• Journal of Life Science
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• v.23 no.5
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• pp.703-709
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• 2013

The aim of this study was to investigate the prevalence of Extended-spectrum ${\beta}$-lactamase (ESBL) gene and quinolone resistance determinant (qnr) and the pattern of antibiotic resistance in the ESBL-producing Escherichia coli clinical isolates. The 42 ESBL-producing strains from total 274 isolates were detected using a double disk synergy test. They were isolated from various specimens, such as urine (28 strains), sputum (6 strains), pus (3 strains), wound (2 strains), blood (2 strains), and tissue (1 strain). Using the PCR with the specific primers ESBL, ESBL and qnr gene types were determined. Thirty-five strains possessed one or two ESBL genes. CTX-M-1 type was the most abundant followed by CTX-M-9 type and TEM, but SHV, CTX-M-2, and CTX-M-8 gene types were not detected. qnr gene types were detected from ten isolates in the order of qnrB4, qnrB1, and qnrS. Coexistence of ESBL and qnr genes was found. ESBL-producing isolates showed high resistance against some antibiotics, such as cefotaxmie (80.0%), levofloxacin (82.9%), and ampicillin (100%). Neither a synergy effect from the coexistence of ESBL and qnr genes on antibiotic resistance nor a correlation between the production of qnr gene and quinolone resistance were found.

• Korean Journal of Veterinary Service
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• v.40 no.3
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• pp.193-200
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• 2017

This study was carried out to investigate the antimicrobial resistance profiles and resistance genes in 62 Escherichia coli isolated from dogs and cats hospitalized at animal hospitals in Daegu. E. coli isolates showed high resistance to nalidixic acid (46.8%) and ampicillin (45.2%). Resistance to the other antimicrobial agents was less than 30%, and no resistant isolates were detected for imipenem and amikacin. Of the 28 ampicillin-resistant isolates, TEM and CTX-M genes were detected in 16 (57.1%) and 11 (39.3%), respectively. The aadA gene was found in 4 (26.7%) of 15 gentamicin-resistant isolates, and strA-strB gene was found in 10 (66.7%) isolates. The sul I and sul II genes were detected in 11 (61.1%) and 14 (77.8%) of 18 trimethoprim/sulfamethoxazole-resistant isolates, and tetB gene in 9 (81.8%) of 11 minocycline-resistant isolates, and cmlA gene in 2 (22.2%) of 8 chloramphenicol-resistant isolates. The qnrB and qnrS genes were found in 3 (10.3%) and 1 (3.4%) of 28 nalidixic acid-resistant isolates, respectively. Whereas, none of the SHV, CMY-2, tetA, dfr Ia and dfr VII, and qnrA genes were found. Our results show a wide variety of resistance genes in E. coli isolates from dogs and cats. This study also represents the first report of qnrB and qnrS gene producing E. coli isolates from dogs in republic of Korea.

• Korean Journal of Veterinary Service
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• v.42 no.1
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• pp.17-24
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• 2019

The aim of this study was to investigate the prevalence and characterization of plasmid-mediated quinolone resistance (PMQR) gene in 79 Enterobacteriaceae isolated from dogs and cats. Of 79 isolates, PMQR genes were found in 10 (12.7%) isolates, including aac(6')-lb-cr, qnrB, qnrS and qnrA detected alone or in combination in 8 (10.1%), 4 (5.1%), 2 (2.5%) and 1 (1.3%) isolates, respectively. Interestingly, two qnrS genes were detected in nalidixic acid and ciprofloxacin susceptible isolates. Extended-spectrum ${\beta}$-lactamase (ESBL) was detected in 90% (9 isolates) of PMQR positives isolates. Among ESBL genes, CTX-M, TEM and SHV were detected in 9, 8 and 3 isolates, respectively. Almost all PMQR genes were detected in co-existence with ESBL genes. All PMQR positives isolates were multidrug resistance (i.e. resistant to five or more antibiotics). qepA, OXA and CMY-2 genes were not found. The six transconjugants were obtained by conjugation experiment. The aac(6')-lb-cr, qnrB and qnrS were co-transferred with CTX-M, TEM and/or SHV, whereas qnrA was not observed among transconugants. This is the first report of the presence of aac(6')-lb-cr and qnrA gene among Enterobacteriaceae isolates from dogs in Korea. The prudent use of antimicrobials and continuous monitoring for companion animals are required.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.3
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• pp.217-224
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• 2021

Fluoroquinolone (FQ) resistant gram-negative pathogens have emerged worldwide, and the recent increase in FQ resistant Escherichia coli is of great concern in Korea. This study investigated FQ resistance determinants and the epidemiological relationship of 56 ciprofloxacin-resistant E. coli isolated from a tertiary hospital in Daejeon, South Korea from June to December 2018. Molecular epidemiology was investigated by multilocus sequence typing (MLST). Polymerase chain reaction (PCR) and sequence analysis were performed to identify chromosomal mutations in the quinolone resistance determining regions (QRDR) of gyrA, gyrB, parC, and parE and to describe the occurrence of the following plasmid-mediated quinolone resistance (PMQR) genes: aac(6)-Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD, and qnrS. MLST analysis showed 12 sequence types (STs) and the most prevalent ST was ST131 (31/56, 55.4%), followed by ST1193 (13/56, 23.2%), and ST405 (3/56, 5.4%). In 56 ciprofloxacin-resistant E. coli isolates, Ser83→Leu and Asp87→Asn in gyrA and Ser80→Ile and Glu84→Val in parC (51.8%, 29/56) were the most frequent amino acid substitutions and aac(6)-Ib-cr (33.9%, 19/56) was the most common PMQR gene. These results of FQ resistance determinants were more frequently observed in ST131 compared with other clones. Continuous monitoring of the epidemiological characteristics of ciprofloxacin-resistant E. coli isolates and further investigation of FQ resistance determinants are necessary.

• Journal of Microbiology
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• v.45 no.2
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• pp.168-170
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• 2007

In this research 26 Shigella isolates were examined by PCR and direct nucleotide sequencing for genetic alterations in the quinolone-resistance determining regions (QRDRs). We tested for the presence of qnr genes by PCR in 91 strains, but no qnr genes were found. The results did show, however, some novel mutations at codon 83 of gyrA ($Ser{\rightarrow}Ile$) and codon 64 of parC ($Ala64{\rightarrow}Cys,\;Ala64{\rightarrow}Asp$), which were related to fluroquinolone resistance.

• Korean Journal of Veterinary Service
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• v.42 no.3
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• pp.153-159
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• 2019

The objective of this study was to identify mutations in the quinolone resistance determining region (QRDR) of the gyrA, gyrB, parC and parE genes, and the presence of plasmid-mediated quinolone resistance (PMQR) genes: qnrA, qnrB, qnrS, aac(6')-lb-cr and qepA in 40 nalidixic acid- resistant ($NA^R$) Salmonella isolates isolated from poultry slaughterhouse. The MIC of NA and ciprofloxacin for 40 $NA^R$ Salmonella isolates was $128{\sim}512{\mu}g/mL$ and < $0.125{\sim}0.25{\mu}g/mL$, respectively. The Salmonella isolates were resistant to NA (100%), gentamicin (5.0%) and ampicillin (2.5%). All $NA^R$ Salmonella isolates represented point mutation in codons Aspartic acid(Asp)-87 (90%) and Serine(Ser)-83 (10%) of QRDR of gyrA gene: $Asp87{\rightarrow}glycine$, $Ser83{\rightarrow}tyrosine$. No mutations were observed in QRDR of the gyrB, parC and parE gene. Moreover PMQR genes was not found in any of the tested isolates. Our findings showed that DNA gyrase is the primary target of quinolone resistance and a single mutation in codon Asp87 and Ser83 of the gyrA gene can confer resistance to NA and reduced susceptibility ciprofloxacin in Salmonella isolates.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.185-190
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• 2014

The aim of this study was to investigate the prevalence of quinolone resistant E. coli from raw bulk milk and to characterize the resistance determinants. In this study, the gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced from quinolone resistant E. coli isolates. Also, the presence of plasmid-mediated quinolone resistance (PMQR) and the expression of efflux pump genes based on quantitative real-time PCR (qRT-PCR) were investigated. Of the 487 coliform bacteria, 9 strains showed nalidixic acid resistance, and 6 of the 9 nalidixic acid resistant isolates were also ciprofloxacin resistant. These 9 strains had a single mutation at codon 83 (S83L) in gyrA, 2 of them had double mutations at codon 83 and 87 (S83L and D87N) in gyrA and 3 of the 9 isolates had single mutations at codon 80 (S80I) in parC. None of the 9 isolates harbored PMQR determinants. Compared with wild-type E. coli ATCC 25922, an over-expression of the acrB gene (2.15-5.74 fold), encoding the pump component of the AcrAB-TolC efflux pump was observed in 4 of 6 ciprofloxacin resistant isolates. This study identified the quinolone resistance mechanism of E. coli isolated from raw milk samples in Gyeonggi-do.

• Korean Journal of Veterinary Service
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• v.46 no.1
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• pp.35-45
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• 2023

Pathogenic Escherichia coli is the cause of a wide range of diseases in pigs, including diarrhea, edema disease, and septicemia. Diarrhea caused E. coli may result in significant economic losses, making pathogenic E. coli an important pathogen for the swine industry. This study investigated the prevalence of virulence factor genes, antimicrobial resistance phenotypes, and resistance genes in E. coli isolated from feces of piglets in Korea between 2017 and 2020. As a result, 119 pathogenic E. coli isolates were obtained from 601 fecal samples. The F4 adhesin gene and the STb enterotoxin gene were commonly present in E. coli isolated from diarrhea samples. The dominant virulotypes of isolates from diarrhea samples were STb, Stx2e, and F4:LT:STb. More than 80% of the screened isolates were resistant to ampicillin, sulfisoxazole, chloramphenicol, or tetracycline. To confirm the resistance mechanisms for β-lactam or quinolone, we investigated the genotypic factors of resistance. Each of the ceftiofur-resistant E. coli produced an extended-spectrum β-lactamase encoded by bla_CTX-M-14, bla_CTX-M-27, and bla_CTX-M-55. And all ciprofloxacin-resistant E. coli harbored mutations in quinoloneresistance-determining-regions. In addition, some of the ciprofloxacin-resistant E. coli contained the plasmid-mediated-quinolone-resistance genes such as qepA, qnrB1, or qnrD. This study has confirmed that the F4 fimbria and the STb enterotoxin are the most predominant in pathogenic E. coli isolated from piglets with diarrhea in Korea and there is a great need for responsible and prudent use of antimicrobials to treat colibacillosis.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.79-88
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• 2020

The aim of this study was to investigate the prevalence of CTX-M β-lactamase and plasmid-mediated quinolone resistance (PMQR) genes, and the pattern of antibiotic resistance in cefotaxime-resistant gramnegative bacteria. A total 126 gram-negative bacteria were isolated from hospitalized dogs and cats between 2018 and 2019. The most predominant isolates were E. coli (n=41), followed by Pseudomonas aeruginosa (n=25), Proteus mirabilis (n=14), Klebsiella pneumoniae (n=9), Sphingomonas paucimobilis (n=7), and Enterobacter cloacae and Serratia marcescens (respectively, n=5). Cefotaxime-resistant isolates were identified in 26.2% (33 isolates) of 126 gram-negative bacteria. CTX-M type β-lactamase were found in 15 isolates (10 E. coli, 1 Ent, cloacae and 4 K. pneumoniae, respectively). Among the CTX-M producing gram-negative bacteria, CTX-M-1 and CTX-M-9 were detected in 10 (66.7%) and 5 (33.3%) isolates, respectively. While, CTX-M-2 and CTX-M-8 were not found. PMQR genes were detected in 12 (36.4%) isolates (4 E. coli, 2 Ent, cloacae and 6 K. pneumoniae, respectively), and the predominant PMQR gene was aac(6')-lb-cr (n=9), followed by qnrB (n=8) and qnrS (n=1) alone or in combination. qnrA and qepA were not found. Additionally, 9 (60%) of 12 PMQR positive isolates were co-existence with CTX-M-1 or CTX-M-9. CTX-M or PMQR producing isolates showed highly resistance to penicillins (100%), cephalosporins (100~66.7%), monobactams (72.2%), and non-β-lactam antibiotics (94.4~61.1%) such as quinolones, trimethoprim/sulfamethoxazole, tetracycline and gentamicin. These findings showed CTX-M-1, CTX-M-9, aac(6')-lb-cr and qnrB were highly prevalent in cefotaxime-resistant Enterobacteriaceae isolates from companion animals in our region. Moreover, PMQR genes were closely associated with CTX-M type β-lactamase.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.4
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• pp.422-430
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• 2018

The inappropriate and widespread use of quinolones in humans and animals may cause accelerated emergence and spread of antimicrobial-resistant determinants. In this study, we investigated quinolone resistance mechanisms in a total of 65 nalidixic acid-resistant E. coli isolated from swine rectal swabs (N=40) and clinical specimens (N=25). Antimicrobial susceptibilities were determined by the disk diffusion method. PCR and DNA sequencing were performed for investigations of genes and mutations associated with quinolone resistance. In our study, 62 of 65 nalidixic acid-resistant E. coli harbored mutations in gyrA, parC, and/or parE genes; of the 65 isolates, 62 (95.4%) had mutations in the gyrA gene, 35 (53.8%) had mutations in the parC gene, 7 (10.8%) had mutations in the parE gene. The 35 isolates harbored mutations in two genes, gyrA and parC. Plasmid-mediated quinolone resistance (PMQR) determinants were investigated in the 65 isolates. Thirteen of 65 nalidixic acid-resistant E. coli contained the qnrS gene and 10 of those isolates had mutations in the gyrA, parC, and/or parE genes. We confirmed that an important mechanism of quinolone resistance in E. coli isolated from human and swine involves chromosomal mutations in the gyrA, parC, and/or parE genes with increasing use of quinolone for treatment or additives.