Screening of Antimicrobial Resistance Genes and Epidemiological Features in Hospital and Community-Associated Carbapenem-Resistant Pseudomonas aeruginosa Infections. - Related Documents




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92401.0000Screening of Antimicrobial Resistance Genes and Epidemiological Features in Hospital and Community-Associated Carbapenem-Resistant Pseudomonas aeruginosa Infections. INTRODUCTION: Researching carbapenem-resistant isolates enables the identification of carbapenemase-producing bacteria and prevents their spread. METHODS: P. aeruginosa isolates were recovered from Medicine Faculty of Recep Tayyip Erdoğan University and identified by conventional methods and the automated Vitek 2 Compact system. Antimicrobial susceptibility experiments were performed in accordance with CLSI criteria and the automated Vitek 2 Compact system. The PCR method was investigated for the presence of β-lactamase resistance genes. PFGE typing was performed to show clonal relation among samples. RESULTS: Seventy P. aeruginosa isolates were isolated from seventy patients. Of the patients, 67.1% had contact with the health service in the last 90 days and 75.7% of the patients had received antimicrobial therapy in the previous 90 days. Twenty-four isolates were carbapenem resistant, 2 isolates were multidrug-resistant except colistin, and none of the samples had colistin resistance. The gene encoding β-lactamase or metallo-β-lactamase was found in a total of 36 isolates. The bla (VEB) and bla (PER) genes were identified in 1 and 5 isolates alone or 17 and 13 isolates in combination with other resistance genes, respectively. The bla (NDM) was the most detected metallo-β-lactamase encoding gene (n=18), followed by bla (KPC) (n=12). bla (IMP) and bla (VIM) were detected in 5 and 1 isolates, respectively. Also, the association of bla (VEB)-bla (PER) and bla (VEB)-bla (KPC)-bla (NDM) was found to be very high. Much more resistance genes and co-occurrence were detected in hospital-acquired samples than community-acquired samples. No difference was found between the community and hospital-associated isolates according to PFGE results. Simultaneously from 6 patients, other microorganisms were also isolated and 5 of them died. CONCLUSION: The average length of stay (days) was found to be significantly higher in HAI group than CAI group. The death of 5 patients with fewer or no resistance genes showed that the co-existence of other microorganisms in addition to resistance genes was important on death.202133907430
92310.9999Prevalence of Oxacillinase Genes in Clinical Multidrug-Resistant Gram-Negative Bacteria. BACKGROUND: The emergence of OXA-type beta-lactamases has become a significant threat to public healthcare systems and may lead to prolonged hospital stays and increased mortality rates among affected patients. This study aimed to determine the prevalence of oxacillinase resistance (OXA) genes in multidrug-resistant (MDR) Gram-negative bacteria. METHODS: One hundred and six clinical isolates were collected from a stock of Gram-negative isolates and were identified and tested for antibiotic susceptibility and presence of OXA genes using polymerase chain reaction (PCR). RESULTS: The most common detected isolate was Klebsiella pneumoniae (36.8%), followed by Escherichia coli (33%), Pseudomonas aeruginosa (16%), and Acinetobacter baumannii (14.2%). Out of these isolates, 97.4%, 87.2%, 84.6%, and 79.5% were resistant to ampicillin/sulbactam, cefotaxime, ceftazidime, and aztreonam, respectively. PCR results confirmed the presence of one or more OXA genes in 34% of the samples studied. The blaOXA-1 and blaOXA-10 genes were the most highly detected genes, followed by blaOXA-4 and blaOXA-51. The total number of Pseudomonas aeruginosa isolates was confirmed to carry at least one OXA gene (70.6%), whereas Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli were confirmed to carry at least one OXA gene (53.3, 28.2, and 22.9%, respectively). There was a significant association (p < 0.05) between the resistance genes and the type of isolate. CONCLUSIONS: Pseudomonas aeruginosa and Acinetobacter baumannii are the most common MDR Gram-negative strains carrying OXA-type beta-lactamase genes. Monitoring of MDR pathogens in Gram-negative bacteria must be continuously undertaken to implement effective measures for infection control and prevention.202540066541
221920.9999Development and validation of a multiplex TaqMan real-time PCR for rapid detection of genes encoding four types of class D carbapenemase in Acinetobacter baumannii. A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D β-lactamases (bla(OXA-23)-like, bla(OXA-24/40)-like, bla(OXA-51)-like and bla(OXA-58)-like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant Acinetobacter baumannii isolates. Well-characterized strains of A. baumannii were used as positive controls and non-Acinetobacter strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMP-resistant A. baumannii isolates were positive for the bla(OXA-23)-like gene, and one isolate (2 %) was positive for the bla(OXA-58)-like gene. The bla(OXA-24/40)-like gene was not detected in any of the 46 IMP-resistant strains and the bla(OXA-51)-like gene was identified in both IMP-resistant and non-resistant A. baumannii. All 11 non-Acinetobacter bacteria produced a negative result for each of the four bla(OXA) genes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in A. baumannii in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in A. baumannii and potentially other clinically significant multidrug-resistant Gram-negative bacteria.201222878252
93130.9999Epidemiological characteristics and antimicrobial susceptibility among carbapenem-resistant non-fermenting bacteria in Brazil. INTRODUCTION: Non-fermenting Gram-negative bacteria such as Pseudomonas aeruginosa and Acinetobacter baumannii are widespread in the environment and are increasingly associated with nosocomial infections. Extensive and indiscriminate use of antibiotics in hospitals has contributed to an increased number of infections caused by these microorganisms, that are resistant to a wide variety of antimicrobials, including β-lactams. This study aimed to isolate and identify carbapenem-resistant Acinetobacter spp. and P. aeruginosa from hospitalized patients, to determine their antimicrobial susceptibility patterns and to screen for blaOXA-23, blaOXA-24, blaOXA-51, blaOXA-58, and blaOXA-143 genes among the isolated bacteria. METHODOLOGY: Antimicrobial resistance patterns were performed using the disk-diffusion method. Genetic markers related to carbapenem resistance were screened by polymerase chain reaction. RESULTS: Carbapenem-resistant Acinetobacter spp. (n = 44) and P. aeruginosa (n = 28) samples were isolated from patients admitted to a tertiary hospital. Polymyxin B was the only effective drug for all isolates. Considering the oxacillinase gene screening, genetic markers were observed only in Acinetobacter isolates. The most frequent genotype observed was blaOXA-23+/blaOXA-51+ (45.5%), followed by blaOXA-51+/blaOXA-143+ (41%). The oxacillinase genes blaOXA-24 and blaOXA-58 were not detected. High mortality rates (> 70%) were observed. CONCLUSIONS: The data suggest the need for rational use of antimicrobials associated with early diagnosis of multidrug-resistant bacteria, especially considering non-fermenting Gram-negative rods, which are widespread in hospitals. The findings of blaoxa-51(-) strains suggest the occurrence and spread of non-A. baumannii species throughout our hospitals. Effective implementation of surveillance programs in hospitals is needed to reduce infectious and resistant intra- and inter-species bacteria.201627367001
103440.9999Detection of metallo-beta-lactamase-producing genes bla(SPM) and bla(NDM) in Pseudomonas aeruginosa isolated from wastewater in Southern Brazil. Pseudomonas aeruginosa is commonly associated with the ability to acquire antimicrobial resistance. The surveillance of resistance genes in various environmental matrices has gained prominence in recent years, being seen as a potential threat to public health. The objective of this study was to investigate genes encoding metallo-beta-lactamases (MBLs), which confer resistance to carbapenems, in wastewater. Fifteen isolates of P. aeruginosa were collected for five months from samples obtained from a municipal wastewater treatment plant in Rio Grande do Sul. These isolates were subjected to disk diffusion testing using 10 different antimicrobials. Phenotypic enzymatic tests for MBLs were conducted, and positive isolates underwent DNA extraction and gene detection using the polymerase chain reaction. The resistance rate to ceftazidime was 100%, cefepime 73.3%, piperacillin-tazobactam 66.67%, imipenem 53.30%, levofloxacin 46.67%, tobramycin 40%, and ciprofloxacin and amikacin 13.33%. Both meropenem and aztreonam resistances were rare accounting for 6.60% of the tested isolates. Among these isolates, 20% were classified as multidrug-resistant and were found to carry the bla(NDM) and bla(SPM) genes. The results suggest that evaluating resistance genes in bacteria from urban raw sewage can provide data that assist in surveillance, as this environment can stimulate increased bacterial resistance.202438678422
217450.9999Frequency of Beta-Lactamase Antibiotic Resistance Genes in Escherichia Coli and Klebsiella pneumoniae. BACKGROUND: This cross-sectional study was performed on isolates of Klebsiella pneumoniae, and E.coli from clinical specimens of patients admitted to Sayyad Shirazi Hospital by census sampling method in 2019. Antibiogram testing was performed using the disk diffusion method as defined by the Clinical and Laboratory Standards Organization for performing this test. Finally, the abundance of genes was evaluated by PCR using specific primers. Frequency, percentage, mean±SD were used to describe the data. Chi-square and Fisher's exact tests were used to compare the presence and absence of the studied genes alone and in the presence of each other. RESULT: This study was performed on 130 positive samples, isolated from 32 (24.6%) males and 98 (65.4%) females with a mean age of 43.78 ± 21.72. From the total number of 130 isolates, 84 (64.6%) consisted of E.coli, and 46 (35.4%) were Klebsiella. Most of the cultures were urine and vaginal (61.5%). The highest antibiotic resistance in isolates was cephalexin and cefazolin (67.9% in E.coli & 63% in Klebsiella). Colistin was identified as the most effective antibiotic (100%) in both. AMPC extendedspectrum β-lactamase genes were present in 40 (30.8%) isolates. The highest frequency about the gene pattern of AMPC positive β-lactamase bacteria was correlated to DHA, FOX, and CIT genes, while none of the samples contained the MOX β-lactamase gene. E.coli and Klebsiella beta-lactamase-producing AMPC isolates were also significantly correlated with antibiotic resistance to the cephalosporin class (P <0.05). CONCLUSION: This study indicated a high percentage of resistance to third and fourth generation cephalosporins. Hence, careful antibiogram tests and prevention of antibiotic overuse in infections caused by AMPC-producing organisms and screening of clinical samples for the resistance mentioned above genes and providing effective strategies to help diagnose and apply appropriate treatments and change antibiotic usage strategies can partially prevent the transmission of this resistance.202134483624
92260.9999Insertion Sequences within Oxacillinases Genes as Molecular Determinants of Acinetobacter baumannii Resistance to Carbapenems-A Pilot Study. Carbapenem-resistant Acinetobacter baumannii is one of the major problems among hospitalized patients. The presence of multiple virulence factors results in bacteria persistence in the hospital environment. It facilitates bacterial transmission between patients, causing various types of infections, mostly ventilator-associated pneumonia and wound and bloodstream infections. A. baumannii has a variable number of resistance mechanisms, but the most commonly produced are carbapenem-hydrolyzing class D β-lactamases (CHDLs). In our study, the presence of bla(OXA-23), bla(OXA-40) and bla(OXA-51) genes was investigated among 88 clinical isolates of A. baumannii, including 53 (60.2%) strains resistant to both carbapenems (meropenem and imipenem) and 35 (39.8%) strains susceptible to at least meropenem. Among these bacteria, all the isolates carried the bla(OXA-51) gene. The bla(OXA-23) and bla(OXA-40) genes were detected in two (5.7%) and three (8.6%) strains, respectively. Among the OXA-23 carbapenemase-producing A. baumannii strains (n = 55), insertion sequences (ISAba1) were detected upstream of the bla(OXA-23) gene in fifty-two (94.5%) carbapenem-resistant and two (3.6%) meropenem-susceptible isolates. A. baumannii clinical strains from Poland have a similar antimicrobial resistance profile as those worldwide, with the presence of ISAba1 among bla(OXA-23)-positive isolates also being quite common. Carbapenem resistance among A. baumannii strains is associated with the presence of CHDLs, especially when insertion sequences are present.202439458366
91770.9999Virulence characterization and clonal analysis of uropathogenic Escherichia coli metallo-beta-lactamase-producing isolates. BACKGROUND: Uropathogenic Escherichia coli (UPEC) is a major cause of urinary tract infection (UTI); however, treatment of UTI has been challenging due to increased antimicrobial resistance (AMR). One of the most important types of AMR is carbapenem resistance (CR). CR bacteria are known as an important threat to global public health today. Class B metallo-beta-lactamases (MBLs) are one of the major factors for resistance against carbapenems. We aimed to investigate the characteristics of UPEC isolates producing MBL. METHODS: A cross-sectional study was conducted from October 2018 to December 2019 in Ahvaz; Iran. UPEC isolates were identified by biochemical and molecular methods. Metallo-beta-lactamase-producing isolates were detected using modified carbapenem inactivation method (mCIM) and EDTA-CIM (eCIM) tests. MBL genes, phylogenetic group, and virulence genes profile of carbapenem resistant isolates were determined. Conjugation assay and plasmid profiling were conducted to evaluate the ability of transferring of CR to other E. coli isolates. Clonal similarity of isolates were assessed using Enterobacterial intergenic repetitive element sequence (ERIC)-PCR. RESULTS: Among 406 UPEC isolates, 12 (2.95%) carbapenem-resistant were detected of which 11 were phenotypically MBL-producing strains. Four isolates were resistant to all investigated antimicrobial agents and were considered possible pandrug-resistant (PDR). bla(NDM), bla(OXA-48), bla(IMP-1), and bla(IMP-2) genes were found in 9, 5, 1, and 1 isolates, respectively. Among 30 virulence genes investigated, the traT, fyuA followed by fimH, and iutA with the frequency of 8 (66.7%), 8 (66.7%), 7 (58.3%), and 7 (58.3%) were the most identified genes, respectively. Siderophore production was the main virulence trait among carbapenem-resistant UPEC isolates. Except for two, all other isolates showed weak to moderate virulence index. In all recovered isolates, CR was readily transmitted via plasmids to other isolates during conjugation experiments. CONCLUSION: MBL and carbapenemase genes, especially bla(NDM) and bla(OXA-48) are spreading rapidly among bacteria, which can be a threat to global public health. Therefore monitoring the emergence and dissemination of new AMR is necessary to continuously refine guidelines for empiric antimicrobial therapy. Understanding the mechanisms of resistance and virulence in this group of bacteria can play an effective role in providing new therapeutic methods.202134344363
212480.9999Evaluation of Phenotypic and Genotypic Characteristics of Carbapnemases-producing Enterobacteriaceae and Its Prevalence in a Referral Hospital in Tehran City. BACKGROUND & OBJECTIVE: Carbapenem-resistant Enterobacteriaceae is a growing concern worldwide including Iran. The emergence of this pathogen is worrying as carbapenem is one of the 'last-line' antibiotics for treatment of infections caused by multi drug resistant gram- negative bacteria. The main objective of this study was to determine the prevalence of carbapenem-resistant Enterobacteriaceae in a referral hospital in Tehran, Iran. METHODS: In this study, all positive isolates of Enterobacteriaceae recorded in blood, urine, and other body fluids were studied during April 2017 to April 2018 in a referral hospital in Tehran. All cases of resistance to carbapenems were first tested by modified Hodge test. All cases with positive or negative test, after gene extraction, were examined genotypically based on the primers designed for the three Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), and OXA-48 genes by conventional PCR method. RESULTS: 108 isolates (13.6%) were resistant to all cephalosporins as well as to imipenem and meropenem. In a genotypic study, including 45 isolates, 13 isolates were positive for OXA-48 gene, 11 isolates for OXA-48 and NDM genes, 11 isolates for OXA-48, NDM and KPC genes, 4 isolates for OXA-48 genes and KPC, 3 isolates for NDM, one isolate for KPC. On the other hand, two isolates were negative for all three genes examined. CONCLUSION: OXA-48 gene was one of the most common genes resistant to carbapenems in Iran. According to studies, the prevalence of antibiotic resistance in Iran is rising dramatically, which reduces the choice of antibiotics to treat severe infections in the future.202032215024
221890.9999Comparison of in-house and commercial real time-PCR based carbapenemase gene detection methods in Enterobacteriaceae and non-fermenting gram-negative bacterial isolates. BACKGROUND: Carbapenemase-producing gram-negative bacteria are increasing globally and have been associated with outbreaks in hospital settings. Thus, the accurate detection of these bacteria in infections is mandatory for administering the adequate therapy and infection control measures. This study aimed to establish and evaluate a multiplex real-time PCR assay for the simultaneous detection of carbapenemase gene variants in gram-negative rods and to compare the performance with a commercial RT-PCR assay (Check-Direct CPE). METHODS: 116 carbapenem-resistant Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii isolates were genotyped for carbapenemase genes by PCR and sequencing. The defined isolates were used for the validation of the in-house RT-PCR by use of designed primer pairs and probes. RESULTS: Among the carbapenem-resistant isolates the genes bla (KPC), bla (VIM), bla (NDM) or bla (OXA) were detected. Both RT-PCR assays detected all bla (KPC), bla (VIM) and bla (NDM) in the isolates. The in-house RT-PCR detected 53 of 67 (79.0%) whereas the commercial assay detected only 29 (43.3%) of the OXA genes. The in-house sufficiently distinguished the most prevalent OXA types (23-like and 48-like) in the melting curve analysis and direct detection of the genes from positive blood culture vials. CONCLUSION: The Check-Direct CPE and the in-house RT-PCR assay detected the carbapenem resistance from solid culture isolates. Moreover, the in-house assay enabled the identification of carbapenemase genes directly from positive blood-culture vials. However, we observed insufficient detection of various OXA genes in both assays. Nevertheless, the in-house RT-PCR detected the majority of the OXA type genes in Enterobacteriaceae and A. baumannii.201728693493
933100.9999Molecular characterization and diversity of carbapenemases in Gram-negative bacteria in Libyan hospitals. INTRODUCTION: Antimicrobial resistance has become a major threat to public health, especially in developing countries, due to the uncontrolled consumption of antibiotics. This study aims to characterize antibiotic resistance genes in different bacteria recovered in different healthcare facilities in Libya. METHODOLOGY: 379 samples were recovered from various sources from different sites. 210 samples were able to grow on culture media. 133 Gram-negative carbapenem-resistant strains were recovered from clinical specimens (n = 64), and hospital environments (n = 69). Antibiotic susceptibility tests were performed to select carbapenem-resistant strains. Colistin resistance was tested by the UMIC method to determine the minimum inhibitory concentration. RT-PCR was conducted to detect the incidence of carbapenemases-encoding genes. RESULTS: Gram-negative bacteria showed a low susceptibility to carbapenems. Molecular investigations indicated that NDM-1 was the most prevalent in Enterobacteriaceae isolated from patients and hospital environment (n = 26, n = 41), followed by blaOXA-48 (n = 16, n = 15) and blaVIM (n = 3) from patients and blaKPC (n = 1) from hospital environment. Concerning A. baumannii, blaOXA-23 was detected in strains isolated from patients (n = 8) and hospital environment (n = 6), followed by blaNDM (n = 9) from patients and one from hospital environment. Carbapenem resistance in P. aeruginosa was encoded by modification in OprD encoding gene, such as IS (ISpa26), polymorphism, and a premature stop codon. CONCLUSIONS: Several carbapenem resistant Gram-negative bacteria were identified by the expression of different carbapenemases and the alteration of OprD.202540720466
980110.9999Phenotypic and Molecular Characterization of Extended-Spectrum β-Lactamase, Plasmid-Mediated- AmpC, and Carbapenemase-Producing Enterobacteriaceae Isolated from Companion and Production Animals in Brazil. The crisis of bacterial resistance is an emerging One Health challenge, driven by the overuse of antimicrobials in medical and agricultural settings. This study aimed to investigate extended-spectrum β-lactamase (ESBL), Ampicillinase (AmpC), and carbapenemase production, and the presence of genes encoding these enzymes in Escherichia coli, Klebsiella spp., and Proteus spp., major contributors to infections and resistance isolates from animals. From 2016 to 2021, 130 multidrug-resistant (MDR) or extensively drug-resistant (XDR) isolates were recovered from the secretions, excretions, and organs of companion and production animals with active infections. Antibacterial sensitivity tests, along with phenotypic and genotypic detection of resistance enzymes, were performed. To the best of our knowledge, this is the first study in Brazil to estimate the prevalence of XDR Enterobacteriales isolated from companion and production animals, which accounted for 13.8% of the strains. Statistically significant differences (P < 0.05) in resistant bacteria between different classes and within the same class of antibacterial bacteria were found. The statistical probability between genotypic detection of ESBL (OR = 3.1) and phenotypic tests for AmpC (OR = 2.3) was also established. Approximately 32.3%, 17.6%, and 16.8% of the strains had positive phenotypic tests for ESBL, AmpC, and carbapenemases, respectively. Genetic analysis revealed the presence of bla(CTX-M) (60.0%), bla(AmpC) (9.18%), bla(KPC-2) (0.76%), and bla(NDM) (1.52%). AmpC genes were identified in 8.46% of the samples, with bla(CMY) being the most frequent (6.92%), followed by bla(DHA) (0.77%), and bla(FOX) (0.77%). The sequenced amplicons were deposited in NCBI. This study reveals critical data on Enterobacteriaceae with antibacterial resistance genes isolated from animals and may pose a significant threat to One health.202539903315
936120.9999Occurrence and Diversity of Intra- and Interhospital Drug-Resistant and Biofilm-Forming Acinetobacter baumannii and Pseudomonas aeruginosa. Acinetobacter baumannii and Pseudomonas aeruginosa are the most relevant Gram-negative bacteria associated with hospital and opportunistic infections. This study aimed to evaluate the dynamics of drug-resistant A. baumannii and P. aeruginosa and biofilm formers from two public hospitals in northeastern Brazil. One hundred isolates (35 from A. baumannii and 65 from P. aeruginosa) were identified using the automated Vitek(®)2 Compact method (bioMérieux) and confirmed using the MALDI-TOF (MS) mass spectrometry technique. Molecular experiments were performed by polymerase chain reaction (PCR) to detect the frequency of bla(KPC), bla(IMP), bla(VIM), and bla(SHV) genes. The biofilm formation potential was evaluated using crystal violet in Luria Bertani Miller and trypticase soy broth culture media under the following conditions: at standard concentration, one quarter (25%) of the standard concentration and supplemented with 1% glucose. In addition, the genetic diversity of the isolates was verified by the ERIC-PCR technique. Isolates presented distinct resistance profiles with a high level of beta-lactam resistance. The highest index of genes detected was bla(KPC) (60%), followed by bla(SHV) (39%), bla(VIM) (8%), and bla(IMP) (1%). All the isolates were sensitive to the polymyxins tested and formed biofilms at different intensities. Twelve clones of A. baumannii and eight of P. aeruginosa were identified, of which few were indicative of intra- and interhospital dissemination. This study reveals the dispersion dynamics of these isolates in the hospital environment. The results demonstrate the importance of monitoring programs to combat the spread of these pathogens.202031916896
1055130.9999Antimicrobial Susceptibility and Molecular Identification of Antibiotic Resistance Enteric Bacteria Isolated From Pigeon Feces in the City of Jeddah, Saudi Arabia. Background Due to their potential to carry a wide range of bacteria, pigeon feces may contribute to the spreading of infectious diseases in urban settings. Objective This study analyzed the presence of enteric bacteria from pigeon feces in Jeddah and their antimicrobial susceptibility and described the molecular characteristics of the carbapenem resistance genes it produced. Method Two hundred twenty-five pigeon feces specimens were collected from eight parks in Jeddah. Conventional microbiology techniques were employed to identify the isolated bacteria, and the automated Vitek2® system (bioMérieux, Marcy-l'Étoile, Lyon, France) provided additional confirmation. Kirby-Bauer disk diffusion method was utilized to screen for antimicrobial resistance. Only 50 antibiotic-resistance isolates further underwent molecular diagnosis for testing groups of carbapenems-encoding genes (blaNDM, blaSIM, and blaAIM), using multiplex polymerase chain reaction (PCR).  Result Of the 50 antibiotic-resistant isolates, 28% (14/50) were Klebsiella pneumoniae, 24% (12/50) were Enterobacter cloacae, and 48% (24/50) were Escherichia coli. Ninety percent (90%) of the isolates showed resistance to cefuroxime, 56% to gentamicin, 52% to amoxicillin/clavulanic acid, and 100% to meropenem. NDM beta-lactamase was the most often discovered gene (26%) and was followed by AIM beta-lactamase (5%) Conclusion According to this study, there may be a chance for resistant K. pneumoniae, E. cloacae, and E. coli to spread amongst several hosts within the same area. Consequently, to prevent the continued occurrence and dissemination of resistant strains among other hosts in the same location, it is essential to monitor the AMR (antimicrobial resistance) of E. coli, E. cloacae, and K. pneumoniae from pigeons.202439310621
1051140.9999Multi-drug Resistance, β-Lactamases Production, and Coexistence of bla (NDM-1) and mcr-1 in Escherichia coli Clinical Isolates From a Referral Hospital in Kathmandu, Nepal. The ability of pathogenic Escherichia coli to produce carbapenemase enzymes is a characteristic that allows them to resist various antibiotics, including last-resort antibiotics like colistin and carbapenem. Our objectives were to identify rapidly developing antibiotic resistance (AR), assess β-lactamases production, and detect mcr-1 and bla (NDM-1) genes in the isolates. A prospective cross-sectional study was carried out in a referral hospital located in Kathmandu from November 2019 to December 2020 using standard laboratory and molecular protocols. Among 77 total E. coli isolates, 64 (83.1%) of them were categorized as MDR. Phenotypically 13 (20.3%) colistin-resistant, 30 (46.9%) ESBL and 8 (12.5%) AmpC producers, and 5 (7.8%) ESBL/AmpC co-producers were distributed among MDR-E. coli. Minimum inhibitory concentrations (MIC) against the majority of MDR isolates were exhibited at 1 g/L. Of these 77 E. coli isolates, 24 (31.2%) were carbapenem-resistant. Among these carbapenem-resistant bacteria, 11 (45.9%) isolates were reported to be colistin-resistant, while 15 (62.5%) and 2 (8.3%) were MBL and KPC producers, respectively. Out of 15 MBL producers, 6 (40%) harbored bla (NDM-1), and 8 (61.5%) out of 13 colistin-resistant pathogens possessed mcr-1. The resistance by colistin- and carbapenem were statistically associated (P < .001). However, only 2 (18.2%) of the co-resistant bacteria were found to have both genes. Our study revealed the highly prevalent MDR and the carbapenem-resistant E. coli and emphasized that the pathogens possess a wide range of capabilities to synthesize β-lactamases. These findings could assist to expand the understanding of AR in terms of enzyme production.202336741474
1033150.9999Antimicrobial Resistance and β-Lactamase Production in Clinically Significant Gram-Negative Bacteria Isolated from Hospital and Municipal Wastewater. Hospital and municipal wastewater contribute to the spread of antibiotic-resistant bacteria and genes in the environment. This study aimed to examine the antibiotic resistance and β-lactamase production in clinically significant Gram-negative bacteria isolated from hospital and municipal wastewater. The susceptibility of bacteria to antibiotics was tested using the disk diffusion method, and the presence of extended-spectrum β-lactamases (ESBL) and carbapenemases was determined using an enzyme inhibitor and standard multiplex PCR. Analysis of antimicrobial resistance of total bacterial strains (n = 23) revealed that most of them were resistant to cefotaxime (69.56%), imipenem (43.47%), meropenem (47.82%) and amoxicillin-clavulanate (43.47%), gentamicin (39.13%), cefepime and ciprofloxacin (34.78%), trimethoprim-sulfamethoxazole (30.43%). A total of 8 of 11 phenotypically confirmed isolates were found to have ESBL genes. The bla(TEM) gene was present in 2 of the isolates, while the bla(SHV) gene was found in 2 of the isolates. Furthermore, the bla(CTX-M) gene was found in 3 of the isolates. In one isolate, both the bla(TEM) and bla(SHV) genes were identified. Furthermore, of the 9 isolates that have been phenotypically confirmed to have carbapenemase, 3 were confirmed by PCR. Specifically, 2 isolates have the bla(OXA-48) type gene and 1 have the bla(NDM-1) gene. In conclusion, our investigation shows that there is a significant rate of bacteria that produce ESBL and carbapenemase, which can promote the spread of bacterial resistance. Identifying ESBL and carbapenemase production genes in wastewater samples and their resistance patterns can provide valuable data and guide the development of pathogen management strategies that could potentially help reduce the occurrence of multidrug resistance.202337107015
2150160.9999Analysis of drug resistance genes of integrons in clinical isolates of Escherichia coli from elderly bloodstream infections. This experiment was carried out to provide a basis for the treatment of clinical bloodstream infections by analyzing the drug resistance characteristics and integrated gene distribution of Escherichia coli in bloodstream infections in elderly patients. For this aim, E. coli were collected for bacterial identification and drug sensitivity testing from bloodstream infections in elderly patients in the hospital from January 2016 to December 2019. ESBLs positive strains were assayed for genotypes and their integron carriage rates by PCR amplification. The characteristics and differences of various genotype rates were compared and analyzed. Results showed that a total of 230 E. coli strains were isolated. The detection rate of ESBLs-producing bacteria was 37.39 %. ESBLs-producing E. coli showed a high rate of resistance to cefepime, levofloxacin, cotrimoxazole, and ticarcillin/clavulanic acid (>40%). The resistance rate of 230 strains of E. coli to meropenem, minocycline, amikacin, gentamicin and cefoxitin was less than 20%. Among the ESBLs-producing E. coli in bloodstream infections in elderly patients, CTX-M-9 accounted for 27.91%, CTX-M-2 for 17.44%, and SHV for 13.95%. The detection rate of type I integrated genes was 41.30%, and type II and III integrated genes were not detected. ESBLs-producing genotyping-positive bacteria were detected with more than 50% of type I integrated genes. It was concluded that type I integrated genes in ESBLs-producing E. coli isolated from elderly patients carried resistance genes such as CTX-M-9 and CTX-M-2 aggravating multi-drug resistance in bacteria.202236227675
916170.9999Prospective multicentre study of rectal carriage of multidrug-resistant Enterobacteriaceae among health-care workers in Spain. OBJECTIVES: To investigate the rectal carriage of multidrug-resistant Enterobacteriaceae (colistin-resistant, extended-spectrum β-lactamase (ESBL) -producers and/or carbapenemase-producers) among health-care workers (HCWs) from six Spanish hospitals. METHODS: Rectal swabs from 258 HCWs, employed in intensive care units, haematology wards and clinical microbiology laboratories from six hospitals in northern Spain were studied. They were cultured in selective media for Gram-negative resistant bacteria. Detection of antimicrobial resistance genes and multilocus sequence typing were performed by PCR and further sequencing. A questionnaire including data related to risk factors of colonization/infection by resistant bacteria (age, gender, chronic diseases, immunosuppressive therapies, invasive procedures or antimicrobial treatments) was given to each participant. RESULTS: No carbapenemase-producing Enterobacteriaceae were recovered. However, 8/258 HCWs (3.1%) were positive for ESBL-producing isolates. This rate was not higher than the colonization rate previously reported in Spain for healthy people in the community. Five isolates showed high-level resistance to colistin (MICs ranging from 8 to 128 mg/L) but all of them were negative for the mcr genes tested. No statistically significant risk factors for gut colonization by ESBL-producing or colistin-resistant Enterobacteriaceae were identified among the HCWs participating in the study. CONCLUSIONS: Our data suggest that working in hospitals does not represent a risk for rectal carriage of multidrug-resistant Enterobacteriaceae.202031972320
860180.9999Investigation 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
1016190.9999Investigation of CTX-M Type Extended-Spectrum β-Lactamase, Carbapenem and Colistin Resistance in Enterobacterales Isolated From Dairy Cattle in Turkey. BACKGROUND: The increasing prevalence of antimicrobial resistance in animals, particularly the spread of multidrug-resistant Enterobacterales, poses a significant zoonotic and public health risk. OBJECTIVE: The aim of this study was to investigate extended-spectrum β-lactamase (ESBL), carbapenem and colistin resistance among Enterobacterales in faecal swabs of dairy cattle. METHODS: A total of 400 samples were cultured on Mac Conkey screening media for ESBL, carbapenem and colistin resistance. The grown Enterobacterales were identified by MALDI-TOF-MS, followed by ceftriaxone, cefotaxime and ceftazidime resistance and double disk synergy. ESBL resistance genes were identified by polymerase chain reaction (PCR) and Sanger sequencing. Bacteria grown on colistin screening media were investigated for colistin resistance by EUCAST microbroth dilution method. RESULTS: A total of 89 (22.25%) of the bacteria grown from 400 samples were identified as potential ESBL-producing Enterobacterales members. A number of 53 (59.5%) of them were identified as ESBL blaCTX-M as a result of PCR, and 10 of them were identified as blaCTX-M-15/28/36/66 as a result of sequencing. None of the samples cultured on carbapenem medium grew. A total of 18 samples grown in colistin medium were found to be colistin sensitive by broth microdilution. Genotypes were not included in the study. All isolated bacteria were identified as Escherichia coli. SOLUTION: In this study, blaCTX-M-15 and its derivatives, which are common in humans, were also found to be the predominant ESBL type in animals. Monitoring resistance in animals together with resistance in human infections may provide more important data on the spread of resistance.202540704983