Determining the Disinfectants Resistance Genes and the Susceptibility to Common Disinfectants of Extensively Drug-Resistant Carbapenem-Resistant Klebsiella pneumoniae Strains at a Tertiary Hospital in China. - Related Documents




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230001.0000Determining the Disinfectants Resistance Genes and the Susceptibility to Common Disinfectants of Extensively Drug-Resistant Carbapenem-Resistant Klebsiella pneumoniae Strains at a Tertiary Hospital in China. Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection has become a significant threat to global health. The application of chemical disinfectants is an effective infection control strategy to prevent the spread of CRKP in hospital environments. However, bacteria have shown reduced sensitivity to clinical disinfectants in recent years. Furthermore, bacteria can acquire antibiotic resistance due to the induction of disinfectants, posing a considerable challenge to hospital infection prevention and control. This study collected 68 CRKP strains from the Fifth Affiliated Hospital of Xinjiang Medical University in China from 2023 to 2024. These strains were isolated from the sputum, urine, and whole blood samples of patients diagnosed with CRKP infection. Antibiotic susceptibility tests were performed on CRKP strains. Concurrently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of disinfectants (benzalkonium bromide, 1% iodophor disinfectant, alcohol, and chlorine-containing disinfectant) against the test isolates were determined by the broth microdilution method. The efflux pump genes (cepA, qacE, qacEΔ1, qacEΔ1-SUL1, oqxA, and oqxB) were detected using polymerase chain reaction. The results showed that 21 out of the 68 CRKP strains exhibited extensive drug resistance, whereas 47 were nonextensively drug-resistant. The MIC value for benzalkonium bromide disinfectants displayed statistically significant differences (p < 0.05) between extensively drug-resistant (XDR) and non-XDR strains. Additionally, the MBC values for benzalkonium bromide disinfectants and 1% iodophor disinfectants displayed statistically significant differences (p < 0.05) between XDR and non-XDR strains. The detection rates for the efflux pump genes were as follows: cepA 52.9%, qacE 39.7%, qacEΔ1 35.2%, qacEΔ1-SUL1 52.9%, oqxA 30.8%, and oqxB 32.3%. The detection rate of the qacEΔ1-SUL1 gene in XDR CRKP strains was significantly higher than in non-XDR CRKP strains (p < 0.05). This indicates a potential link between CRKP bacterial disinfectant efflux pump genes and CRKP bacterial resistance patterns. Ongoing monitoring of the declining sensitivity of XDR strains against disinfectants is essential for the effective control and prevention of superbug.202439166283
229910.9998Determining the resistance of carbapenem-resistant Klebsiella pneumoniae to common disinfectants and elucidating the underlying resistance mechanisms. INTRODUCTION: Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is a serious problem in hospitals worldwide, posing a particular risk to immunocompromised patients. Elimination strategies may prevent these drug-resistant bacteria from spreading within hospital environments. Here, the susceptibility of patient-derived CRKP strains to common chemical disinfectants and possible correlations between the presence of drug-resistance genes and increased resistance to disinfectants were investigated. METHODS: The minimum inhibitory (MIC) and the minimum bactericidal concentrations (MBC) of common chemical disinfectants against each CRKP strain were determined using agar dilution; K. pneumoniae ATCC700603 served as a standard. The presence of the drug-resistance genes qacΔE, qacA, acrA and qacE was determined using PCR. RESULTS: A total of 27 clinically isolated CRKP strains collected in our hospital from 2011 to 2013 exhibited sensitivity to the following common chemical disinfectants in decreasing order of sensitivity: 75% ethyl alcohol > 2% glutaraldehyde > "84" disinfectant > 0.2% benzalkonium bromide > 2% iodine tincture > 1% iodophor > 0.1% chlorhexidine acetate. Of the 27 strains, 59, 41, 19 and 15% contained qacΔE, qacA, acrA and qacE resistance genes; 15% carried acrA, qacΔE and qacA, and 26% carried both qacA and qacΔE. Comparative analysis indicated that drug-resistance genes were correlated with higher MIC values. CONCLUSION: These pan-resistant pathogenic CRKP strains contained various drug-resistance genes and exhibited relatively high resistance to ethyl alcohol, chlorhexidine acetate and iodophor. Monitoring the drug-resistance rates of CRKP strains displaying disinfectant resistance may facilitate appropriate and effective sterilisation and thus preventing the spread of these pan-resistant strains.201526184804
215820.9998Relationship of OqxAB efflux pump to antibiotic resistance, mainly fluoroquinolones in Klebsiella pneumoniae, isolated from hospitalized patients. OBJECTIVES: This research was designed to study the prevalence of OqxAB efflux pump genes and also to investigate the relationship between efflux pump and resistance to antibiotics, especially to fluoroquinolones, evaluate the expression levels of OqxAB genes, and molecular typing of Klebsiella pneumoniae isolated from hospitalized patients in Hamadan hospitals, west of Iran. MATERIALS AND METHODS: In a cross-sectional study, 100 clinical strains of K. pneumoniae were isolated from hospitalized patients in three major teaching hospitals from January to June 2021. The antibiotic susceptibility of isolates was evaluated by the disk-diffusion agar method. The frequency of genes encoding oqxA and oqxB of efflux pump genes was investigated by PCR, and the expression of the oqxA efflux pump gene was investigated by the Real-time PCR method. The genetic relationship of K. pneumoniae isolates was analyzed by the Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR technique. RESULTS: According to our results, the multi-drug resistance phenotype (MDR) in 65% and high prevalence resistance to ciprofloxacin in 89% of K. pneumoniae isolates was detected. The higher prevalence of oqxA (95%) and oqxB (98%) was also detected. There was a significant relationship between ciprofloxacin resistance and the oqxB gene as well as between ceftriaxone and chloramphenicol resistance and the oqxA gene. The expression of the oqxA gene was higher in ciprofloxacin-resistant isolates. CONCLUSION: The results of this study suggest a potential reservoir for the spread of OqxAB genes among hospital-acquired bacteria. Infection control strategies should be used prudently to reduce the spread of resistant strains of K. pneumoniae in hospitals.202336594055
230630.9997Resistance to nitrofurantoin is an indicator of extensive drug-resistant (XDR) Enterobacteriaceae. Introduction. Nitrofurantoin is one of the preferred antibiotics in the treatment of uropathogenic multidrug-resistant (MDR) infections. However, resistance to nitrofurantoin in extensively drug-resistant (XDR) bacteria has severely limited the treatment options.Gap statement. Information related to co-resistance or collateral sensitivity (CS) with reference to nitrofurantoin resistant bacteria is limited.Aim. To study the potential of nitrofurantoin resistance as an indicator of the XDR phenotype in Enterobacteriaceae.Methods. One hundred (45 nitrofurantoin-resistant, 21 intermediately resistant and 34 nitrofurantoin-susceptible) Enterobacteriaceae were analysed in this study. Antibiotic susceptibility testing (AST) against nitrofurantoin and 17 other antimicrobial agents across eight different classes was performed by using the Vitek 2.0 system. The isolates were screened for the prevalence of acquired antimicrobial resistance (AMR) and efflux pump genes by PCR.Results. In total, 51 % of nitrofurantoin-resistant and 28 % of intermediately nitrofurantoin resistant isolates exhibited XDR characteristics, while only 3 % of nitrofurantoin-sensitive isolates were XDR (P=0.0001). Significant co-resistance was observed between nitrofurantoin and other tested antibiotics (β-lactam, cephalosporin, carbapenem, aminoglycoside and tetracycline). Further, the prevalence of AMR and efflux pump genes was higher in the nitrofurantoin-resistant strains compared to the susceptible isolates. A strong association was observed between nitrofurantoin resistance and the presence of bla (PER-1), bla (NDM-1), bla (OXA-48), ant(2) and oqxA-oqxB genes. Tigecycline (84 %) and colistin (95 %) were the only antibiotics to which the majority of the isolates were susceptible.Conclusion. Nitrofurantoin resistance could be an indicator of the XDR phenotype among Enterobacteriaceae, harbouring multiple AMR and efflux pump genes. Tigecycline and colistin are the only antibiotics that could be used in the treatment of such XDR infections. A deeper understanding of the co-resistance mechanisms in XDR pathogens and prescription of AST-based appropriate combination therapy may help mitigate this problem.202133830906
85840.9997Minocycline and Omadacycline Resistance Among Carbapenem-Resistant Gram-Negative Bacteria: Antimicrobial Susceptibility Testing and Molecular Characterization. Increasing prevalence of multidrug-resistant infections has rendered the healthcare systems ineffective in managing infectious diseases. Drugs of "last resort" like carbapenems and polymyxins are becoming less effective in the management of antibiotic-resistant Gram-negative bacterial infections, leaving the clinicians with limited choices. Evaluation of the efficacy of other available broad-spectrum antibiotics (belonging to a different class) is warranted as a treatment alternative. The current study was undertaken to evaluate the in vitro antibacterial activity of minocycline and a new drug, omadacycline among carbapenem-resistant Gram-negative bacteria (GNB), isolated from clinical samples (pus and sputum) and to genotypically analyze them. A prospective cross-sectional study was conducted in a 3,200-bedded tertiary care medical center, located in Lucknow in the northern part of India. All the clinical isolates recovered from pus and sputum samples of patients admitted in intensive care units were processed according to the standard protocols. Identification and antibiotic susceptibility testing were performed, and carbapenem-resistant Gram-negative bacteria (CRGNB) showing resistance to minocycline were included in the study. Molecular screening of β-lactamase and tetracycline resistance genes was done by the conventional polymerase chain reaction method. Minimum inhibitory concentration analysis was performed using the broth microdilution technique. Among 700 CRGNB, 15.29% (n = 107/700) were minocycline resistant by disk diffusion method. Genetic analysis demonstrated the presence of tetracycline-resistant genes in about one-third isolates, among which the tet(B) gene was present in 41.12% (n = 44/107). Upon broth microdilution analysis, the overall minimum inhibitory concentration for minocycline was raised, wherein 4.76% (n = 5/107) of our clinical Gram-negative isolates were inhibited at ≤8 mg/L and 15.23% (n = 28/107) were inhibited at ≤16 mg/L. Omadacycline was able to inhibit 13.08% (n = 14/107) of the minocycline-resistant isolates at ≤4 mg/L (susceptible breakpoint for Enterobacterales). Based on the cut-off value proposed, 15.09% (n = 16/107) isolates resistant to minocycline were inhibited by omadacycline. High prevalence of multidrug-resistant bugs entails judicious use of minocycline and omadacycline. The presence of tet genes coexisting with bla(NDM) and bla(OXA) in our bacterial isolates shows that the resistance pattern in Gram-negative bacilli is regularly evolving, and a fully functional surveillance program across the health care system is needed to prevent the emergence and spread of antimicrobial resistance.202540126171
86050.9997Investigation of Plasmid-Mediated Colistin Resistance Genes (mcr-1-8) in Enterobacterales Isolates. Background The escalating global rise in multidrug-resistant gram-negative bacteria presents an increasingly substantial threat to patient safety. Over the past decade, carbapenem-resistant Enterobacterales (CRE) have emerged as one of the most critical pathogens in hospital-acquired infections, notably within intensive care units. Colistin has become one of the last-resort antimicrobial agents utilized to combat infections caused by CRE. However, the use of colistin has been accompanied by a notable increase in the prevalence of colistin-resistant bacteria. This study aimed to investigate plasmid-mediated colistin resistance genes ranging from mcr-1 to mcr-8 among members of the Enterobacterales order. Materials and methods This prospective study was conducted in the microbiology laboratory of Afyonkarahisar Health Sciences University Health Research and Practice Center between May 1, 2021 and July 31, 2022. A total of 2,646 Enterobacterales isolates were obtained from all culture-positive clinical samples sent from various clinics. Of these, 79 isolates exhibiting resistance to carbapenem antibiotics were included in the study. Among the 79 isolates, the presence of mcr-1 to mcr-8 genes was investigated in 27 isolates that were shown to be resistant to colistin. The identification of bacteria at the species level and antibiotic susceptibility tests were conducted using the VITEK 2 automated system (bioMérieux, USA). Colistin resistance among Enterobacterales strains exhibiting carbapenem resistance was evaluated using the broth microdilution technique (ComASP™ Colistin, Liofilchem, Italy), in accordance with the manufacturer's instructions. Results In our in vitro investigations, the minimum inhibitory concentration (MIC) values for meropenem were determined to be >8 µg/ml, whereas for colistin, the MIC50 value was >16 µg/ml and the MIC90 value was 8 µg/ml. A total of 27 colistin-resistant strains were identified among the 79 carbapenem-resistant Enterobacterales strains analyzed. The most prevalent agent among colistin-resistant strains was Klebsiella pneumoniae (K. pneumoniae), representing 66.7% of the isolates. This was followed by Proteus mirabilis (P. mirabilis) with 29.6% and Escherichia coli (E. coli) with 3.7%. The colistin resistance rate among carbapenem-resistant strains was found to be 34.2%, with colistin MIC values in strains tested by the broth microdilution method ranging from 4 to >16 µg/ml concentrations. In polymerase chain reaction (PCR) studies, the mcr-1 gene region was successfully detected by real-time PCR in the positive control isolate. Nevertheless, none of the gene regions from mcr-1 to mcr-8 were identified in our study investigating the presence of plasmid-mediated genes using a multiplex PCR kit. Conclusion Although our study demonstrated the presence of increased colistin resistance rates in carbapenem-resistant Enterobacterales isolates, it resulted in the failure to detect genes from mcr-1 to mcr-8 by the multiplex PCR method. Therefore, it is concluded that the colistin resistance observed in Enterobacteriaceae isolates in our region is not due to the mcr genes screened, but to different resistance development mechanisms.202438957246
86160.9997The Prevalence Rates of Colistin Resistance Among Third-Generation Cephalosporin-Resistant E. coli Isolates From Thai Patients. Drug-resistant infections, such as colistin resistance, are reportedly increasing due to the inappropriate use of antibiotics, lack of good control, and the use of excessive quantities of antibiotics. Colistin resistance has been observed in bacteria of the order Enterobacterales exhibiting resistance to third-generation cephalosporins or carbapenems. Currently, the prevalence of colistin resistance in third-generation cephalosporin-resistant E. coli obtained from patients at Nopparat Rajathanee Hospital, Thailand, in 2023 was determined by broth microdilution. Among 343 isolates, 1.45% (95% CI: 0.19%-2.73%, n = 5/343) exhibited a minimum inhibitory concentration (MIC) of colistin between 4 and 8 μg/mL, indicating colistin-resistant E. coli. Nevertheless, five isolates were positive for resistant genes by multiplex PCR: two isolates for mcr-1, two isolates for mcr-3, and one isolate for both mcr-1 and mcr-3. This study reveals crucial data on resistance genes, informing surveillance of antibiotic resistance, treatment decisions, and public health initiatives to combat resistant bacteria.202541170286
230570.9997In-vitro activity of tigecycline against multidrug-resistant Gram negative bacteria: The experience of a university hospital. The emergence of multidrug-resistant Gram negative bacteria has given rise to significant therapeutic challenges. These pathogens may have developed resistance to tigecycline, which is an alternative antibiotic used empirically in the treatment of serious infections. The objectives of this study were to identify the in-vitro activity of tigecycline against multidrug-resistant Gram negative strains isolated from clinical specimens and their related genes, at a university hospital. For this, 150 clinical isolates of multidrug-resistant Gram negative cultures from various clinical specimens were collected. Bacterial isolates were cultured, identified and their antibiotic susceptibilities were determined. Polymerase chain reaction was performed to amplify AcrB, AmpC, RamR, MexR, AdeB, TetA genes. Results revealed that all isolates were multidrug-resistant. The resistance of isolates was 91.4% to aztreonam, 94.6% to piperacillin, 34% to imipenem, 38.7% to meropenem, 71.3% to levofloxacin, 97.3% to ceftriaxone, 94.7% to cefepime, 9.3% to colistin, 78% to tetracycline, 21.4% to tigecycline and 68% to trimethoprim. AcrB, AmpC, RamR, MexR, AdeB, TetA genes were present in multidrug-resistant Gram negative bacteria. AcrB, RamR, TetA genes were related to tigecycline resistance. It is concluded that infections caused by multidrug-resistant Gram negative bacteria occur at a high rate. Most isolates were multi drug resistant, with 21.4% being resistant to tigecycline.202133743369
265080.9997Prevalence of azithromycin resistance after the COVID-19 era in clinical bacterial isolates from a tertiary care hospital in Gurugram, India. The increasing prevalence of antibiotic resistance in pathogenic bacteria poses a great healthcare problem worldwide. Azithromycin (AZM) is a very effective macrolide antibiotic to treat many bacterial infections, but increasing azithromycin resistance in clinical bacteria decreases the effectiveness of this vital antibiotic, which is a major concern. The primary aim of the present study was to investigate the prevalence of azithromycin resistance and the occurrence of mphA gene in bacteria isolated from various clinical samples in Gurugram, India. For this, 138 pure bacterial isolates were obtained from the Department of Microbiology, Faculty of Medicine and Health Sciences, SGT Medical College, Hospital & Research Institute, Gurugram, India, from February to June 2024. All the isolates were identified by VITEK 2 system, and E. coli (22.5%) was found to be the most common pathogen in urine samples. Screening for azithromycin resistance by agar dilution and minimum inhibitory concentration (MIC) method found 30 azithromycin-resistant bacterial isolates. The present study found the prevalence of azithromycin resistance in pathogenic bacteria from clinical samples is 22%, indicating an increase in prevalence after the COVID-19 era, which is a major concern. Antibiotic profiling data revealed that 100% of the azithromycin-resistant isolates were multidrug-resistant, which is a serious issue. Furthermore, plasmid-mediated mphA gene was successfully amplified by the PCR method from 11 bacterial isolates, which may be responsible for azithromycin resistance. Our findings indicate the rapid emergence of azithromycin resistance in pathogenic bacteria, highlighting the urgency of stringent surveillance and control measures.202540365063
231390.9997Evaluated gene expressions of Metallo beta lactamase genes GIM and , VIM, SPM in Pseudomonas aeruginosa clinical isolates. Pseudomonas aeruginosa is considered as one of the human health care problems, P. aeruginosa's carbapenem resistance emerges by several different mechanisms, some of which include carbapenems genes. P. aeruginosa's carbapenem resistance is a significant health concern, So this study aims to evaluate MBL gene expressions. The study was conducted at the Department of Microbiology, AL-Mahmoodia Hospital, over one year from January to December 2022. The samples were collected from patients with different clinical sources (Burn, Urine, Wound, Sputum, Ear, and Blood), from different ages while. Samples were collected from three hospitals in Baghdad including Al-Yarmouk Teaching Hospital, AL-Mahmmodiya Hospital, and Child's Central Teaching Hospital. A study analyzed 55 P. aeruginosa strains from various clinical sources, the study utilizes the chemical characterization, VITEK 2 system, 16s rRNA, antibiogram sensitivity tests, antibiotic susceptibility using eight antibiotics, including Amikacin, Ciprofloxacin, Levofloxacin, Imipenem Meropenem, Piperacillin, Cefepim and Aztreonam. The test of bacterial susceptibility revealed that each isolate was highly resistant to piperacillin, which are 96.36%, and lower resistance to Ciprofloxacin, which are 32%. Phenotypic screening carbapenem resistance methods combined the disk synergy test and conventional PCR that were used to detect isolates by using 16 S rRNA. This proves that the bacteria is P. aeruginosa and computed by measuring gene expression of the target genes (GIM, VIM, SPM) by using the real-time PCR, which is employed for twenty-five isolates. The result indicates that the expression level of the VIM gene is highly regulated in carbapenem-resistance isolates compared to control isolates that is 1.00. While the expression level of gene GIM and SPM is downregulated in carbapenem-resistance isolates compared to control isolates that is 6. The carbapenem VIM and GIM, SPM (class B) genes are essential for resistance in P. aeruginosa induced by chromosomal changes that modify membrane permeability efflux pump overexpression for genes. As a result, many studies require for discovering new strategies to reduce the threat to public health through preventing the spread of these isolates via tight infections, control measures, and the reduction of the danger to public health.202337917414
1140100.9997High abundance of the colistin resistance gene mcr-1 in chicken gut-bacteria in Bangladesh. Colistin is considered a last-resort reserved drug for the treatment of critical human infections by Gram-negative bacteria. Phenotypic colistin-resistance is strongly associated with plasmid-mediated mobile colistin resistance (mcr) genes. The mcr-bearing Enterobacteriaceae have been detected in many countries from environments, animals, and humans. This study investigated phenotypic colistin-resistance and the distribution of mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes in chicken-gut bacteria in Bangladesh. Bacteria were isolated from poultry- and native-chicken droppings, and their susceptibilities to colistin were determined by agar dilution and E-test minimal inhibitory concentration (MIC) measurements. Multiplex polymerase chain reactions detected mcr-1 to mcr-5 genes. Overall, 61.7% (92/149) of the isolates showed colistin resistance by agar dilution assessment (MIC > 2.0 μg/mL). The phenotypic resistance was observed considerably higher in poultry-chicken isolates (64.6%, 64/99) than in native-chicken isolates (56%, 28/50; p = 0.373). All the resistant isolates showed MIC levels between > 2 and > 128 μg/mL. The mcr-genes (mcr-1and mcr-2 combined) were detected more in poultry gut bacteria (36.4%) than native-chicken isolates (20%, p = 0.06). Despite bacteria sources, mcr-genes appeared to be significantly associated with phenotypic colistin-resistance phenomena (p < 0.001). Prior colistin usage led to a substantial increase in the proportion of bacteria with mcr-genes and phenotypic resistance (p < 0.001).202033057111
875110.9997Characteristics of Carbapenem-resistant Klebsiella pneumoniae in sewage from a tertiary hospital in Jilin Province, China. Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is a serious problem in hospitals worldwide. We monitored a tertiary hospital in Changchun, Jilin Province, China, and found that CRKP was the major species among the carbapenem-resistant isolates in sewage. Subsequently, we evaluated the drug susceptibility, resistance genes, virulence genes, outer pore membrane protein-related genes (OmpK35 & OmpK 36), multi-locus sequence typing and replicons, biofilm formation capabilities, and resistance to chlorine-containing disinfectants among KP isolates. Identification of drug sensitivity, multiple resistance profiles were observed including 77 (82.80%) multidrug resistant (MDR), 16 (17.20%) extensive drug resistant (XDR). Some antibiotic resistance genes were detected, the most prevalent carbapenemase gene was blaKPC, and 16 resistance genes were associated with other antibiotics. In addition, 3 (3.23%) CRKP isolates demonstrated loss of OmpK-35 and 2 (2.15%) demonstrated loss of OmpK-36. In the detection of multi-locus sequence typing (MLST), 11 ST11 isolates carried virulence genes. The most common replicon type was IncFII. Biofilm-forming capabilities were demonstrated by 68.8% of the isolates, all of which were resistant to chlorine-containing disinfectants. The results of the study showed that antibiotic-resistant isolates, especially CRKP, could resist disinfectants in hospital wastewater, and improper treatment of hospital wastewater may lead to the spread of drug-resistant bacteria and their genes. Thus, these bacteria must be eliminated before being discharged into the municipal sewage system.202337195919
2160120.9997Detection of AdeAB, TetA, and TetB efflux pump genes in clinical isolates of tetracycline-resistant Acinetobacter baumannii from patients of Suez Canal University Hospitals. BACKGROUND: Acinetobacter baumannii is an opportunistic bacteria associated primarily with hospital-acquired infections. Its tendency to acquire or donate resistance genes to neighboring bacteria is a major concern. Tetracyclines have shown promise in treating A. baumannii infections, but tetracycline resistance is growing globally in A. baumannii isolates. OBJECTIVES: The study aimed to study (1) the prevalence of multidrug-resistant (MDR) A. baumannii infections at Suez Canal University Hospitals, (2) the distribution of efflux pump genes AdeA &B, TetA, and TetB, and (3) the effect of efflux pump inhibitor (CCCP) on tetracycline-resistant isolates. METHODS: Clinical samples (457) were collected (blood, urine, sputum, ETA, pus, and pleural fluid), followed by A. baumannii isolation and identification, PCR detection of efflux pump genes, and detection of tetracycline susceptibility and its MIC before and after treatment with the efflux pump inhibitor (CCCP). RESULTS: A total of 31 A. baumannii isolates were recovered (6.78%). The highest rate of isolation was from the ICU (48.3%) from the ET aspirate samples (48.3%). The efflux system AdeA and TetB genes were distributed in 100% of isolates, whereas AdeB was found in 93.5% of isolates and the TetA gene in 87.1% of isolates. All A. baumannii isolates were MDR showing resistance to three or more classes of antibiotics. 45% of the isolates showed a 4-fold reduction of MIC and 12.9% showed a 2-fold reduction in the MIC. CONCLUSIONS: Efflux pump is an important mechanism for tetracycline resistance among A. baumannii isolates.202539905304
2308130.9997Trends of Antibiotic Resistance in Multidrug-Resistant Pathogens Isolated from Blood Cultures in a Four-Year Period. BACKGROUND: Multidrug-resistant organisms cause serious infections with significant morbidity and mortality in the worldwide. These organisms have been identified as urgent and serious threats by CDC. The aim of this study was to determine the prevalence and changes of antibiotic resistance of multidrug-resistant pathogens isolated from blood cultures over a four-year period in a tertiary-care hospital. METHODS: Blood cultures were incubated in a blood culture system. Positive signalling blood cultures were subcultured on 5% sheep-blood agar. Identification of isolated bacteria was performed using conventional or automated identification systems. Antibiotic susceptibility tests were performed by disc diffusion and/or gradient test methods, if necessary, by automated systems. The CLSI guidelines were used for interpretation of antibiotic susceptibility testing of bacteria. RESULTS: The most frequently isolated Gram-negative bacteria was Escherichia coli (33.4%) followed by Klebsiella pneumoniae (21.5%). ESBL positivity was 47% for E. coli, 66% for K. pneumoniae. Among E. coli, K. pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii isolates, carbapenem resistance was 4%, 41%, 37%, and 62%, respectively. Carbapenem resistance of K. pneumoniae isolates has increased from 25% to 57% over the years, and the highest rate (57%) occured during the pandemic period. It is noteworthy that the aminoglycoside resistance in E. coli isolates gradually increased from 2017 to 2021. The rate of methicillin-resistant S. aureus (MRSA) was found to be 35.5%. CONCLUSIONS: Increased carbapenem resistance in K. pneumoniae and A. baumannii isolates is noteworthy, but carbapenem resistance in P. aeruginosa decreased. It is of great importance for each hospital to monitor the increase in resistance in clinically important bacteria, especially isolated from invasive samples, in order to take the necessary precautions in a timely manner. Future studies involving clinical data of patients and bacterial resistance genes are warranted.202337307126
2332140.9997Detection and characterization of heteroresistance to chloramphenicol in Klebsiella pneumoniae isolates. BACKGROUND: Heteroresistance represents a significant pathway through which sensitive bacteria evolve into resistant strains, posing challenges for current clinical laboratory detection methods. OBJECTIVES: This study aimed to investigate the differences in resistance among K. pneumoniae isolates from various sources, assess the prevalence of chloramphenicol heteroresistance (CHR), and explore the potential causes and key genes associated with CHR. METHODS: K. pneumoniae was isolated from 801 samples obtained from various sources, and its susceptibility to antibacterial agents was assessed. The modified Kirby-Bauer disk diffusion method, population analysis profiling (PAP), and bactericidal curve assays were employed to identify heteroresistant bacteria. Additionally, the growth curve and stability of CHR strains were measured. To analyze the factors influencing the formation of CHR, we detected the resistance genes cmlA, cat1, and floR across 17 resistant subpopulations, along with virulence genes such as fimH, wabG, kfu, uge, and aerobactin. RESULTS: Among the 198 K. pneumoniae tested, resistance rates to nitrofurantoin, tetracycline, and chloramphenicol were found to be 73.74%, 57.58%, and 51.01%, respectively. The prevalence of CHR was determined to be 8.59% (17 out of 198), which significantly diminished the in vitro bactericidal efficacy of chloramphenicol. Notably, 76.47% (13/17) of the isolates harbored the cat1 and/or floR genes, while the prevalence of the virulence genes wabG, fimH, uge, and kfu was 100%, 100%, 76.47%, and 47.06%, respectively. CONCLUSION: The floR and/or cat1 genes are pivotal in the mechanism underlying heteroresistance to chloramphenicol, and the presence of virulence genes could further contribute to the development of CHR.202540993538
2298150.9997Burden of biocide resistance among multidrug-resistant bacteria isolated from various clinical specimens in a tertiary care hospital. BACKGROUND: Most studies on biocide resistance and its genetic determinants arise from environmental or food-borne microbial isolates and only a few from clinically relevant isolates. OBJECTIVES: This study determines the proportion of biocide resistance against five commonly used biocides and detects biocide resistance genes among MDR bacterial isolates using PCR. METHODS: Consecutive MDR isolates (n ​= ​180) were included (30 each of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, and Enterococcus species) from clinical specimens of various inpatient units at JIPMER. The isolates were challenged at 0.5,1 and 2 Macfarland (McF) inoculum with discrete dilutions of disinfectants. The minimum bactericidal concentrations (MBCs) for 70% Ethanol, 1.5% Cresol, 2% Glutaraldehyde, 1% Cetrimide, and 1% Chlorhexidine were determined for the isolates using ATCC reference strains as controls. PCR was performed targeting qac A/B, G; smr; and nfx B genes. RESULTS: For all biocides, MDR isolates had MBCs less than the maximum MBCs of ATCC strains. For MDR K. pneumoniae, A. baumannii, and P. aeruginosa, the highest MBCs of chlorhexidine and cetrimide were ≥75 and ​≥ ​150 ​μg/ml respectively at 0.5 McF inoculum; whereas these organisms grew at higher inoculum (2McF) even at commercially recommended biocidal concentration (1%) corresponding to 750 and 1500 ​μg/ml of chlorhexidine and cetrimide respectively. Meanwhile, the highest MBCs of MDR E. coli were 75 and 150 ​μg/ml for chlorhexidine and cetrimide respectively. Interestingly, the Gram-positive cocci survived the action of up to 35% ethanol. The nfxB and qacG genes were detected in 87% and 6.67% of MDR P. aeruginosa isolates respectively with no biocide resistance genes detected among the other organisms. CONCLUSIONS: Biocide dilutions challenged with higher inoculum indicated a narrow margin of effectiveness for certain biocides. Although a significant proportion of clinical MDR isolates of P. aeruginosa harbored biocide resistance genes, this finding had no phenotypic correlation.202337769586
865160.9997High Prevalence of bla(NDM-1), bla(VIM), qacE, and qacEΔ1 Genes and Their Association with Decreased Susceptibility to Antibiotics and Common Hospital Biocides in Clinical Isolates of Acinetobacter baumannii. The objective of this study was to evaluate the susceptibility of metallo-β-lactamase (MBL)-producing Acinetobacter baumannii (A. baumannii) clinical isolates to biocides. We also determined the prevalence and correlation of efflux pump genes, class 1 integron and MBL encoding genes. In addition, bla(VIM), bla(NDM-1), qacE and qacEΔ1 nucleotide sequence analysis was performed and compared to sequences retrieved from GenBank at the National Center for Biotechnology Information database. A. baumannii had a resistance rate to carbapenem of 71.4% and 39.3% and was found to be a MBL producer. The minimum inhibitory concentrations (MICs) of chlorhexidine and cetrimide were higher than the recommended concentrations for disinfection in 54.5% and 77.3% of MBL-positive isolates respectively and their MICs were significantly higher among qac gene-positive isolates. Coexistence of qac genes was detected in 68.1% and 50% of the isolates with bla(VIM) and bla(NDM-1) respectively. There was a significant correlation between the presence of qac genes and MBL-encoding bla(VIM) and bla(NDM-1) genes. Each of the bla(NDM-1), bla(VIM), qacE and qacEΔ1 DNA sequences showed homology with each other and with similar sequences reported from other countries. The high incidence of Verona integron-encoded metallo-β-lactamases (VIM) and New-Delhi-metallo-β-lactamase (NDM) and qac genes in A.baumannii highlights emerging therapeutic challenges for being readily transferable between clinically relevant bacteria. In addition reduced susceptibility to chlorhexidine and cetrimide and the potential for cross resistance to some antibiotics necessitates the urgent need for healthcare facilities to periodically evaluate biocides efficacy, to address the issue of antiseptic resistance and to initiate a "biocidal stewardship".201728417918
859170.9997Analysis of mcr family of colistin resistance genes in Gram-negative isolates from a tertiary care hospital in India. AIM: Colistin serves as the drug of last resort for combating numerous multidrug-resistant (MDR) Gram-negative infections. Its efficacy is hampered by the prevalent issue of colistin resistance, which severely limits treatment options for critically ill patients. Identifying resistance genes is crucial for controlling resistance spread, with horizontal gene transfer being the primary mechanism among bacteria. This study aimed to assess the prevalence of plasmid-mediated mcr genes associated with colistin resistance in Gram-negative bacteria, utilizing both genotypic and phenotypic tests. METHODS AND RESULTS: The clinical isolates (n = 913) were obtained from a tertiary care center in Chennai, India. Colistin resistance was seen among Gram-negative isolates. These strains underwent screening for mcr-1, mcr-3, mcr-4, and mcr-5 genes via conventional PCR. Additionally, mcr-positive isolates were confirmed through Sanger sequencing and phenotypic testing. The bacterial isolates predominantly comprised Klebsiella pneumoniae (62.43%), Escherichia coli (19.71%), Pseudomonas aeruginosa (10.73%), and Acinetobacter baumannii (4.81%), along with other species. All isolates exhibited multidrug resistance to three or more antibiotic classes. Colistin resistance, determined via broth microdilution (BMD) using CLSI guidelines, was observed in 13.08% of the isolates studied. Notably, mcr-5 was detected in K. pneumoniae in PCR, despite its absence in Sanger sequencing and phenotypic tests (including the combined-disk test, colistin MIC in the presence of EDTA, and Zeta potential assays). This finding underscores the importance of employing multiple diagnostic approaches to accurately identify colistin resistance mechanisms.202438986507
2317180.9997Molecular Detection of Adefg Efflux Pump Genes and their Contribution to Antibiotic Resistance in Acinetobacter baumannii Clinical Isolates. BACKGROUND: Acinetobacter baumannii (A. baumannii) is one of the most important bacteria causing nosocomial infections worldwide. Over the past few years, several strains of A. baumannii have shown antibiotic resistance, which may be due to the activity of efflux pumps. This study was aimed to detect AdeFG efflux pump genes and their contribution to antibiotic resistance in A. baumannii clinical isolates. METHODS: A total of 200 A. baumannii clinical isolates were collected from clinical specimens of ulcers, pus, sputum, and blood. All isolates were identified using standard biochemical tests. After identifying and cleaving the genome by boiling, PCR was performed on samples using specific primers. The antimicrobial susceptibility patterns were determined by disk diffusion, with and without CCCP efflux pump inhibitor were determined according to CLSI guidelines. RESULTS: We identified 60 clinical isolates of A. baumannii using biochemical differential tests. Identification of all A. baumannii isolates was confirmed by blaOXA-51-like PCR. According to the results of our study, 98.37% of A. baumannii isolates were resistant to ciprofloxacin, norfloxacin, and levofloxacin. PCR results indicated that all 60 A. baumannii isolates contained the AdeF and 76.66% contained AdeG. CONCLUSION: the results of this study demonstrated that most of the A. baumannii isolates contained AdeF and AdeG efflux pump genes, and more than 98% of the isolates were resistant to ciprofloxacin, norfloxacin, and levofloxacin. This reflected the significant contribution of efflux pumps to the development of resistance to these antibiotics.202032582800
2316190.9997Clinical Klebsiella pneumoniae isolates and their efflux pump mechanism for antibiotic resistance challenge. BACKGROUND: Klebsiella pneumoniae is a serious pathogen that causes many disorders in humans and animals. Klebsiella pneumoniae, which is one of the most important pathogens in hospitals, often causes many clinical manifestations, including pneumonia, urinary tract infections, and meningitis. Interest in this bacterium has increased due to the increasing incidence of infection caused by it, as well as its high resistance to antibiotics, especially broad-spectrum antibiotics. AIM: This study showed the efflux pump mechanism of clinical K. pneumoniae isolates and antibiotic resistance in samples collected from sheep and human respiratory tract infection in southern Iraq. METHODS: Three hundred samples were collected, and the samples included: 150 nasal swabs from sheep and 150 sputum samples from humans. Through bacteriological and biochemical examinations. The isolates were identified K. pneumoniae isolates were also confirmed by 16S rRNA. Susceptibility testing of the antibiotics used in the study. To determine the phenotypic efflux pump activity, the agar ethidium bromide cartwheel method was used. RESULTS: Of 150 sputum human specimens and 150 nasal swabs from sheep were tested, 25 and 17 K. pneumoniae species isolates from patients and sheep, respectively, for the resistance of the bacteria isolated from humans to antibiotics. The highest rate of resistance was to piperacillin (88%), and the lowest rate was to antibiotics (36%), imipenem. The highest of bacterial susceptibility to the antibiotic imipenem was (44%) and (36%) for levofloxacin, respectively. For the bacterial isolates from sheep, the highest percentage of resistance to rifampin was (82.3%), and the highest percentage of sensitivity was to imipenem and Levofloxacin antibiotics. The results showed that most of the 39 bacterial isolates (92.8%) possessed an efflux pump mechanism. The result of genotyping to identify the efflux pump genes tolC and acrAB revealed that all isolates carried the genes. CONCLUSION: All the isolates were resistant to antibiotics, and the bacterial isolates under study most possess the efflux pump mechanism. All bacteria also have efflux pump genes, and this gives the bacteria more resistance against many antibiotics.202541036356