The prevalence of the
Campylobacter multi-drug efflux pump (CmeABC) was evaluated in
Campylobacter isolates recovered from freshly processed turkeys at two Midwestern processing plants. A total of 94
...Campylobacter isolates recovered from processed turkeys were examined using polymerase chain reaction (PCR) to determine the presence of the multi-drug efflux pump genes
cmeA,
cmeB, and
cmeC.
Results from this study found that 51% of all isolates tested were positive for CmeABC. 46.6% of these positive isolates were from plant A and 55.1% from plant B. Differences were observed in the prevalence of individual genes found among
Campylobacter isolates from each plant. Additional analysis found that among the isolates positive for CmeABC, 85.5% were identified as
C. jejuni and 14.5% identified as
C. coli.
There was a relatively high occurrence of the
Campylobacter multi-drug efflux pump genes in
Campylobacter spp. recovered from processed turkeys, however, the presence of the genes could not be significantly linked to antimicrobial resistance observed in the test strains and suggests that the CmeABC genes are only one factor associated with antimicrobial resistance in
Campylobacter spp.
Aims: In view of recent findings that a multidrug efflux pump CmeABC exists in Campylobacter jejuni, 391 C. jejuni and 52 Campylobacter coli of human and animal origin were examined for a multidrug ...resistance phenotype. Materials and methods: The MICs of ampicillin, chloramphenicol, ciprofloxacin, erythromycin, kanamycin, tetracycline, cetrimide, triclosan, acridine orange, paraquat and ethidium bromide were determined. Resistance to organic solvents and the effect of salicylate (known inducer of the marRAB operon in Escherichia coli and Salmonella) were also examined. Results: Two C. coli and 13 C. jejuni isolates, mainly from pigs or poultry, were resistant to three or more antibiotics and 12 of these strains had reduced susceptibility to acridine orange and/or ethidium bromide. Strains (n = 20) that were less susceptible to acridine orange, ethidium bromide and triclosan were significantly more resistant (P < 0.05) to ampicillin, chloramphenicol, ciprofloxacin, erythromycin, nalidixic acid and tetracycline, with two- to four-fold increases in MIC values compared with strains (n = 20) most susceptible to acridine orange, ethidium bromide and triclosan. Growth of strains with 1 mM salicylate caused a small (up to two-fold) but statistically significant (P ≤ 0.005) increase in the MICs of chloramphenicol, ciprofloxacin, erythromycin and tetracycline. Conclusions: These data indicate that multiple antibiotic resistant (MAR)-like Campylobacter strains occur and it may be postulated that these may overexpress cmeABC or another efflux system.
A multidrug-resistant mutant of Campylobacter jejuni was selected in vitro using increasing concentrations of enrofloxacin. This mutant accumulated less ethidium bromide than the parental strain, ...suggesting the participation of active efflux as a resistance mechanism. Inactivation of the cmeB gene confirmed active efflux and indicated the involvement of the CmeABC efflux pump in the multidrug resistance of the mutant. Sequencing of the cmeR-cmeA intergenic region revealed a point mutation in the binding site of the CmeR repressor. Transcriptional lacZ fusions showed an increase of transcription of the cmeABC operon in the multidrug-resistant mutant. Gel mobility shift assays and Surface Plasmon Resonance experiments further indicated a decrease in the affinity of the CmeR for the promoting region of the cmeABC operon consecutive to this mutation. Thus, these results showed that the point mutation was responsible, via a lack of binding of the CmeR repressor, for increased expression of the CmeABC efflux pump and consecutive multidrug resistance.
Objectives: This study was conducted to examine the genetic variation occurring in the cmeB gene encoding the transporter component of the CmeABC efflux pump. Methods: Expression of the CmeABC pump ...in 21 strains of Campylobacter jejuni and Campylobacter coli was studied by western-blot analysis. MIC determination was conducted in the presence or absence of an efflux pump inhibitor (EPI). Inactivation of the cmeB gene and sequencing of the cmeABC operon were performed for a single strain. The remaining strains were compared by RFLP analysis of the cmeB-specific PCR amplicon. The cmeB genes of two C. coli strains with different RFLP patterns were sequenced completely. Results: Conflicting results were obtained in the western-blot analysis with anti-CmeB and anti-CmeC antibodies for one strain, whereas MIC determinations with EPI and cmeB gene inactivation confirmed the efflux pump's activity. The cmeB gene of this isolate showed only 78% nucleotide sequence identity with the sequence of reference strains. PCR–RFLP analysis identified 4 different patterns among the 5 C. jejuni and 14 different patterns among the 16 C. coli strains investigated. At the amino acid sequence level, variation was higher in the periplasmic loops of the transporter. Conclusions: A total of 18 different cmeB-specific PCR–RFLP patterns were detected among the 21 C. jejuni and C. coli strains. These sequence variations might have an impact on the function and substrate recognition of this transporter. The sequence data obtained in this study will help to design suitable tools to study the presence or the expression of the gene cmeB.
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