it is important to explore mortality through two lenses, mortality rate and case fatality risk.

A pair-matched longitudinal study conducted on three dairy farms in the U.S. High-Plains explored the temporal effects of two-dose ceftiofur crystalline-free acid (CCFA) treatment for metritis on third-generation cephalosporin (3GC) resistance among enteric E. coli in Holstein-Friesian cows. The current 13-day slaughter withholding period does not account for rising populations of third-generation cephalosporin (3GC) resistant bacteria in feces of animals following CCFA treatment. A total of 124 matched-pairs of cows were enrolled in the study. Cows diagnosed with postpartum metritis received the product twice at the labeled dose of 6.6 mg/kg subcutaneously at the base of alternating ears. Untreated cows–absent clinical metritis–were matched on lactation number and calving date. Feces were collected per rectum on days 0 (baseline), 6, 16, 28, and 56. Environmental samples, from watering troughs as well as surface manure from fresh-cow, hospital, maternity, and milking pens, and from the compost pile were collected prior to the animal sample collection period. Historical data on metritis rates and CCFA use were compiled from herd records. On day 0, cows exhibited an overall mean difference of over 4 log10 colony forming units (CFU) comparing 3GC resistant E. coli to the general E. coli population. At the first eligible slaughter date, the difference declined to 3.31 log10 CFU among cows in the CCFA group (P<0.01 compared to control cows). Such differences were no longer observed between the treated and control groups by day 28. Results suggest a 13-day withholding period following the final treatment is insufficient to allow levels of 3GC resistant E. coli to return to baseline. This effect varied by farm and was dependent upon the starting level of resistance. A farm-specific extended slaughter-withholding period could reduce the microbial risk to food products at slaughter.

Epidemiology and mortality outcome of carbapenem- and colistin-resistant Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, and Pseudomonas aeruginosa bloodstream infections

Background Treatment options for Carbapenem-resistant (CR) Gram-negative infections due to metallo-beta-lactamase (MBL) enzymes are limited. The clinical impact of MBLs vs. other mechanisms of carbapenem resistance in Enterobacterales and non-fermenting bacteria remains unclear.Table 1.Demographics, isolate characteristics, and outcomes of patients with carbapenem-resistant Gram-negative infections, stratified by MBL statusFigure 1.30-day desirability of outcome ranking (DOOR) outcomes of patients with carbapenem-resistant Gram-negative infections, stratified by MBL statusCRPA – carbapenem-resistant Pseudomonas aeruginosa, CRAb – carbapenem-resistant Acinetobacter baumannii, CRE – carbapenem-resistant Enterobacterales, DOOR – desirability of outcome ranking. MBL – metallo-beta-lactamase. DOOR events assessed at 30 days include: unsuccessful discharge, lack of clinical response, and C. difficile infection and/or renal failure. Not all rows total to 100 due to rounding. P-value calculated using Wilcoxon test. Methods We conducted a matched cohort study of patients enrolled in one of three MDRO Network studies, POP (CR Pseudomonas aeruginosa [CRPA]), SNAP (CR Acinetobacter baumannii [CRAb]), or CRACKLE-2 (CR-Enterobacterales [CRE]) with isolates that met infection criteria. Patients with MBL-producing isolates (blaVIM, blaIMP, or blaNDM present) were matched 1:2 to patients with non-MBL CR isolates (a different carbapenemase or CR without a carbapenemase) based on study, region, and anatomical source. We compared baseline characteristics, 30- and 90-day mortality, and 30-day desirability of outcome ranking (DOOR) scores.30-day Desirability of Outcome Ranking (DOOR) Probability by MBL StatusLegend: CI – confidence interval, CRPA – carbapenem-resistant Pseudomonas aeruginosa, CRAb – carbapenem-resistant Acinetobacter baumannii, CRE – carbapenem-resistant Enterobacterales, DOOR – desirability of outcome ranking. MBL – metallo-beta-lactamase. The DOOR probability was calculated as the probability of a more desirable result in the presence of MBL as compared to non-MBL isolate. Confidence intervals were calculated using the method in Halperin et al (Biometric 1989; 45:500-521), and CI’s that do not include 50% are considered statistically significant. Estimates less than 50% signify a less favorable outcome for the MBL group, while estimates greater than 50% signify more favorable outcomes for the MBL group. Results In total, 170 MBL isolates were matched to 340 non-MBL isolates from 10 countries (Table 1). The cohort included 42% CRPA (216/510), 5% CRAb (24/510), and 53% CRE (270/510). Demographics were balanced between groups; median age at culture was 61 (Q1, 44, Q3 73) years. Common infection sources were respiratory (151/510, 30%), urine (141/510, 28%), and blood (105/510, 21%). Of the MBL isolates, 92/170 harbored blaNDM (54%), 62/170 harbored blaVIM (36%), and 20/170 harbored blaIMP (12%); four isolates co-harbored two distinct MBL enzymes. All-cause 30-day mortality was 19% (33/170) for MBL vs 18% (61/340) for non-MBL (p=0.69); MBL presence was not associated with 30- or 90-day mortality. DOOR outcomes at 30-days (Figure 1) did not differ by MBL status in the full cohort or the CRE subgroup, but did differ in the CRPA/CRAb subgroup (p=0.037). Among CRPA/CRAb infections, MBL presence was associated with less desirable outcomes (DOOR probability 42.1%; 95% Halperin confidence interval: 35.0%-49.5%, Figure 2). Conclusion MBL presence was not associated with increased 30- or 90-day mortality compared to matched non-MBL isolates. However, in non-fermenter infections (CRPA/CRAb), MBL presence was linked to less desirable outcomes, an association not seen in CRE. These findings may inform prioritization of anti-MBL agents in future drug development. Disclosures Angelique E. Boutzoukas, MD, MPH, Elion Therapeutics: Advisor/Consultant|Innoviva Speciality Therapeutics: DSMB Participant Souha S. Kanj, MD, Menarini: Honoraria|pfizer: Honoraria Vance G. Fowler, MD, MHS, Affinergy, Janssen, Contrafect: Advisor/Consultant|AstraZeneca; EDE; Basilea: Grant/Research Support|Debiopharm, GSK; Affinium, Basilea,: Advisor/Consultant|Destiny, Amphliphi, Armata, Akagera: Advisor/Consultant|Merck; Contrafect; Karius; Janssen: Grant/Research Support|UpToDate: Royalties|Valanbio: Stock options Yohei Doi, MD, PHD, GSK: Advisor/Consultant|Meiji Seika Pharma: Advisor/Consultant|Shionogi: Advisor/Consultant|Shionogi: Honoraria Michael Satlin, MD, MS, AbbVie: DSMB Participant|bioMerieux: Grant/Research Support|Merck: Grant/Research Support|SNIPRBiome: Grant/Research Support Robert A. Bonomo, MD, Merck: Grant/Research Support|Shinogi: Grant/Research Support|VenatoRx: Grant/Research Support David van Duin, MD, PhD, British Society for Antimicrobial Chemotherapy: Editor stipend|Merck: Advisor/Consultant|Merck: Grant/Research Support|Pfizer: Advisor/Consultant|Roche: Advisor/Consultant|Shionogi: Advisor/Consultant

The emergence of carbapenem-resistant Enterobacteriaceae (CRE) and other multidrug-resistant Gram-negative bacteria (GNB) presents a serious threat to infection control, particularly in high-risk environments such as Intensive Care Units (ICUs). ICU surfaces and healthcare workers (HCWs) may serve as reservoirs and transmission routes for these pathogens. This study aimed to assess the prevalence and distribution of CRE, and other Gram-negative bacteria isolated from ICU environmental surfaces and from the nasal swab of healthcare workers. A descriptive cross-sectional study was used to study 127 collected samples: 36 from healthcare workers and 91 from surface smears. Samples were collected from four units of intensive care: Intensive Care Unit (ICU), Cardiac Care Unit (CCU), Pediatric Care Unit (PICU), and Neonatal Intensive Care Unit (NICU). Identified Enterobacteriaceae were confirmed by biochemical methods, and tested for carbapenemase production using three discs (EME, IMP, ETP). Resistance to Ertapenem, Imipenem, and Meropenem was considered carbapenem resistance. A total of 13 (36.1%) out of 36 HCW samples yielded clinical isolates: 3 Pantoea sp., 3 Klebsiella pneumoniae, 2 Acinetobacter baumannii, 2 Escherichia coli, 2 Flavimonas oriyzihabitans, and 1 Serratia marcescens. One (7.69%) isolate showed carbapenem resistance: 1 Klebsiella pneumonia. From 91 surface samples, 39 (43%) yielded isolates: 13 Flavimonas oriyzihabitans, 6 Acinetobacter baumannii, 5 Pantoea sp., 4 Escherichia coli, 3 Enterobacter cloacae, 2 Serratia marcescens, 2 Shigella sp., 1 Klebsiella pneumonia, 1 Leclercia adecarboxylata, and 1 Salmonella sp. Of these, 11 (28.2%) were carbapenem-resistant: 3 Escherichia coli, 3 Acinetobacter baumannii, 2 Pantoea sp., 1 Klebsiella pneumonia, 1 Citrobacter freundii, and 1 Enterobacter cloacae

Globally, intensive care units (ICUs) are encountering the emergence and spread of antibiotic-resistant pathogens, and for some pathogens, there are few therapeutic options available. This study tries to assess the prevalence, susceptibility pattern, and risk factors of Carbapenem-Resistant Enterobacteriaceae (CRE) infections among ICU patients, healthcare workers, and surface swabs in the medical center of Tobruk, Libya. Also, to assess the incidence and prevalence rate of CRE in the medical center of Tobruk. A descriptive cross-sectional study was used to study 119 collected samples. Of which, 119 samples were collected from patients. The above samples were collected from four units of intensive care were intensive care unit (ICU) and cardiac care unit (CCU), Pediatric intensive care unit (PICU), Neonatal intensive care unit (NICU). The identified Enterobacteriaceae were first confirmed by biochemical methods, while the confirmed isolates were tested for carbapenemase production using three discs (EME, IMP, and ETP). The resistance to (Ertapenem, Imipenem, and Meropenem) was considered as CRE. A total of 79 (66.4%) out of the 119 samples yielded clinical isolates of Enterobacteriaceae and gram negative bacteria from patients as follows: 23 (29.1%) Klebsiella pneumoniae, 15 (19%) Acinetobacter baumannii, 12 (15.2%) Escherichia coli, 7 (9%) Pantoea sp., 5 (6.3%) Pseudomonas aeruginosa, 5 (6.3%) Citrobacter freundii, 4 (5.1%) Enterobacter cloacae, 2 (2.6%) Citrobacter freundii, 3 (3.8%) Chryseomonas luteola, 2 (2.5%) Proteus vulgaris, 2 (2.5%) Flavimonas oriyzihabitans, and 1 (1.3%) Serratia marcecens. Screening the Enterobacteriaceae-positive samples for carbapenem resistance showed 48 samples of 79 (60.8%) as carbapenem resistant Enterobacteriaceae as follows: 18 (22.8%) Klebsiella pneumonia, 9 (11.4%) Acinetobacter baumannii, 6 (7.6%) Escherichia coli, 4 (5.1%) Citrobacter freundii, 3 (3.8%) Pseudomonas aeruginosa, 3 (3.8%) Pantoea sp., 2 (2.5%) Enterobacter cloacae, 2 (2.5%) Chryseomonas luteola, 1 (1.3%) Serratia marcecens. The study provided evidence of the presence of CRE infections among patients admitted to ICUs in the study centers. This underscores the need for effective infection prevention and control measures to avoid the spread of CRE in a hospital setting.

Globally, intensive care units (ICUs) are encountering the emergence and spread of antibiotic-resistant pathogens, and for some pathogens, there are few therapeutic options available. This study tries to assess the prevalence, susceptibility pattern, and risk factors of Carbapenem-Resistant Enterobacteriaceae (CRE) infections among ICU patients, healthcare workers, and surface swabs in the medical center of Tobruk, Libya. Also, to assess the incidence and prevalence rate of CRE in the medical center of Tobruk. A descriptive cross-sectional study was used to study 119 collected samples. Of which, 119 samples were collected from patients. The above samples were collected from four units of intensive care were intensive care unit (ICU) and cardiac care unit (CCU), Pediatric intensive care unit (PICU), Neonatal intensive care unit (NICU). The identified Enterobacteriaceae were first confirmed by biochemical methods, while the confirmed isolates were tested for carbapenemase production using three discs (EME, IMP, and ETP). The resistance to (Ertapenem, Imipenem, and Meropenem) was considered as CRE. A total of 79 (66.4%) out of the 119 samples yielded clinical isolates of Enterobacteriaceae and gram negative bacteria from patients as follows: 23 (29.1%) Klebsiella pneumoniae, 15 (19%) Acinetobacter baumannii, 12 (15.2%) Escherichia coli, 7 (9%) Pantoea sp., 5 (6.3%) Pseudomonas aeruginosa, 5 (6.3%) Citrobacter freundii, 4 (5.1%) Enterobacter cloacae, 2 (2.6%) Citrobacter freundii, 3 (3.8%) Chryseomonas luteola, 2 (2.5%) Proteus vulgaris, 2 (2.5%) Flavimonas oriyzihabitans, and 1 (1.3%) Serratia marcecens. Screening the Enterobacteriaceae-positive samples for carbapenem resistance showed 48 samples of 79 (60.8%) as carbapenem resistant Enterobacteriaceae as follows: 18 (22.8%) Klebsiella pneumonia, 9 (11.4%) Acinetobacter baumannii, 6 (7.6%) Escherichia coli, 4 (5.1%) Citrobacter freundii, 3 (3.8%) Pseudomonas aeruginosa, 3 (3.8%) Pantoea sp., 2 (2.5%) Enterobacter cloacae, 2 (2.5%) Chryseomonas luteola, 1 (1.3%) Serratia marcecens. The study provided evidence of the presence of CRE infections among patients admitted to ICUs in the study centers. This underscores the need for effective infection prevention and control measures to avoid the spread of CRE in a hospital setting.