Molecular characterization of extended-spectrum β-lactamases in clinical Escherichia coli and Klebsiella pneumoniae isolates from Surabaya, Indonesia (original) (raw)
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Journal of Antimicrobial Chemotherapy, 2010
The presence of broad-spectrum-cephalosporin-resistant Escherichia coli isolates and the implicated mechanisms of resistance and virulence factor genes were investigated in red fox (Vulpes vulpes) in Portugal. Cefotaxime-resistant E. coli isolates were isolated from two of 52 fecal samples (4 %), being both ESBL producers. The b-lactamase genes found in the two isolates were bla SHV-12 ? bla TEM-1b . The tet(A) and sul2 genes were also detected in these isolates, together with the non-classical class 1 integron (intI1-dfrA12-orfF-aadA2-cmlA1-aadA1-qacH-IS440-sul3) with the PcH1 promoter. The two isolates belonged to the phylogroup A. Amino acid changes in GyrA (S83L ? D87G) and ParC (S80I) proteins were identified in our study. Concerning MLST typing, both isolates were assigned to ST1086, never found before in wild animals, and they presented closely related PFGE patterns. This study reveals the presence of ESBL-producing E. coli isolates, in a wild ecosystem, which could be disseminated through the environment to other niches.
Journal of Medical Microbiology, 2010
The presence of broad-spectrum-cephalosporin-resistant Escherichia coli isolates and the implicated mechanisms of resistance and virulence factor genes were investigated in red fox (Vulpes vulpes) in Portugal. Cefotaxime-resistant E. coli isolates were isolated from two of 52 fecal samples (4 %), being both ESBL producers. The b-lactamase genes found in the two isolates were bla SHV-12 ? bla TEM-1b . The tet(A) and sul2 genes were also detected in these isolates, together with the non-classical class 1 integron (intI1-dfrA12-orfF-aadA2-cmlA1-aadA1-qacH-IS440-sul3) with the PcH1 promoter. The two isolates belonged to the phylogroup A. Amino acid changes in GyrA (S83L ? D87G) and ParC (S80I) proteins were identified in our study. Concerning MLST typing, both isolates were assigned to ST1086, never found before in wild animals, and they presented closely related PFGE patterns. This study reveals the presence of ESBL-producing E. coli isolates, in a wild ecosystem, which could be disseminated through the environment to other niches.
Archives of Microbiology, 2012
The presence of broad-spectrum-cephalosporin-resistant Escherichia coli isolates and the implicated mechanisms of resistance and virulence factor genes were investigated in red fox (Vulpes vulpes) in Portugal. Cefotaxime-resistant E. coli isolates were isolated from two of 52 fecal samples (4 %), being both ESBL producers. The β-lactamase genes found in the two isolates were blaSHV-12 + blaTEM-1b. The tet(A) and sul2 genes were also detected in these isolates, together with the non-classical class 1 integron (intI1-dfrA12-orfF-aadA2-cmlA1-aadA1-qacH-IS440-sul3) with the PcH1 promoter. The two isolates belonged to the phylogroup A. Amino acid changes in GyrA (S83L + D87G) and ParC (S80I) proteins were identified in our study. Concerning MLST typing, both isolates were assigned to ST1086, never found before in wild animals, and they presented closely related PFGE patterns. This study reveals the presence of ESBL-producing E. coli isolates, in a wild ecosystem, which could be dissemina...
2020
Extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae has become a major threat globally. Here we have characterized ESBL producing E. coli and K. pneumoniae from various sources, studied antibiogram and resistance gene profiles. Out of 385 samples, 31 (8.05%) were positive for ESBL producing E. coli. Such isolates could be recovered from 10.05, 8.33, 15.63, 6.67 and 4.35 per cent of cattle milk, curd, chicken, pork and cattle faeces samples, respectively. A total of 59 (15.32%) samples were positive for ESBL producing K. pneumoniae, which were isolated from 14.35, 6.25, 21.43 and 34.78 per cent cattle milk, chicken, beef and cattle faeces, respectively. All the 90 isolates were confirmed as ESBL producers by CDT and ESBL-E strip tests. Antibiogram revealed that 74.19% and 69.49% of the ESBL producing E. coli and K. pneumoniae isolates, respectively showed resistance to ceftizoxime, 25.81% and 23.73% to both co-trimoxazole and tetracycline, 19.35% and 25.42% to ciprof...
Journal of Antimicrobial Chemotherapy, 2013
The aims of this study were (i) to detect extended-spectrum b-lactamase (ESBL) genes among 1378 Escherichia coli isolates from defined disease conditions of companion and farm animals and (ii) to determine the localization and organization of ESBL genes. Methods: E. coli isolates from the German resistance monitoring programme GERM-Vet were included in the study. Plasmids were transferred by conjugation or transformation and typed by PCR-based replicon typing. ESBL genes were detected by PCR; the complete ESBL genes and their flanking regions were sequenced by primer walking. Phylogenetic grouping and multilocus sequence typing (MLST) were performed for all ESBL-producing E. coli isolates. Results: Of the 27 ESBL-producing E. coli isolates detected, 22 carried bla CTX-M-1 genes on IncN (n¼ 16), IncF (n¼ 3), IncI1 (n¼ 2) or multireplicon (n¼ 1) plasmids. A bla CTX-M-3 gene was located on an IncN plasmid and a bla CTX-M-15 gene was located on an IncF plasmid. A multireplicon plasmid and an IncHI1 plasmid harboured bla CTX-M-2. A bla TEM-52c gene was identified within Tn2 on an IncI1 plasmid. The bla CTX-M genes located within the same or related genetic contexts showed differences due to the integration of insertion sequences. Various MLST types were detected, with ST10 (n¼ 7), ST167 (n¼ 4) and ST100 (n¼ 3) being the most common. Conclusions: This study showed that the bla CTX-M-1 gene is the predominant ESBL gene among E. coli isolates from diseased animals in Germany and a considerable structural heterogeneity was found in the regions flanking the bla CTX-M-1 gene. Insertion sequences, transposons and recombination events are likely to be involved in alterations of the ESBL gene regions.
Applied and Environmental Microbiology, 2012
ABSTRACTIn line with recent reports of extended-spectrum beta-lactamases (ESBLs) inEscherichia coliisolates of highly virulent serotypes, such as O104:H4, we investigated the distribution of phylogroups (A, B1, B2, D) and virulence factor (VF)-encoding genes in 204 ESBL-producingE. coliisolates from diarrheic cattle. ESBL genes, VFs, and phylogroups were identified by PCR and a commercial DNA array (Alere, France). ESBL genes belonged mostly to the CTX-M-1 (65.7%) and CTX-M-9 (27.0%) groups, whereas those of the CTX-M-2 and TEM groups were much less represented (3.9% and 3.4%, respectively). One ESBL isolate wasstx1andeaepositive and belonged to a major enterohemorrhagicE. coli(EHEC) serotype (O111:H8). Two other isolates wereeaepositive butstxnegative; one of these had serotype O26:H11. ESBL isolates belonged mainly to phylogroup A (55.4%) and, to lesser extents, to phylogroups D (25.5%) and B1 (15.6%), whereas B2 strains were quasi-absent (1/204). The number of VFs was significant...
Foodborne Pathogens and Disease, 2012
The prevalence of extended-spectrum beta-lactamase (ESBL)- and plasmidic AmpC-beta-lactamase (pAmpC-BL)-producing Escherichia coli isolates has been studied in food-producing animals at the farm level in Tunisia, and recovered isolates were characterized for the presence of other resistance genes and integrons. Eighty fecal samples of food-producing animals (23 sheep, 22 chickens, 22 cattle, six horses, five rabbits, and two dromedaries) were obtained from 35 different farms in Tunisia in 2011. Samples were inoculated onto MacConkey agar plates supplemented with cefotaxime (2 mg/L) for cefotaxime-resistant (CTX(R)) E. coli recovery. CTX(R) E. coli isolates were detected in 11 out of 80 samples (13.8%), and one isolate per sample was further characterized (10 from chickens and one from a dromedary). The 11 CTX(R) isolates were distributed into phylogroups: B1 (five isolates), A (two isolates), D (three isolates), and B2 (one isolate). The following beta-lactamase genes were detected: bla(CTX-M-1) (seven isolates), bla(CTX-M-1)+bla(TEM-135) (one isolate), bla(CTX-M-1)+bla(TEM-1b) (one isolate), and bla(CMY-2) (two isolates). All ESBL- and pAmpC-BL-producing E. coli strains showed unrelated pulsed-field gel electrophoresis patterns. Seven isolates contained class 1 integrons with four gene cassette arrangements: dfrA17-aadA5 (three isolates), dfrA1-aadA1 (two isolates), dfrA15-aadA1 (one isolate), and aadA1 (one isolate). All isolates showed tetracycline resistance and contained the tet(A) +/- tet(B) genes. Virulence genes detected were as follows (number of isolates in parentheses): fimA (10); aer (eight); papC (two); and papGIII, hly, cnf, and bfp (none). Chicken farms constitute a reservoir of ESBL- and pAmpC-BL-producing E. coli isolates of the CTX-M-1 and CMY-2 types that potentially could be transmitted to humans via the food chain or by direct contact.
Microbial Drug Resistance, 2003
A total of 1439 Escherichia coli isolates from sick animals were received from the Spanish Network of Veterinary Antimicrobial Resistance Surveillance (VAV) from 1997 to 2001. Antimicrobial susceptibility tests were performed and diminished susceptibility to cefotaxime and ceftazidime was identified in 2.5% and 2.8% of the isolates, respectively. b-lactamase characterization was carried out in the group of 20 E. coli isolates with both characteristics. The MIC ranges of different b-lactams showed by these 20 isolates were as follows (in mg/ml): ampicillin (64-.256), amoxicillin-clavulanic acid (4-64), ticarcillin (8-.128), cefazolin (32-.256), cefoxitin (4-.128), cefotaxime (1-64), ceftazidime (2-.64), ceftriaxone (0.5-64), imipenem (#0.06-0.25), and aztreonam (2-.32). TEM, SHV, CMY, and FOX b-lactamase genes were analyzed by PCR and sequencing. The b-lactamase genes detected were the following ones (number of isolates): bla TEM-1b (3), bla TEM-1a (1), bla TEM-30f (2), bla TEM-1b 1 bla CMY-2 (2), and bla SHV-12 (1). Sequences of the promoter and/or attenuator region of the chromosomal ampC gene were studied in all the 20 isolates. Mutations at position 242 or 232 were detected in 16 isolates and these mutations were associated with the presence of a TEM type b-lactamase in 6 isolates. Besides, a high variety of plasmidic b-lactamases was detected including TEM-30 and CMY-2. To our knowledge, this is the first time that TEM-30 b-lactamase has been detected in E. coli isolates of animal origin.
Frontiers in microbiology, 2017
Background: The emergence and spread of antimicrobial resistance has become a major global public health concern. A component element of this is the spread of the plasmid-encoded extended-spectrum b-lactamase (ESBL) genes, conferring resistance to third-generation cephalosporins. The purpose of this study was to investigate the molecular characteristics of ESBL-encoding genes identified in Escherichia coli cultured from diarrheic patients in China from 2013 to 2014. Materials and Methods: A total of 51 E. coli were confirmed as ESBL producers by double-disk synergy testing of 912 E. coli isolates studied. Polymerase chain reaction (PCR) and DNA sequencing were performed to identify the corresponding ESBL genes. Susceptibility testing was tested by the disk diffusion method. Plasmids were typed by PCR-based replicon typing and their sizes were determined by S1-nuclease pulsed-field gel electrophoresis. Multi-locus sequence typing (MLST) and phylogrouping were also performed. Broth ma...