• Journal of Practical Agriculture & Fisheries Research
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• v.22 no.2
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• pp.51-58
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• 2020

Continuous mortality in commercially cultured Japanese eel (Anguilla japonica), showing symptoms of dermal ulcerations and focal hemorrhages on the body, occurred on a private farm in November, 2019 in Korea. A series of mortality had been described in one local eel culture farm from November to December in 2019. From the three cases, three isolates of Vibrio spp. were recovered from the blood, ascitic fluid, and kidney of the dead fish, respectively. Based on the 16S rRNA sequence comparisons, the Vibrio isolates from the 1^st and 3^rd cases (strain named 1E1-2 and 3K1-2) were identified as V. fluvialis and the isolate from the 2^nd case was identified as V. plantisponsor (strain named 2A3-1). Moreover, the 16S rRNA-based phylogenetic analysis revealed that strain 1E1-2 and 3K1-2 were most similar to V. fluvialis NBRC 103150^T, and strain 2A3-1 was most similar to V. plantisponsor NBRC103148^T. According to the results of the antibiotic resistance determination, V. fluvialis 1E1-2 showed intermediate resistance to tetracycline and chloramphenicol, and was resistant to trimethoprim-sulfamethoxazole. V. plantisponsor 2A3-1 showed intermediate resistance to ciprofloxacin and levofloxacin, and was resistant to trimethoprim-sulfamethoxazole. V. fluvialis 3K1-2 showed intermediate resistance to tetracycline, and was resistant to ampicillin and trimethoprim-sulfamethoxazole. These results have provided the evidences on the occurrence of antibiotic-resistant Vibrio infection in commercially cultured Japanese eels are present in Korea.

• Fisheries and Aquatic Sciences
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• v.18 no.3
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• pp.241-247
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• 2015

In an effort to discover alternative phytotherapeutic antimicrobial agents to combat Streptococcus parauberis, a fish pathogenic bacterium, we evaluated the antibacterial activity of seaweed extracts in vitro. A methanolic extract of Ecklonia cava exhibited strong antibacterial activity against S. parauberis isolated from olive flounder Paralichthys olivaceus. Furthermore, the n-hexane soluble (Hexane) fraction of the E. cava methanolic extract exhibited the greatest antibacterial effect on S. parauberis strains with a minimum inhibitory concentration (MIC) ranging from 256 to $1,024{\mu}g/mL$. In addition, the MIC values of oxytetracycline against antibiotic-resistant S. parauberis were markedly reduced up to 64-fold in combination with the Hexane fraction, suggesting that the antibacterial activity of the antibiotic was restored when combined with the Hexane fraction. The interaction between both antibiotics and the Hexane fraction was assessed by the fractional inhibitory concentration (FIC) index. The Hexane fraction and oxytetracycline combination against antibiotic-resistant S. parauberis strains resulted in a median ${\sum}FIC$ range of 0.502 to 0.516. Thus, the synergistic ranges of median ${\sum}FIC$ < 1 were observed for all combinations of the Hexane fraction and oxytetracycline against S. parauberis. To the best of our knowledge, this is the first report indicating the efficacy of an E. cava extract against fish pathogenic bacterium S. parauberis.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.394-401
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• 2016

Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) system, a genome editing technology, was shown to be versatile in treating several antibiotic-resistant bacteria. In the present study, we applied the CRISPR/Cas9 technology to kill extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. ESBL bacteria are mostly multidrug resistant (MDR), and have plasmid-mediated antibiotic resistance genes that can be easily transferred to other members of the bacterial community by horizontal gene transfer. To restore sensitivity to antibiotics in these bacteria, we searched for a CRISPR/Cas9 target sequence that was conserved among >1,000 ESBL mutants. There was only one target sequence for each TEM- and SHV-type ESBL, with each of these sequences found in ~200 ESBL strains of each type. Furthermore, we showed that these target sequences can be exploited to re-sensitize MDR cells in which resistance is mediated by genes that are not the target of the CRISPR/Cas9 system, but by genes that are present on the same plasmid as target genes. We believe our Re-Sensitization to Antibiotics from Resistance (ReSAFR) technology, which enhances the practical value of the CRISPR/Cas9 system, will be an effective method of treatment against plasmid-carrying MDR bacteria.

• Korean Journal of Plant Resources
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• v.33 no.6
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• pp.578-585
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• 2020

This study was designed to assess the possibility of using medicinal plant extracts as β-lactamase inhibitors to control antibiotic-resistant Staphylococcus aureus. The susceptibilities of S. aureus ATCC 15564 (SA^WT), ciprofloxacininduced S. aureus ATCC 15564 (SA^CIP), oxacillin-induced S. aureus ATCC 15564 (SA^OXA), and clinically-isolated S. aureus CCARM 3008 (SA^CLI) to ampicillin were determined in the absence and presence of medicinal plant extracts, including Cleyera japonica (CJ), Carpinus laxiflora (CL), Euphorbia helioscopia (EH), Euscaphis japonica (EJ), Oenothera erythrosepala (OE), and Rosa multiflora (RM). The phenotypic change in the clear inhibition zones around ampicillin disc was observed for SA^WT, SA^CIP, and SA^OXA, indicating the production of ampicillinase. Compared to the controls, the MICs of ampicillin against SA^WT, SA^CIP, and SA^OXA were decreased from 4 to 0.5 ㎍/mL in the presence of CL, 16 to 4 ㎍/mL in the presence of RM, and 32 to 2 ㎍/mL in the presence of CL, EH, and RM, respectively. The medicinal plant extracts, OE, EJ, and CL, effectively inhibited the β-lactamase activities of SA^WT (78%), SA^CIP (57%), and SA^OXA (76%) when compared to the control. This results suggest that the medicinal plant extracts can be used as BLIs to control the antibiotic-resistant S. aureus.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.386-393
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• 2014

Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n=465) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.

• Korean Journal of Clinical Laboratory Science
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• v.37 no.3
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• pp.155-163
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• 2005

The purpose of this study is to investigate the carrier rate of S. aureus in the community, antibiotic susceptibility patterns of the organism, detection of MRSA and mecA gene in MRSA. Identification and antibiotic resistance patterns of S. aureus and MRSA were done by MicroScan Panels. MRSA strain was confirmed by disk diffusion method using oxacillin disk. The mecA gene in MRSA was detected by PCR. Eighty-four strains (27.4%) of S. aureus were isolated from the nasal specimens of 307 university students in Busan in 2004. Sixty-eight strains (81.9%) of 83 S. aureus were resistant to penicllin, 16 strains(19.3%) to erythromycin, 15 strains (18.1%) to gentamicin, 12 strains (14.5%) to tetracycline, 6 strains (7.2%) to chloramphenicol, 3 strains (3.6%) to ofloxacin, 2 strains (2.4%) to cefepime, clindamycin, imipenem, meropenem, norfloxacin, respectively. One strain (1.2%) was resistant to ciprofloxacin, cefazolin, cefotaxime, cefuroxime, and oxacillin. And all the strains (100%) of 84 S. aureus were susceptible to amoxicilin/K clavulanate, ticarcillin/K clavulanate, trimethoprim/sulfamethoxazole, rifampin, syncroid, teicoplanin, and vancomycin. One strain of 84 S. aureus isolates was methicillin-resistant Staphylococcus aureus (MRSA). The mecA gene was detected from the MRSA strain.

• Journal of Pharmacopuncture
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• v.22 no.1
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• pp.49-54
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• 2019

Objective: Antibiotic resistance is a global health problem and threatens health of societies. These problems have led to a search for alternative approaches such as combination therapy. The aim of the present study was to investigate the effect of caffeine and omeprazole in combination with gentamicin or ciprofloxacin against standard and clinically resistant isolates of Staphylococcus aureus and Escherichia coli. Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of different agents against bacterial strains were determined. The interaction of non- antibiotic drugs with gentamicin and ciprofloxacin was studied in vitro using a checkerboard method and calculating fraction inhibitory concentration index (FICI). Verapamil as efflux pump inhibitor was used to evaluate the possible mechanism of bacterial resistance to antibiotics. Results: The MIC and MBC values of gentamicin against bacterial strains were in the range of $20-80{\mu}g/ml$ and $40-200{\mu}g/ml$, respectively. Caffeine and omeprazole had no intrinsic inhibitory activity against tested microorganisms. However, upon combination of caffeine with antibiotics, the synergistic effects were observed. Verapamil was able to reduce the MIC values of gentamicin (4 folds) only in some bacterial strains. Conclusion: These findings indicated that caffeine was effective in removing bacterial infection caused by S. aureus and E. coli. The relevant mechanisms of antibiotic resistance were not related to the drug efflux.

• Journal of Practical Agriculture & Fisheries Research
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• v.23 no.1
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• pp.81-87
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• 2021

Olive flounders which were cultured in commercial fish farm showed abnormal swimming behavior in November 2020. Then, gradual mortality was observed in the fish farm. During the diagnosis, bacterial strain KNCFKW-PN1 was isolated from the kidney of the dead fish. Based on the sequence of gyrase B subunit gene, KNCFKW-PN1 was proved to be Pseudoalteromonas nigrifaciens, showing 99.59% nucleotide identity with that of P. nigrifaciens LMG 2227^T. According to the result of antibiotic susceptibility test, P. nigrifaciens KNCFKW-PN1 showed intermediate resistance to ciprofloxacin, and was resistant to ampicillin, cefepime, cefotaxime, ceftazidime, and amikacin. This is the first report of the isolation of multiple-antibiotic-resistant P. nigrifaciens from olive flounder.

• Journal of the Korean Wood Science and Technology
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• v.52 no.2
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• pp.145-156
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• 2024

Unlike resistant cells, persister cells resist antibiotics due to a decreased cellular metabolic rate and can transition back to normal susceptible cells when the antibiotic is removed. These persister cells contribute to the chronic symptoms of infectious diseases and promote the emergence of resistant strains with continuous antibiotic exposure. Therefore, eliminating persister cells represents a promising approach to significantly enhance antibiotic efficacy. Here, we found that Coicis Semen extract reduced Staphylococcus aureus persister cells at a concentration of 0.5 g/L. Linoleic acid and oleic acid, the major components of Coicis Semen extract, exhibited a comparable reduction in persister cells when combined with three antibiotics: ciprofloxacin, oxacillin, and tobramycin. Conversely, these effects were nullified in the presence of the surfactant Tween 80 (1%), suggesting that the hydrophobic characteristics of linoleic acid and oleic acids play a pivotal role in reducing the number of S. aureus persister cells. Considering the concentration-dependent effects of linoleic acid and oleic acid, the persister-reducing activity of Coicis Semen extract was primarily attributed to these fatty acids. Moreover, Coicis Semen extract, linoleic acid, and oleic acid increased the cell membrane permeability of S. aureus. Interestingly, this effect was counteracted by 1% Tween 80, indicating a close association between the reduction of persister cells and the increase in cell membrane permeability. The identified compounds could thus be used to eliminate persister cells, thereby enhancing therapeutic efficacy and shortening treatment duration. When used in conjunction with antibiotics, they may also mitigate chronic symptoms and significantly reduce the emergence of antibiotic-resistant bacteria.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.5
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• pp.626-633
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• 2023

Vibrio spp. are aquatic bacteria that are ubiquitous in warm estuarine and marine environments. Especially, V. vulnificus and V. cholerae are currently known to cause potentially fatal infections in humans. This study investigated the distribution and antibiotic resistance of V. vulnificus and V. cholerae isolated from coastal areas of Gyeongsangnam-do in 2022. A total of 252 samples of water, shellfish and coastal sediment were collected from 7 locations along the coast, and 124 samples of fishery products were collected from markets. Among the 252 samples, forty-four V. vulnificus (11.7%) and fourteen V. cholerae non-O1/non-O139 (3.7%), none of which carried the ctx gene, were isolated. Out of the 124 samples, 6 (4.8%) tested positive for V. vulnificus and V. cholerae was not detected. The isolation rates of V. vulnificus and V. cholerae showed a significant correlation with environmental factors such as seawater temperature and salinity. In an antibiotic resistance test, V. vulnificus was susceptible to amikacin, gentamicin, imipenem trimethoprim/sulfamethoxazole, and ciprofloxacin, but resistant to cefoxitin (100.0%), followed by tetracycline (9.1%). Multidrug resistance was also observed. Continuous monitoring of Vibrio pathogens with water temperature and salinity is expected to help reduce the outbreaks, and rational use of antibiotic agents is needed to prevent the accession of antibiotic-resistant microorganisms in aquatic ecosystems.