Gram-negative bacteria as causative agents of ventilator-associated pneumonia and their respective resistance mechanisms. - Related Documents




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211801.0000Gram-negative bacteria as causative agents of ventilator-associated pneumonia and their respective resistance mechanisms. Ventilator-associated pneumonia (VAP) is a serious and common complication in patients admitted to intensive care unit (ICU) and contributes to mortality. Multidrug Gram-negative bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae are frequently associated with VAP in ICU. A prospective study was set up in three ICUs of the University Hospital Center Zagreb and one ICU in General Hospital Pula from September 2017 to March 2018. Antibiotic susceptibility was determined by broth microdilution method. Production of extended-spectrum β-lactamases (ESBLs) was determined by double-disk synergy test and carbapenemases by Hodge and carbapenem inactivation method (CIM). The genes encoding ESBLs, carbapenemases of class A, B and D and qnr genes were determined by PCR. In total 97 Gram-negative bacteria isolates were analyzed. P. aeruginosa demonstrated high resistance rates for imipenem and meropenem with 74% and 68% of resistant strains, respectively. Moderate resistance rates were observed for ceftazidime andpiperacillin/tazobactam, ciprofloxacin and gentamicin (44%). All except three A. baumannii isolates, were resistant to carbapenems and to all other antibiotics apart from colistin and amikacin. Eight A. baumannii isolates were positive for bla(OXA-23) and 12 for bla(OXA-24) genes. Four K. pneumoniae and two E. cloacae strains were ESBL positive and harboured group 1 of CTX-M β-lactamases. Three P. mirabilis strains were positive for plasmid-mediated ampC β-lactamase of CMY family. Two carbapenem-resistant K. pneumoniae harboured OXA-48 and one carbapenem-resistant E. cloacae VIM-1. A high proportion of multidrug-resistant P. aeruginosa, K. pneumoniae and extensively resistant A. baumannii was reported. Acquired resistance mechanisms, mainly production of carbapenemases and ESBLs were dominant in A. baumannii and K. pneumoniae, respectively. Resistance of P. aeruginosa isolates was more likely due to upregulation of efflux pumps or porin loss. A marked diversity of β-lactamases was identified in Enterobacteriaceae.202032729399
211910.9999Detection of bla(IMP) and bla(VIM) metallo-β-lactamases genes among Pseudomonas aeruginosa strains. Acquired Metallo-β-Lactamases (MBLs) are emerging resistance determinants in Pseudomonas aeruginosa and other gram-negative bacteria.Using Combination Disk Diffusion test, it was found that among 83 imipenem non-susceptible P. aeruginosa strains, 48 (57.9%) were MBL producers. PCR and Sequencing methods proved that these isolates were positive for blaIMP-1 genes, whereas none were positive for bla(VIM) genes. The mortality rate due to MBL-producing Pseudomonas infection was 4 (8.3%) among the hospitalized patients. Therefore, identification of drug resistance patterns in P. aeruginosa and detection of MBLs producing isolates are of great importance in the prevention and control of infections.201323638331
212120.9999Investigation of VIM, IMP, NDM-1, KPC AND OXA-48 enzymes in Enterobacteriaceae strains. Gram-negative bacteria especially Enterobacteriaceae species have become an increasing etiologic agent of nosocomial infections. The development of resistance to carbapenems have become an increasing problem in the treatment of nosocomial infections. Especially carbapenamases are common for Enterobacteriaceae strains. This study was performed to detect the types of carbapenemases in Enterobacteriaceae strains isolated from various clinical samples. Enterobacteriaceae species were isolated from urine, blood, tracheal aspirates, wound, and other respiratory samples. Susceptibility of isolates to imipenem, meropenem and ertapenem was tested. Carbapenemase genes were studied using HyplexSuperBug ID kit. VIM (1-13), IMP (1-22), NDM-1, KPC(1-10) and OXA-48 genes were investigated. Ninety-five isolates of Enterobacteriaceae spp. were included in the study. Sixty isolates were resistant to imipenem, meropenem and ertapenem and 20 isolates were found resistant to imipenem or ertapenem while 15 were susceptible to all carbapenems. Among the isolates with carbapenem resistance, 57 were positive for one carbapenemase gene and susceptible isolates did not have carbapenemase gene. OXA-48 was found in 49 of the isolates (86%), NDM-1 in 6 (10.5%) isolates, VIM in 2 isolates. IMP and KPC gene loci were not identified. Carbapenemase genes play a crucial role in the development and spread of resistant strains.201526051720
212030.9999Antimicrobial Resistance Patterns of Gram-negative Bacteria in an Iranian Referral Pediatric Hospital: A Present Danger of New Delhi Metallo-β- lactamase. BACKGROUND: Antimicrobial resistance among gram-negative bacteria has been growing, particularly in developing countries, like Iran. The emergence and spread of carbapenem-resistance mechanisms is a major public health concern because no definite treatments have yet been established for this problem. This study aimed to evaluate antibiotic susceptibility of gram-negative bacteria, metallo-β-lactamases (MBLs) and carbapenemase-producing genes, including bla (NDM), bla (VIM), and bla (IMP) in patients referred to Children's Medical Center, Tehran, Iran. MATERIAL AND METHODS: In this cross-sectional study, a total of 944 gram-negative isolates were tested in the study, and antimicrobial susceptibility testing was performed. Moreover, MBL production of carbapenem-resistant isolates, as well as the presence of bla (NDM), bla (VIM), and bla (IMP), was investigated. RESULTS: The most common gram-negative isolated bacteria were Escherichia coli (489 samples, 52%), followed by Klebsiella pneumoniae (167 samples, 18%), Pseudomonas aeruginosa (101 samples, 11%), Enterobacter spp. (64 samples, 7%), Pseudomonas spp. (35 samples, 4%), Acinetobacter baumannii (18 samples, 2%), and Burkholderia cepacia (17 samples, 2%). Imipenemresistant was found in 75%, 61%, and 60% of Stenotrophomonas maltophilia, Enterobacter spp., and A. baumannii isolates, respectively. Moreover, the highest resistance to meropenem was observed in S. maltophilia, A. baumannii, P. aeruginosa, and B. cepacia (100%, 96%, 83%, and 61.5%, respectively). Double disk synergy test (DDST) results showed that 112 out of 255 carbapenem- resistant isolates (44%) were MBL-producing ones. The presence of the bla (NDM) gene was identified in 32 (29%) of MBL-producing isolates, 13 of which were K. pneumoniae, 7 P. aeruginosa, and 7 E. coli, 3 Enterobacter spp., and 2 Klebsiella spp., respectively. The presence of the bla (IMP) and bla (VIM) genes was detected in 2 (2%) and 1 (1%) of MBL-producing isolates. These genes were detected in only MBL-producing P. aeruginosa isolates. CONCLUSION: Our findings suggest the emergence of NDM-producing strains in our hospital, and bla NDM was the most frequently detected carbapenemase gene in MBL-producing P. aeruginosa, K. pneumoniae, and Klebsiella spp. Since such bacteria can easily spread among patients in the hospital, a strong infection control and prevention plan is highly recommended.202337106518
143140.9999The using of the polymerase chain reaction for the detection of resistance genes in gram-negative bacteria in routine practice in a pediatric hospital. Objective - assessment of RT-PCR for the detection of carbapenem-resistance genes in gram-negative bacteria. A total, 499 strains of gram-negative microorganisms isolated in two pediatric hospitals in 2019-2020 were studied. Species identification was performed using MALDI-ToF mass-spectrometry (Bruker Daltonics, Germany). Meropenem and imipenem minimal inhibitory concentration (MIC) was determined by E-test method (BioMerieux, France). The presence of acquired carbapenemase genes of IMP, NDM, VIM, KPC, OXA-48, OXA-23, OXA-40, OXA-58-groups was determined by RT-PCR. Klebsiella pneumoniae (34%), Escherichia coli (4%), Serratia marcescens (6%) and other members of Enterobacterales (6%), also gram-negative non-glucose-fermenting bacteria Acinetobacter baumannii (14%), Pseudomonas aeruginosa (36%) were found among selected strains. Carbapenemase production was found in 385 isolates (77%). The main mechanism determining carbapenem resistance in P. aeruginosa was the production of blaVIM (100%). A. baumanii strains harbored OXA-23 (55%) and OXA-40 (45%) carbapenemases. The major determinant of carbapenem resistance in K. pneumoniae isolates was OXA-48 carbapenemase, detected in 63% strains, 13% of the strains possessed blaNDM-group, 16% isolates had a combination of blaNDM-group and blaOXA-48-like. Carbapenemase of KPC-group was found in 8% K. pneumoniae strains. OXA-48 carbapenemase prevailed (95%) among S. marcescens strains. Most of E. coli isolates harbored metallo-beta-lactamase NDM (89%). Other members of Enterobacterales most often had OXA-48 carbapenemase (57%), 39% of the isolates carried blaNDM-group. In one strain, a combination of blaNDM-group and blaOXA-48-like was discovered. RT-PCR is a fast and reliable method for the detection of acquired carbapenemases and can be recommended for routine use in bacteriological laboratories.202235320635
212650.9999Carbapenemase genes among multidrug resistant gram negative clinical isolates from a tertiary hospital in Mwanza, Tanzania. The burden of antimicrobial resistance (AMR) is rapidly growing across antibiotic classes, with increased detection of isolates resistant to carbapenems. Data on the prevalence of carbapenem resistance in developing countries is limited; therefore, in this study, we determined the prevalence of carbapenemase genes among multidrug resistant gram negative bacteria (MDR-GNB) isolated from clinical specimens in a tertiary hospital in Mwanza, Tanzania. A total of 227 MDR-GNB isolates were analyzed for carbapenem resistance genes. For each isolate, five different PCR assays were performed, allowing for the detection of the major carbapenemase genes, including those encoding the VIM-, IMP-, and NDM-type metallo-beta-lactamases, the class A KPC-type carbapenemases, and the class D OXA-48 enzyme. Of 227 isolates, 80 (35%) were positive for one or more carbapenemase gene. IMP-types were the most predominant gene followed by VIM, in 49 (21.59%) and 28 (12%) isolates, respectively. Carbapenemase genes were most detected in K. pneumoniae 24 (11%), followed by P. aeruginosa 23 (10%), and E. coli with 19 isolates (8%). We have demonstrated for the first time a high prevalence of MDR-GNB clinical isolates having carbapenem resistance genes in Tanzania. We recommend routine testing for carbapenem resistance among the MDR-GNB particularly in systemic infections.201424707481
212760.9999Molecular characterization of carbapenem-resistant Klebsiella pneumoniae in a tertiary university hospital in Turkey. The aim of this study was to identify the resistance genes and genetic relationship of carbapenemase-resistant Klebsiella pneumoniae (CRKP) identified in a tertiary university hospital in Turkey. During the study, CRKP was isolated from 137 patients. Resistance genes were studied in 94 isolates. Among these isolates, most of the CRKP produced only oxacillinase (OXA)-48 (91.5%); however, 4.3% of the isolates produced only New Delhi metallo-beta-lactamase 1 (NDM-1), 1% produced both OXA-48 and NDM-1, and 3.2% produced imipenem. This study adds Turkey to the growing list of countries with NDM-1-producing bacteria and shows that NDM-1 may easily spread worldwide.201323623803
99870.9998Extended spectrum beta-lactamases among Gram-negative bacteria of nosocomial origin from an intensive care unit of a tertiary health facility in Tanzania. BACKGROUND: Resistance to third generation cephalosporins due to acquisition and expression of extended spectrum beta-lactamase (ESBL) enzymes among Gram-negative bacteria is on the increase. Presence of ESBL producing organisms has been reported to significantly affect the course and outcome of an infection. Therefore infections due to ESBL isolates continue to pose a challenge to infection management worldwide. The aim of this study was to determine the existence and to describe phenotypic and genotypic characteristics of ESBLs in an Intensive Care Unit (ICU) setting in Tanzania. METHODS: Between October 2002 and April 2003, clinical information and samples were collected from patients suspected to have nosocomial infections in an Intensive Care Unit of a tertiary hospital in Tanzania. The isolates were identified, tested for antimicrobial susceptibility and analysed for presence of ESBL genes. RESULTS: Thirty-nine Gram-negative bacteria were isolated from clinical samples of 39 patients. These isolates included 13 Escherichia coli, 12 Enterobacter spp, 5 Pseudomonas spp, 4 Proteus spp, 2 Klebsiella. pneumoniae, 2 Citrobacter freundii and 1 Chryseomonas luteola. Eleven (28.2%) of these isolates were ESBL producing. The ESBL genes characterised were SHV-12, SHV-28 and CTX-M-15. The ESBL producing isolates were more resistant to gentamicin and ciprofloxacin than non-ESBL producing isolates. CONCLUSION: This study shows the presence of ESBL genes among Gram-negative bacteria in the ICU setting in Tanzania. There is a need to institute strict hospital infection control policy and a regular surveillance of resistance to antimicrobial agents.200516225701
143080.9998Prevalence of multidrug-resistant Gram-negative bacteria from blood cultures and rapid detection of beta-lactamase-encoding genes by multiplex PCR assay. INTRODUCTION: This study aimed to determine the prevalence of multidrug-resistant Gram-negative bacteria (GNB) from blood cultures in a tertiary-care hospital and the multiplex PCR assay's ability to detect resistance genes. METHODS: A total of 388 GNB isolates obtained from hospitalized patients between November 2019 and November 2021 were included in the study. Antimicrobial susceptibility testing was done by VITEK 2 system and broth microdilution method. Beta-lactamase-encoding genes were detected by multiplex PCR assays, BioFire-Blood Culture Identification 2 (BCID2) panel (bioMérieux, France). Extended-spectrum beta-lactamases (ESBLs) were detected phenotypically with VITEK AST-GN71 card (bioMérieux, France). The isolates of GNB were classified into multidrug-resistant, extensively-drug-resistant, and pandrug-resistant categories, and their prevalence and distribution in different wards, including coronavirus diseases 2019 (COVID-19) intensive care units (ICU), were calculated. RESULTS: Results revealed that all isolates of Acinetobacter baumannii and Pseudomonas aeruginosa were multidrug-resistant as well as 91.6% of Enterobacter cloacae, 80.6% of Proteus mirabilis, and 76.1% of Klebsiella pneumoniae, respectively. In fermentative bacteria, bla(OXA-48-like) (58.1%), bla(NDM) (16.1%), bla(KPC) (9.7%) and bla(VIM) (6.5%) genes were detected. More than half of Enterobacter cloacae (58.3%) and Klebsiella pneumoniae (53.7%) produced ESBLs. Among non-fermenters, the bla(NDM) gene was carried by 55% of Pseudomonas aeruginosa and 19.5% of Acinetobacter baumannii. In the COVID-19 ICU, Acinetobacter baumannii was the most common isolate (86.1%). CONCLUSIONS: This study revealed high proportions of multidrug-resistant blood isolates and various underlying resistance genes in Gram-negative strains. The BCID2 panel seems to be helpful for the detection of the most prevalent resistance genes of fermentative bacteria.202238021186
212590.9998Emergence of Carbapenem-Resistant Gram-Negative Isolates in Hospital Settings in Djibouti. Introduction: The antimicrobial resistance (AMR) of bacteria is increasing rapidly against all classes of antibiotics, with the increasing detection of carbapenem-resistant isolates. However, while growing prevalence has been reported around the world, data on the prevalence of carbapenem resistance in developing countries are fairly limited. In this study, we investigated and determined the resistance rate to carbapenems among multidrug-resistant Gram-negative bacteria (MDR-GNB) isolated in Djibouti and characterized their resistance mechanisms. Results: Of the 256 isolates, 235 (91.8%) were identified as Gram-negative bacteria (GNB). Of these GNBs, 225 (95.7%) isolates exhibited a multidrug resistance phenotype, and 20 (8.5%) isolates were resistant to carbapenems, including 13 Escherichia coli, 4 Acinetobacter baumannii, 2 Klebsiella pneumoniae and 1 Proteus mirabilis. The most predominant GNB in this hospital setting were E. coli and K. pneumoniae species. Carbapenemase genes such as bla(OXA-48) and bla(NDM-5) were identified, respectively, in six and four E. coli isolates, whereas the carbapenemase bla(NDM-1) was identified in three E. coli, two K. pneumoniae, one P. mirabilis and one A. baumannii. Moreover, three A. baumannii isolates co-hosted bla(OXA-23) and bla(NDM-1). Materials and Methods: A total of 256 clinical strains collected between 2019 and 2020 were identified using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF). Antibiotic susceptibility testing was performed using disk diffusion and E-test methods. Real-time polymerase chain reaction (RT-PCR), standard PCR and sequencing were used to investigate genes encoding for extended-spectrum-β-lactamases, carbapenemases and colistin resistance genes. Conclusions: We report, for the first time, the presence of MDR-GNB clinical isolates and the emergence of carbapenem-resistant isolates in Djibouti. In addition to performing antimicrobial susceptibility testing, we recommend phenotypic and molecular screening to track the spread of carbapenemase genes among clinical GNB isolates.202337508230
2122100.9998Phenotypic and genotypic characteristics of carbapenemase- and extended spectrum β-lactamase-producing Klebsiella pneumoniae ozaenae clinical isolates within a hospital in Panama City. Klebsiella pneumoniae spp ozaenae is a versatile bacterial species able to acquire antimicrobial resistance; the species presents a higher antimicrobial resistance profile compared to Klebsiella pneumoniae spp pneumoniae. Carbapenemase and extended spectrum β-lactamase (ESBL)-producing bacteria commonly arise in clinical settings where antimicrobial stewardship is limited. Our study aims to report the phenotypical and genetic characteristics of nosocomial Klebsiella pneumoniae spp ozaenae isolates associated with mortality collected from a tertiary-level hospital in Panama City. In October 2020, 11 consecutive multidrug-resistant Gram-negative isolates were recovered from secretions and blood cultures from hospitalized patients. Nearly 90% (10/11) of these patients died, and bacteria was obtained from six patients for investigation. Biochemical evaluation of the six isolates revealed the presence of multidrug-resistant Klebsiella pneumoniae spp ozaenae. Phenotypic evaluation indicated resistance to carbapenemase and EBSL. In contrast, genetic evaluation by PCR showed that only 30% (2/6) were resistant to CTX-M-1 (CTX-M group 1), whereas 60.7% (4/6) presented carbapenemase resistance genes, and 33.3% (2/6) presented New Delhi metallo-β-lactamase (NDM) resistance genes. Klebsiella pneumoniae ST258 was identified in 83.3% (5/6) of the isolates. Phylogenetic analysis using 16S revealed low homology among the six isolates. These results suggest that antibiotic resistance genes may have been incorporated into these Klebsiella pneumoniae spp ozaenae isolates within the hospital environment. We recommend strengthening the antimicrobial stewardship program and antibiotic control policy, as well as heightened infection control and prevention measures, such as ward sanitation and increased hand washing frequency.202134733509
2124110.9998Evaluation 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
922120.9998Insertion 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
837130.9998Diversity of Carbapenem Resistance Mechanisms in Clinical Gram-Negative Bacteria in Pakistan. Antibiotic resistance is a health challenge worldwide. Carbapenem resistance in Gram-negative bacteria is a major problem since treatment options are very limited. Tigecycline and colistin are drugs of choice in this case, but resistance to these drugs is also high. The aim of this study was to describe the diversity of resistance mechanisms in carbapenem-resistant clinical Gram-negative bacteria from Pakistan. Carbapenem-hydrolyzing enzyme-encoding genes were detected using PCR and DNA sequencing and clonal types determined by multilocus sequence typing (MLST). Forty-four carbapenem-resistant isolates were collected from the microbiology laboratory of Fauji Foundation Hospital and Al-Syed Hospital, Rawalpindi, Pakistan, including Klebsiella spp., Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae, and Achromobacter xylosoxidans. bla(NDM-1), bla(NDM-4,) bla(NDM-5,) bla(NDM-7), bla(OXA-48), and bla(OXA-181) were detected in Enterobacteriaceae; bla(OXA-23,) bla(OXA-72), and bla(NDM-1) in A. baumannii, and bla(VIM-6) and bla(VIM-11) in P. aeruginosa. MLST analysis revealed several predominant clonal types: ST167 in E. coli, ST147 in Klebsiella pneumoniae, ST2 in Acinetobacter, and ST664 in P. aeruginosa. In Acinetobacter, a new clonal type was observed for the first time. To the best of our knowledge, this is the first study describing the clonality and resistance mechanisms of carbapenem-resistant Gram-negative bacteria in Pakistan.202133211640
2123140.9998Phenotypic and genotypic detection of resistance mechanisms in carbapenem-resistant gram-negative bacteria isolated from Egyptian ICU patients with first emergence of NDM-1 producing Klebsiella oxytoca. BACKGROUND AND OBJECTIVES: Carbapenems are considered the last resort to treat several infections, particularly in intensive care units (ICUs). However, increasing carbapenem resistance is problematic because it leads to high morbidity and mortality rates. This study aimed to determine the rate of carbapenem resistance among Gram-negative bacteria collected from patients in ICUs and to identify their resistance mechanisms using phenotypic and genotypic methods. MATERIALS AND METHODS: Antimicrobial susceptibility testing was carried out using the disc diffusion method among 180 Gram-negative bacterial isolates. Productions of carbapenemases, metallo-beta-lactamases (MBLs) and the harboring of carbapenemase-encoding genes, were detected in 40 selected carbapenem-resistant Gram-negative bacteria (CR-GNB). RESULTS: Of 40 selected CR-GNB isolates, 28 (70%), and 20 (50%) isolates were phenotypically positive for carbapenemase, and MBL production, respectively. Furthermore, 22 (55%) showed amplification of one or more of the carbapenemase-encoding genes, including bla (NDM-1), bla (VIM-2), and bla (OXA-48). This study described the first emergence of NDM-1 producing Klebsiella oxytoca in Egyptian ICUs. CONCLUSION: High incidence of CR-GNB detected in the ICUs in our study area may be attributed to the overuse of antibiotics, including carbapenems, and improper application of infection control measures. These findings confirm the need for the application of a strict antibiotic stewardship program.202236721446
992150.9998Phenotypic and genotypic evaluation of beta-lactamases (ESBL and KPC) among enterobacteria isolated from community-acquired monomicrobial urinary tract infections. Beta-lactamases enzymes such as extended-spectrum beta-lactamases (ESBL) and carbapenemase type beta-lactamases (KPC) confer resistance to beta-lactam drugs among Gram-negative rods, mainly Enterobacteriaceae, as those frequently related to urinary tract infections (UTI). The aim of this study was to evaluate ESBL and KPC among enterobacteria isolated from monomicrobial UTI and to establish correlations between the presence of genetic markers and the phenotypic resistance to beta-lactam antibiotics. Out of 12 304 urine samples collected during 2009, 93 enterobacteria showing an ESBL phenotype were recovered. Imipenem was used for KPC screening and modified disk approximation assay was used for detection of ESBL phenotype. Polymerase chain reaction was used for screening of bla(SHV), bla(TEM), bla(CTX-M), and bla(KPC). Considering the isolated bacteria showing ESBL phenotype 56% of the isolates were positive for two genes. The bla(TEM) was the most frequent (87·1%). Neither KPC phenotype nor bla(KPC)-harboring bacteria were observed. Monitoring the antimicrobial resistance is extremely important to sustain empirical therapy of community-acquired urinary tract infections (Co-UTI).201424621159
1435160.9998Epidemiology, Phenotypic and Genotypic Characterization of Carbapenem-Resistant Gram-Negative Bacteria from a Libyan Hospital. Antimicrobial resistance, particularly resistance to carbapenems, has become one of the major threats to public health. Seventy-two isolates were collected from patients and hospital environment of Ibn Sina Hospital, Sirte, Libya. Antibiotic susceptibility tests, using the disc diffusion method and E-Test strips, were performed to select carbapenem-resistant strains. The colistin (CT) resistance was also tested by determining the minimum inhibitory concentration (MIC). RT-PCR was conducted to identify the presence of carbapenemase encoding genes and plasmid-mediated mcr CT resistance genes. Standard PCR was performed for positive RT-PCR and the chromosome-mediated CT resistance genes (mgrB, pmrA, pmrB, phoP, phoQ). Gram-negative bacteria showed a low susceptibility to carbapenems. Molecular investigations indicated that the metallo-β-lactamase New Delhi metallo-beta-lactamases-1 was the most prevalent (n = 13), followed by Verona integron-encoded metallo-beta-lactamase (VIM) enzyme (VIM-2 [n = 6], VIM-1 [n = 1], and VIM-4 [n = 1]) that mainly detected among Pseudomonas spp. The oxacillinase enzyme OXA-23 was detected among six Acinetobacter baumannii, and OXA-48 was detected among one Citrobacter freundii and three Klebsiella pneumoniae, in which one coharbored the Klebsiella pneumoniae carbapenemase enzyme and showed resistance to CT (MIC = 64 μg/mL) by modification in pmrB genes. In this study, we report for the first time the emergence of Pseudomonas aeruginosa carrying the bla(NDM-1) gene and belonging to sequence type773 in Libya. Our study reported also for the first time CT resistance by mutation in the pmrB gene among Enterobacteriaceae isolates in Libya.202337145891
997170.9998Prevalence and antibacterial resistance patterns of extended-spectrum beta-lactamase producing Gram-negative bacteria isolated from ocular infections. PURPOSE: Extended-spectrum beta-lactamases (ESBLs) mediated resistance is more prevalent worldwide, especially among Gram-negative bacterial isolates, conferring resistance to the expanded spectrum cephalosporins. As limited data were available on the prevalence of ESBLs in this area, the current study was undertaken to determine the prevalence, antibacterial resistance patterns, and molecular detection and characterization of ESBL encoding resistance genes among ocular Gram-negative bacterial isolates from ocular infections. MATERIALS AND METHODS: A prospective study was done on 252 ocular Gram-negative bacterial isolates recovered from ocular infections during a study period from February 2011 to January 2014. All isolates were subjected to detection of ESBLs by cephalosporin/clavulanate combination disc test and their antibacterial resistance pattern was studied. Molecular detection and characterization of ESBL encoding blaTEM -, blaSHV , blaOXA -, and blaCTX-M (phylogenetic groups 1, 2, 9, and 8/25) resistance genes by multiplex polymerase chain reaction and DNA sequence analysis. RESULTS: Of all Gram-negative bacteria, Pseudomonas aeruginosa (44%) was the most common strain, followed by Enterobacter agglomerans and Klebsiella pneumoniae each (10%). Among the 252, 42 (17%) were ESBL producers. The major source of ESBL producers were corneal scraping specimens, highest ESBL production was observed in P. aeruginosa 16 (38%) and Escherichia coli 7 (16.6%). Among ESBL-producing genes, the prevalence of blaTEM -gene was the highest (83%) followed by blaOXA -gene (35%), blaSHV -gene (18.5%), and blaCTX-M-1 -gene (18.5%) alone or together. CONCLUSION: The higher rate of prevalence of ESBLs-encoding genes among ocular Gram-negative bacteria is of great concern, as it causes limitation to therapeutic options. This regional knowledge will help in guiding appropriate antibiotic use which is highly warranted.201627221683
996180.9998Rapid Detection of New Delhi Metallo-β-Lactamase Gene Using Recombinase-Aided Amplification Directly on Clinical Samples From Children. New Delhi metallo-β-lactamase, a metallo-β-lactamase carbapenemase type, mediates resistance to most β-lactam antibiotics including penicillins, cephalosporins, and carbapenems. Therefore, it is important to detect bla (NDM) genes in children's clinical samples as quickly as possible and analyze their characteristics. Here, a recombinase-aided amplification (RAA) assay, which operates in a single one-step reaction tube at 39°C in 5-15 min, was established to target bla (NDM) genes in children's clinical samples. The analytical sensitivity of the RAA assay was 20 copies, and the various bacterial types without bla (NDM) genes did not amplify. This method was used to detect bla (NDM) genes in 112 children's stool samples, 10 of which were tested positive by both RAA and standard PCR. To further investigate the characteristics of carbapenem-resistant bacteria carrying bla (NDM) in children, 15 carbapenem-resistant bacteria (Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Citrobacter freundii, Klebsiella oxytoca, Acinetobacter junii, and Proteus mirabilis) were isolated from the 10 samples. Notably, more than one bacterial type was isolated from three samples. Most of these isolates were resistant to cephalosporins, cefoperazone-sulbactam, piperacillin-tazobactam, ticarcillin-clavulanic acid, aztreonam, co-trimoxazole, and carbapenems. bla (NDM) (-) (1) and bla (NDM) (-) (5) were the two main types in these samples. These data show that the RAA assay has potential to be a sensitive and rapid bla (NDM) gene screening test for clinical samples. The common existence of bla (NDM) and multi-drug resistance genes presents major challenges for pediatric treatment.202134367092
1432190.9998Prevalence of difficult-to-treat resistance in ESKAPE pathogens in a third level hospital in Mexico. BACKGROUND: Antimicrobial resistance and difficult-to-treat resistance (DTR) in ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) is a threat to human health. The aim of this study was to determine the prevalence of antimicrobial resistance and DTR rates in ESKAPE pathogens over six years in a third-level hospital from Monterrey, Mexico. METHODS: Antimicrobial susceptibility testing was determined by either disk diffusion or broth microdilution in strains from 2018 to 2023. Isolates were screened for carbapenemase genes. Multidrug resistance (MDR), extensively drug resistance (XDR), carbapenem resistance (CR), extended-spectrum cephalosporin-resistance (ESCR), fluoroquinolone resistance (FQR), and DTR were determined. RESULTS: From 3,239 strains, 48.5% were from respiratory infections, resistance was 87.5% to meticillin in Staphylococcus spp. and 39.8% in S. aureus, and 13.9% to vancomycin in Enterococcus spp. MDR, FQR and ESCR rates were between 54-90% in A. baumannii, 20-60% in Enterobacterales and 17-25% in P. aeruginosa. CR was 85.7% in A. baumannii, 33.3% in P. aeruginosa and <5% in Enterobacterales. Most frequent CR genes were OXA-24/40-like in A. baumannii and NDM and OXA-48 in carbapenem-resistant Enterobacterales. DTR rates were 59.7% in A. baumannii (49.2% in 2018 vs 62.9% in 2023), 8.9% in P. aeruginosa and <3% in Enterobacterales. XDR in A. baumannii was 14.4%. CONCLUSIONS: Antimicrobial resistance rates were high in Gram-negative pathogens. CR and DTR rates were higher in A. baumannii than P. aeruginosa and Enterobacterales. DTR surveillance in healthcare providers should be continuous updating local and regional DTR trends among Gram-negative bacteria.202539758683