• Journal of Life Science
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• 제26권11호
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• pp.1259-1268
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• 2016

In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by $C_{18}$ reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], $20.8{\mu}g/ml$), Bacillus subtilis (MEC, $0.2{\mu}g/ml$), Streptococcus mutans (MEC, $0.2{\mu}g/ml$), gram-negative bacteria including Pseudomonas aeruginosa (MEC, $5.7{\mu}g/ml$), Escherichia coli (MEC, $2.6{\mu}g/ml$) and fungi, Candida albicans (MEC, $56.3{\mu}g/ml$). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.

• Korean Journal of Agricultural Science
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• 제2권1호
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• pp.199-231
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• 1975

It is well known fact that antibiotic residues in milk of cows create significant problem for the fermented dairy industry and public health because of inhibition of starter activity and of creation of allergic disease. It can be assumed that antibiotic residues in milk of other aniimals also can create some problems for their infants as in the case of humen. For the above mentioned reasons, present studies were undertaken to determine concentration and duration of antibiotic residues in milk of cows, goats and dogs following intramuscular or intravenous injection and intramammary infusion of penicillin, streptomycin and oxytetracycline at usual dosage. The cylinder-plate method was used for their assay. The results obtained were summerized as follows: 1) Following the intramuscular injection of penicillin, the antibiotic was detected in milk of cows up to 72 hours, in milk of goats 48 hours and in milk of dogs 60 hours of postinjection. The mean peak concentrations were recorded at 12 hours as 0.136 I.U./ml in cows. 6 hours as 0.773 I.U./ml in goats and 3 hours as 1.192 I.U./ml in dogs. 2) Following the intramuscular injection of streptomycin, the antibiotic was detected in milk of cows and goats up to 36 hours and in milk of dogs 24 hours of post-injection. The mean peak concentration were recorded at 6 hours as $0.26{\mu}g/ml$ in cows and at 3 hours in goats and dogs $0.45{\mu}g/ml$ and $0.36{\mu}g/ml$ respectively. 3) Following the intra venous injection of oxytetracycline, the antibiotic was detectable in milk of all the test animals up to 48 hours of postinjection. The mean peak concentrations were recorded at 6 hours as $3.5{\mu}g/ml$ in cows $2.4{\mu}g/ml$ in goats and $2.0{\mu}g/ml$ in dogs respectively. 4) Following intrarnammary infusion of penicillin in amounts of 100,000 I.U. for cows, 20.000 I.U. for goats and 10,000 I.U. for dogs, the penicillin residues in milk of the infused quarter perssisted to 72 hours in cows and 84 hours in goats and dogs. 5) Following intramammary infusion of streptomycin in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the streptomycin residues in milk of the infused quarter persisted to 72 hours in cows and goats and 60 hours in dogs. 6) Following intramammary infusion of oxytetracycline in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the oxytetracycline residues in milk of the infused quarter persisted to 72 hours in cows and 60 hours in goats and dogs. 7) A corelation between the residual antibiotic concentration and milk yield in cows and goats was observed; That is, the lower in the milk production showed a higher the concentration of an antibiotic residues and a longer the time in persistance. 8) Intramammary transfer of the antibiotic from an infused to non infused quarters, in dogs, was observed following the intramammary infusion of penicillin. streptomycin and oxytetracyclne in amounts of 10.000 I.U. 25mg and 25mg respectively. However, no transfer by 100.000 I.U. or 20.000 I.U. of penicillin. 500mg of streptomycin and 100mg of oxytetracyline was observed in cows and goats. 9) In dogs, minimum dosage of antibiotics for transfer fro in treated to untreated quarters following intramammary infusion were 2,500 I.U. of penicillin and 5mg each of streptomycin and oxytetracycline.

• Journal of Food Hygiene and Safety
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• 제19권4호
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• pp.198-202
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• 2004

It was carried out to compare the relationship of EEC 4-plate test method and Charm II test method by HPLC. 46 only tetracyclines, 4 only sulfonamides, 4 sulfonamides+tetracyclines, one amphenicols+tetracyclines, and one amphenicols+sulfonarnides+tetracyclines, it was detected on 71 porks of EEC 4-plate positive or Charm II positive by HPLC. In case of sulfonamides and chloramphenicols, two test methods had related with each other, but had no relation in tetracyclines. Also, it was $10.5\%(6/57)$ tetracyclines, $81.8\%(9/11)$ chloramphenicols that were EEC 4 plate positive or Charm II positive, but not detectrd by HPLC. Consequently, EEC 4-plate test method and Charm II test method were suitable by screening test on antibiotics and sulfonamides etc.

• Korean Journal of Microbiology
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• 제42권3호
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• pp.165-171
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• 2006

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

• Korean Journal of Food Science and Technology
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• 제42권3호
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• pp.287-291
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• 2010

In this study, a simple and rapid pre-treatment method based on liquid extraction was applied for the simultaneous determination of three macrolides (spiramycin, tylosin, and tilmicosin) residues. In these studies, the stock farm products was used as a matrix sample. When the liquid extraction method was compared with the solid phase extraction (SPE) method, the former showed higher recovery percentages and simpler steps than the latter. The macrolids were separated using a reverse-phase C18 ($250\;mm{\times}4.6\;mm$, $5\;{\mu}m$) column and a gradient elution with mobile phases consisting of phosphate buffer (pH 2.5) and acetonitrile. Tylosin and tilmicosin were detected at 288 nm and spiramycin was detected at 232 nm. The average recovery percentage ranged between 83.0-90.2% for samples spiked with the three macrolids at 50 and 100 ng/g The validation results showed that the limit of detection (7 (spiramycin), 12 (tilmiconsin), 12 (tylosin) ng/g)) was under the regulatory tolerances and the linearity from calibration curves was satisfactory for determining the multi-residue of three macrolids in farm products. Monitoring samples were collected at the main cities in Korea as Seoul, Busan, Deajeon, Incheon, Deagu, and Gwangju. Microlide antibiotics were not detected in most samples.

• Analytical Science and Technology
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• 제25권4호
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• pp.257-264
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• 2012

Penicillins belong to the ${\beta}$-lactam class of antibiotics, and are frequently used in human and veterinary medicine. Despite the positive effects of these drugs, improper use of penicillins poses a potential health risk to consumers. This study has been undertaken to determinate multi-residues of penicillins, including amoxicillin, ampicillin, oxacillin, bezylpenicillin, cloxacillin, dicloxacillin, and nafcillin, using liquid chromatographic tandem mass spectrometer (LC-MS/MS). The developed method was validated for specificity, precision, recovery, and linearity in livestock and marine products. The analytes were extracted with 80% acetonitrile and clean-up by a single reversed-phase solid-phase extraction step. Six penicillins presented recoveries higher than 76% with the exception of Amoxicillin. Relative standard deviations (RSDs) were not more than 10%. The method was applied to 225 real samples. Benzylpenicillin was detected in 12 livestock products and 7 marine products. Amoxicillin, ampicillin, cloxacilllin, dicloxacillin, nafcillin and oxacillin were not detected. The detected levels were 0.001~0.009 mg/kg in livestock products excluding eggs and milk. In marine products, the detected levels were under 0.03 mg/kg. They were under the MRL levels. As monitoring results, it is identified to be safe but it is considered that safety management of antibiotics should continue by monitoring.