• Korean Journal of Fisheries and Aquatic Sciences
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• v.38 no.6
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• pp.365-371
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• 2005

The antimicrobial resistance patterns to the 10 antimicrobial agents of potential pathogenic vibrios isolated from seawater and a variety of farmed fishes, including oliver flounder (Payalichthys olivaceus), black rock fish (Sebastes schlegeli), red sea bream (Pagyus major) and sea bass (Lateolabyax japonicus), were investigated from May to October, 2004. A total of 314 strains of the genus vibrios were isolated from 126 collected samples, and the number of isolated strains of Vibrio payahaemolyticus and Vibrio alginolyticus were 194 and 120, respectively. Apparently $98.5\%$ of V. parahaemolyticus and $100\%$ of V. alginolyticus isolates demonstrated antimicrobial resistance against at least one antimicrobial agent. The resistance of V. parahaernolyticus isolates to ampicillin ($97.9\%$) was highest, followed by oxolinic acid ($26.8\%$), amikacin ($19.1\%$) and tetracycline and amoxicillin/clavulanic acid ($6.7\%$). V. alginolyticus isolates were resistant to ampicillin ($100\%$), solfamethoxazoleit,imethopenem ($25\%$), amikacln ($21.7\%$), amoxicillin/clavulanic acid ($15.8\%$), ciprofloxacin ($13.3\%$), and tetracycline and doxycycline ($11.7\%$). The rate of multiple antimicrobial resistance to at least four antimicrobials was higher in the V. alginolyticus isolates ($20.8\%$) than in the V. parahaemolyticus ($6.7\%$).

• Korean Journal of Veterinary Research
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• v.47 no.3
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• pp.303-308
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• 2007

The high mortality (65.5%) of young olive flounder (Paralichthys olivaceus) showing skin ulcers occurred in fish obtained from a commercial cultured farm at Donghae on the eastern coast of Korea at March 2006. The dorsal skin showed ulcer lesions and large amount of mucus. We isolated Ichthyobodo sp. from the gill, and Vibrio anguillarum from the liver and kidney of dead and diseased fish. All of the fish infected with the V anguillarum and Ichthyobodo sp. were treated with 100 ppm formalin for 1 h, and treated with oxolinic acid for 4 h at $22^{\circ}C$, respectively. The formalin treated group was observed the low mortality (2.9%) when compared with high mortality of non-treated (82.9%), and of antibiotic treated group (85.9%) against mixed infections by Vibrio anguillarum and Ichthyobodo sp. organisms. We confirmed complete elimination against mixed infections of Vibrio anguillarum and Ichthyobodo sp. with 100 ppm formalin treatment.

• Fisheries and Aquatic Sciences
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• v.22 no.12
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• pp.29.1-29.7
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• 2019

Veterinary drugs are widely used to protect production-related diseases and promote the growth of farmed fish. The use of large amounts of veterinary drugs may have potential risk and cause adverse effects on both humans and the environment. In this study, we developed risk-based ranking based on a scoring system to be applied in the national residue program. In this approach, the following three factors of veterinary drugs that may occur as residues in fishery products were considered: potency (acceptable daily intake), usage (number of dose and withdrawal period), and residue occurrence. The overall ranking score was calculated using the following equation: potency × usage (sum of the number of sales and withdrawal period) × residue occurrence. The veterinary drugs that were assigned high score by applying this approach were enrofloxacin, amoxicillin, oxolinic acid, erythromycin, and trimethoprim. The risk-based approach for monitoring veterinary drugs can provide a reliable inspection priority in fishery products. The developed ranking system can be applied in web-based systems and residuemonitoring programs and to ensure safe management of fishery products in Korea.

• Journal of fish pathology
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• v.16 no.1
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• pp.61-68
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• 2003

MICs of 8 chemotherapeutic agents against forty streptococcal isolates were determined. These strains were isolated from diseased olive flounder, Paralichthys olivaceus, and showed 2-5 multiple drug resistance against different antibacterial agents including ampicillin, doxycycline (DOXY), erythromycin, florfenicol, flumequine, norfloxacin, oxolinic acid, and oxytetracycline (OTC). In conjugation experiment, we found transferable R plasmids carrying OTC and DOXY resistance determinant in 3 drug resistance strains analyzed. Six out of 40 isolates showed positive signal in colony hybridization with the R plasmid DNA (pST9) as a probe. It suggests that other types R plasmid different from pST9 is also involving in multiple drug resistance of streptococci isolated from olive flounder.

• Korean Journal of Veterinary Research
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• v.20 no.2
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• pp.79-84
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• 1980

One hundred and eighteen cultures of Gram-negative rods isolated from cases of clinical bovine mastitis during lactation were examined for distribution of specific types, and activity of several antimicrobial agents to the isolates was determined by two-fold tube dilution method employing sterile whole milk as fluid medium. Of the isolates, 59.2% were Escherichia coli. Most of the remaining isolates were Klebsiella pneumoniae and Enterobacter aerogenes. Most Gram-negative rods(89.8%) were isolated from acute local and chronic mastitis. The cases of peracute systemic form with a marked symptoms of toxemia were associated with Escherichia coli and Klebsiella pneumoniae. The minimal lethal concentrations (MLC) of gentamicin and oxolinic acid in sterile whole milk were 16-128 times higher than the MLC obtained in trypticase soy broth (TSB), while the MLC of ampicillin and tetracycline in milk increased 2-4 times compared with TSB. Of the drugs tested, gentamicin was the most active antibiotics with MLC of $100{\mu}g/ml$ in sterile whole milk against all of Gram-negative rods isolated from bovine udder infections.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.6
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• pp.838-844
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• 2016

In this study, epidemiological cut-off values were estimated for 44 Photobacterium damselae isolates, since clinical breakpoints have not been established for this pathogenic bacterium. The susceptibility of the isolates to 10 antibiotics was evaluated using internationally standardized disc diffusion protocols. Normalized resistance interpretation was used to generate statistically valid epidemiological cut-off values for the susceptibility data. There were not enough strains exhibiting full sensitivity to ampicillin and amoxicillin to allow analysis of these antibiotics. Because there were only a marginally sufficient number of strains exhibiting full sensitivity to oxytetracycline, the cut-off value generated provided only a provisional estimate. The valid wild-type cut-off values were <13, 13, 9, 22, 25, 27, and 28 mm for gentamicin, cephalexin, erythromycin, oxolinic acid, flumequine, florfenicol, and sulfamethoxazole/trimethoprim, respectively. The application of these cut-off values should greatly facilitate the rational selection of antibiotics for use in commercial fish farms.

• Journal of Veterinary Clinics
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• v.19 no.1
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• pp.19-22
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• 2002

The examination with 96 bacteria isolates for antibiotics susceptibility was resulted in exploring many antibiotics resistance isolates from the diseased fishes. Vibrio sp. isolates were demonstrated over 90% resistances for Penicillin (P), Amoxicillin (Amc), Erythromycin (I), Colistin (CL). Streptomycin (S). Novobiocin (NV) and Neomycin(N), these isolates were, however, appeared over 80% susceptibilities for Norfloxacin (Nor), CE and UB. In Edwardsiella tarda case, 90 to 100% high resistance was observed for P, Doxycycline (Do), Tetracyclin (Te), Oxytetracycline (T), E, CL, Sulfamethoxasoletrimetoprim (SXT), S, Oxolinic acid (OA), W, Neomycin(N), on the contrary, 90 to 100% high susceptibilities was found for Amc, Nor, Ciprofloxacin (Cip), Orbifloxacin (ORB), Enrofloxacin (ENR), Flumeguine (UB) and NA. CL, Pefloxacin (PEF),S, Flumeguine (UB), OA, NA, NV N was exhibited 90 to 100% resistances for Streptococcus sp., on the other hand, 100% susceptible to AMC and 80% susceptible to Do, Te, ENR and UB was recognised. Lastly, Photobacterium damsela subsp. piscicida was showed 100fe susceptible to Amc and 86% susceptible to NOR, CIP ENR and UB. As a consequence, fish bacterial pathogens isolated from Kyeongnam area, especially Tongyeong-si, Geoje-si, and Goseong-gun, were showed highly resistant to a variety of antibiotics available in the field.

• Korean Journal of Veterinary Research
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• v.21 no.1
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• pp.25-31
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• 1981

A total of 250 enteric bacteria (148 Escherichia coli, 41 Klebsiella pneumoniae, 46 Enterobacter spp. and 15 Proteus spp.) isolated from bovine udder infections in 1979 through 1980 were examined for drug resistance and prevalence of R. plasmids. The drug tested were streptomycin (SM), kanamycin (KM), ampicillin (AP), chloramphenicol (CP), tetracycline (TC), gentamicin (GM), oxolinic acid (OA) and nalidixic acid (NA). The detection of R. plasmids was performed with Escherichia coli ML 1410 NAr as the recipient. Of the 148 Escherichia coli isolated, 68(45.9%) were found to be resistant to one or more drugs tested, and about 50% of the resistant strains were multiply resistant. of the 68 drugresistant strains, 13(19.1%) were found to carry R. plasmids which were capable of performing a conjugal transfer. CP resistance was transfered together with the other resistance. Of 41 strains of Klebsiella pneumoniae isolated, 90.2% were resistant to the drugs, alone or in combination thereof. Strains resistant to AP and TC were 63.4%, and 48.8%, respectively. R. plasmids were detected in 78.4% of the drug-resistant strains, and these strains transfered all or a part of their drug resistance pattern. AP and CP resistance were transfered in 100% of AP and CP-resistant strains. Eleven (37.9%) of 29 R. plasmids showed a thermosensitive transfer. Of the 46 strains of Enterobacter spp. isolated, 37(80.4%) were resistant to the drugs tested. A high percentage of resistance was noted for AP(65.2%). All strains resistant to four or more drugs transferred their resistances to Escherichia coli ML 1410, but strains resistant to three or fewer drugs did not transfer the resistances. All of the 15 Proteus strains isolated were resistant to more than two drugs. of them, 6 were quadruple resistance to SM, KM, CP and TC, and 9 were double one to AP and TC. Three (20.0%) of the drug-resistant isolates had R.plasmids conferring AP and TC resistance. GM, OA and NA of the drugs tested were very active to all of 250 Gram-negative enteric bacteria isolated from bovine udder infections.

• Journal of fish pathology
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• v.10 no.2
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• pp.125-135
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• 1997

The possibility of identification of families of antibacterial agent residues in fish tissue was studied by disk assay using three test organisms, Bacillus subtilis BGA, Micrococcus luteus ATCC 9341, and Bacillus cereus var. mycoides ATCC 11778. In the present method, a simple clean-up procedure was performed to obtain the aqueous solution from homogenized flounder muscle sample(10g) in Mcilvaine buffer. Then, aqueous solution was fractionated into A and B to be used in disk assay by choloroform and Sep-Pak $C_{18}$ cartridge column after being defatted in hexane. The chloroform layer of fraction A was used for the analysis of macrolide antibiotics(ML), sulfa drugs(SA), chloramphenicol(CP), and quinolone antibiotics(QN). Adsorbed materials to Sep-Pak $C_{18}$ of fraction B were also employed for the analysis of penicillins(PC), tetracyclines(TC), and nitrofuran derivatives(NF) Minimun-detectable concentrations by the present method were, $0.1{\mu}g$/g for oxytetracycline, tetracycline, doxycycline, spiramycin and ciprofloxacin, $0.025{\mu}g$/g for erythromycin and ampicillin, $1.0{\mu}g$/g for sodium nifurstyrenate and florfenical, $0.25{\mu}g$/g for sulfamonomethoxie and sulfadimethoxine, $2.5{\mu}g$/g for oxolinic acid and flumequine, and $15{\mu}g$/g for piromidic acid, respectively. Three test organisms showed different sensitivity patterns for each family of antibacterial agent. Sensitivity patterns were B. cereus > B. subtilis > M. luteus for TC and NF, M. luteus > B, subtilis > B. cereus for ML and PC, B. cereus = B. subtilis > M. luteus for CP and QN, and B. subtilis > B. cereus＝M. luteus for SA. The present method utilizing these characteristics could be useful as a routine screening test for the determination of family of antibacterial agent residues in fish tissue.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.