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Objectives: This study was conducted to measure the prevalence of antibiotic resistance, and corresponding resistance genes among Bacteroides and related genera in a tertiary hospital., Methods: We examined 138 clinical strains of Bacteroides, Phocaeicola and Parabacteroides species isolated between July 2018 and June 2022. Antibiotic susceptibility tests were conducted using agar dilution. The bft gene and antibiotic resistance genes were targeted by real-time PCR., Results: Resistance rates of all strains against ampicillin, cefoxitin, piperacillin-tazobactam, meropenem, imipenem, clindamycin, metronidazole, and tigecycline were 97.8 %, 28.3 %, 11.6 %, 7.9 %, 5.1 %, 47.8 %, 0 % and 4.3 %, respectively. Non-fragilis Bacteroidales spp. (NFB) exhibited lower susceptibility rates compared to B. fragilis for cefoxitin, clindamycin, and piperacillin-tazobactam. The prevalence of meropenem resistance was higher in B. fragilis (15.5 %) than in NFB (0 %). Among all strains, the rates of cepA, cfxA, cfiA, ermF, ermG, ermB, nim, linA, mefA, msrSA, tetQ, tetX, tetX1 and bft genes were 42.8 %, 44.9 %, 8.7 %, 44.2 %, 10.9 %, 2.2 %, 0.7 %, 29.0 %, 17.4 %, 7.2 %, 76.1 %, 8.0 %, 37.7 % and 16.7 %, respectively. In five B. fragilis strains, insertion sequences [IS1187(n = 3), ISBf6(n = 1), IS612B(n = 1)] were detected in the upstream region of cfiA. NimE with ISBf6 on plasmid pBFM29b was detected in one B. fragilis strain, intermediate to metronidazole (MIC = 16 μg/mL). ErmF was the most abundant gene responsible for clindamycin resistance. TetQ and tetX1 genes exhibited a higher frequency in strains that were not susceptible to tigecycline (MIC ≥8 μg/ml)., Conclusions: Monitoring the resistance trends of Bacteroides and related genera is crucial given the observed resistance to all classes of antibiotics and the presence of various resistance mechanisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

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 bla KPC , bla NDM , bla VIM , bla OXA-48 , bla IMP , and bla GES . 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 bla NDM , 2 carrying bla GES , 2 carrying bla VIM , and 1 carrying bla IMP . 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.

The increasing antibiotic resistance in Pseudomonas aeruginosa, responsible for both community-acquired and hospital-acquired infections, is of global significance. The primary mechanisms contributing to resistance development in P.aeruginosa include the increased activity of efflux pumps, decreased permeability of outer membrane porins and the production of carbapenemases. This study aimed to determine the effects of resistance nodulation division (RND) efflux pumps, outer membrane porin D (OprD) outer membrane protein and carbapenemase production on the development of resistance to different antibiotics in P.aeruginosa isolates. Eighty P.aeruginosa isolates obtained from clinical samples in our hospital between 2019 and 2021 were included in the study. Species-level identification of the isolates was performed using MALDI-TOF MS (Bruker Daltonics, Germany). Antibiotic susceptibilities were determined using the VITEK® 2 (bioMérieux, France) system according to the criteria of the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The expression levels of the outer membrane porin protein OprD and the regulatory genes of efflux pumps (mexB, mexC, mexE, and mexX) were investigated using real-time quantitative reverse transcriptase polymerase chain reaction (Rt-qPCR). The rpsL gene was used as the reference gene and P.aeruginosa PAO1 strain was used as the control strain in Rt-qPCR. Comparative expression analysis was calculated using the delta-delta cycle threshold (ΔΔCt) method. The presence of blaOXA-48, blaNDM, blaVIM, blaIMP, blaKPC and blaOXA-10 was investigated by PCR. Clonal relationships among the isolates were determined by AP-PCR using the M13 primer. Band profiles were analyzed using GelCompar II (Applied Maths) software. Decreased OprD expression was detected in 63.7% of the isolates and 74% of carbapenem-resistant isolates. The decrease in OprD expression was found to be significant only between the carbapenem-susceptible (n= 30) and carbapenem-resistant (n= 50) groups (p= 0.014). The overexpression rate of mexB was 82% in carbapenem-resistant isolates and 33% in carbapenem-susceptible isolates; 76.6% in multidrug-resistant (MDR) isolates and 45.5% in non-MDR isolates (p= 0.001 and p= 0.004, respectively). The overexpression rate of mexX was 68% and 40% in the amikacin-resistant and susceptible groups, respectively; and 64.3% and 40.4% in the gentamicin-resistant and susceptible groups, respectively (p= 0.02 and p= 0.041, respectively). Overexpression of mexX was found in 61.7% of MDR isolates and 30.3% of non-MDR isolates (p= 0.006). No significant difference was found in the expression of mexC and mexE between antibiotic-susceptible and resistant groups. While blaOXA-10 was detected in 32% (16/50) of carbapenem-resistant isolates, blaOXA-48, blaNDM, blaVIM, blaIMP, blaKPC were not detected in any of the isolates. AP-PCR analysis identified 64 different genotypes and no dominant genotype was observed. The isolates were grouped into 14 different clusters with a clustering rate of 36%. RND efflux systems play a crucial role in the development of antibiotic resistance in P.aeruginosa. This study showed that overexpression of mexB and decreased expression of OprD contributed to the development of carbapenem resistance, while overexpression of mexX contributed to the development of amikacin and gentamicin resistance. Overexpression of mexB and mexX has significant correlation with MDR isolates. The detection of blaOXA-10 in 16 isolates suggests that the presence of this resistance gene may contribute to the development of carbapenem resistance.

Introduction: To ensure the appropriate usage of ceftazidime-avibactam (CAZ-AVI), recently introduced in our hospital, we aimed to determine susceptibility rates, enzyme analysis, and clonal relationship among strains, together with clinical data., Methodology: Between June 1 and September 30, 2021, demographic and microbiological data of the patients were recorded. In the obtained samples, meropenem and colistin minimal inhibitory concentration (MIC) levels, carbapenem resistance genes, and the clonal relationship were studied by molecular methods. CAZ-AVI was not used in any of the patients., Results: 140 carbapenem-resistant Klebsiella pneumoniae were isolated from 57 patients. Resistance to CAZ-AVI was found in 76 (54.3%) strains. Out of 57 patients, 31 (54.4%) isolates could be reached. Meropenem MIC level was ≥ 32 µg/mL in 26 (83.9%), and colistin MIC level was ≥ 4 µg/mL in 17 (54.8%) isolates. Enzyme analysis revealed NDM in 20 (64.5%), OXA-48 in 17 (54.8%), and KPC in seven (22.6%). NDM + OXA-48 was determined in 10 (32.2%) strains. NDM was determined in all CAZ-AVI resistant strains, OXA-48 in 16.1% (2/5) strains. Seven genotypes were detected. The largest cluster was genotype 3 clusters (11 isolates). Of 31 patients, 22 (71.0%) died. CAZ-AVI was susceptible in one of the patients who survived and four who died., Conclusions: Before using a new antibiotic, each center should determine the basal data and phenotypic/genotypic resistance ratios specific to that antibiotic. While a high NDM rate and low CAZ-AVI sensitivity limit the use of the drug in our center, it is clear that CAZ-AVI use in sensitive strains will decrease mortality., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2024 Oya Özlem Eren-Kutsoylu, Hacer Ceylan-Çimendağ, Ayşe Nur Sari-Kaygisiz, Elif Seren Tanriverdi, Özgen Alpay Özbek, Ibrahim Mehmet Ali Öktem, Barış Otlu, Vildan Avkan-Oğuz.)

Listeria (L.) innocua is typically considered a non-pathogenic bacterium that can sometimes act as an opportunistic pathogen in severely immunocompromised patients. However, it plays an important role in food safety because it acts as an indicator organism for potential contamination and the effectiveness of sanitation methods. The aim of this study was to determine the prevalence, virulence genes, antibiotic resistance profiles, heavy metal and disinfectant resistance genes of L. innocua isolates from animal-derived foods. In this study, we isolated and characterized 39 L. innocua strains recovered from commonly 400 consumed beef meat, fresh fish meat, raw cow milk, and traditional cheese samples collected in Giresun, Turkey. The occurrence of virulence-associated genes was detected, such as plcA (97.4%), iap (35.8%), and hlyA (15.3%). A high incidence of resistance was recorded for fusidic acid (100%), followed by oxacillin (97.4%), clindamycin (82%), trimethoprim/sulfamethoxazole (69.2%), benzylpenicillin (41%), nitrofurantoin (35.8%), and fosfomycin (35.8%). Overall, 100% (39/39) of the isolates were resistant to at least one antibiotic, while 92.3% (36/39) of the isolate strains were multidrug resistant in the antimicrobial susceptibility tested. Among the L. innocua isolates (n = 39), 51.2%, 38.4%, 20.5%, 7.6%, 5.1%, 2.5%, and 2.5% were positive for qacH, cadA1, qacE, qacEΔ1-sul, qacJ, qacF, and qacG heavy metal and disinfectant resistance genes, respectively. The results highlight the need for more comprehensive studies to understand the monitoring and surveillance of L. innocua and their potential hazards to both humans and animals. [ABSTRACT FROM AUTHOR]

Background: Emerging carbapenem-resistant Klebsiella pneumoniae (K. pneumoniae) (CRKP) bacteremias are presenting significant public health risks due to limited treatment options and increased mortality. K. pneumoniae isolates exhibit carbapenem resistance rates that vary from 25% to 50% throughout the European continent, including our country. Aims: To assess the characteristics of CRKP bacteremia, a condition that has recently demonstrated an increasing prevalence in our center. We sought to ascertain the resistance rates of isolated strains to antibiotics other than carbapenems, identify the responsible carbapenemase genes, evaluate the efficacy of antibiotics, determine mortality rates, explore clonality among strains, and investigate the influence of the COVID-19 pandemic on all these factors. Study Design: Retrospective observational study. Methods: This study included patients aged 18 and older who had experienced meropenem-resistant K. pneumoniae bacteremia. Meropenem resistance was confirmed by employing the Kirby-Bauer disk diffusion method. Meropenem minimum inhibitory concentration (MIC) levels were determined using the gradient test, while colistin MIC levels were ascertained using the disk elution technique. Carbapenemase genes were evaluated via colony polymerase chain reaction (PCR), and clonality analysis was performed using the arbitrarily primed PCR technique. Results: The study comprised 230 patients, with a mean age of 63.1 ± 15.9 years, of whom 58.7% were male. Oxacillinase-48 (OXA-48) was detected in 74.8% of the patients, New Delhi metallo-beta-lactamase (NDM) in 12.6%, OXA-48 + NDM in 7.8%, and KPC in 4.8%. The 14-day and 30-day mortality rates were 57% and 69.6%, respectively. Multivariate analysis of the 30-day mortality revealed several crucial factors, including bacteremia development in the intensive care unit, the occurrence of bacteremia during the COVID-19 pandemic, polymicrobial bacteremia, the use of indwelling intravenous catheters, a platelet count of ≤ 140,000/µl, procalcitonin levels of ≥ 6 µg/l, and a Charlson comorbidity score ≥ 3. Notably, the OXA-48 and KPC genes were upregulated significantly during the COVID-19 pandemic, while the NDM gene groups were downregulated. Additionally, both 14-day and 30-day mortality rates increased significantly. Conclusion: In this study, the most prevalent carbapenemase gene was OXA-48; however, there has been a recent increase in KPC genes. No dominant epidemic strain was identified through clonality analysis. The clustering rate was 68% before the pandemic, increasing to 85.7% during the pandemic. The significance of infection control measures is underscored by the rise in both clustering and mortality rates during the COVID-19 pandemic. [ABSTRACT FROM AUTHOR]

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Our objective was to evaluate the performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification of the Chryseobacterium and Elizabethkingia spp. isolated from pediatric patients at Hacettepe University Hospital using 16S rRNA gene sequencing as the gold standard and to determine the antimicrobial susceptibility patterns of the isolates and baseline characteristics of patients. All stored Chryseobacterium and Elizabethkingia spp. isolated from various clinical specimens (sputum, blood, and urine) of pediatric patients at Hacettepe University Hospital between 2012 and 2016 were included in this study. Minimum inhibitory concentrations of 10 antimicrobial agents were determined by Etest for all isolates. To determine the baseline characteristics of patients, medical records of all patients were retrospectively reviewed. In total, 18 isolates of Chryseobacterium spp. (16 C. indologenes , 2 C. gleum ) and 5 isolates of Elizabethkingia spp. (3 E. meningoseptica , 2 E. anophelis ) were identified by 16S rRNA sequencing. MALDI-TOF MS correctly identified 19 (82.6%) isolates to the species level. The quinolones (ciprofloxacin and levofloxacin), trimethoprim/sulfamethoxazole and piperacillin/tazobactam showed the highest spectrum of activity against the overall collection of isolates. Cystic fibrosis (CF) was the underlying disease in 81.8% of patients. To our knowledge, this study includes the largest number of Chryseobacterium spp. isolated from clinical specimens of pediatric patients in Turkey. In this study, we also report the first clinical isolate of E. anophelis in Turkey. Since, the majority of strains were isolated from patients with CF; larger, prospective clinical studies are needed to establish whether chryseobacteria could be considered as an emerging opportunistic pathogen in patients with CF. [ABSTRACT FROM AUTHOR]