Extended Spectrum Beta-Lactamase-Producing Gram-Negative Bacteria Recovered From an Amazonian Lake Near the City of Belém, Brazil. - Related Documents




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107501.0000Extended Spectrum Beta-Lactamase-Producing Gram-Negative Bacteria Recovered From an Amazonian Lake Near the City of Belém, Brazil. Aquatic systems have been described as antibiotic resistance reservoirs, where water may act as a vehicle for the spread of resistant bacteria and resistance genes. We evaluated the occurrence and diversity of third generation cephalosporin-resistant gram-negative bacteria in a lake in the Amazonia region. This water is used for human activities, including consumption after appropriate treatment. Eighteen samples were obtained from six sites in October 2014. Water quality parameters were generally within the legislation limits. Thirty-three bacterial isolates were identified as Escherichia (n = 7 isolates), Acinetobacter, Enterobacter, and Klebsiella (n = 5 each), Pseudomonas (n = 4), Shigella (n = 3), and Chromobacterium, Citrobacter, Leclercia, Phytobacter (1 isolate each). Twenty nine out of 33 isolates (88%) were resistant to most beta-lactams, except carbapenems, and 88% (n = 29) were resistant to antibiotics included in at least three different classes. Among the beta-lactamase genes inspected, the bla (CTX-M) was the most prevalent (n = 12 positive isolates), followed by bla (TEM) (n = 5) and bla (SHV) (n = 4). bla (CTX-M-15) (n = 5), bla (CTX-M-14) (n = 1) and bla (CTX-M-2) (n = 1) variants were detected in conserved genomic contexts: bla (CTX-M-15) flanked by ISEcp1 and Orf477; bla (CTX-M-14) flanked by ISEcp1 and IS903; and bla (CTX-M-2) associated to an ISCR element. For 4 strains the transfer of bla (CTX-M) was confirmed by conjugation assays. Compared with the recipient, the transconjugants showed more than 500-fold increases in the MICs of cefotaxime and 16 to 32-fold increases in the MICs of ceftazidime. Two isolates (Escherichia coli APC43A and Acinetobacter baumannii APC25) were selected for whole genome analysis. APC43A was predicted as a E. coli pathogen of the high-risk clone ST471 and serotype O154:H18. bla (CTX-M-15) as well as determinants related to efflux of antibiotics, were noted in APC43A genome. A. baumannii APC25 was susceptible to carbapenems and antibiotic resistance genes detected in its genome were intrinsic determinants (e.g., bla (OXA-208) and bla (ADC-like)). The strain was not predicted as a human pathogen and belongs to a new sequence type. Operons related to metal resistance were predicted in both genomes as well as pathogenicity and resistance islands. Results suggest a high dissemination of ESBL-producing bacteria in Lake Água Preta which, although not presenting characteristics of a strongly impacted environment, contains multi-drug resistant pathogenic strains.201930873145
104910.9998Multiple Antibiotic-Resistant, Extended Spectrum-β-Lactamase (ESBL)-Producing Enterobacteria in Fresh Seafood. Members of the family Enterobacteriaceae include several human pathogens that can be acquired through contaminated food and water. In this study, the incidence of extended spectrum β-lactamase (ESBL)-producing enterobacteria was investigated in fresh seafood sold in retail markets. The ESBL-positive phenotype was detected in 169 (78.60%) isolates, with Escherichia coli being the predominant species (53), followed by Klebsiella oxytoca (27), and K. pneumoniae (23). More than 90% of the isolates were resistant to third generation cephalosporins, cefotaxime, ceftazidime, and cefpodoxime. Sixty-five percent of the isolates were resistant to the monobactam drug aztreonam, 40.82% to ertapenem, and 31.36% to meropenem. Resistance to at least five antibiotics was observed in 38.46% of the isolates. Polymerase Chain Reaction (PCR) analysis of ESBL-encoding genes detected bla(CTX), bla(SHV), and bla(TEM) genes in 76.92%, 63.3%, and 44.37% of the isolates, respectively. Multiple ESBL genes were detected in majority of the isolates. The recently discovered New Delhi metallo-β-lactamase gene (bla(NDM-1)) was detected in two ESBL⁺ isolates. Our study shows that secondary contamination of fresh seafood with enteric bacteria resistant to multiple antibiotics may implicate seafood as a potential carrier of antibiotic resistant bacteria and emphasizes an urgent need to prevent environmental contamination and dissemination of such bacteria.201728867789
102520.9998Detection of Extended Spectrum Beta-Lactamases Resistance Genes among Bacteria Isolated from Selected Drinking Water Distribution Channels in Southwestern Nigeria. Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include bla TEM, bla SHV, and bla CTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with bla TEM being the most abundant (50/61) and bla CTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, bla SHV + bla TEM or bla CTX + bla TEM, were observed in 11 and 1 isolate, respectively. In conclusion, classic bla TEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria.201627563674
107930.9998CTX-M-15-producing Escherichia coli clone B2-O25b-ST131 and Klebsiella spp. isolates in municipal wastewater treatment plant effluents. OBJECTIVES: The global occurrence of antibiotic resistance genes in bacteria in water environments is an increasing concern. Treated wastewater was sampled daily over a 45 day period from the outflow of a municipal wastewater treatment plant in Brno, Czech Republic, and examined for extended-spectrum β-lactamase (ESBL)-producing bacteria. METHODS: Water samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) and individual colonies were examined for ESBL production. Phenotypic ESBL-positive bacteria identified as Escherichia coli or Klebsiella spp. were tested for the presence of antibiotic resistance genes, the virulence gene afa/dra and the bla(CTX-M) upstream region. Genetic relatedness was analysed by PFGE, multilocus sequence typing and plasmid analysis. RESULTS: A total of 68 ESBL-producing Enterobacteriaceae isolates were detected in 34 out of 45 wastewater samples. ESBL-producing isolates included 26 E. coli isolates, 4 Klebsiella pneumoniae isolates and 1 Klebsiella oxytoca isolate. The pandemic and multiresistant B2-O25b-ST131 clone was predominant, being detected among 19 E. coli isolates, and 17 of the B2-O25b-ST131 isolates were positive for the FIA replicon and the afa/dra operon and had an IS26 element flanking bla(CTX-M-15). Seventeen of the B2-O25b-ST131 isolates showed closely related PFGE profiles (defined by 84% band similarity) and belonged to identical clonal groups. CONCLUSIONS: The results highlight the inadequacy of the treatment process in removing multiresistant bacteria from municipal wastewater and point to a risk of transmission of clinically important multiresistant strains, such as the pandemic ST131 clone, to the environment. This is the first study demonstrating the pandemic ST131 clone in wastewater.201121954457
90240.9998Occurrence of IMP-8, IMP-10, and IMP-13 metallo-β-lactamases located on class 1 integrons and other extended-spectrum β-lactamases in bacterial isolates from Tunisian rivers. BACKGROUND: Antibiotic-resistant bacteria have been surveyed widely in water bodies, but few studies have determined the diversity of antibiotic-resistant bacteria in river waters. This study was undertaken to investigate the origin of resistance among polluted river bacterial isolates in Tunisia. METHODS: In this study 128 isolates resistant to β-lactam antibiotics were obtained from 2 polluted rivers in the north of Tunisia. Isolates were identified using Phoenix phenotyping criteria. The occurrence of bla(TEM), bla(SHV), bla(CTX-M), bla(CMY), bla(VIM), and bla(IMP) was studied by polymerase chain reaction (PCR) amplification and sequencing, and the genetic relatedness of the 16 IMP-producing Klebsiella pneumoniae isolates was analyzed by comparison of XbaI pulsed-field gel electrophoresis (PFGE) profiles. RESULTS: Using Phoenix phenotyping criteria, diverse genera of bacteria were identified with different rates of prevalence and with different minimum inhibitory concentrations against different antibiotics. The occurrence of bla(TEM), bla(SHV), bla(CTXM), bla(CMY), bla(VIM), and bla(IMP) genes was confirmed. The DNA sequences upstream and downstream of bla(IMP) genes were determined, revealing that all IMP-encoding genes constituted the first cassette of class 1 integrons, followed by aacA gene cassettes encoding aminoglycoside resistance. Comparison of PFGE profiles showed that only 2 of the isolates were clonal, the other 14 displaying unique profiles. The bla(CTX-M) gene was the most dominant of the extended-spectrum β-lactamase (ESBL) genes, while the bla(TEM) gene was the second-most dominant. CONCLUSION: The discovery of highly diverse ESBL-producing bacteria and metallo-β-lactamases, particularly bla(IMP), in polluted river water raises alarms with regard to the potential dissemination of antibiotic-resistant bacteria in communities through river environments.201322992193
99050.9998Resistance phenotype-genotype correlation and molecular epidemiology of Citrobacter, Enterobacter, Proteus, Providencia, Salmonella and Serratia that carry extended-spectrum β-lactamases with or without plasmid-mediated AmpC β-lactamase genes in Thailand. Extended-spectrum β-lactamases (ESBLs) and plasmid-mediated AmpC β-lactamases (pAmpCs) have been increasingly reported among less commonly encountered genera of Enterobacteriaceae. However, little is known regarding the genetic characteristics of resistance genes and epidemiology of these genera. Lack of accurate ESBL and pAmpC detection may adversely affect therapeutic outcomes. This study investigated resistance phenotype-genotype correlation and molecular epidemiology among six genera of Enterobacteriaceae (Citrobacter, Enterobacter, Proteus, Providencia, Salmonella and Serratia) that carried ESBL with or without pAmpC genes at a university hospital in Thailand. From a total of 562 isolates, 105 isolates (18.7%) had ESBL-positive phenotype whilst 140 isolates (24.9%) harboured one or more ESBL genes. CTX-M and TEM were common ESBL-related bla genes among these isolates. The sensitivity and specificity of ESBL phenotypic detection as opposed to ESBL gene detection were 70.7% and 98.6%, respectively. pAmpC genes were detected in 96 ESBL gene-carrying isolates (68.6%) and significantly caused false negative detection of ESBL. Molecular typing based on pulsed-field gel electrophoresis revealed several clones that may be endemic in this hospital. This study indicated a high prevalence of ESBLs and pAmpCs among less common members of the family Enterobacteriaceae in Thailand and these resistant bacteria need to be monitored.201120880563
102260.9997Characterization of Beta-lactamases in Faecal Enterobacteriaceae Recovered from Healthy Humans in Spain: Focusing on AmpC Polymorphisms. The intestinal tract is a huge reservoir of Enterobacteriaceae, some of which are opportunist pathogens. Several genera of these bacteria harbour intrinsic antibiotic resistance genes, such as ampC genes in species of Citrobacter, Enterobacter or Escherichia genera. In this work, beta-lactamases and other resistance mechanisms have been characterized in Enterobacteriaceae isolates recovered from healthy human faecal samples, focusing on the ampC beta-lactamase genes. Fifty human faecal samples were obtained, and 70 Enterobacteriaceae bacteria were isolated: 44 Escherichia coli, 4 Citrobacter braakii, 9 Citrobacter freundii, 8 Enterobacter cloacae, 1 Proteus mirabilis, 1 Proteus vulgaris, 1 Klebsiella oxytoca, 1 Serratia sp. and 1 Cronobacter sp. A high percentage of resistance to ampicillin was detected (57%), observing the AmpC phenotype in 22 isolates (31%) and the ESBL phenotype in 3 isolates. AmpC molecular characterization showed high diversity into bla CMY and bla ACT genes from Citrobacter and Enterobacter species, respectively, and the pulsed-field gel electrophoresis (PFGE) analysis demonstrated low clonality among them. The prevalence of people colonized by strains carrying plasmid-mediated ampC genes obtained in this study was 2%. The unique plasmid-mediated bla AmpC identified in this study was the bla CMY-2 gene, detected in an E. coli isolate ascribed to the sequence type ST405 which belonged to phylogenetic group D. The hybridization and conjugation experiments demonstrated that the ISEcp1-bla CMY-2-blc structure was carried by a ~78-kb self-transferable IncK plasmid. This study shows a high polymorphism among beta-lactamase genes in Enterobacteriaceae from healthy people microbiota. Extensive AmpC-carrier studies would provide important information and could allow the anticipation of future global health problems.201525501887
99170.9997Characterization of extended-spectrum beta-lactamases in Enterobacteriaceae causing nosocomial infections in a Zagreb University Hospital. The bacteria producing extended-spectrum beta-lactamases (ESBLs) are increasingly reported. production of ESBLs by Gram-negative bacteria is the major mechanism of resistance to oxymino-cephalosporins and aztreonam. the aim of the present study was to characterize ESBLs produced by Enterobacteriaceae, collected during 2003-2005 in a University Hospital in Zagreb, and to determine the risk factors associated with nosocomial infections due to them. 76 isolates of Enterobacteriaceae were included in the study. Antibiotic susceptibility testing was performed by disk-diffusion and broth microdilution method according to CLSI. beta-lactamases were characterized by PCR and sequencing of bla(ESBL )genes. plasmids were extracted by alkaline lysis method and digested with EcoRI enzyme. Most of the strains displayed CAZ phenotype meaning a higher level of resistance to ceftazidime compared to cefotaxime and ceftriaxone. 50 strains produced SHV-ESBL, 28 tem and 8 CTX-M beta-lactamase. Sequencing of bla(SHV )genes from representative strains revealed SHV-5 beta-lactamase in 6 strains whereas sequencing of bla(CTX-M )genes identified CTX-M-3 beta-lactamase in 3 and CTX-M-15 in 5 strains. Strains were assigned to groups from A to f according to plasmid fingerprinting. The spread of SHV-5-producing strains throughout the hospital units could be due to selective pressure of ceftazidime which is widely prescribed in our hospital thus favoring survival of strains possessing a mutation at the Ambler position 240 responsible for ceftazidime and aztreonam resistance.200919567348
95580.9997Occurrence and characteristics of extended-spectrum β-lactamase- and carbapenemase- producing bacteria from hospital effluents in Singapore. One of the most important resistance mechanisms in Gram-negative bacteria today is the production of enzymes causing resistance to cephalosporin and carbapenem antibiotics. The spread of extended-spectrum β-lactamases (ESBL)- and carbapenemase- producing Gram-negative bacteria is an emerging global public health problem. The aim of the present study was to (i) assess the prevalence of carbapenem-resistant bacteria (CRB) and ESBL-producing strains in sewage effluents from two major hospitals in Singapore, (ii) characterize the isolated strains and (iii) identify some of the ESBL and carbapenemase genes responsible for the resistance. CHROMagar ESBL and KPC plates were used to rapidly screen for ESBL-producing bacteria and those expressing reduced susceptibility to carbapenems, respectively. The abundance of ESBL-producers and CRB in hospital wastewater ranged between 10(3) and 10(6)CFU/mL. Out of the 66 isolates picked from ESBL and KPC plates, 95%, 82%, 82% and 76% were resistant to ceftriaxone, ceftazidime (3rd generation cephalosporin family), ertapenem and meropenem (carbapenem family), respectively. Among the resistant isolates, the most predominant taxa identified were Pseudomonas spp. (28.2%), Klebsiella spp. (28.2%), Enterobacter spp. (18.3%) and Citrobacter spp. (11.3%). PCR and sequencing analysis showed that the predominant β-lactamase genes were bla(SHV) (41.1%) followed by bla(NDM-1) (35.6%), bla(CTX) (35.6%) and bla(KPC) (28.8%). The results of this study show a high prevalence of bacteria resistant to modern extended-spectrum cephalosporins and carbapenems and the presence of ESBL- and carbapenemase producers in hospital effluents. These findings support the need to improve management of hospital wastewater in order to minimize the spread of antimicrobial resistant microorganisms from this source.201829751417
90390.9997Carbapenemase and ESBL genes with class 1 integron among fermenting and nonfermenting bacteria isolated from water sources from India. The present study was aimed to detect the carbapenemase, extended-spectrum β-lactamase (ESBL), and intI1 gene of class 1 integron among fermenting (n = 61) and nonfermenting (n = 10) bacterial isolates recovered from water samples (n = 128). Isolates were identified by 16S rRNA sequencing. These isolates showed reduced-susceptibility to third-generation cephalosporins and carbapenems. The isolates varied in number and size of plasmids (2 kb to >20 kb). Plasmid DNA screening showed 5·6, 7, 11·2 and 26·7% prevalence of bla(KPC) , bla(NDM) , bla(SHV) and bla(TEM) genes respectively. Diverse bla(NDM) (bla(NDM-1) and bla(NDM-4) ) and bla(SHV) subtypes (bla(SHV-2) and bla(SHV-11) ) were recorded, unlike the single allelic bla(KPC) (bla(KPC-2) ) and bla(TEM) (bla(TEM-1) ) gene. Of the total 27 bla-gene-producing bacterial isolates, seven isolates co-harboured the carbapenemase genes (bla(NDM) or bla(KPC) or the both) along with the ESBL genes (bla(SHV) or bla(TEM) ). The intI1 gene of class 1 integron was detected among 12 (44·4%) of ESBL- and/or carbapenemase-harbouring isolates. Gene transferability was seen among four of the 10 Enterobacteriaceae donors. Carbapenemases and ESBLs with class 1 integron among aquatic environmental isolates raise the serious issue of the biosecurity and health of the ecosystem. SIGNIFICANCE AND IMPACT OF THE STUDY: Anthropologically affected and polluted environment harbours the resistance threats, where a diverse bacterial species maintain, develop and exchange genetic determinants that constitute a risk to human and ecological health. The antimicrobial resistance (AMR) in Enterobacteriaceae and non-Enterobacteriaceae bacteria caused the failure of the therapy of last resort (carbapenems) and thus lead to life-threatening infections affecting public health. Surveillance and monitoring of AMR could be important for epidemiological, diagnostic testing and control of pathogens. This is a point-prevalence study reporting the comparative occurrence and co-occurrence of carbapenemase and extended-spectrum β-lactamase genes among fermenting and nonfermenting bacteria isolated from the aquatic environment in India.202031587338
901100.9997Emergence of plasmid-borne bla (oxa-181) gene in Ochrobactrum intermedium: first report from India. Wastewater has become a potential habitat for multi-drug-resistant bacteria. The present study aims to screen for the presence of carbapenem-resistant bacteria in sewage water samples collected from hospital and non-hospital sources. From a total of 19 sewage water samples collected, 100 carbapenem-resistant non-lactose-fermenting Gram-negative bacteria (CR-NF-GNB) were isolated using MacConkey agar cultured with 8 mg l(-1) of meropenem. On screening for beta-lactamase resistance genes (bla (NDM), bla (OXA-48-like), bla (IMP), bla (VIM) and bla (KPC)), one isolate, Ochrobactrum intermedium , was found to carry the plasmid-borne bla (OXA-48-like) gene. To the best of our knowledge, we provide the first report of the rare and emerging opportunistic pathogen Ochrobactrum intermedium encoding the OXA-181 gene in its plasmid.201932974517
1017110.9997Evaluation of canine raw food products for the presence of extended-spectrum beta-lactamase- and carbapenemase-producing bacteria of the order Enterobacterales. OBJECTIVE: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity. SAMPLES: A total of 200 canine raw food products. METHODS: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order. RESULTS: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%). CLINICAL RELEVANCE: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.202235895774
1070120.9997Survey for beta-lactamase among bacterial isolates from Guangzhou, China hospitals between 2005-2006. The beta-lactamase genes, which confer multi-drug resistance, are spreading among clinical bacterial isolates. As part of a routine surveillance program, we collected 302 bacilli isolates between June 2005 and October 2006 from four hospitals in Guangzhou, China. The isolates were screened for multidrug resistance and for the presence of beta-lactamases. In all, 80 isolates were identified as multidrug-resistant with the K-B method. These isolates were phenotypically screened for beta-lactamase activity by disk diffusion prescreening, diffusion confirmation, the cefoxitin three-dimensional test and the metallo-beta-lactamase (MBL) synergy test. Bacteria were genotypically screened for beta-lactamase genes by PCR and DNA sequencing. Among the 80 strains, drug resistance was lowest to amikacin (18.75%) and highest to ampicillin (97.50%), 26.49% had a beta-lactamase phenotype, 16.56% had the extended-spectrum beta-lactamase (ESBL) phenotype, 24.83% had a beta-lactamase genotype, 51 carried integrons, 30 carried class I integrons and 18.75% had the ISEcp1B insertion sequence. Sequencing also detected a new CTX-M ESBL gene subtype, which had an ISEcp1B element upstream of bla(CTX-M-Like), and an IS903 element downstream, forming a composite transposon. Multidrug resistance and beta-lactamases continue to be prevalent in Guangzhou. Our results suggest that resistance genes are evolving and being horizontally transmitted between species.201020339396
1072130.9997Characterization of carbapenem-resistant gram-negative bacterial isolates from Nigeria by whole genome sequencing. This study characterized the mechanisms of carbapenem resistance in gram-negative bacteria isolated from patients in Yola, Nigeria. Whole genome sequencing (WGS) was performed on 66 isolates previously identified phenotypically as carbapenem-non-susceptible. The patterns of beta-lactamase resistance genes identified were primarily species-specific. However, bla(NDM-7) and bla(CMY-4) were detected in all Escherichia coli and most Providencia rettgeri isolates; bla(NDM-7) was also detected in 1 Enterobacter cloacae. The E. coli and E. cloacae isolates also shared bla(OXA-1,) while bla(OXA-10) was found in all P. rettgeri, one Pseudomonas aeruginosa and 1 E. coli. Except for Stenotrophomonas maltophilia isolates, which only contained bla(L1), most species carried multiple beta-lactamase genes, including those encoding extended-spectrum beta-lactamases, AmpC and OXA in addition to a carbapenemase gene. Carbapenemase genes were either class B or class D beta-lactamases. No carbapenemase gene was detected by WGS in 13.6% of isolates.202134111650
1103140.9997Characterization of β-Lactamases and Multidrug Resistance Mechanisms in Enterobacterales from Hospital Effluents and Wastewater Treatment Plant. Antimicrobials in wastewater promote the emergence of antibiotic resistance, facilitated by selective pressure and transfer of resistant genes. Enteric bacteria belonging to Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Citrobacter species (n = 126) from hospital effluents and proximate wastewater treatment plant were assayed for susceptibility to four antimicrobial classes. The β-lactamase encoding genes harbored in plasmids were genotyped and the plasmids were sequenced. A multidrug resistance phenotype was found in 72% (n = 58) of E. coli isolates, 70% (n = 43) of Klebsiella species isolates, and 40% (n = 25) of Enterobacter and Citrobacter species. Moreover, 86% (n = 50) of E. coli, 77% (n = 33) of Klebsiella species, and 25% (n = 4) of Citrobacter species isolates phenotypically expressed extended spectrum β-lactamase. Regarding ESBL genes, bla(CTX-M-27) and bla(TEM-1) were found in E. coli, while Klebsiella species harbored bla(CTX-M-15), bla(CTX-M-30), or bla(SHV-12). Genes coding for aminoglycoside modifying enzymes, adenylyltransferases (aadA1, aadA5), phosphotransferases (aph(6)-1d, aph(3″)-Ib), acetyltransferases (aac(3)-IIa), (aac(6)-Ib), sulfonamide/trimethoprim resistant dihydropteroate synthase (sul), dihydrofolate reductase (dfrA), and quinolone resistance protein (qnrB1) were also identified. Monitoring wastewater from human sources for acquired resistance in clinically important bacteria may provide a cheaper alternative in regions facing challenges that limit clinical surveillance.202235740182
1051150.9997Multi-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
1038160.9997Isolation of Extended Spectrum β-lactamase (ESBL) Producing Bacteria from Urban Surface Waters in Malaysia. BACKGROUND: This was a preliminary study to test for the presence of multiple antibiotic-resistant extended spectrum β-lactamase (ESBL) producing bacteria in Malaysian urban surface waters. Although the literature review revealed several published papers on clinical ESBL isolates in Malaysia, none were found on ESBL isolates obtained from local surface waters. METHODS: Isolated bacterial species were tested for resistance to cefotaxime, amoxicillin/clavulanate and aztreonam, and susceptibility to imipenem and meropenem using antibiotic susceptibility testing (AST) by disc diffusion. This served as a screening step to detect bacteria that could be potential ESBL species. 16S ribose ribonucleic acid (rRNA) polymerase chain reaction (PCR) testing with two clusters of bla (β-lactamase) gene primers was used to test for the bla genes CTX-M (Groups 1, 2, 9), OXA-1, SHV and TEM. RESULTS: A total of 19 isolates were found, possessing at least one of the bla genes tested for. There was a relatively high occurrence of CTX-M genes (84.2%) among these, followed by TEM genes (47.4%). The isolates were identified as Enterobacteriaceae (89.5%), predominantly Escherichia coli and Klebsiella pneumoniae. CONCLUSION: There appears to be a high occurrence of ESBL-bacteria in local surface waters, among these being opportunistic pathogens. The persistence and spread of these species in the environment poses a threat to exposed human populations.201323966820
979170.9997Integrative phenotypic and genomic analysis of extended-spectrum Beta-lactamase (ESBL) and carbapenemase genes in Enterobacteriaceae and Pseudomonaceae strains isolated from animals in a Spanish Veterinary Teaching Hospital. Antimicrobial resistance (AMR) is a major global health threat, exacerbated by globalization which facilitates the spread of resistant bacteria. Addressing this issue requires a One Health perspective, involving humans, animals, and the environment. This study aims to compare the phenotypic resistance profiles of 69 clinical bacterial isolates (Enterobacteriaceae and Pseudomonaceae) from a Veterinary Teaching Hospital in Spain with their genotypic resistance profiles based on the presence of Extended-Spectrum Beta-Lactamases (ESBLs), AmpC and carbapenemases -enconding genes. For the genotypical analysis, whole genome sequencing (WGS) was used. Phenotypic characterization revealed that 37 isolates (53.6 %) grew on ESBL-selective medium. Phenotypic confirmatory tests showed that 12 strains (17.4 %) had some type of ESBL and 21 (30.4 %) could have an AmpC. Also, 24 isolates (34.8 %) grew in selective media for carbapenemases-producing bacteria, and 2 of these had a class A carbapenemase based on the KPC&MBL&OXA-48 disc kit. The genotypic analysis revealed 20 isolates (29 %) had bla(TEM), 8 (11.6 %) had bla(CTX-M) and 7 (10.1 %) bla(SHV). 27 (39.1 %) isolates had class C beta-lactamase genes. 35 isolates (50.7 %) had bla(OXA), class D beta-lactamase. 37 strains (53.6 %) had an Inc. plasmid replicon associated with the spread of AMR genes, including beta-lactamases and carbapenemases. This study emphasizes the value of combining phenotypic and genomic analyses to better understand and address antibiotic resistance, especially in veterinary contexts. Integrating these approaches enhances diagnostic accuracy by identifying strains with resistance genes that may not show phenotypically, helping clinicians in anticipating resistance under selective pressure.202539808975
1009180.9997The resistance patterns and molecular characteristics of ESBL/AmpC-producing Escherichia coli from captive panda ecosystem in China. Escherichia coli (E. coli) plays an important ecological role, and is a useful bioindicator to recognize the evolution of resistance in human, animal and environment. Recently, extended-spectrum β-lactamases (ESBL) producing E.coli has posed a threat to public health. Generally, captive healthy giant pandas are not exposed to antibiotics; however, they still acquire antimicrobial resistant bacteria. In order to understand whether there is an exchange of resistance genes within the ecosystems of captive giant pandas, this study explored resistance characteristics of 330 commensal E. coli isolates from feces of giant pandas, the surroundings, and breeders. Isolates from different sources showed similar resistance phenotype, and ESBL/AmpC-producing isolates showed more profound resistance to antibiotics than non-ESBL/AmpC-producing isolates (P<0.05). Furthermore, the occurrence of broad-spectrum β-lactamase related resistance genes and colistin resistance genes was detected, and isolates phylogenetic typing and multilocus sequence typing (MLST) were applied in this study. Seven different β-lactamase resistance genes (bla(CTX-M-55), bla(CTX-M-15), bla(CTX-M-27), bla(CTX-M-65), bla(TEM-1), bla(OXA-1) and bla(CMY)) and mcr-1 were found in 68 ESBL/AmpC-producing isolates. bla(CTX-M-55) (48.53 %) was found the most predominant resistance genes, followed by bla(TEM-1) (19.12 %) and bla(CTX-M-27) (16.18 %). Nonetheless, bla(CTX-M-55) was commonly detected in the isolates from giant pandas (63.16 %), the surroundings (43.48 %), and breeders (33.33 %). However, there were no carbapenemase genes detected in this study. mcr-1 was harbored in only one isolate from giant panda. Forty-five tansconjugants were successfully obtained in the conjugation experiments. The presence of antimicrobial resistance and related resistance genes tested were observed in the transconjugants. The results indicated that 52.63 % of the isolates from giant panda 73.91 % of the isolates from surroundings, and 100 % of the isolates from breeders were phylogroup A. Total of 27 sequence types (ST) were recognized from the isolate by MLST and found that ST48 (19/68; 27.94 %) was the predominant ST type, especially in the isolates from giant pandas and the surroundings. In conclusion, commensal ESBL/AmpC-producing E. coli becomes a reservoir of ESBL resistance genes, which is a potential threaten to health of giant pandas. The interaction between giant pandas, surroundings and breeders contribute to development of resistant phenotypes and genotypes which might transfer across species or the surroundings easily; hence, strict monitoring based on a "One Health" approach is recommended.202438728939
1074190.9997Extended-Spectrum β-Lactamase-Producing Klebsiella pneumoniae from Pharmaceutical Wastewaters in South-Western Nigeria. Emergence and spread of Klebsiella pneumoniae isolates producing extended-spectrum β-lactamases (ESBLs) present a major threat to public health. In this study, we characterized β-lactam-resistant K. pneumoniae isolates from six wastewater samples obtained from two pharmaceutical industries located in Lagos and Ogun States, Nigeria. Bacteria were isolated by using MacConkey agar; species identification and antibacterial susceptibility testing were performed by Vitek 2. Etest was used for ESBL phenotype confirmation. The presence of β-lactamase genes was investigated by PCR and sequencing. Bacterial strain typing was done by XbaI-macrorestriction and subsequent pulsed-field gel electrophoresis (PFGE) as well as multilocus sequence typing (MLST). Thirty-five bacterial species were isolated from the six samples; among them, we identified seven K. pneumoniae isolates with resistance to β-lactams and co-resistance to fluoroquinolones, aminoglycosides, and folate pathway inhibitors. The ESBL phenotype was confirmed in six K. pneumoniae isolates that harbored ESBL genes bla(CTX-M-15) (n = 5), bla(SHV-2) (n = 1), and bla(SHV-12) (n = 1). PFGE and MLST analysis revealed five clones belonging to four sequence types (ST11, ST15, ST37, ST101), and clone K. pneumoniae-ST101 was present in the wastewater samples from two different pharmaceutical industries. Additionally performed conjugation assays confirmed the location of β-lactamase genes on conjugative plasmids. This is the first confirmation of K. pneumoniae isolates producing CTX-M-15-ESBL from pharmaceutical wastewaters in Nigeria. The co-resistance observed might be a reflection of the different drugs produced by these industries. Continuous surveillance of the environmental reservoirs of multidrug-resistant bacteria is necessary to prevent their further spread.201728375698