Dataset Information

Genetic Basis and Clonal Population Structure of Antibiotic Resistance in Campylobacter jejuni Isolated From Broiler Carcasses in Belgium.

ABSTRACT: Human campylobacteriosis is the leading food-borne zoonosis in industrialized countries. This study characterized the clonal population structure, antimicrobial resistance profiles and occurrence of antimicrobial resistance determinants of a set of Campylobacter jejuni strains isolated from broiler carcasses in Belgium. Minimum inhibitory concentrations (MICs) against five commonly-used antibiotics (ciprofloxacin, nalidixic acid, tetracycline, gentamicin, and erythromycin) were determined for 204 C. jejuni isolates. More than half of the isolates were resistant to ciprofloxacin or nalidixic acid. In contrast, a lower percentage of screened isolates were resistant to gentamicin or erythromycin. C. jejuni isolates resistant to ciprofloxacin and/or nalidixic acid were screened for the substitution T86I in the quinolone resistance determining region (QRDR) of the gyrA gene, while C. jejuni isolates resistant to tetracycline were screened for the presence of the tet(O) gene. These resistance determinants were observed in most but not all resistant isolates. Regarding resistance to erythromycin, different mutations occurred in diverse genetic loci, including mutations in the 23S rRNA gene, the rplD and rplV ribosomal genes, and the intergenic region between cmeR and cmeABC. Interestingly, and contrary to previous reports, the A2075G transition mutation in the 23S rRNA gene was only found in one strain displaying a high level of resistance to erythromycin. Ultimately, molecular typing by multilocus sequence typing revealed that two sequence types (ST-824 and ST-2274) were associated to quinolones resistance by the presence of mutations in the gene gyrA (p = 0.01). In addition, ST-2274 was linked to the CIP-NAL-TET-AMR multidrug resistant phenotype. In contrast, clonal complex CC-45 was linked to increased susceptibility to the tested antibiotics. The results obtained in this study provide better understanding of the phenotypic and the molecular basis of antibiotic resistance in C. jejuni, unraveling some the mechanisms which confer antimicrobial resistance and particular clones associated to the carriage and spread of resistance genes.

SUBMITTER: Elhadidy M

PROVIDER: S-EPMC5966580 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Genetic Basis and Clonal Population Structure of Antibiotic Resistance in <i>Campylobacter jejuni</i> Isolated From Broiler Carcasses in Belgium.

Elhadidy Mohamed M Miller William G WG Arguello Hector H Álvarez-Ordóñez Avelino A Duarte Alexandra A Dierick Katelijne K Botteldoorn Nadine N

Frontiers in microbiology 20180517

Human campylobacteriosis is the leading food-borne zoonosis in industrialized countries. This study characterized the clonal population structure, antimicrobial resistance profiles and occurrence of antimicrobial resistance determinants of a set of <i>Campylobacter jejuni</i> strains isolated from broiler carcasses in Belgium. Minimum inhibitory concentrations (MICs) against five commonly-used antibiotics (ciprofloxacin, nalidixic acid, tetracycline, gentamicin, and erythromycin) were determined ...[more]

PMID: 29867900

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Project description:BACKGROUND:Campylobacter is one of the leading bacterial species causing foodborne illnesses in humans. Antimicrobial agents have been extensively used for treatment of Campylobacter infections; but in the recent years, both animal and human isolates of this bacterium have shown resistance to several antibiotics such as tetracycline. OBJECTIVES:The aim of this study was to investigate the presence of genetic determinants of tetracycline resistance in Campylobacter spp. recovered from poultry carcasses in Shiraz, Iran. MATERIALS AND METHODS:Eighty-three thermophilic Campylobacter spp. Isolates were first identified based on multiplex polymerase chain reaction (PCR) and then screened for presence of tetracycline resistance genes (tet (A), tet (B), tet (O) and te (S)) by PCR. RESULTS:The overall prevalence of Campylobacter jejuni and C. coli among the examined isolates was 51.8% and 48.2%, respectively. Tetracycline resistance genes of tet (B) and tet (S) were not seen among these Campylobacter spp. Isolates, whereas the most common tet gene identi?ed was tet (O), found in 83.1% (69/83) of all the isolates. The tet (O) gene sequence comparison between C. jejuni and C. coli showed 100% similarity and these sequences (JX853721and JX853722) were also identical to the homologous sequences of other strains of Campylobacter spp. existing in the GenBank databases. In addition, tet (A) was found in 18% (15/83) of Campylobacter spp. isolates. To our knowledge, this represents the first report of tet (A) in Campylobacter spp. There was 100% homology between the sequences of tet (A) from this study (JX891463 and JX891464) and the tet (A) sequences mentioned for other bacteria in the GenBank databases. CONCLUSIONS:The high prevalence of tet (O) resistance gene along with new detection of tet (A) resistance gene in Campylobacter spp. isolated from poultry carcasses revealed an extensive tetracycline resistance among Campylobacter isolates from poultry in Iran. It emphasized the need for cautious use of tetracycline in poultry production to decrease the extension of tetracycline-resistant Campylobacter spp.

Project description:Antimicrobial resistance was determined for 341 thermophilic Campylobacter jejuni isolates obtained from human clinical cases (n = 101), broiler products (n = 98), dairy cattle (n = 41) and wild birds (n = 101) with known multilocus sequence types (MLST) in Lithuania. The minimum inhibitory concentration (MIC) values for ciprofloxacin, tetracycline, gentamicin, ceftriaxone and erythromycin were determined with the agar dilution method. MIC values were compared with MLST types to find possible associations among isolation source, sequence type and resistance to antibiotics. The proportions of resistant strains were 94.2% (human), 95% (wild birds), 100% (broiler products) and 100% (dairy cattle) for one of the tested antibiotics. Most frequently, resistance to ciprofloxacin was observed (91.5%), followed by ceftriaxone with 60.4%, and tetracycline (37.8%). However only three C. jejuni strains were resistant to erythromycin (0.9%) and all tested thermophilic Campylobacter strains were sensitive to gentamicin. Most of the examined C. jejuni isolates (80.6%) showed resistance to at least one of three profiles: CIP+AXO (28.1%), TET+CIP+AXO (26.7%) and CIP (25.8%). Statistically significant differences in resistance to tetracycline were found between C. jejuni strains obtained from cattle (85.4%) and broiler products (64.3%) (P < 0.05). The majority (87.1%) of the tested strains from wild birds were resistant to ciprofloxacin (P < 0.05). The results showed that strains of novel ST's showed significantly lower resistance to ceftriaxone (P < 0.05). The ST-21 (CC21) (78.8%) was identified with significantly higher multidrug resistance relatively to other tested ST's in this study. Our results emphasize the high antimicrobial resistance of phylogenetically diverse C. jejuni strains isolated from different sources including specific genotypes of wild bird's strains in Lithuania. The results support the opinion that not only broiler products but cattle and wild birds may be a reservoir of resistant C. jejuni and stipulate a risk of spread or resistant bacteria. There is increasing need for broad surveillance and control measures to track changes and pathways of antimicrobial resistance of C. jejuni in epidemiologically distinct populations.

Project description:The aim of the study was to investigate the mode of action of (-)-α-pinene in terms of its modulation of antibiotic resistance in Campylobacter jejuni. Broth microdilution and ethidium bromide accumulation assays were used to evaluate the (-)-α-pinene antimicrobial activity, modulation of antimicrobial resistance, and inhibition of antimicrobial efflux. The target antimicrobial efflux systems were identified using an insertion mutagenesis approach, and C. jejuni adaptation to (-)-α-pinene was evaluated using DNA microarrays. Knock-out mutants of the key up-regulated transcriptional regulators hspR and hrcA were constructed to investigate their roles in C. jejuni adaptation to several stress factors, including osmolytes, and pH, using Biolog phenotypical microarrays. Our data demonstrate that (-)-α-pinene efficiently modulates antibiotic resistance in C. jejuni by decreasing the minimum inhibitory concentrations of ciprofloxacin, erythromycin and triclosan by up to 512-fold. Furthermore, (-)-α-pinene promotes increased expression of cmeABC and another putative antimicrobial efflux gene, Cj1687. The ethidium bromide accumulation was greater in the wild-type strain than in the antimicrobial efflux mutant strains, which indicates that these antimicrobial efflux systems are a target of action of (-)-α-pinene. Additionally, (-)-α-pinene decreases membrane integrity, which suggests that enhanced microbial influx is a secondary mode of action of (-)-α-pinene. Transcriptomic analysis indicated that (-)-α-pinene disrupts multiple metabolic pathways, and particularly those involved in heat-shock responses. Thus, (-)-α-pinene has significant activity in the modulation of antibiotic resistance in C. jejuni, which appears to be mediated by multiple mechanisms that include inhibition of microbial efflux, decreased membrane integrity, and metabolic disruption. These data warrant further studies on (-)-α-pinene to develop its use in the control of antibiotic resistance in Campylobacter.

Dataset Information

Genetic Basis and Clonal Population Structure of Antibiotic Resistance in Campylobacter jejuni Isolated From Broiler Carcasses in Belgium.

Publications

Genetic Basis and Clonal Population Structure of Antibiotic Resistance in <i>Campylobacter jejuni</i> Isolated From Broiler Carcasses in Belgium.

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