• Food Science of Animal Resources
• /
• v.43 no.5
• /
• pp.792-804
• /
• 2023

Non-aureus staphylococci (NAS), particularly antimicrobial-resistant NAS, have a substantial impact on human and animal health. In the current study, we investigated (1) the species profiles of NAS isolates collected from healthy broilers, farm environments, and farm workers in Korea, (2) the occurrence of antimicrobial-resistant NAS isolates, especially methicillin resistance, and (3) the genetic factors involved in the methicillin and fluoroquinolone resistance. In total, 216 NAS isolates of 16 different species were collected from healthy broilers (n=178), broiler farm environments (n=18), and farm workers (n=20) of 20 different broiler farms. The two most dominant broiler-associated NAS species were Staphylococcus agnetis (23.6%) and Staphylococcus xylosus (22.9%). Six NAS isolates were mecA-positive carrying staphylococcal cassette chromosome mec (SCCmec) II (n=1), SCCmec IV (n=1), SCCmec V (n=2), or nontypeable SCCmec element (n=2). While two mecA-positive Staphylococcus epidermidis isolates from farm workers had SCCmec II and IV, a mecA-positive S. epidermidis isolate from broiler and a Staphylococcus haemolyticus isolate farm environment carried SCCmec V. The occurrence of multidrug resistance was observed in 48.1% (104/216 isolates) of NAS isolates with high resistance rates to β-lactams (>40%) and fusidic acid (59.7%). Fluoroquinolone resistance was confirmed in 59 NAS isolates (27.3%), and diverse mutations in the quinolone resistance determining regions of gyrA, gyrB, parC, and parE were identified. These findings suggest that NAS in broiler farms may have a potential role in the acquisition, amplification, and transmission of antimicrobial resistance.

• Korean Journal of Clinical Laboratory Science
• /
• v.48 no.2
• /
• pp.118-123
• /
• 2016

Antimicrobial agents have been used in poultry for treatment of bacterial infections or additives over the past half century. However, increasing antimicrobial resistance has led to selective pressure for therapeutic use in humans and made treatment of bacterial infection more difficult. In this study, we examined the prevalence of plasmid mediated antimicrobial resistant determinants for resistance to ${\beta}-lactam$, quinolone, and aminoglycoside in Enterobacteriaceae isolates obtained from chickens in Gyeongsang provinces, and correlation between the resistant genes and antimicrobial resistance rate was also assessed. A total of 43 Enterobacteriaceae isolates were recovered from 40 chickens at Gyeongsang provinces in Korea. Antimicrobial susceptibility was determined by disk diffusion method. PCR and DNA sequencing were performed to characterize the antimicrobial resistant genes. Of the 43 Enterobacteriaceae isolates tested, 2 isolates harbored $bla_{CTX-M-14}$ gene, and 2 and 5 strains contained qnrS and aac(6')-Ib-cr genes, respectively. A total of 43 isolates displayed a relatively lower susceptible rate ranging between 0.0 and 23.3% to most of the antimicrobial agents, except cefepime, ceftazidime, and cefaclor. We confirmed that plasmid mediated antimicrobial resistant determinants were distributed in Enterobacteriaceae isolates from chickens. Investigation of the genes and monitoring of antimicrobial resistance rate is required to prevent further spreading of antimicrobial resistant genes among Enterobacteriaceae isolates.

• Biomedical Science Letters
• /
• v.18 no.1
• /
• pp.79-82
• /
• 2012

Acinetobacter infections are of great concern in clinical settings because of multi-drug resistance (MDR) and high mortality of the infected patients. The MDR Acinetobacter baumannii has emerged as a significant infectious agent in hospitals worldwide. The purpose of this study was to determine for molecular characterization of MDR A. baumannii clinical isolates obtained from the Wonju Christian Hospital in Gangwon province of Korea. A total of seventy nonduplicate A. baumannii isolates were collected from the Wonju Christian Hospital in Korea from March to April in 2011. All of the MDR A. baumannii isolates were encoded by $bla_{OXA-23-like}$ gene and all isolates with the $bla_{OXA-23-like}$ gene had the upstream element ISAba1 to promote increased gene expression and subsequent resistance to carbapenem. 16S rRNA methylase gene (armA) was detected in 44 clinical isolates which were resistant to amikacin, and phosphotransferase genes encoding aac(3)-Ia and aac(6')-Ib were the most prevalent. A combination of 16S rRNA methylase and aminoglycoside-modifying enzyme genes (armA, aac(3)-Ia, aac(6')-Ib, and aph(3')-Ia) were found in 31 isolates. The sequencing results for the quinolone resistance-determining region (QRDR) of gyrA and parC revealed the presence of Ser (TCA) 83 Leu (TTA) and Ser (TCG) 80 Leu (TTG) substitutions in the respective enzymes for all MDR. Molecular typing for MDR A. baumannii could be helpful in confirming the identification of a common source or cross-contamination. This is an important step in enabling epidemiological tracing of these strains.

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

• Journal of Life Science
• /
• v.17 no.3 s.83
• /
• pp.435-443
• /
• 2007

Antimicrobial susceptibility test of the 116 strains of Mycoplasma pneumoniae isolates were performed by a broth micro-dilution method against to moxifloxacin, levofloxacin, sparfloxacin, ofloxacin, ciprofloxacin, clarithromycin minocycline, erythromycin, josamycin, and tetracycline. The initial-minimum inhibitory concentration (I-MIC) was evaluated as the lowest concentration of antimicrobial agents that prevented a color change in the medium at that time when the drug-free growth control, about 7 days after incubation, and the final-minimum inhibitory concentration (F-MIC) was defined a color change about 14 days after incubation. The evaluation to the drug-resistant M. pneumoniae isolates were determined the $MIC{\pm}1.0$ ${\mu}g/ml$ of each antimicrobial agent. According to the I-MIC, single drug-resistant M. pneumoniae strains to ciprofloxacin, ofloxacin, clarithromycin and erythromycin were 79.3, 53.5, 10.3, and 7.8%, respectively. Two kinds of drug-resistant M. pneumoniae strains to ofloxacin and ciprofloxacin, or ciprofloxacin and clarithromycin were 42.2 and 9.5%. Three kinds of drug-resistant M. pneumoniae strains to erythromycin, ofloxacin, and ciprofloxacin, or ofloxacin, ciprofloxacin and clarithromycin were 6.9 and 6.0% . According to the F-MIC, single drug-resistant M. pneumoniae strains to tetracycline, ciprofloxacin, ofloxacin, minocycline,erythromycin, josamycin, clarithromycin and sparfloxacin were 91.4, 91.4, 91.4, 89.7, 68.1, 52.6, 28.5, and 11.2%, respectively. The incidence of two kinds of drug-resistant M. pneumoniae strains were from 20.7% to 91.4%, three kinds of drug-resistant M. pneumoniae strains were from 28.5% to 89.7%, four kinds of drug-resistant M. pneumoniae strains were 2.6%, five kinds of drug-resistant M. pneumoniae were from 2.6% to 21.6%, six kinds of drug-resistant M. pneumoniae strains were from 0.9% to 24.1%, seven kinds of drug-resistant M. pneumoniae strains were from 0.9% to 2.6%, and eight kinds of drug-resistant M. pneumoniae strains were 1.7%. These results suggest that sparfloxacin, moxifloxacin and levofloxacin might be promising antimicrobial agents for the treatment of M. pneumoniae infection in Korea. However, most strains of M. pneumoniae isolates were single or multi-resistance pattern to the other tested antimicrobial agents. Therefore, tetracycline, minocycline, erythromycin, clarithromycin, and second-generation quinolones are more carefully used to patients with M. pneumoniae infection in Korea.

• Journal of Microbiology
• /
• v.40 no.1
• /
• pp.63-69
• /
• 2002

A total of twenty-five norfloxacin resistant Escherichia coli were isolated from Joongrang-chun stream, a branch of the Han River in Seoul, Korea from May to July in 2000 and their norfloxacin resistance mechanism was characterized for target site mutation, permeability, and efflux pump. Fourteen iso- lates contained the same three mutations, Ser83→Leu and Asp87→Asn in GyrA and Ser90→ lle in ParC. Six isolates had Ser83→Leu and Asp87→Tyr in GyrA and Ser87→lle in ParC while one isolate had Ser83→Leu and Va1103→Ala in GyrA and Ser80→lle in ParC. Two isolates had mutation(s) in GyrA without any mutation in ParC. Two isolates had Ser80→Arg in ParC instead of the commonly found Ser80→lle. Every norfloxacin resistant isolate had an efflux system but the correlation between the efflux activity and MIC was not observed. The amount of OmpF for norfloxacin permeability decreased in resistant isolates compared to the susceptible strains. When amplified polymorphic DNA (RAPD) and pulse field gel electrophoresis (PFGE) were performed, these isolates showed no similarity to each other or clinical isolates.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.1
• /
• pp.155-158
• /
• 2003

In early 2002, over 200 people in the city of Pusan. Korea suffered from paratyphoid fever resulting from Salmonella Paratyphi A Infection. Antimicrobial susceptibility tests and Xbal pulsed-field gel electrophoresis (PFCE) were conducted to 54 Salmonella Paratyphi A isolated from humans during the period of 1998 to 2002. Most of the isolates ($83\%$) were only nalidixic acid-resistant and $78\%$ were X 1 PFGE patterns. Also, we measured the MIC of ciprofloxacin and screened gyrA mutation(5) using allele- specific PCR and restriction fragment length polymorphism (AS-PCR-RFLP). The representative 5 isolates in 2002 and 1 isolate in 2000 were $1{\mu}g/ml$ of MIC and had mutation at the 83rd codon in gyrA. These data suggest that the outbreak in the early 2002 might have been due to dissemination of the strain present In 2000. Also, decreased susceptibility to ciprofloxacin was partly due to the mutation at the 83rd codon in gyrA.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.10a
• /
• pp.113-113
• /
• 2018

Endophthalmitis is a disease that causes ocular inflammation and has a catastrophic effect on eyesight. Recent studies show that Enterococcus faecalis is rapidly increasing causative bacterium of endophthalmitis. It is predicted that the increased endophthalmitis by E. faecalis is presumable due to the high resistance of E. faecalis to moxifloxacin (MFX), which is a common antibiotic used for eye drop. Because of the need for therapeutic agents to overcome this problem, this study sought to explore the feasibility of developing a combination therapy using Orostachys japonicus. The ethylacetate fraction of O. japonicus (OJA) used in this study. Antimicrobial activity was tested 13 E. faecalis strains including one E. faecalis standard strain, eight clinically isolated E. faecalis strains and four quinolone resistant E. faecalis strains using CLSI antibiotic susceptibility test method. Minimal Inhibitory Concentration (MIC) of OJA was confirmed to be $500{\mu}g/ml$ for all 13 strains. Then we tested for the synergistic effect of OJA to MFX using checkboard test method. The MIC of MFX was $0.25{\mu}g/ml$ for the standard strain and 8 for the clinical isolates, and $16{\sim}64{\mu}g/ml$ for the quinolone - resistant strains. When OJA was mixed with MFX, no synergistic effect was observed in all strains, but the antibacterial activity of OJA remained unchanged. Most ocular other strains can be removed by MFX except the MFX resistant E. faecalis, which can be removed by OJA in combination therapy. Therefore, OJA can be a potential candidate for the combined treatment endophthalmitis.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.9
• /
• pp.1293-1303
• /
• 2013

Antibiotic resistance of soilborne Acinetobacter species has been poorly explored. In this study, norfloxacin resistance of a soil bacterium, Acinetobacter oleivorans DR1, was investigated. The frequencies of mutant appearance of all tested non-pathogenic Acinetobacter strains were lower than those of pathogenic strains under minimum inhibitory concentration (MIC). When the quinolone-resistance-determining region of the gyrA gene was examined, only one mutant (His78Asn) out of 10 resistant variants had a mutation. Whole transcriptome analysis using a RNA-Seq demonstrated that genes involved in SOS response and DNA repair were significantly up-regulated by norfloxacin. Determining the MICs of survival cells after norfloxacin treatment confirmed some of those cells were indeed persister cells. Ten colonies, randomly selected from among those that survived in the presence of norfloxacin, did not exhibit increased MIC. Thus, both the low mutation frequency of the target gene and SOS response under norfloxacin suggested that persister formation might contribute to the resistance of DR1 against norfloxacin. The persister frequency increased without a change in MIC when stationary phase cells, low growth rates conditions, and growth-deficient dnaJ mutant were used. Taken together, our comprehensive approach, which included mutational analysis of the target gene, persister formation assays, and RNA sequencing, indicated that DR1 survival when exposed to norfloxacin is related not only to target gene mutation but also to persister formation, possibly through up-regulation of the SOS response and DNA repair genes.

• The Plant Pathology Journal
• /
• v.38 no.5
• /
• pp.482-489
• /
• 2022

Fire blight caused by Erwinia amylovora (Ea) is a devastating disease in apple and pear trees. Oxolinic acid (OA), a quinolone family antibiotic that inhibits DNA gyrase, has been employed to control fire blight in South Korea since 2015. The continuous use of this bactericide has resulted in the emergence of OA-resistant strains in bacterial pathogens in other countries. To investigate the occurrence of OA-resistant Ea strains in South Korea, we collected a total of 516 Ea isolates from diseased apple and pear trees in 2020-2021 and assessed their sensitivities to OA. We found that all isolates were susceptible to OA. To explore the possibility of emerging OA-resistant Ea by continuous application of OA, we exposed Ea stains to a range of OA concentrations and constructed OA-resistant mutant strains. Resistance was associated with mutations in the GyrA at codons 81 and 83, which result in glycine to cysteine and serine to arginine amino acid substitutions, respectively. The in vitro growth of the mutants in nutrient media and their virulence in immature apple fruits were lower than those of wild-type. Our results suggest that OA-resistance decreases the fitness of Ea. Future work should clarify the mechanisms by which OA-resistance decreases virulence of this plant pathogen. Continuous monitoring of OA-resistance in Ea is required to maintain the efficacy of this potent bactericide.