• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.10a
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• pp.196-203
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• 2005

축산식품 고기내의 잔류항생물질을 신속하고 간편하고 정확하게 분석하기 위한 시험법 개발 을 목적으로 하였다. 축산식품내의 일반적인 잔류항생물질에 대한 지금까지의 분석법으로는 Bioassay법, TLC법, ELISA법, GC법 및 HPLC법 등이 있지만 Streptomycin/Dihydro streptomycin, Neomycin에 대한 HPLC법은 거의 확립되어 있지 않은 실정이다. 우리나라의 공인 검사법으로는 Bioassay법 및 HPLC법 등이 있지만 그러나 지금까지의 방법으로는 검출감도가 낮은 것이 큰 문제점으로 되어 왔다. 본 연구에서는 DST에 대한 HPLC법에 대한 보고한 P. Edder 방법 중에 clean-up과정 및 이동상 조건을 대폭 수정하여 DST의 분리 및 검출감도를 낮추려고 시도하였다. 본 연구에 사용된 유도체화 장치 Post-Column Derivatization Instrument PCX 5100 (Pickering Laboratories, Inc.)의 컬럼온도는 $40^{\circ}C$, 오븐온도 $55^{\circ}C$, reagent 유속 0.6ml/min mobile phase 유속 0.8ml/min으로 검출기는 형광검출기를 이용하여 DST 검출에 대해 만족할 만한 결과를 얻었다. 이때의 분석소요시간은 약 15분이었다. 표준시료 DST의 검량선은 넓은 농도범위(0.02${\sim}$1.0ppm)에서 양호한 직선성을 나타냈다. 본 시험법에 의한 검출한계는 limit of detection (LOD)은 0.02ppm이었으며, 적어도 고기에서의 MRL이 0.6ppm임을 감안하면 DST를 정량적으로 정도 좋게 측정할 수 있다는 것을 확인했다. 상기의 조건하에서 실제시료인 고기에 표준 DST를 1ppm을 spiking한 후 SPE상에서 SCX(Strong cation exchange column)을 통한 clean-up과정을 거친 후의 DST의 limit of quantification(LOQ)는 약 0.47ppm이었으며, 이에 대한 회수율은 97.7%(n= 8)를 나타냈다. 실제 codex에서 권장한 고기의 MRL이 0.6ppm인 점을 감안하면 codex 권고치에 도달할 수 있는 것으로 판단되었다. 따라서 본 연구에서 개발된 시험법은 지금까지 국내적으로 DST에 대한 시험법이 확립되어 있지 않은 것으로 이와 아울러 간편한 parallux와 병용해 DST에 대한 정량 및 정성 분석을 유도체화 장치 및 형광검출기를 이용해 잔류항생물질 DST에 대한 분석시험법의 개발이 가능하다고 여겨진다.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.10a
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• pp.310-316
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• 2005

축산식품(우유)내의 잔류항생물질을 신속하고 간편하고 정확하게 분석하기 위한 시험법 개발을 목적으로 하였다. 축산식품내의 일반적인 잔류항생물질에 대한 지금까지의 분석법으로는 Bioassay법, TLC법, ELISA법, GC법 및 HPLC법 등이 있지만 Streptomycin/dihydrostreptomycin, neomycin에 대한 HPLC법은 거의 확립되어 있지 않은 실정이다. 우리나라의 공인 검사법으로는 Bioassay법 및 HPLC법등이 있지만 그러나 지금까지의 방법으로는 검출감도가 낮은 것이 큰 문제점으로 되어 왔다. 본 연구에서는 STP에 대한 HPLC법에 대한 보고한 Edder 방법 중에 clean-up 과정 및 이동상 조건을 대폭 수정하여 STP의 분리 및 검출감도를 낮추려고 시도하였다. 본 연구에 사용된 유도체화 장치 Post-Column Derivatization Instrument PCX 5100 (Pickering La-boratories, Inc.)의 컬럼 온도는 $40^{\circ}C$, 오븐온도 $55^{\circ}C$, 유도체화 용매 유속 0.6ml/min 이동상 유속 0.8ml/min으로 검출기는 형광검출기를 이용하여 STP 검출에 대해 만족할 만한 결과를 얻었다. 이때의 분석소요시간은 약 15분이었다. 표준시료 STP의 검량선은 넓은 농도범위(0.02${\sim}$1.0ppm)에서 양호한 직선성을 나타냈다. 본 시험법에 의한 검출한계는 limit of detection(LOD)은 0.02ppm이었으며, 적어도 우유에서의 MRL이 0.6ppm임을 감안하면 STP를 정량적으로 정도 좋게 측정할 수 있다는 것을 확인했다. 상기의 조건하에서 실제시료인 우유에 표준 STP를 0.5ppm을 spiking한 후 SPE상에서 SCX(Strong cation exchange column)을 통한 clean-up과정을 거친 후의 STP의 limit of quantification(LOQ)는 약 0.44ppm이었으며, 이에 대한 회수율은 89.7${\pm}$2.3%(n=6)를 나타냈다. 실제 CODEX에서 권장한 우유의 MRL이 0.6ppm인 점을 감안하면 CODEX권고치에 도달할 수 있는 것으로 판단되었다. 따라서 본 연구에서 개발 된 시험법은 지금까지 국내적으로 STP에 대한 시험법이 확립되어 있지 않은 것으로 이와 아울러 간편한 parallux와 병용해 STP에 대한 정량 및 정성 분석을 유도체화 장치 및 형광검출기를 이용해 잔류항생물질 STP에 대한 분석시험법을 개발하였다.

• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.6-10
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• 2010

Bacterial pathogens were isolated from dogs with urinary tract infection (UTI) in local animal hospitals between August 2003 and December 2009. Bacteria were isolated from urine of 47 dogs. The isolated pathogens were Escherichia coli (n = 27), Streptococcus spp. (n = 7), Staphylococcus spp. (n = 5), Enterobacter spp. (n = 3), Proteus spp. (n = 2), other species were 3 strains, respectively. E. coli were susceptible to imimpenem, polymyxin B, amikacin, cephalosporins, aztreonam, amoxicillin clavulate, cephalosporins, tricarcillin, and amoxicillin clavulate, while were resistant bacitracin, erythromycin, lincomycin, oxacillin, penicillin, and novobiocin. Streptococcus spp. were susceptible to bacitracin, imimpenem, and trimethoprime-sulfa, while were highly resistant amikacin, cefotaxim, cefoxitin, cloxacillin, gentamicin, lincomycin, oxacillin, penicillin, streptomycin, and tobramycin. Staphylococcus spp. were susceptible to cefoxitin, doxycycline, enrofloxacin, imimpenem, and tobramycin, but were resistant aztreonam and tetracycline.

• Journal of Veterinary Clinics
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• v.25 no.5
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• pp.346-354
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• 2008

The combined effects of EDTA-Tris and eighteen antimicrobial agents have been evaluated in eight clinically isolated methicillin-resistant bacteria (Staphylococcus aureus, Escherichia coli, Streptococcus uberis and Streptococcus agalactiae) from bovine mastitis. The antimicrobial activity was evaluated by measuring the minimal bactericidal concentration (MBC) for the antibiotics alone or in combination with EDTA-Tris in Mueller-Hilton broth and milk. Combined use of EDTA-Tris and antibiotics potentiated or antagonized activity of antibiotics against mastitic pathogens. Milk increased the antibiotic potency of erythromycin and spiramycin on S. aureus. Culture in milk changed patterns of EDTA-Tris combinational effects compared with that in standard Mueller-Hilton broth. Combined with EDTA-Tris in milk, synergic effects were observed in colistin, dihydrostreptomycin, kanamycin, erythromycin, gentamycin, oxytetracycline, streptomycin to E. coli, Str. uberis, and Str. agalactiae. However, significant antagonistic effects of milk on antibiotic susceptibility in combination with EDTA-Tris were noted in neomycin, streptomycin, penicillin, roxithromycin, and amoxicillin. This study indicates that combination therapy of EDTA-Tris with antibiotics in bovine mastitis should be used with caution because of the possible antagonistic effects of antibiotic combination with EDTA-Tris on mastitic pathogens. In addition, antibiotic susceptibility test in combination with EDTA-Tris in milk culture condition can be benefit in search of effective treatment regimen for some antibiotic-resistant bacteria of mastitis.

• Food Science of Animal Resources
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• v.33 no.4
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• pp.481-486
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• 2013

This study evaluates the effects of fat contents on the thermal resistance, antibiotic sensitivity, and Caco-2 cell invasion of Listeria monocytogenes. Ten strain mixture of L. monocytogenes in milk (0, 1, and 4% fat) and pork sausage patties (10, 20, and 30% fat) were exposed to $63^{\circ}C$. To evaluate effects of fat on the antibiotic sensitivity of L. monocytogenes, the L. monocytogenes strains NCCP10811 (most antibiotic resistant to streptomycin) and NCCP10943 (most antibiotic sensitive to streptomycin) were exposed to different fat contents in milk and pork sausage patties, and L. monocytogenes from the foods were used for antibiotic sensitivity assays. The most invasive L. monocytogenes strains (NCCP10943) was exposed to different fat contents in milk or pork sausage patties, and L. monocytogenes from the foods were used for the Caco-2 cell invasion assays. The reductions of L. monocytogenes populations were not generally influenced by fat contents. The L. monocytogenes subjected to milk fat had increased sensitivities (p<0.05) due to some antibiotics. In addition, Caco-2 cell invasion efficiency of L. monocytogenes NCCP10943 increased (p<0.05) as fat contents increased. These results indicated that higher fat contents may be related to L. monocytogenes invasions and heat resistances in pork sausage patties, but the relationship between fat and antibiotic sensitivity varied according to antibiotics, strains, and fat contents.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.47-52
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• 2005

This study was carried out to identify bacterial strains adhered to human scalp hair and to investigate the antibiotic susceptibility of them. A total of 39 isolates were obtained from patients in intensive care units and healthy persons. The most common species isolated was Staphylococcus epidermidis (19 isolates), followed by S. aureus (14 isolates), S. waneri (5 isolates), and S. pasteuri (1 isolate). The susceptibility of isolates to amikacin, ampicillin, bacitracin, carbenicillin, cefazolin, cefoperazone, chloramphenicol, erythromycin, gentamicin, methicillin, nalidixic acid, neomycin, oxacillin, penicillin, streptomycin, tetracycline and vancomycin was determined by the disk diffusion method. All of the antibiotic resistant isolates were obtained from patient scalp hair. To examine the effect of conventional shampoo and detergent SDS on removing of bacteria from hair, we treated hair with culture solution of S. aureus. The bacteria attached to hair were not removed even by repeated washing with detergents. These results suggested that hair could be a source of bacterial contamination in hospital.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.163-167
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• 1985

To investigate the condition for the protoplast fusion of Streptococcus lactis, streptomycin(200 $\mu\textrm{g}$/$m\ell$) resistant and rifampicin (200$\mu\textrm{g}$/$m\ell$) resistant mutants were isolated. By using these markers, protoplast fusion was carried out in the presence of CaCl$_2$ and polyethylene glycol. The optimal conditions for the protoplast fusion were obtained by treatment of protoplasts with 150 mM CaCl$_2$ (final concentration; 25 mM) and 40％ (w/v) PEG 4, 000 for 2 min. At the optimal conditions, the fusion frequency was 6.26$\times$10$^{-5}$ . On the other hand, genetic recombination between the antibiotic resistant mutants by mating was not observed.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.289-294
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• 1992

Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.66-73
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• 1993

Psychrotrophic bacterial strains utilizing crude oil as their sole carbon and energy sources were isolated from Antarctic soil and sea sediments. One of the strains named AI-I showed the hightest activity for emulsification of crude oil and the best growth. This strain was identified as Acinetobacter calcoaceticus. A. calcoaceticus AI-I strain contains a plasmid (OCT plasmid) which was related to the utilization of alkane compounds. The molecular weight of this plasmid was estimated to be about 110 Md by agarose gel electrophoresis. The cured strain of A. calcoaceticus AI-I strain (OCT ) was not able to utilize normal hydrocarbon compounds ($C_6C_{17}$) as carbon and energy sources. A. ca/coaceticus AI-1 was resistant to ampicillin and sensitive to streptomycin, kanamycin, chloramphenicol, tetracycline. The results suggested that this strain carries a plasmid (OCT) responsible for oil utilization which is quite stable and might be concerned with antibiotics resistancy.

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.112-117
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• 2007

Protein kinases play very important role for maintaining viability in prokaryote and eukaryote. The metabolism of prokaryotic cell is generally regulated by bacterial two-component regulatory systems that are composed of histidine and asparitic acid kinases, however, some eukaryotic signal transduction system such as, serine and threonine kinases, have been also found to be involved in the regulation of morphogenesis and physiological differentiation in Streptomyces. Streptomyces griseus, a streptomycin producer, was expected to have varlous types of eukaryotic-type serine/threonine protein kinases, controlling morphogenesis. Thus, many steps of chromatographies were applied to isolate serine and threonine kinases from S. griseus IFO13350. The immunoaffinity steps using anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine agarose column chramatographies were successfully introduced to identify eukaryotic protein kinases from S. griseus IFO13350. Eight proteins with the expected molecular weight of 14, 29, 31, 35, 40, 52, 56, and 60 kDa, were identified on SDS-PAGE, and the their kination activity was confirmed by nonradioactive protein kination assay using FITC-labeled peptide as the substrate.