Frequency of conjugative transfer of plasmid-encoded ISEcp1 - blaCTX-M-15 and aac(6')-lb-cr genes in Enterobacteriaceae at a tertiary care center in Lebanon - role of transferases

Harajly, Mohamad; Khairallah, Marie-Therese; Corkill, J. E.; Araj, George F.; Matar, Ghassan M.

doi:10.1186/1476-0711-9-19

Cited by 18 publications

(15 citation statements)

References 21 publications

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“…Those expressing an ESBL phenotype frequently carry genes encoding resistance to other commonly used antimicrobial drug classes (such as aminoglycosides, chloramphenicols, fluoroquinolones, or tetracycline; Harajly et al, 2010; Rogers et al, 2011). Plasmid-mediated transfer of drug-resistance genes among various bacterial species is considered to be one of the most important mechanisms driving the spread of multi-drug resistance (MDR) markers.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of blaESBL–harboring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food-producing animals and healthy humans

Juan¹,

Stephan²,

Karczmarczyk³

et al. 2013

Front. Microbiol.

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Background: Extended-spectrum β-lactamase (ESBL)-encoding genes are frequently mapped to plasmids, yet few of these structures have been characterized at the molecular level, to date.Methods: Eighty-seven ESBL-producing Escherichia coli were isolated from fecal samples of food-producing animals and healthy humans in Switzerland from 2009 to 2011. Plasmid DNA of all isolates was purified. Broth mating assays were carried out individually for 32 isolates to determine if the ESBL marker could be transferred by conjugation. The plasmid sizes were determined by S1-nuclease pulsed-field gel electrophoresis (PFGE) and the plasmids were typed by PCR-based replicon typing. Susceptibility tests by disk diffusion followed with a re-analysis S1-nuclease PFGE and PCRs were performed to confirm plasmid transfer. Microarray was performed to detect additional antibiotic resistance markers and multi-locus sequence typing was also performed in selected donor strains. The phylotypes were identified by triplex PCR.Results: About half (n = 46) of the 87 isolates carried small (<20-kb) plasmids. All selected 32 isolates contained large plasmids (ranging in sizes from 20- to 600-kb). Eleven plasmid replicon types were detected. Of these, IncFIA (n = 5), IncFIB (n = 9), and IncK/B (n = 4) were common. Nine isolates demonstrated the ability to transfer their cefotaxime resistance marker at high transfer rates. Plasmid profile re-analysis of these transconjugants identified 16 plasmids. IncFIB and IncI1 were the most prevalent replicon types. Phylogenetic grouping showed that five of the nine donor strains belonged to phylogroup B1. Nine different sequence types were identified in nine tested donor strains.Conclusion: Characterization of these ESBL-encoding conjugative plasmids extends our understanding on these resistance markers in multi-drug resistant E. coli cultured from healthy human and animal sources.

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Section: Introductionmentioning

confidence: 99%

Molecular characterization of blaESBL–harboring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food-producing animals and healthy humans

Juan¹,

Stephan²,

Karczmarczyk³

et al. 2013

Front. Microbiol.

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“…Indeed, aac(69)-Ib-cr alone confers a degree of resistance sufficiently low that the MIC might not distinguish organisms that carry it from those that do not (Robicsek et al, 2006). Recently, Harajly et al (2010) described transconjugants with no elevation of MIC after the transfer of aac(69)-Ib-cr.…”

Section: A Barguigua and Others 1348mentioning

confidence: 99%

Characterization of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates from the community in Morocco

Barguigua

Otmani

Talmi

et al. 2011

Journal of Medical Microbiology

104

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Of 803 community Escherichia coli (n5767) and Klebsiella pneumoniae (n536) isolates collected from patients with urinary tract infections in three Moroccan cities, 10 E. coli (1.3%) and 2 K. pneumoniae (5.6 %) isolates were shown to produce extended-spectrum b-lactamases (ESBLs). PFGE revealed that the E. coli isolates comprised seven distinct genotypes. The presence of plasmids in the 12 isolates was revealed by conjugation experiments of plasmids from these Enterobacteriaceae strains with E. coli K 12 J 5 , with further isolation of the plasmids in the transconjugants. Subsequent nucleotide sequencing indicated that the plasmids encoded the bla CTX-M , bla OXA , bla TEM and bla SHV genes, including genes for CTX-M-15 (n511), OXA-1 (n511), TEM-1b (n54), SHV-5 (n51) and SHV-1 (n52). Identification of plasmid-mediated quinolone-resistance genes was performed by PCR. The aac(69)Ib-cr variant was detected in all strains, and two strains co-expressed qnrS1, bla CTX-M-15 and bla OXA-1 genes. The presence of ESBLs in the Enterobacteriaceae strains studied was probably due to the dissemination of resistance plasmids with the predominant genotype of bla CTX-M-15.

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“…In the United Kingdom, Woodford et al (24) found that a CTX-M-15 epidemic strain had ceftazidime MICs consistently lower than those seen with other CTX-M-15 producers (MICs Յ 2 g/ml), placing them in the "susceptible" category by British Society of Antimicrobial Chemotherapy (BSAC) criteria (24). CTX-M-15 strains with variable levels of resistance to cefotaxime (MIC of 2 g/ml), ceftazidime (MIC 1.5 g/ml), and cefepime (MIC 1 to 2 g/ml) have also been isolated recently in hospitals in Belgium (11) and Lebanon (6). Previous studies have characterized the bla CTX-M-15 -containing plasmids (2,16) and have shown that bla CTX-M-15 alleles are located on plasmids downstream of an ISEcp1 element.…”

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confidence: 99%

Detection of Extended-Spectrum-β-Lactamase-Positive Escherichia coli in Bile Isolates from Two Dogs with Bacterial Cholangiohepatitis

et al. 2011

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This is the first report of Escherichia coli isolates producing CTX-M-15, the predominant type of extendedspectrum ␤-lactamase (ESBL) associated with clinical disease in humans in the United Kingdom, in a United Kingdom pet dog. This report also describes the first isolation of CTX-M/Tem ESBL-positive E. coli from bile in dogs with hepatobiliary disease. CASE REPORTS Case 1. A 7-year-old female spayed golden retriever was referred for exercise intolerance, vomiting, diarrhea, and panting. Previous treatment had included administration of potentiated amoxicillin, ranitidine, and maropitant. One episode of icterus was also reported before the dog was referred. Abdominal discomfort was noted upon physical examination, and in abdominal ultrasound analysis, the liver appeared normal with respect to size and echogenicity, but the gallbladder showed mildly thickened walls and sludge content. A true cut of the liver for histology and culture and bile aspiration for culture were performed. A minimal amount of inflammatory cells were present on bile cytology, but free bacteria were noted. Histology results were consistent with moderate lymphoplasmacytic and neutrophilic hepatitis (with negative copper staining results); a bile culture revealed heavy growth of two culture variants of nonhemolytic Escherichia coli, but no bacterial growth was obtained from liver tissue.Treatment was started with S-adenosyl-methionine (SAMe) and marbofloxacin (3 mg/kg of body weight orally once daily [p.o. SID]) for 6 weeks. One week after administration of the antibiotics was stopped, the dog had clinically improved and gained two kilograms in weight. Episodic lethargy was still reported. The appearance of the gallbladder remained unchanged in the ultrasound analysis. Bile culture revealed the same two culture variants (0784A and 0784B) of E. coli, which were both shown to be sensitive to imipenem and were therefore treated with that antibiotic (10 mg/kg subcutaneously [SQ] every 8 hours [q8h]) for 14 days. A follow-up 2 weeks into treatment had been scheduled to ensure efficacy of the treatment, but the owner was not able to attend. The dog was treated for a total of 4 weeks and rechecked 1 week after the end of the antibiotic treatment. Although marked clinical improvement was reported during treatment, lethargy and panting had recommenced within days of stopping it. Repeat bile cultures yielded a mixed growth consisting of the same two culture variants of E. coli (0784A and 0784B) and also yielding Enterococcus faecium. It was decided to perform a cholecystectomy at that stage. The gallbladder was sent for histology, and cystic mucinous hyperplasia was diagnosed. The dog recovered from the surgery uneventfully. Two months after surgery, the abdominal ultrasound analysis was repeated and a gastric mass was found to be present (suspected neoplasia) that had not been previously noted. The owner elected no further diagnosis or treatment at that point.Case 2. A 4.5-year-old female spayed standard poodle presented with acute onset jaundice. Th...

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Frequency of conjugative transfer of plasmid-encoded ISEcp1 - blaCTX-M-15 and aac(6')-lb-cr genes in Enterobacteriaceae at a tertiary care center in Lebanon - role of transferases

Cited by 18 publications

References 21 publications

Molecular characterization of blaESBL–harboring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food-producing animals and healthy humans

Molecular characterization of blaESBL–harboring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food-producing animals and healthy humans

Characterization of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates from the community in Morocco

Detection of Extended-Spectrum-β-Lactamase-Positive Escherichia coli in Bile Isolates from Two Dogs with Bacterial Cholangiohepatitis

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