[논문]Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea

Cho, Hyun-Ho; Kim, Sang-Hyun; Min, Wongi; Ku, Bok-Kyung; Kim, Jong-Hyun; Kim, Yong-Hwan

doi:10.14405/kjvr.2014.54.2.91

[국내논문] Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea 원문보기

大韓獸醫學會誌 = Korean journal of veterinary research, v.54 no.2 = no.155, 2014년, pp.91 - 99

Cho, Hyun-Ho (Animal, Plant and Fisheries Quarantine and Inspection Agency) , Kim, Sang-Hyun (Viral Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology) , Min, Wongi (College of Veterinary Medicine, Research Institute of Life Science, Institute of animal medicine, Gyeongsang National University) , Ku, Bok-Kyung (Animal, Plant and Fisheries Quarantine and Inspection Agency) , Kim, Jong-Hyun (Division of Enteric Bacteria Infections, Center for Infectious Disease, National Institute of Health) , Kim, Yong-Hwan (College of Veterinary Medicine, Research Institute of Life Science, Institute of animal medicine, Gyeongsang National University)

Abstract ▼ AI-Helper

An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

주제어

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문제 정의

Accordingly, the aim of this study was to investigate the prevalence of antimicrobial resistant Campylobacter spp. isolated from dogs and humans and to confirm the relationship between fluoroquinolone-resistance and the gyrA mutation by DNA sequencing of the gyrA gene.

제안 방법

0 Kb DNA ladder (Takara Bio). Matching and the dendrogram of the fingerprints determined by the unweighted pair group method with averages were performed using the Dice coefficient with 1% tolerance windows.
The amplification reaction was carried out in a DNA thermal cycler using forward primer (5’-GGATTTCGTATTAACACAAATGGTGC-3’) and reverse primer (5’-CTGTAGTA ATCTTAAACATTTG-3’).
According to the antimicrobial susceptibility test, a large number of Campylobacter spp. isolates used in this study showed a multidrug resistance phenotype. A total of 26 antimicrobial resistance pattern was determined (Table 3).
Ninety-four strains of Campylobacter spp. were selected at random for the analysis of flaA-RFLP after digestion with DdeI restriction enzyme to compare phenotype and genotype of the Campylobacter isolates from dogs and humans (Fig. 1). The fla-RFLP typing distributed 73 strains of C.

대상 데이터

Antimicrobial susceptibility test was performed by agar dilution method with Mueller-Hinton agar supplement with 5% sheep blood, according to the protocols of Clinical and Laboratory Standards Institute (CLSI) [5]. C. jejuni ATCC 33560 and C. coli ATCC 33559 were used as the quality control organisms for the test. Erythromycin (EM 0.

데이터처리

Data were analyzed by Student’s t tests.

이론/모형

The flaA-PCR-RFLP has been applied as one of the tools of epidemiological study to differentiate isolates below the species level [10, 13]. Accordingly, flaA-RFLP typing method was used to analyze the genotype. Ninety four strains of Campylobacter isolates were subtyped by flaA-PCR-RFLP into 12 major clusters.
Antimicrobial susceptibility test was performed by agar dilution method with Mueller-Hinton agar supplement with 5% sheep blood, according to the protocols of Clinical and Laboratory Standards Institute (CLSI) [5]. C.

성능/효과

Also sequencing analysis showed that 100% of NAR and CIPR C. jejuni isolates (n = 63) from dogs and humans have the mutation Thr-86th-Ile, which is known to be associated with quinolone resistance, and the mutations were not exhibited in all fluorquinolone-susceptible C. jejuni at the 86th codon (Table 5).
C. jejuni isolates from humans showed a higher resistance to tetracycline than those isolated from dogs (p ≤ 0.01).
C. jejuni isolates from humans showed higher resistance to ampicillin than those isolated from dogs (p ≤ 0.05).
C. jejuni isolates from humans showed higher resistance to tetracycline than those isolated from dogs (p ≤ 0.01).
coli isolated from pigs and broilers (62 and 52%, respectively) [30]. In this study, the resistance of C. jejuni to tetracycline ranged from 21.7~40% in dog isolates and 83.3% in human isolates. C.
The silent mutations at the codons 78, 81, 110, 117, 118, and 119 were assessed (Tables 4 and 5). The results showed that the silent mutation at the codons 81 (26.8%), 119 (19.5%) and 81 (24%), 110 (2.2%), 117 (2.2%), 119 (4.3%) in the QRDR of the gyrA genes in C. jejuni were isolated from dogs and humans respectively. Also sequencing analysis showed that 100% of NA^Rand CIP^R C.

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