• Fisheries and Aquatic Sciences
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• 제21권2호
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• pp.6.1-6.10
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• 2018

The intention of this study was to identify the bacterial pathogens infecting Oreochromis niloticus (Nile tilapia) and Clarias gariepinus (African catfish), and to establish the antibiotic susceptibility of fish bacteria in Uganda. A total of 288 fish samples from 40 fish farms (ponds, cages, and tanks) and 8 wild water sites were aseptically collected and bacteria isolated from the head kidney, liver, brain and spleen. The isolates were identified by their morphological characteristics, conventional biochemical tests and Analytical Profile Index test kits. Antibiotic susceptibility of selected bacteria was determined by the Kirby-Bauer disc diffusion method. The following well-known fish pathogens were identified at a farm prevalence of; Aeromonas hydrophila (43.8%), Aeromonas sobria (20.8%), Edwardsiella tarda (8.3%), Flavobacterium spp. (4.2%) and Streptococcus spp. (6.3%). Other bacteria with varying significance as fish pathogens were also identified including Plesiomonas shigelloides (25.0%), Chryseobacterium indoligenes (12.5%), Pseudomonas fluorescens (10.4%), Pseudomonas aeruginosa (4.2%), Pseudomonas stutzeri (2.1%), Vibrio cholerae (10.4%), Proteus spp. (6.3%), Citrobacter spp. (4.2%), Klebsiella spp. (4.2%) Serratia marcescens (4.2%), Burkholderia cepacia (2.1%), Comamonas testosteroni (8.3%) and Ralstonia picketti (2.1%). Aeromonas spp., Edwardsiella tarda and Streptococcus spp. were commonly isolated from diseased fish. Aeromonas spp. (n = 82) and Plesiomonas shigelloides (n = 73) were evaluated for antibiotic susceptibility. All isolates tested were susceptible to at-least ten (10) of the fourteen antibiotics evaluated. High levels of resistance were however expressed by all isolates to penicillin, oxacillin and ampicillin. This observed resistance is most probably intrinsic to those bacteria, suggesting minimal levels of acquired antibiotic resistance in fish bacteria from the study area. To our knowledge, this is the first study to establish the occurrence of several bacteria species infecting fish; and to determine antibiotic susceptibility of fish bacteria in Uganda. The current study provides baseline information for future reference and fish disease management in the country.

• KSBB Journal
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• 제14권4호
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• pp.460-464
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• 1999

Methicillin-resistant Staphlococcus aureus(MRSA)는 다양한 항생제에 저항성을 나타내며 제 3세대 cephalosporin계가 보급된 1980년대 이후 원내 감염의 중요 원인균으로 대두되고있다. 그러나 MRSA 동정의 오류나 실수로 인한 부적절한 치료에 문제가 있으며 항생제의 오용으로 인한 다제약제 내성획득이 임상에서 심각한 문제가 되거 있다. 따라서 MRSA 감염의 적절한 치료는 이 원인 균의 신속하고 믿을 수 있는 동정을 필요로 한다.본 연구에서는 분자 생물학적인 동정법을 확립하기 위하여 항생제의 내성 기구인 페니실린 결합 단백질 2‘(PBP2')를 암호화하는 mec A 유전자를 PCR 방법으로 증폭하여 MIC test 결과와 비교하여 MRSA를 동정하는데 PCR법이 유용한지를 판명하였다.각 환자의 다종 검체로부터 S. aureus 120균주를 분리하여 oxcillin으로 MIC test를 한 결과 MRSA의 비율로는 농에서는 MRSA가 61.9%로 가장 높게 판명되었다. 120균주 중 MRSA 40균주, MSSA 40균주를 성별하여 PCR 방법으로 동정하여 MIC test와 비교 한 결과, MRSA 1균주(2.5%), MSSA는 2균주(5%)의 차이만이 MIC test와 상이성을 나타내었다. 이러한 결과로 볼 때 PCR법으로 MRSA를 동정하는 것은 유용하며 일상검사 업무로서 활용성이 높을 것으로 판단되었다.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• 대한본초학회지
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• 제31권5호
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• pp.93-98
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• 2016

Objectives : Hwangheuk-san is a complex prescription composed of oriental traditional medicine and has been reported for antioxidant, antimicrobial and anticancer effects in the recent study. Methicillin-resistant Staphylococcus aureus (MRSA) is one of important causes of fatal infectious diseases such as septicemia, endocarditis, toxic shock syndrome, pneumonia, skin and soft tissue infections (SSTIs). S. aureus is reported as being for a variety of human diseases and its epidemiological relevance is mainly due to their ability of becoming highly resistant to common antimicrobials such as tetracycline, penicillin, cphalosporin and aminoglycoside. The objective of this study is to determine the antimicrobial effect of Hwangheuk-san ethanol extracts (HHS) and synergistic effects with antibiotics oxacillin against MRSA.Methods : The antimicrobial activity of HHS was measured by the disc diffusion method, broth microdilution method and the checkerboard dilution test, time-kill curve assay was performed to investigate synergistic effects with antibiotics oxacillin against MRSA.Results : HHS showed antimicrobial activity against MRSA with a MIC value of 125 ㎍/㎖. In the checkerboard test, the interaction of HHS with antibiotics oxacillin produced almost synergy or partial synergy against MRSA. This study showed that HHS reduced the MICs of oxacillin tested, and a remarkable antibacterial effect of HHS, with membrane permeability enhancers.Conclusions : These results suggest that HHS has the antimicrobial effect and synergistic effects with antibiotics oxacillin against MRSA. This study thus can be a valuable source for the development of a new drug with low MRSA resistance.

• 대한수의학회지
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• 제26권2호
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• pp.283-292
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• 1986

Isolation and identification of Mycoplasma were performed to clarify Mycoplasma infection of mice fed by conventional feeding at two ($K_1$, $K_2$) institutes in Korea. The twenty mice to be tested were randomly sampled from each of 10 breeding colonies in respective institute. Identification of the Mycoplasma strains isolated from the nasal cavity, lung and synovia of mice was made according to the morphology of colonies, biological and biochemical properties with special reference to M. pulmonis, M. arthrotodis and M. neurolyticum. In addition, growth inhibition test was performed using hyperimmune rabbit antisera to the strain PG-22 of M. pulmonis, the strain PG-6 of M, arthritidis and the strain PG-28 of M. neurolyticum and also differentiation of isolates from L-form bacteria was dont by Dieses staining and culture method with passage of the isolates on liquid media eliminated antibacterial drug. On the other hand, a total of 13 strains out of the 44 isolated M. pulmonis from mice was investigated for their susceptibility against 16 antibiotics in vitro. The antibiotic sensitivity test was made using $3{\times}10^4$ organisms/0.3ml on each plate(90mm diameter) with antibiotic mono-or tri-disk. The results obtained are summarized as follows: 1. Out of 20 mice from 10 breeding colonies in Kl institute, mycoplasma-like strains from the nasal cavity of 16 mice(80%) and from the lung of 8 mice(40%) were isolated, while out of 20 mice in K2 institute, M-like strains were isolated from the nasal cavity of 14 mice(70%) and from the lung of 6 mice(30%). However, no mycoplasma-like organisms were isolated from the synovia of the 40 mice examined. All the 44 strains isolated were identified as the organisms of M. pulmonis. 2. Out of the 16 antibiotics tested, penicillin, oleandomycin and bacitracin showed no activity against all the 13 M. pulmonis strains. On the contrary, lincomycin, clindamycin, chloramphenicol, tetracycline, minocycline, kanamycin, gentamycin and tobramycin showed high activity with three different antibiotic concentration of tridisk, but amikasin and spiramycin showed intermediate activity. Other antibiotics such as polymyxin B and colistin showed low activity, while erythromycin showed lower activity than others.

• 대한수의학회지
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• 제40권2호
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• pp.345-360
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• 2000

The effects on CTC (T1) and CTC, Sulfathiazole and Penicillin Combination (T2) medication in feed through one life cycle of pigs, namely, weaning, mating, farrowing, lactation, growing, finishing and slaughter, were tested under local condition. In sow phase, productivity and the number of microflora in urine before and after medication of CTC were studied and average daily gain and feed conversion rate were checked during growing and finishing period. All pigs reached at 155 days old were slaughtered for pathological examination. 1. Litter size, farrowing rate and survival rate at birth were improved by CTC medication from weaning to 21th day after mating and mortality of piglet at weaning, 25 days after farrowing, was reduced in the CTC medication group, but no siginificant. 2. The number of microflora in the sow urine was changed with the medication at 200ppm of CTC in feed. In particular, the number of E coli, Samonella and Staphylococci were reduced by CTC medication. 3. The average daily gain and feed conversion rate of grower and finisher pigs was improved significantly in both treated groups, most in the high level CTC (T1) medicated group and was lowest in the control group. 4. The number of infected lungs was reduced not significant by both treatments (as % pneumonic lesions Co 66.7%, T1 47.1%, T2 31.4%) and the severity of lung lesions was significantly reduced by both high level of CTC and CTC combination medication in feed. 5. Although there were no statistical differences in atrophic rhinitis based on turbinate scores among the 3 groups, the number of mild and moderate (Grade 2 and 3) infections was higher in the control group (9/36) than in the treated groups (T1 2/34 & T2 4/35).

• Journal of Periodontal and Implant Science
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• 제28권2호
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• pp.291-310
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• 1998

This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.

• 대한화학회지
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• 제57권1호
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• pp.81-87
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• 2013

황색포도상구균은 병원에 의한 감염, 혈류 감염을 포함하는 중요한 병원체이다. 특히, 포도상구균의 혈액 수집의 신속한 동정과 메치실린 내성이 일어나게 되면 폐혈증으로 추측되어 진다. 본 저자는 적은 양의 핵산을 등온증폭반응에 적응시켜 증폭산물을 SYBR Green I을 결합하여 염기서열에 특이적으로 검출할 수 있는 새로운 방법을 제시하고 있다. 그리고 이 독특한 유전자 증폭방법은 등온의 상태에서 하나의 효소만으로 증폭이 가능하다. 포도상구균-등온증폭반응은 황색포도상구균에 특이적인 protein A를 암호화하는 spa 유전자와, 메치실린 내성인 pennicillin-binding protein-2를 암호화하는 mecA 유전자를 타겟으로 하여 MRSA와 MRSE를 검출하였다. 본 연구에서는 등온증폭법을 사용하여 황색포도상구균과 표피포도상구균의 임상샘플을 검출하였다. 황색포도상구균과 표피포도상구균을 10배위 정량 희석하여 시리즈별로 샘플을 만들어 실험을 수행하였다. 칩을 기반으로 하는 LAMP법은 포도상구균 감염여부를 쉽고, 빠르고, 정확하게 민감도 있는 검출을 가능하게 해 주었고, 샘플을 측정할 수 있는 한계값을 넘는 상황에 특이적으로 적용할 수 있다.

• 생명과학회지
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• 제20권5호
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• pp.649-654
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• 2010

Streptococcus equi subsp. zooepidemicus는 말의 생식기 질환을 유발하는 중요한 병원체 중 하나로서 국내 말의 생산성 향상을 도모할 목적으로 제주도에서 사육중인 더러브렛 암말의 유산태아 및 그 말의 자궁내용물과 장수목장에서 암말의 교배전 생식기 질환이 의심되는 말의 질 내용물로부터 시료를 채취하여 Streptococcus equi subsp. zooepidemicus을 분리하여 그 분리균의 특성 및 항균제 감수성 검사를 실시한 결과, RAPD typing에서는 크게 4개의 cluster로 구분되었다. 특히 제주도에서 분리한 균주와 장수목장에서 분리한 하나의 균주가 동일한 pattern을 나타내었다. 항균제 감수성 검사에서는 대부분의 균주가 ampicillin 등의 약제에 감수성이 있음을 확인하였다. 본 연구의 결과는 국내에서 사육중인 암말의 생식기 질병의 예방 및 치료에 유용할 것으로 생각된다.