• Journal of Applied Biological Chemistry
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• v.57 no.3
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• pp.189-196
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• 2014

In Korea, over 4,700 animal carcass disposal sites were installed until 2011 due to the outbreak of foot and mouth disease. Due to the putrefaction of buried animals, the leachate containing veterinary antibiotics may release into surrounding environments. Antibiotic residues in the environment cause the formation of antibiotic resistance bacteria threatening human and ecosystem health. This study reports the concentrations of five antibiotics, including tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC), sulfamethazine (SMZ), and sulfamethoxazole (SMX), in soils from animal carcass disposal site and adjacent agricultural field. Concentrations of antibiotics at animal carcass disposal sites (TC: $144.26-350.73{\mu}g/kg$, SMZ: $17.72-44.94{\mu}g/kg$) were higher than those at agricultural field (TC: $134.16-320.73{\mu}g/kg$, SMZ: $6.48-8.85{\mu}g/kg$) whereas the concentrations of CTC, OTC, and SMX were below detection limit in both sites. Results showed that the antibiotics in animal carcass site might leach to the soil and possibly contaminating the groundwater. Future studies will focus on the transfer of antibiotics residues into food crops.

• Analytical Science and Technology
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• v.18 no.3
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• pp.250-256
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• 2005

A selective method of high performance liquid chromatography with UV detector has been applied to determine 4 tetracycline antibiotics in the animal food, simultaneously. The targets were chlortetracycline (CTC), doxycycline (DC), oxytetracycline (OTC), and tetracycline (TC) that are used routinely in veterinary medicine for prevention and control of disease. Food samples were beef, pork, chicken, milk, whole egg, flatfish (Limanda yokohamae), jacopever (Sebastes hubbsi), seabream (Chrysophrys major), eel (Anguilla japonica) and lobster (Hommarus americanus). After extracting food samples with 20% trichloroacetic acid and McIlvaine buffer, they were purified by a $C_18$ SPE cartridge with 0.01M methanolic oxalic acid solution. The concentrated residue was re-dissolved in methanol, filtered, cleaned up and analyzed on a $C_18$ column. The mobile phase was a mixture of 0.01M oxalic acid and acetonitrile with a gradient ratio from 85:15 to 60:40. The UV wavelength was 365 nm. The overall recoveries were ranged from 71% to 98% and the limit of detections were 0.022 for CTC, 0.012 for DC and OTC and 0.009 mg/kg for TC at signal/noise > 3, respectively. As results, CTC, DC and TC were not detected in all selected food samples, however, OTC was detected in meat and fishes. The determined level of OTC was 0.04 ppm for pork, 0.17 ppm for flatfish and 0.05 and 0.08 ppm for jacopever, that were within the Maximum Residue Limits (MRLs) in the food.

• Journal of Fisheries and Marine Sciences Education
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• v.29 no.3
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• pp.834-846
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• 2017

Aquaculture practices to ensure greater production, such as high density breeding and excessive feeding, are become stressors that raise the prevalence of diseases. Accordingly, increasingly large volumes of antibiotics are used more frequently each year. Long term use antibiotics can generate resistant bacteria, which interrupt treatments and cause a potential transfer to human bodies. Thus, antibiotic resistance is of importance in public health. Tetracycline (Tc) is one of the typical medicines used in the aquaculture drugs, which has a wide range of application including gram-positive and gram-negative bacteria. In the examination of 153 strains isolated from olive flounder (Paralichthys olivaceus) farms located in Jeju in 2016, it turned out that a total of 84 strains were resistant to Tc or oxytetracycline (OTC). The extent to which the strains are resistant to Tc and OTC was confirmed through MIC test, mostly within the range of 25 to $100{\mu}g/m{\ell}$. Twelve different types of tet genes were detected using single and multiplex PCR in the 84 Tc-resistant strains. The PCR was used to find tet(K), tet(M), tet(O), and tet(S), which are known to exist primarily in gram positive strains. According to the results, - tet(S) is the most dominant gene in 49 strains of Streptococcus parauberis, accounting for 63.2%. And there were two strains that have two different types of resistant genes. The multiplex PCR was used to detect tet(A), tet(B), tet(C), tet(D), tet(E), and tet(G), which are commonly found in gram-negative strains. Each of tet(B), tet(D), and tet(B)&(M) was found in a strain presumed to be Vibrio sp., and only tet(D) was found in 10 Edwardsiella tarda strains.

• Journal of environmental and Sanitary engineering
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• v.5 no.2
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• pp.63-80
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• 1990

A sensitive and simple analytical system for the simultaneous determination of residual oxytetracycline, tetracycline, and doxycycline in honey was described, and that the analytical method for determination of residual propionic acid in honey was established. Experimental subjects were purchased four kinds of honey, native kind honey, acaccia honey, mixed floral honey, chestnut honey in Kyung Sang Nam Do. Several microbiological methods are available to determine tetracycline antibiotecs(TCs) in foods but their precision apears to be variable and the specificity is questionable. These methods are considered to be not suitable for analysis of tetracycline antibiotics in honey because honey itself has bacteriostatic action. For determination of tetracycline antibiotics in honey, therefore the High Performance Liquid Chromatography(HPLC) method was applied, and the propionic acid were determined by Gas Chromatography(5.C). Ethylacetate, as an extract solvent, was found to be suitable for seperation of TCs in honey, but methanol and acetone were not. The recoverly rate of Oxytetracycline(OTC), Tetracycline(TC), Doxycycline(DC) from honey spiked at a level of 10 $\mu $g/g were 97%, 89%, and 91%, respectively. The cailbration curve in TCs was linear expression from 2$\mu $g/ml to 10$\mu $g/ml. As the results of analysis, the residual tetracycline antibiotics were not detected in the 100 samples of honey. The recovery rate of propionic acid from honey spiked at level of 10$\mu $g/g was 98.3% , and the calibra lion curves were linear expression from 21$\mu $g/ml to 101$\mu $g/ml. As the results of analysis, the residual propionic acid was not detected in the 100 samples of honey. Retention time(min) of OTC, DC, and TC were 3.35, 4.61, and 5.30 minutes at the conditions of table 2, respectively, and retention time(min) of propionic acid was 3.50 minutes at the conditions of table 3. The residual TCs and propionic acid were not detected in the 100 samples of honey, but there is a possibility that antibiotics or propionic acid will be to remain in honey if they are used during product period in order to prevent putrefaction of honey-bee.

• Korean Journal of Environmental Agriculture
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• v.30 no.1
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• pp.82-88
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• 2011

BACKGROUND: Tetracyclines (TCs) are mainly regulated as parent compounds by bioactivity-based screening methods in food. Especially with respect to antimicrobial residues, their metabolites/epimers are also highly concerning chemicals and traditionally applied microbial detection methods are needed to improve with validation for regulatory control. METHODS AND RESULTS: Detection capability and biological activity of tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC) and their epimers; anhydrotetracycline (ATC), epianhydrotetracycline (EATC), epitetracycline (ETC), 4-epi-chlortetracycline (ECTC), 4-epianydrochlotetra-cycline (EACTC), 4-epioxychlortetracycline (EOTC), were measured by microbial growth inhibition screening method of Korea Food Code. CONCLUSION(S): Limited detection capabilities were found, B. megarerium and B. subtilis showed for TC and CTC, and B. subtilis for OTC. Biological potency of each epimer was also presented against various microorganisms, at the level from 50% to 96%, comparing with parent TCs. It is recommended that more advanced microbial screening methods with validation are needed, and biologically active epimers are to be considered as marker residues for MRL setting of regulatory control purpose.

• Journal of fish pathology
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• v.11 no.1
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• pp.77-81
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• 1998

By standardized method, Bacillus subtilis BGA, Bacillus cereus var. mycoides ATCC 11778, and Micrococcus luteus ATCC 9341 were seeded on the muller hinton agar (Difco) plate, and pH was adjusted to 6.0, 7.2, and 8.0. Five agar plates, B. subtilis (pH 6.0), B. cereus (pH 6.0), B. subtilis (pH 7.2), B. subtilis (pH 8.0), and M. luteus (pH 8.0), were employed as test plates of modified EEC 4-plate method. Oxytetracycline (OTC) with a diet was orally administered to flounder, Paralichthys olivaceus, at 100 mg/kg once a day. After oral administration, modified EEC 4-plate method by the three screening test using muscle-direct, extraction-disk and direct-disk methods was conducted for 3 fish at 1, 3, 5, 10, 15, 20, 25 and 30 days. Muscle-direct treatment of B. subtilis (pH 6.0) was found to be dubious positive (${\pm}$) at the 1st day after the administration; thereafter, it was found to be negative to the last day of the experiment. Extraction-disk and direct-disk treatment of B. subtilis (pH 6.0) were found to be negative from the 1st day to the last day after the administration. B. subtilis (pH 7.2), B. subtilis (pH 8.0), and M. luteus (pH 8.0) by the three screening tests, were found to be negative all the way after the administration. On the other hand, B. cereus (pH 6.0) by the three screening tests was clearly found to be positive for the first 15 days after the administration, and then muscle-direct and direct-disk treatment of B. cereus (pH 6.0) were found to be dubious positive at 20th days after the administration. However extraction-disk treatment of B. cereus (pH 6.0) was clearly found to be negative at the same stage; thereafter, the three screening tests of B. cereus (pH 6.0) were found negative to the last of the experiment. These findings showed that to have equal sensitivity to those determination for the residual detection of OTC, and also confirmed that B. cereus was effective test organism for the monitoring of OTC.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.2
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• pp.100-104
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• 2015

Adverse effect of veterinary antibiotics (VAs) released into environment has been issued recently and concerns about analysis and management for VAs in the environment were increased. Main objective of this research was to calculate predicted environmental concentration (PEC) of the VAs in soil based on avaiable statistical data and result of previous study such as consumption rate and physiological properties of VAs. Total of 5 VAs, Chlortetracycline (CTC), Oxytetracycline (OTC), Sulfadimethoxine (SDX), Sulfamethazine (SMT), and Tylosin (TYL) were examined. Result showed that calculated PEC value in manure and soil was ordered as SMT > TYL > SDX > CTC > OTC. Range of calculated value for manure and soil was 0.50-67.04 and $0.48-64.45mg\;kg^{-1}$ respectively. Comparing to measured concentration of VAs in manure and soil, lower concentration of VAs in manure and soil was evaluated due to fate and degradation of VAs in manure and soil. Overall, evaluated simple modeling for calculating PEC of VAs in manure and soil can be adapted for preliminary screening purpose in environmental risk assessment and more refined modeling is necessary to examine detailed assessment of VAs in manure and soil.

• 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 Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.910-916
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• 2022

The purpose of this study is to evaluate the effect of underwater non-thermal dielectric barrier discharge plasma (DBD plasma) on the sterilization of three types of pathogenic bacteria that cause diseases in freshwater fish and the reduction of a tetracycline antibiotics. This experiment was conducted in the DBD plasma generator, and the voltages used to generate plasma were 11.6 kV and 23.1 kV. The measurement intervals were 0, 1, 5, 10 and 15 min. As a result of DBD plasma treatment, Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens were removed 93-99% after 5 min at 23.1 kV, and the tetracycline antibiotics were reduced 70-95% after 15 min at 23.1 kV. In this study, as a result of treating the effluent with DBD plasma at a fish farm where the medicinal bath was conducted with oxytetracycline-HCl (OTC-HCl) products, OTC-HCl decreased by 62% after 10 min at 23.1 kV.

• Journal of fish pathology
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• v.19 no.3
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• pp.267-276
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• 2006

Seventy-five streptococci were isolated from diseased olive flounder, Paralichthys olivaceus in Jeju. Their drug susceptibility and transferable multiple drug resistance were characterized. All isolates were resistant to flumequine (AR) and oxolinic acid (OA) and 26 isolates (34.7%) showed 4～6 multiple resistance of ampicillin (ABPC), AR, doxycycline (DOXY), erythromycin (EM), norfloxacin(NOR), OA and oxytetracycline (OTC) in various combinations. pST9 of a transferable R plasmid was detected from a multiple drug resistance strain, Streptococcus sp., ST9 originated from diseased flounder in Jeju, previously. We performed DNA hybridization to know the distribution of plasmid with the same DNA structure as pST9 in streptococci. Thirteen out of 60 isolates analyzed were positive in colony DNA hybridization and the part of bacteria isolated from raw meal was also hybridized with pST9. It suggested that raw meal is one of the origin of the resistance plasmid and R plasmid with DNA structure differing from pST9 is also involving in multiple drug resistance of the streptococci. In conjugation experiment, we found transferable R plasmid carrying OTC, DOXY and/or EM resistance determinant in the 13 resistance strains. all of the streptococci carrying the transferable R plasmid were similar in RAPD patterns. However, pST -type R plasmid was rare in S. iniae most frequently appearing in flounder farm.