Extended-spectrum Β-lactamase Production Research Articles

Genetic Insights on Meropenem Resistance Concerning Klebsiella pneumoniae Clinical Isolates

The transferable genetic elements are associated with the dissemination of virulence determinants amongst Klebsiella pneumoniae. Thus, we assessed the correlated antimicrobial resistance in carbapenem-resistant Klebsiella pneumoniae clinical isolates. Each isolate’s ability to biosynthesize biofilm, carbapenemase, and extended-spectrum β-lactamase were examined. Genotypically, the biofilm-, outer membrane porin-, and some plasmid-correlated antimicrobial resistance genes were screened. About 50% of the isolates were multidrug-resistant while 98.4% were extended-spectrum β-lactamase producers and 89.3% were carbapenem-resistant. Unfortunately, 93.1% of the multidrug-resistant isolates produced different biofilm levels. Additionally, fimD and mrkD genes encoding adhesins were detected in 100% and 55.2% of the tested isolates, respectively. Also, the blaKPC, blaOXA-48-like, and blaNDM-encoding carbapenemases were observed in 16.1%, 53.6%, and 55.4% of the tested isolates, respectively. Moreover, the blaSHV and blaCTX-M extended-spectrum β-lactamase-associated genes were detected at 95.2% and 61.3%, respectively. Furthermore, aac(3)IIa, qnrB, and tetB resistance-correlated genes were observed in 38.1%, 46%, and 7.9% of the tested isolates, respectively. Certainly, the tested antimicrobial resistance-encoding genes were concurrently observed in 3.2% of the tested isolates. These findings confirmed the elevated prevalence of various antimicrobial resistance-associated genes in Klebsiella pneumoniae. The concurrent transferring of plasmid-encoding antimicrobial resistance-related genes could be associated with the possible acquisition of multidrug-resistant Klebsiella pneumoniae phenotypes.

A worrisome prevalence of extended-spectrum β-lactamase producers in patients with biliary obstruction and cholangitis: Phenotypic and molecular characterization of biliary Escherichia coli and Klebsiella pneumoniae isolates

Background & aimsThe alarming rise of antibiotic resistance presents a substantial and worrisome issue within the context of biliary obstruction, specifically in the treatment of cholangitis. This abovementioned scenario underscores the critical importance of addressing extended spectrum β-lactamase (ESBL) producers in the biliary system to adequately tackle cholangitis using third-generation cephalosporins. Hence, we aimed to determine the frequency of ESBL and carbapenemases among biliary Escherichia coli and Klebsiella pneumoniae isolated from patients with biliary obstruction. MethodsIn this cross-sectional study, bile samples were collected via aspiration from patients diagnosed with biliary obstruction during the endoscopic retrograde cholangiopancreatography (ERCP) procedure. Subsequent culturing of these samples was performed, followed by phenotypic and molecular assessments for the detection of ESBL- and carbapenemase-producing strains of E. coli and K. pneumoniae. ResultsApproximately 23.5 % of patients with biliary obstruction harbored biliary ESBL-producers, with the majority (70.2 %) being diagnosed with cholangitis. Moreover, 2.1 % of patients had biliary carbapenemase-producing K. pneumoniae strains. Molecular analysis confirmed the high prevalence of blaCTX-M and blaTEM in E. coli, and blaTEM and blaSHV in K. pneumoniae. Additionally, the presence of biliary K. pneumoniae harboring blaKPC, blaNDM, and blaIMP was observed. ConclusionOur study reveals a noteworthy observation that over half of patients experiencing biliary obstruction harbor ESBL-producing bacteria in their biliary tract. Notably, we discovered a significant link between ESBL producers and the risk of cholangitis. These findings raise important concerns regarding the suitability of employing third-generation cephalosporins as initial treatment for cholangitis and other similar biliary infections.

Molecular Characterization of Multidrug-Resistant Escherichia coli from Fecal Samples of Wild Animals.

Antimicrobial resistance (AMR) surveillance in fecal Escherichia coli isolates from wildlife is crucial for monitoring the spread of this microorganism in the environment and for developing effective AMR control strategies. Wildlife can act as carriers of AMR bacteria and spread them to other wildlife, domestic animals, and humans; thus, they have public health implications. A total of 128 Escherichia coli isolates were obtained from 66 of 217 fecal samples obtained from different wild animals using media without antibiotic supplementation. Antibiograms were performed for 17 antibiotics to determine the phenotypic resistance profile in these isolates. Extended-spectrum β-lactamase (ESBL) production was tested using the double-disc synergy test, and 29 E. coli strains were selected for whole genome sequencing. In total, 22.1% of the wild animals tested carried multidrug-resistant E. coli isolates, and 0.93% (2/217) of these wild animals carried E. coli isolates with ESBL-encoding genes (blaCTX-M-65, blaCTX-M-55, and blaEC-1982). The E. coli isolates showed the highest resistance rates to ampicillin and were fully susceptible to amikacin, meropenem, ertapenem, and imipenem. Multiple resistance and virulence genes were detected, as well as different plasmids. The relatively high frequency of multidrug-resistant E. coli isolates in wildlife, with some of them being ESBL producers, raises some concern regarding the potential transmission of antibiotic-resistant bacteria among these animals. Gaining insights into antibiotic resistance patterns in wildlife can be vital in shaping conservation initiatives and developing effective strategies for responsible antibiotic use.

In vitro activity of cefepime-enmetazobactam on carbapenem-resistant gram negatives

ObjectivesCefepime-enmetazobactam is a new β-lactam/βlactamase inhibitor combination with broad-spectrum activity against multidrug-resistant Enterobacterales, including extended-spectrum β-lactamase producers. This study evaluated the in vitro activity of cefepime-enmetazobactam towards a collection of carbapenem-resistant Enterobacterales (CRE), Pseudomonas aeruginosa and Acinetobacter baumannii compared to the other β-lactam/β-lactamase inhibitor combinations. MethodsThe MIC of cefepime, cefepime-enmetazobactam, ceftazidime, ceftazidime-avibactam, meropenem, meropenem-vaborbactam, imipenem, imipenem-relebactam, and ertapenem were determined by broth microdilution on 2212 CRE, including 2089 carbapenemase producers (1000 OXA-48-like, 49 KPC, 697 NDM, 180 VIM, 1 IMP, 9 IMI, and 158 multiple carbapenemases) and 123 CRE that do not produce carbapenemase received at the French National Reference Centre (from March 1, 2023 to August 31, 2023), 50 P. aeruginosa, and 30 A. baumannii. All strains were fully sequenced. ResultsWe confirmed the absence of inhibitory activity of enmetazobactam towards metallo-β-lactamases. Cefepime-enmetazobactam and ceftazidime-avibactam exhibited a similar susceptibility (96.7% vs. 99.5%, respectively) on OXA-48-producers. Cefepime-enmetazobactam exhibited 66.9% and 63.3% susceptibility for CRE non-EPC and KPC, whereas those rates rose to 96.7%/95.9%, 93.4%/95.9%, and 95.9%/98.0% for ceftazidime-avibactam, imipenem-relebactam, and meropenem-vaborbactam, respectively. Low MICs (≤0.25 mg/L) were obtained for ceftazidime-avibactam-resistant KPC variants.Cefepime-enmetazobactam did not display a significant added value when compared with cefepime alone on Pseudomonas aeruginosa and Acinetobacter baumannii. DiscussionOXA-48 producers displayed high susceptibility to cefepime-enmetazobactam, which is similar to ceftazidime-avibactam, including for OXA-48 producers that coproduce a ceftazidime hydrolyzing enzyme (extended-spectrum β-lactamases or AmpC). In vivo experiments have to be implemented to confirm if cefepime-enmetazobactam might be a relevant alternative to ceftazidime-avibactam for the treatment of infections caused by OXA-48 producers.

Enteropathogenic and Multidrug-Resistant blaCTX-M-Carrying E. coli Isolates from Dogs and Cats.

Enteropathogenic Escherichia coli (EPEC) are pathogens associated with gastrointestinal illnesses. Dogs and cats can harbor EPEC, and antimicrobial resistance may impair necessary treatments. This study characterized E. coli strains from dogs and cats, focusing on phylogroup classification, virulence factors, and antimicrobial resistance profiles. Ninety-seven E. coli isolates from fecal samples of 31 dogs and 3 cats were obtained from a private diagnostic laboratory in Botucatu, Brazil, from March to October 2021. The antimicrobial susceptibility was assessed using the disk diffusion method. Polymerase chain reaction (PCR) was employed to screen for blaCTX-M and genes encoding virulence factors, as well as to classify the isolates into phylogroups. Twenty isolates were positive for intimin encoding gene eae and, consequently, these isolates were classified as EPEC (20.62%). Notably, 5.1% (5/97) of the isolates exhibited extended-spectrum β-lactamase (ESBL) production and 13.4% (13/97) were identified as multidrug-resistant bacteria. Phylogroups A and B2 were the most prevalent, comprising 29.9% (29/97) and 26.8% (26/97) of the bacterial isolates, respectively. This characterization highlights the prevalence of EPEC in domestic animals, emphasizing the potential risk they pose to public health and highlighting the urgency of responsible antimicrobial use in veterinary practices and the important role of laboratories in the surveillance of pathogenic multidrug-resistant bacteria.

Evaluation of extended-spectrum β-lactamase production and plasmid-mediated quinolone resistance in Escherichia coli isolated from raw milk

This study aimed to determine the antibiotic resistance and extended-spectrum β-lactamase production profiles of Escherichia coli isolates obtained from raw milk samples. To this end, the study used a chromogenic-based culture method, MALDI-TOF-MS, 16S rRNA gene sequencing, disc diffusion, and combined disc techniques. PCR was also performed to detect ESBL (blaSHV, blaTEM, and blaCTX-M), quinolone (qnrA, qnrB, and qnrS) genes, and the presence of class I integrons. A total of 47 (78.33%) E. coli isolates were identified by morphological, biochemical, and molecular tests. All isolates exhibited susceptibility to gentamicin and tobramycin. Resistance rates to different antibiotics were observed as follows: ampicillin, 51.06%; nalidixic acid, 23.40%; trimethoprim/sulfamethoxazole, 17.02%; meropenem, 14.89%; ceftazidime, 8.51%; cefotaxime, 8.51%; piperacillin-tazobactam, 4.25%; aztreonam, 4.25%; imipenem, 4.25%; and levofloxacin, 2.12%. ESBL production was positive in 3 (6.38%) of the isolates. Bla genes were detected in 11 isolates (blaSHV = 9 [19.14%], blaTEM = 4 [8.5%], and blaCTX = 3 [6.38%]). The qnrB, qnrS, and qnrA genes were found in 5 (10.63%), 4 (8.51%), and 1 (2.12%) isolates, respectively. The intI1 gene was detected in 14 (29.78%) isolates.

Identification of the Genotypes of Extended-spectrum β-lactamase Producing Escherichia coli in the Small Intestine of Broiler Chickens at a Traditional Market in Surabaya, Indonesia

Background: Escherichia coli infection from inside the chicken’s body may be present if the previously slaughtered chicken was contaminated with pathogens or the cage’s hygiene is inadequate. One of the resistance mechanisms against antibiotics used by Gram-negative bacteria of the Enterobacteriaceae family is the production of extended-spectrum β-lactamases (ESBL). Research is required to determine which E. coli genotypes in broiler chicken samples collected from Surabaya’s traditional marketplaces produce ESBL. Methods: A total of 100 small intestine samples collected from broiler chickens were used in this study. After initial processing all the samples were separately injected into Eosin Methylene Blue Agar (EMBA) and Indole, Methyl Red, Voges Proskauer and Citrate (IMViC) tests were used to confirm the results. A test for antibiotic sensitivity based on the Kirby-Bauer method called the Double Disk Synergy Test (DDST) was used to analyze isolates that showed resistance to aztreonam and multidrug resistance (MDR). PCR testing was performed on ESBL-verified isolates to detect the TEM and CTX-M genes. Result: The results of the test for E. coli antibiotic resistance revealed that 14 isolates were proven to be multidrug-resistant (MDR) and six of them were found to be E. coli that produced ESBL. Four isolates were found to carry the TEM and CTX-M genes after genotyping analysis. Understanding the dangers of using antibiotics as feed additives and growth promoters is crucial for farmers to address the problem of ESBL-producing E. coli bacteria in chickens. Furthermore, the government needs to take action to monitor and enforce stronger controls on the use of antibiotics, which are even readily available on the market.

Trends in the Prevalence of Antimicrobial Resistance in Escherichia coli Isolated from Outpatient Urine Cultures in French Amazonia.

Antimicrobial resistance (AMR) in the community is increasing worldwide. We aimed to assess AMR trends in Escherichia coli from the community urine isolates in French Amazonia. We conducted a retrospective study from January 2016 to December 2022 in the Cayenne General Hospital microbiology laboratory (French Guiana). It included all urine samples positive for E. coli collected from adult outpatients. During the study period, 3,443 urinalyses positive for E. coli were studied. In 46% of cases, patients were women. In 64.4% of cases, E. coli were β-lactamase producers. The most frequently diagnosed resistance mechanisms were penicillinase production and sparing third-generation cephalosporins. Isolated E. coli were extended-spectrum β-lactamase (ESBL) producers in 6.1% of cases. Overall, E. coli was susceptible to amoxicillin in 35.9% [95% CI: 34.3-37.5], to amoxicillin/clavulanic acid in 62.2% [95% CI: 60.6-63.9], to cefotaxime in 94% [95% CI: 93.1-94.7], to gentamicin in 92.1% [95% CI: 89.1-92.6], to ofloxacin in 76.8% [95% CI: 75.3-78.2], to trimethoprim/sulfamethoxazole (SXT) in 58.8% [95% CI: 57.1-60.5], to fosfomycin in 99.1% [95% CI: 98.6-99.4], and to nitrofurantoin in 99% of cases [95% CI: 98.6-99.3]. We have observed a gradual decline in the susceptibility profile of E. coli for amoxicillin/clavulanic acid (P <0.001), piperacillin/tazobactam (P = 0.003), and temocillin (P = 0.006). However, susceptibility to ciprofloxacin was increasing (P = 0.001). In contrast, the susceptibility trends for amoxicillin, third-generation cephalosporins, gentamicin, SXT, nitrofurantoin, and fosfomycin remained stable over the 28 quarters of the study. In conclusion, isolated E. coli from outpatient urinalyses showed increased resistance profiles involving penicillinase and ESBL production. Close monitoring and strategies to decrease antibiotic consumption in the community are needed.

Resistance to ceftazidime-avibactam and other new β-lactams in Pseudomonas aeruginosa clinical isolates: a multi-center surveillance study.

Multidrug-resistant Pseudomonas aeruginosa infections are a growing threat due to the limited therapeutic options available. Ceftazidime-avibactam (CZA) is among the last-resort antibiotics for the treatment of difficult-to-treat P. aeruginosa infections, although resistance due to the acquisition of transferable β-lactamase genes is increasing. With this work, we report that CZA represents a highly active antipseudomonal β-lactam compound (after cefiderocol), and that metallo-β-lactamases (VIM-type) and extended-spectrum β-lactamases (GES and PER-type) production is the major factor underlying CZA resistance in isolates from Southern Italian hospitals. In addition, we reported that such resistance mechanisms were mainly carried by the international high-risk clones ST111 and ST235.

Investigation of plasmid-mediated quinolone resistance among extended-spectrum β-lactamase isolates of E. coli and K. pneumoniae.

Escherichia coli and Klebsiella pneumoniae are important members of the Enterobacteriaceae family, involved in many infections. The increased resistance rate towards β-lactams and fluoroquinolones -which are the main therapeutic options- limits their treatment options. This study aimed to assess the local resistance patterns against different antimicrobials and to determine the extended-spectrum β-lactamase (ESBLs) producers. The study revealed that 36% of clinical isolates were ESBL producers, showing high resistance rates towards β-lactams and non-β-lactams, especially sulphamethoxazole-trimethoprim and fluoroquinolones. However, they were susceptible to chloramphenicol and doxycycline (33% and 20%; respectively). Also, the investigation aimed to screen the plasmid profile of quinolone-resistant ESBLs-producers and to detect the plasmid-mediated quinolone resistance genes including qnrA, qnrS, qnrB, qnrC, qnrD, and qnrVC. Moreover, the conjugative plasmid among the quinolone-resistant isolates was elucidated. The results showed that extracted plasmids of sizes ranging from ≈0.9 to 21.23 Kb, divided into 7 plasmid patterns were detected. A plasmid of approximately 21.23 Kb was found in all isolates and the QnrS gene was the most predominant gene. Moreover, the frequency of transconjugation within the same genus was higher than that recorded between different genera; where 68% of E. coli isolates transferred the resistance genes compared to Klebsiella isolates (36.6%). Plasmid profiles of transconjugants demonstrated great similarity, where 21.23 Kb plasmid was detected in all transconjugants. Since these transconjugants were quinolone-resistant ESBL producers, it has been suggested that quinolone resistance determinants might be carried on that plasmid.

Monitoring Multi-Drug Resistant Klebsiella pneumoniae in Kitagata Hot Spring, Southwestern Uganda: A Public Health Implication.

The concerning frequency of K. pneumoniae in various recreational settings, is noteworthy, especially regarding multi-drug resistant (MDR) strains. This superbug is linked to the rapid spread of plasmids carrying these resistance genes. The objective of this study was to evaluate the spatiotemporal prevalence of MDR-K. pneumoniae in the Kitagata hot spring, Southwestern Uganda. A laboratory-based descriptive longitudinal study was conducted between May and July 2023. During rainy and dry seasons, we collected eighty water samples in the morning and evening from the hot spring. The temperature at each point was measured prior to sample collection, and two samples were obtained at varying depths. 5 mL of each homogenized sample were pre-enriched in brain heart infusion broth, and subsequently in both blood and violet red bile agar. The Kirby-Bauer disk diffusion method was performed, followed by the detection of carbapenemase (CR) and extended-spectrum β-lactamase (ESBL) production. Polymerase chain reaction showed resistance genes viz. bla TEM, bla CTX-M and bla KPC. Data were analyzed using SPSS-20 to obtain chi-square tests and regression analysis. K. pneumoniae accounted for 30.0% of isolates obtained from Kitagata hot springs, with all isolates classified as multi-drug resistant. All isolates were resistant to ampicillin, rifampicin, ceftazidime, and azithromycin (79.2%). Additionally, 95.8% of isolates harbored bla TEM gene alone and both bla TEM and bla CTX genes, followed by bla KPC alone (33.3%), with 25% harboring all three resistance genes. During the dry season, K. pneumoniae had a higher prevalence (35.0%) compared to the wet season (25.0%). The prevalence of MDR-K. pneumoniae significantly increased over the course of the study. The presence of the three studied resistance genes in the isolates showed a positive correlation with the second phase of sample collection and the dry season but exhibited a negative correlation with temperature, except for isolates harboring either bla TEM alone or bla TEM+KPC+CTX genes. Kitagata hot spring serves as a hotspot for continuous dissemination and acquisition of MDR-K. pneumoniae harboring resistance genes that encode for ESBL and CR production. The healthcare sector ought to implement an ongoing monitoring and surveillance system as well as robust antimicrobial resistance stewardship programs aimed at delivering health education to the community.

Culture-based bacterial evaluation of the appendix lumen and antibiotic susceptibility of acute appendicitis in Japan: A single-center retrospective analysis.

The question of whether to perform an appendectomy or conservative treatment for acute appendicitis can differ depending on the facility or surgeon, but antibiotic treatment is administered regardless of whether an appendectomy or conservative treatment is selected. We investigated the contemporary bacteriology for acute appendicitis and evaluated the antibiotic sensitivity of the bacteria that are currently associated with appendicitis. We retrospectively analyzed the bacterial culture results and antibiotic susceptibility of 141 patients who underwent appendicitis surgery, including the identification and antimicrobial susceptibility of the cultured bacteria within the excised appendices. Bacterial cultures were positive in 131 cases (92.9%). The most commonly isolated bacteria were Escherichia coli (90 isolates, 66.7%), followed by Enterococcus species (n = 19, 14.5%), Pseudomonas aeruginosa (n = 18, 13.7%), Streptococcus species (n = 15, 11.5%), and Klebsiella species (n = 8, 6.1%). Eight strains (8.8%) of E coli were extended-spectrum β-lactamase producers, and ten strains (11.1%) were fluoroquinolone-resistant. Tazobactam/piperacillin and meropenem inhibited the growth of 100% of the major identified bacteria. The patients with appendicoliths had a significantly higher bacterial culture rate. Enterococcus species were frequently isolated from the patients with complicated appendicitis. For the antibiotic treatment of appendicitis, it is essential to understand the patient's microbiological profile and antibiotic susceptibilities. Research from Asian countries such as Japan can enhance our knowledge of regional antibiotic resistance patterns and inform effective treatment strategies.

Impact of the COVID-19 pandemic on extended-spectrum β-lactamase producing Escherichia coli in urinary tract and blood stream infections: results from a nationwide surveillance network, Finland, 2018 to 2022

BackgroundBefore the COVID-19 pandemic there has been a constant increase in antimicrobial resistance (AMR) of Escherichia coli, the most common cause of urinary tract infections and bloodstream infections. The aim of this study was to investigate the impact of the COVID-19 pandemic on extended-spectrum β-lactamase (ESBL) production in urine and blood E. coli isolates in Finland to improve our understanding on the source attribution of this major multidrug-resistant pathogen.MethodsSusceptibility test results of 564,233 urine (88.3% from females) and 23,860 blood E. coli isolates (58.8% from females) were obtained from the nationwide surveillance database of Finnish clinical microbiology laboratories. Susceptibility testing was performed according to EUCAST guidelines. We compared ESBL-producing E. coli proportions and incidence before (2018–2019), during (2020–2021), and after (2022) the pandemic and stratified these by age groups and sex.ResultsThe annual number of urine E. coli isolates tested for antimicrobial susceptibility decreased 23.3% during 2018–2022 whereas the number of blood E. coli isolates increased 1.1%. The annual proportion of ESBL-producing E. coli in urine E. coli isolates decreased 28.7% among males, from 6.9% (average during 2018–2019) to 4.9% in 2022, and 28.7% among females, from 3.0 to 2.1%. In blood E. coli isolates, the proportion decreased 32.9% among males, from 9.3 to 6.2%, and 26.6% among females, from 6.2 to 4.6%. A significant decreasing trend was also observed in most age groups, but risk remained highest among persons aged ≥ 60 years.ConclusionsThe reduction in the proportions of ESBL-producing E. coli was comprehensive, covering both specimen types, both sexes, and all age groups, showing that the continuously increasing trends could be reversed. Decrease in international travel and antimicrobial use were likely behind this reduction, suggesting that informing travellers about the risk of multidrug-resistant bacteria, hygiene measures, and appropriate antimicrobial use is crucial in prevention. Evaluation of infection control measures in healthcare settings could be beneficial, especially in long-term care.

Antimicrobial Resistance in Uropathogenic Escherichia coli Strains Isolated from Relapses from Recurrent Urinary Tract Infections.

Little is known about the characteristics of uropathogenic Escherichia coli (UPEC) associated with recurrent urinary tract infections (RUTIs). The present study aimed to analyze the phenotypic antimicrobial resistance of recurrent UPEC isolates attributable to either relapse or reinfection. A total of 140 E. coli strains were isolated from 70 outpatients with RUTIs. All isolates were analyzed by random amplified polymorphic DNA-polymerase chain reaction to evaluate genetic similarity between the first and second isolates. We found that 64.2% (45/70) of outpatients had a relapse with the primary infecting E. coli strain and 35.7% (25/70) had reinfection with a new E. coli strain. Compared with reinfecting strains, relapse UPEC isolates exhibited much higher antimicrobial resistance; 89% of these isolates were multidrug-resistant and 46.6% were extended-spectrum β-lactamase producers. Our study provides evidence that RUTIs are mainly driven by the persistence of the original strain in the host (relapses) despite appropriate antibiotic treatments, and only RUTIs attributed to relapses seem to favor multidrug resistance in UPEC isolates.

Genomic insights into resistome, virulome, and mobilome as organic contaminants of ESKAPE pathogens and E. coli recovered from milk, farm workers, and environmental settings in Hainan, China

The ESKAPE pathogens and E. coli are of significant public health concern and are listed among the top priorities of the World Health Organization. These pathogens are equally important for animal and public health, but their interaction with animal, human, and environmental aspects needs to be sufficiently studied. We investigated the comprehensive molecular characterization of three ESKAPE pathogens and E. coli recovered from milk, farm workers, and environmental settings in Hainan province. Minimum inhibitory concentration (MIC), extended-spectrum β-lactamase production, and biofilm production were performed per set standard protocols. The isolates were further processed by PCR and sequencing to detect resistome, virulome, and mobilome. The study found four Enterobacteriaceae species (E. coli, n=32; K. pneumoniae, n=13; E. cloacae, n=3; E. aerogenes, n=1) among n=49 bacterial isolates from the study area. Most of the strains exhibited resistance against tetracycline (71.4%), ampicillin (61.2%), florfenicol (44.9%), and ciprofloxacin (42.9%), while none of the strains was resistant against meropenem and amikacin. The MDR and ESBL-production percentages were 55.1% and 34.7 %, respectively. The resistance determinants for β-lactams (blaTEM, blaOXA-2, blaSHV, blaCTX-M), aminoglycosides (aac(3ʹ)-IIa), quinolones (qnrB, qnrS), tetracyclines (tetA, tetD), florfenicol (floR), sulfonamides (sul1, sul2), trimethoprim (dfrA14), and MDR efflux pumps (oqxA, oqxB) were detected along with diverse plasmid replicon types and integrons (intl3, 55.1%; intl1, 18.4%). The detection of virulence determinants was noted at a higher rate, and most of the strains were identified as biofilm producers by genotype (85.7%). The current study findings revealed diversified resistance and virulence determinants along with mobilome in ESKAPE pathogens and E. coli, which may pose a significant threat to human health through the food chain and environmental exposure and need immediate attention.

Molecular Characterization and Antibacterial Resistance Determination of Escherichia coli Isolated from Fresh Raw Mussels and Ready-to-Eat Stuffed Mussels: A Major Public Health Concern.

Our study focused exclusively on analyzing Escherichia coli (E. coli) contamination in fresh raw mussels and ready-to-eat (RTE) stuffed mussels obtained from authorized and regulated facilities. However, it is critical to recognize that such contamination represents a significant public health threat in regions where unauthorized harvesting and sales practices are prevalent. This study aimed to comprehensively assess the prevalence, molecular characteristics, and antibacterial resistance profiles of E. coli in fresh raw mussels and RTE stuffed mussels. E. coli counts in fresh raw mussel samples ranged from 1 to 2.89 log CFU/g before cooking, with a significant reduction observed post-cooking. RTE stuffed mussel samples predominantly exhibited negligible E. coli presence (<1 log CFU/g). A phylogenetic analysis revealed a dominance of phylogroup A, with variations in the distribution observed across different sampling months. Antibacterial resistance was prevalent among the E. coli isolates, notably showing resistance to ampicillin, streptomycin, and cefotaxime. Extended-spectrum β-lactamase (ESβL) production was rare, with only one positive isolate detected. A variety of antibacterial resistance genes, including tetB and sul1, were identified among the isolates. Notably, virulence factor genes associated with pathogenicity were absent. In light of these findings, it is imperative to maintain rigorous compliance with quality and safety standards at all stages of the mussel production process, encompassing harvesting, processing, cooking, and consumption. Continuous monitoring, implementation of rigorous hygiene protocols, and responsible antibacterial drug use are crucial measures in mitigating food safety risks and combating antibacterial resistance. Stakeholders, including seafood industry players, regulatory agencies, and healthcare professionals, are essential to ensure effective risk mitigation and safeguard public health in the context of seafood consumption.

Antibiotyping, RAPD- and ERIC-PCR fingerprinting of Klebsiella pneumoniae clinical isolates at a tertiary reference hospital in Denpasar, Bali, Indonesia.

Klebsiella pneumoniae is a healthcare-associated infections agent and could be an extended spectrum β-lactamase (ESBL) producer. Understanding the transmission of this bacterium in a hospital setting needs accurate typing methods. An antibiogram is used to detect the resistance pattern of the isolates. Random Amplified Polymorphic DNA (RAPD) and Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR are rapid, technically simple, and easy-to-interpret DNA typing methods. This study aimed to evaluate the use of antibiotyping, RAPD-, and ERIC-PCR to investigate the heterogeneity of K. pneumoniae isolated from clinical specimens. The antibiograms of 46 K. pneumoniae clinical isolates were determined by Vitek® 2 Compact. All isolates underwent RAPD-PCR using AP4 primer and ERIC-PCR using ERIC-2 primer. The dendrogram was generated using the GelJ software and analyzed to determine its similarity. The analysis of antibiogram and the molecular typing diversity index was calculated using the formula of the Simpson's diversity index. About 71.7% of the isolates were ESBL-producers, and more than 80% of isolates were susceptible to amikacin, ertapenem, and meropenem. The antibiotyping produced 32 diverse types with DI = 0.964. In addition, the RAPD-PCR produced 47 different types with DI = 1, while ERIC-PCR was 46 (DI=0.999). Antibiotyping, RAPD- and ERIC-PCR showed powerful discrimination power among the isolates, supported the diversity of K. pneumoniae isolates in current study. These combination could be promising tools for clonal relationship determination, including in tracking the transmission of the outbreak's agent in hospital setting.

Surgical Site Infections in Gastrointestinal Surgeries: Estimation of Prevalence, Risk Factors and Bacteriological Profile.

Surgical site infections (SSIs) are one of the leading causes of operation-related adverse effects.To create an effective hospital infection program, information about a local pattern is essential. The ever-changing pattern of infections and inappropriate use of antibiotics has predisposed to the development of drug-resistant strains and has made the management of SSIs arduous. The purpose of this study is to estimate prevalence and identify risk factors and commonest organisms associated with SSIs. In this analytical cross-sectional study, the relationship between various risk factors and the development of SSIswas evaluated in patients undergoing elective surgery and staying at least seven days postoperatively in the Department of Surgical Gastroenterology and Liver transplant for a study duration of two months. Diagnosis of SSIs was based on modified CDC criteria. Wound pus was followed by blood collection in suspected secondary septicemia. MacConkey and blood agar were used to culture pus; brain heart infusion broth was used for blood samples. Antimicrobial susceptibility testing was carried out using Mueller-Hinton agar by the Kirby-Bauermethod. Twelve out of 50 had developed postsurgical wound infections where Gram-negative organisms prevailed over Gram-positive ones. The associated risk factors identified in this study are age, BMI, wound class, American Society of Anesthesiologists (ASA) score, preprocedural WBC count >10,000, and the duration of surgery. Escherichia coli is the causative microbe for the majority of infections (35.7%). Gram-negative bacteria isolated in this study were extended-spectrum β-lactamase (ESBL) producers. Multidrug-resistant organisms were predominant. Conclusion: The present study identified an SSI rate of 24% in gastrointestinal surgeries. The sensitivity and resistance pattern of the organisms isolated will help in measures to be taken to devise a proper and effective current hospital antibiotic prophylaxis policy.

Assessment of Airborne Extended Spectrum β-Lactamase Producing Enterobacteriaceae Isolates from Motor Parks in Benin City, Nigeria

Aim: The study was aimed at assessing airborne bacteria and extended spectrum β-lactamase producing Enterobacteriaceae isolated from the motor parks in Benin City. Methodology: Airborne bacteria samples were collected in the motor parks (Central, Ugbowo, Agip, 19th Street, and Ekosodin Park), in Benin City using the settled plate method on Nutrient and MacConkey agar. Temperature, relative humidity, and airborne bacteria of the different parks were determined. The bacterial isolates were identified using phenotypic method. The antibiotic susceptibility profile was investigated by the disc diffusion method. The Enterobacteriaceae isolates were screened for Extended-spectrum β-lactamase (ESBL) production potentials. Virulence factor analysis and detection of ESBL genes (TEM, CTX-M, SHV, and OXA) were carried out on the selected ESBL producers. Results: The relative humidity recorded at the parks at the point of agar plate exposure ranged from 82 ̶ 90%. The airborne heterotrophic bacterial and coliform counts from the motor parks ranged from 9.50±0.63 × 102 ̶ 1.18±0.02 ×103 cfu/m3 and 2,48±0.41×102 ̶ 3.30±0.41×102 cfu/m3 respectively. The identified Enterobacteriaceae were Citrobacter freundii, Serratia marcescens, Citrobacter koseri, Klebsiella pneumoniae, Morganella morganii, Klebsiella oxytoca, Shigella dysenteriae, Proteus mirabilis, Escherichia coli, Enterobacter cloacae and Citrobacter diversus. The isolate with highest frequency of occurrence was E. coli with values of 30.48 %. The isolate K. pneumoniae had the highest MAR index of 0.8. The isolate S. marcescens had the highest virulent factor of 60% and possessed all the ESBL genes detected. Conclusion: Strong infection control measures should be put in place because more ESBL-producing Enterobacteriaceae have emerged in the surroundings of specific parks in Benin City.

Association Between Biofilm Formation and Extended-Spectrum Beta-Lactamase Production in Klebsiella pneumoniae Isolated from Fresh Fruits and Vegetables.

The presence of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae in fresh fruits and vegetables is a growing public health concern. The primary objective of this study was to investigate the relationship between biofilm formation and extended-spectrum β-lactamase (ESBL) production in K. pneumoniae strains obtained from fresh fruits and vegetables. Out of 120 samples analysed, 94 samples (78%) were found to be positive for K. pneumoniae. Among the K. pneumoniae strains isolated, 74.5% were from vegetables, whereas the remaining (25.5%) were from fresh fruits. K. pneumoniae isolates were resistant to at least three different classes of antibiotics, with ceftazidime (90%) and cefotaxime (70%) showing the highest resistance rates. While the high occurrence of ESBL-producing and biofilm-forming K. pneumoniae strains were detected in vegetables (73.5% and 73.7%, respectively), considerable amounts of the same were also found in fresh fruits (26.5% and 26.3%, respectively). The results further showed a statistically significant (P < 0.001) association between biofilm formation and ESBL production in K. pneumoniae strains isolated from fresh fruits and vegetables. Furthermore, the majority (81%) of the ESBL-producing strains harbored the blaCTX-M gene, while a smaller proportion of strains carried the blaTEM gene (30%), blaSHV gene (11%) or blaOXA (8%). This study highlights the potential public health threat posed by K. pneumoniae in fresh fruits and vegetables and emphasizes the need for strict surveillance and control measures.