• 한국동물위생학회지
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• 제24권3호
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• pp.223-241
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• 2001

From September 1996 to September 1999, 419 Korean-native calves with diseases under 6-month old collected from Kyonggi, Chungcheong, Chonlla and Kyongsang were examined by clinical, microbiological, parasitic, hematologic and histopathological mean. Among them, 124 cases were tested about the neutralization antibodies against infectious bovine rhinotracheitis virus(IBRV), Parainfluenza-3 virus(PI-3V), bovine uiral diarrhea virus(BVDV), bovine ephemeral fever virus(BEFV). In calf diseases in the survey, enteric diseases(72.8%) were most frequently involved and the following orders were taken by respiratory(17.4%) and reproductive (5.0%) disorders. In the causative pathogens associated with calf diseases and motality, 48.4% was induced by bacteria origin and also 35.6% by viral agents. Calf mortality was up to 76.3% in the cae of bacterial diseases and 55.7% in viral diseases. In bacterial diseases, frequent disorders were composed of colibacillosis(52.7%), salmonellosis(13.8%), pasteurellosis(12.8%) and campylobacteriosis(3.9%) and their mortalities showed 73.8% in colibacillosis, 73.0% in pasteurellosis, 67.9% in salmonellosis and 50.0% in campylobacteriosis (50.0%). Among the outbreaks of viral diseases, there were BVD(22.8%), bovine rotavirus infection(20.8%), bovine coronavirus infection(16.8%), bovine respiratory syncytial virus infection(15.4%), IBR(15.4%). Akabane disease(4.7%) and Chuzan diseases(3.4%). Interesting results through this studies were obtained the first isolate to Chuzan virus and Ainovirus in Korea which could be promised the development for diagnostic method and vaccines soon. Calf mortality to Akabane and Chuzan diseases was 100%. Main parasitic diseases were occupied by coccidiosis and babesiosis and their mortality of babesiosis was 20.0%. Other diseases were abomasal impaction(6.7%) and toxicosis(4.5%). The mortality of abomasal impaction was 89.3%. In some causes with malformations(1.9%) were confirmed as anasarca, derodidymus, polymelia, humerus hypoplasia, and tracheal collapse. Calf diseases had mostly been occurred in one month old grout (52.5%) and its prevalence was 25.1% in two to three month old group and 22.4% in four to six month old group. In calf mortality by age, there were 37.9% in one month old group, 18.1% in two and three month old group, and 13.8% in four to six month old group, respectively. The older the age of calf, the less the prevalence of calf enteric diseases. Respiratory diseases in calves to be tested frequently occurred in one to two month old group (41.4%). In one month old calves, the prevalence of enteric disease was 80.0%(p<0.05) and that of reproductive and respiratory disease was 9.5% and 8.2%, respectively. In two month old and four to six month old, enteric disease was 65.7% and 63.8% and respiratory disease was 28.6% and 26.6%. Seasonal prevalence and mortality of Korean-native calf diseases were not a significant difference. Prevalence of calf diseases in summer(31.5%) frequently occurred to compare that in winter(20.3%). Abortion and malformation in calves frequently occurred in spring. Hematological values in 84 calves with clinical signs showed mild to marked leukocytosis. Also, there was slight increase in hematocrit, platelet, mean corpuscular volume and mean plasma volume, but all of those were included the higher level to normal ranges. Calves with respiratory signs showed slightly erythrocytosis. One hundred seventy three calves without clinical signs were not significant different to ill cases in hematological values, but number of platelets was in higher normal range. In 125 calves, 84.8% was taken the antibody to IBRV, but 72% with the antibody had recorded the titer level lower than log$_2$5. The neutralizing antibody levels of higher than $log_{2}5$ to PI-3V and BVD virus were 60.8% and 67.2% cases, respectively. There were the cases of 57.6% had the neutralizing antibody level lower than log$_2$5 to BEFV.

• 한국미생물·생명공학회지
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• 제38권2호
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• pp.168-176
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• 2010

사체 피부에서 면역반응을 일으킬 수 있는 세포들을 제거한 무세포 진피는 다양한 의료용 소재로 사용되고 있다. Trin-butyl phospahate(TnBP)와 deoxycholic acids를 세포제거 용액으로 사용하여 피부조직 내 진피층의 3차원적 구조를 손상시키지 않고 다양한 구조 단백질 및 성분들을 유지한 상태에서 면역반응의 대상인 세포성 항원만을 선별적으로 제거한 이식용 동종 무세포 진피인 $SureDerm^{TM}$을 개발하였다. $SureDerm^{TM}$ 제조공정은 감염성 위해인자 불활화 공정으로 70% 에탄올 처리와 산화에틸렌 가스 처리 공정을 포함하고 있다. 본 연구에서는 SureDermTM 제조공정 중 70% 에탄올 소독 공정, 세포제거용액(0.1% TnBP와 2% deoxycholic acids) 처리 공정, 산화에틸렌 가스 멸균의 바이러스 불활화 효과를 검증하기 위해 국제적 가이드에 따라 5종의 바이러스 [human immunodeficiency virus type 1(HIV-1), bovine herpes virus(BHV), Bovine viral diarrhoea virus(BVDV), hepatitis A virus(HAV), porcine parvovirus(PPV)]를 생물학적 지표로 사용하였다. 피부조직에 각 생물학적 지표를 첨가한 후 불활화 공정을 실시한 다음 각 바이러스를 회수하여 정량한 후 불활화 정도를 비교하였다. 70% 에탄올 20분처리 공정에서 HIV-1, BHV, BVDV 같은 외피 바이러스는 처리 시간 2.5분 안에 불활화되었지만, HAV와 PPV 같은 비-외피 바이러스는 에탄올에 저항성을 나타내어 20분 처리 후 log 바이러스 감소인수가 각각 1.85와 1.15였다. 세포제거용액 처리 공정에서 HIV-1, BHV, BVDV는 각각 5분, 30분, 5분 안에 검출한계 이하로 불활화되었다. 산화에틸렌 가스처리에 의해 본 연구에 사용한 모든 바이러스가 검출한계 이하로 불활화되었다. 3가지 공정에서 HIV-1, BHV, BVDV, HAV, PPV에 대한 log 바이러스 감소인수 합은 각각 $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, $\geq7.18$이었다. 이와 같은 결과에서 $SureDerm^{TM}$ 제조공정은 바이러스 안전성을 보증할 수 있는 충분한 바이러스 불활화 능력을 갖고 있는 것으로 판단된다.

• International Journal of Oral Biology
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• 제35권1호
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• pp.1-5
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• 2010

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

• 대한수의학회지
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• 제52권3호
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• pp.169-176
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• 2012

Probiotics colonize the intestines and exert an antibacterial effect on pathogens. Therefore, probiotics could be used as a preventive agent against lethal infections. To isolate probiotic microorganisms, 116 bacterial strains were isolated from healthy cow's milk and were subjected to Gram-stain, morphology and biochemical analyses, Vitek analysis, and 16S rRNA analysis. One of the strains identified as Bacillus (B.) thuringiensis 87 was found to grow very well at pH 4.0~7.0 and to be resistant to high concentrations of bile salts (0.3~0.9% w/v). B. thuringiensis was susceptible to the antibiotics used in the treatment of bovine mastitis, yet it exhibited an antimicrobial effect against Staphylococcus (S.) aureus 305. Moreover, it protected mice from experimental lethal infections of E. coli O55, Salmonella typhimurium 01D, and S. aureus 305 through a significant induction of interferon-${\gamma}$, even at four-week post-administration of B. thuringiensis. Although oral administration of B. thuringiensis 87 did not provide significant protection against these lethal challenges, these results suggest that B. thuringiensis 87 could be a feasible candidate as a probiotic strain.

• 한국동물위생학회지
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• 제20권3호
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• pp.261-279
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• 1997

The study for a effect of monitoring on bovine mastitis was conduced for improvement of raw milk from Jan. to Dec. in 1996. Sampling the milk of 367 cows(1, 406 quarters) from 5 herds in Inchon and were carried out California mastitis test(CMT), somatic cell count(SCC), isolation of pathogens and antibiotic sensitivity tests. The results were summarized as follows, 1. The number of bovine mastitis was 177 cows(48.2%) and 371 quarters(26.4%) : clinical mastitis : 25 cows(6.8%), 32 quarters(2.3%) and subclinicsl mastitis : 152 cows(41.4% ), 339 quarters(24.1%). Incidence rate of mastitis by season were Summer 52.0%, Fall and Winter 48% and Spring 41%. Incidence rate of mastitis by quarters were Summer 30%, Fall 28%, Winter 25% and Spring 21%, respectively. 2. In the distribution of CMT degree by quarter, CMT positive(CMT$\pm$) of 1, 406 quarters milk were 50.1% (704 quarters). The ratio of CMT positivity by quarter were left front quarter 55.8%, right front quarter 48.9%, right hind quarter 48.6% and left hind quarter 47% The ratio of CMT positivity by season were Summer 54.1%, Fall 49.7%, Spring 48.5% and Winter 48% 3. The highest mean SCC by season among 5 herds was "A" herd. Mean SCC (cell/ml) of A herd were Summer 2, 032, 000cells/ml, Fall 1, 109, 000cells/ml, Winter 782, 000cells/ml and Spring 577, 000cells/ml. The lowest mean SCC by season among 5 herds was "E" herds. Mean SCC of E herd were Summer 1, 064, 000cells/ml, Spring 795, 000cells/m1, Fall 429, 000cells/ml and Winter 400, 000cells/ml. Mean SCC of the other herds by season were little difference. 4. The milk samples of "A" herd were collected from 10 cows. In 3 seasons, mean SCC of No. 2 and 3 cows were than 1, 000, 000cells/ml. In 1 season, mean SCC of No. 6, 7 and 8 cows were than 1, 000, 000cells/ml. The more than mean SCC 1, 000, 000cells/ml of cows by season were distributed Summer 4 cows, Winter 3 cows, Spring and Fall 1 cow respectively. The milk samples of "B" herd were collected from 14 cows. In 3 seasons, mean SCC of No. 1 cow was more than 1, 000, 000cells/ml. In 2 seasons, mean SCC of No. 5, 9 and 14 cows were more than 1, 000, 000cells/ml. In 1 season, No. 3, 6 and 7 cows were more than 1, 000, 000cells/ml. The more than mean SCC 1, 000, 000cells/ml of cows by season were distributed Fall and Winter 4 cows respectively, Summer 3 cows and Spring 1 cow. The milk samples of "C" herd were collected from 18 cows. In 2 seasons, mean SCC of No. 16 cow was more than 1, 000, 000cells/ml. In 1 season, mean SCC of No. 1, 2, 6, 7, 13, 15 and 18 cows were more than 1, 000, 000cells/ml respectively. The more than mean SCC 1, 000, 000cells/ml of cows by season were distributed Summer 5 cows, Fall 3 cows, Spring 2 cows and Winter 1 COW. The milk sampes of "D" herd were collected 24 cows. In 3 season, mean SCC of No. 14 cow was more than 1, 000, 000cells/ml. In 2 seasons, mean SCC of No. 14 and 18 cows were more than 1, 000, 000cells/ml. In 1 season, mean SCC of No. 1, 2, 3, 8, 12, 17, 19, 20 and 21 cows were more than 1, 000, 000cells/ml. The more than mean SCC 1, 000, 000cells/ml of cows were distributed Fall 15 cows, Spring and Winter 4 cows respectively and Summer 3 cows. The milk samples of "E" herd were collected from 27 cows. In 2 seasons, mean SCC of No. 6, 7 and 21 cows were more than 1, 000, 000cells/ml. In 1 season, mean SCC of No. 2, 4, 7, 11, 14, 16 and 23 cows were more than 1, 000, 000cells/ml. The more than mean SCC 1, 000, 000cells/ml of cows were distributed Spring and Fall 5 cows respectively, Summer and Winter 2 cows, respectively. 5. The rate of isolated pathogenic microorganisms from bovine mastitis were summarized as follows : Staphylococcus sp 168 strains(45.8%), Streptococcus sp 82 strains(22.3%), Gram(-) sp 45 strains(12.3%), Gram(＋) sp 51 strains and the other sp 21 strains(5.7%). 6. The highest of antibiotic sensitivity test of each microorganism was summarized as follows : Staphyolcoccus sp - cephalosporin 76%, gentamicin 55%, Streptococcus sp - ampicillin 61%, cephalosporin 63%, Gram(-) sp - gentamicin 58%, Gram(＋) sp - cephalosporin 63%, The other sp - cephalosporin 90%. Microorganisms showed the highest sensitivity(68%) to cephalospsorin. Microorganisms showed the highest sensitivity(68%) to cephalospsorin.

• 대한의생명과학회지
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• 제6권1호
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• pp.65-72
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• 2000

반추수에서 발생하는 Johne병의 조기 진단 방법을 제시하고 이 질병의 원인체와 미생물학적 특징 이 유사한 M. bovis, M. avium 등의 mycobacteria 감염증을 감별 진단하는 방법 을 개발하기 위하여 Mycobacterium 균속의 표준균주를 사용하여 중합효소 연쇄반응을 확립하였다. Johne병으로 의심되는 소의 혈액과 유즙을 채취하여 분리한 단핵구 및 거식세포로부터 genomic DNA를 추출하였다. 각 시료로부터 추출한 DNA를 template로 이용하여 Mycobacterium spp.에 특이적인 16S rDNA primer set를 이용한 PCR을 수행하여 시료내의 mycobacterial DNA 보유 여부를 확인하였다. 한편 mycobacteria 양성으로 확인된 시료는 M. avium complex 균종에 특이한 16S rDNA 염기서열을 기초로 하여 제작한 primer set와 M. paratuberculosis의 IS900 sequence에 특이한 primer set를 이용하여 duplex PCR을 수행하여 Johne병 원인체의 보균 여부를 조사하였으며, 이 결과를 oligonucleotide probe를 사용한 Southern blot hybridization을 통하여 다시 확인하였다. 이와 같은 duplex PCR기법을 실제 축산 현장에서 수집한 유즙과 말초혈액으로부터 분리한 단핵구 및 거식세포 시료에 적용한 결과 본 연구에서 확립한 duplex PCR기법 유용성을 확인할 수 있었다.

• 방사선산업학회지
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• 제5권1호
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• pp.75-80
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• 2011

This study was to determine the microbiological safety and tensile strength of gammairradiated porcine tendon for the development of safe xenografts. Escherichia coli and Bacillus subtilis were used as model pathogens and inoculated as $10^6{\sim}10^7log$ colonies forming unit $(CFU)g^{-1}$. As model virus from porcine, porcine parvovirus (PPV), bovine viral diarrhoea virus (BVDV) and poliovirus were inoculated as $10^5{\sim}10^6$ tissue culture infectious dose $(TCID)_{50}g^{-1}$ into porcine skin. The $D_{10}$ value of E. coli and B. subtilis was measured as $0.32{\pm}0.082kGy$ and $4.0{\pm}0.312kGy$, respectively. Additionally, the $D_{10}$ values of PPV, BVDV and poliovirus were also shown as $1.75{\pm}0.131kGy$, $3.70{\pm}0.212kGy$ and $6.26{\pm}0.332kGy$, respectively. Gamma irradiation decreased the tensile strength of porcine tendon. Results indicate that microbiological safety of porcine tendon can be improved significantly by gamma irradiation. However, further studies are needed to improve the tensile strength of gamma-irradiated porcine tendon.