• 한국동물위생학회지
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• 제34권2호
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• pp.159-166
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• 2011

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

• 한국동물위생학회지
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• 제30권3호
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• pp.375-384
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• 2007

This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.

• 한국동물위생학회지
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• 제35권4호
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• pp.263-268
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• 2012

From the end of July 2012, several cases of abortion have been happened at the Korean indigenous cattle farm with 124 heads in Chungbuk province, Korea. Serological tests such as Rose-bengal test (RBT) and standard tube agglutination test (STAT) have been performed according to the standard official protocols of bovine brucellosis and 41 cattle turned out to be brucellosis-positive simultaneously. To find out the main factors of brucellosis outbreaks and spreads, additional serological, etiological and molecular investigation were applied. Totally, 11 B. abortus were isolated from 10 cattle's specimens including lymph-nodes and/or testis, and drinking water in cowhouse. In genotyping by multi-locus VNTR assay (MLVA) using 17 loci markers, the present B. abortus isolates were shown all the same pattern, D1 genotype, which has been reported in Gyeonggi and Gangwon province, Korea. These results suggest that the input of brucellosis might come from neighboring farms directly or indirectly, even if by unknown factor and expansion within farm would accelerate by materials related with aborting cows.

• 한국동물위생학회지
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• 제32권4호
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• pp.335-341
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• 2009

Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.

• 한국동물위생학회지
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• 제28권2호
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• pp.99-106
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• 2005

This survey was carried out to investigate the seroprevalence of antibodies to Neospora caninum in Korean indigenous cattle that was representative livestock raised in Jeongeup province Jeonbuk Korea. A total of 1,162 sera were tested for N caninum antibodies using ELISA (Herdcheck anti-Neospora. IDEXX Laboratories Inc., Westbrook, Maine USA). $15(1.3\%)$ sera were positive by ELISA. A total of 290 farms were tested. $11(3.8\%)$ farms were positive. Among the seroprevalence of cattle according to the areas, 8 of 44 counties were infected N caninum. Sero-positive 11 farms of N caninum antibodies using ELISA test and 62 of 301 sera $(20.6\%)$ were infected to N caninum. We found that cattle be raised in Jeongeup province was slightly infected to N caninum. It seems to be infected of N caninum an early stage. but the positive rates was variable that it was $6\%\~62\%$. It was supposed to the highest positive rate of 3 farms had some factors to be infected. The factors were supposed to keep dogs yearly, located the base of hills, and almost feed with imported dried grass. Farmers suppose to the cause of abortion was not the neosporosis but also broke out an accident or was infected Brucella spp. The results of Brucella Rose Bengal Test (RBT) were all negative.

• 한국안광학회지
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• 제9권2호
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• pp.333-343
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• 2004

Impression cytology는 결막상피의 표층을 떼어내기 위해 안구 표면에 cellulose acetate filter 재질을 같이 이용한다. 이 방법은 비 침습적이고 쉽게 시행할 수 있으며 환자에게 최소한의 불편감을 주며 결막표면에 반복해서 시행하여 변화를 관찰하기 위해 사용될 수 있다. 이 방법으로 결막 표면의 형태학적 연구와 squmaous metaplasia의 정도를 평가할 수 있다. 본 연구는 건성안 환자에서 impression cytology를 이용하여 결막표면을 평가하기 위해 수행되었다. 70명의 콘택트렌즈 비착용 학생을 대상으로 조사하였는데 먼저 조사 대상자들에게 McMonnies dry eye symptom questionnaire을 작성하도록 하였다. 그리고 비침습성 tear thinning time(TTT) 검사를 시행하고 다음에 침습성 검사인 Schirmer tear test(SIT), tear film break-up time(TBUT) tests 그리고 Rose-bengal staining올 시행하여 건성안 의심자를 분류하였다. 하 안구결막으로부터 결막 상피세포는 impression cytology기법으로 수집하여 PAS(Periodic Acid Schift)-haematoxylin 염색을 시행하였다. 배상세포와 결막상피세포는 광학현미경 400배에서 관찰하여 squamous metaplasia의 정도에 근거한 Nelson Grading scale에 따라 분류하였는데 이는 배상세포 밀도, 세포의 크기와 형태, 핵:세포질 비 등을 포함한다. 건성안 의심자는 상피세포의 형태적 변화, 핵의 변화, 배상세포 밀도 감소를 보였다. 세포학적 변화 정도는 건성안 조건의 정도에 관련이 있었다. 상피세포 형태를 Nelson의 분류시스템으로 단계를 나눈 결과 대조군의 91.43%가 grade 0 이었고 8.57%가 약간의 이상형태인 grade 1이었다. 반면에 건성안 의심자에서는 20%가 grade 0, 42.86%가 grade 1, 34.29%가 grade 2 그리고 2.86%가 심한 형태적 변화를 보인 grade 3를 나타냈다. Impression cytology는 부작용이나 거부반응이 없는 안구 표면 상피의 비 침습적 또는 최소 침습적 생검법으로 생각되며 안구표면을 포함하는 광범위한 과정을 위한 유용한 진단 기법으로 안전하고 간단하며 건성안 의심자의 다양한 안구 표면 변화의 이해를 증진시키는데 도움을 줄 수 있을 것으로 사료된다.

• 대한수의학회지
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• 제56권3호
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• pp.147-153
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• 2016

A total of 782 blood and 465 tissue samples from 1,039 wild animals and 127 dairy goats were collected from January 2011 to December 2013 in 10 provinces of South Korea and tested for the presence of brucellosis. The Rose Bengal test revealed that 8.0% (52/650) of the serum samples were seropositive, while 4.2% (33/782) of the serum samples were positive for Brucella antibodies by competitive enzyme-linked immunosorbent assay. Of the 650 sera examined, only 16 (2.5%) were positive by both serological tests. Direct polymerase chain reaction (PCR) assay using B4/B5 primers for Brucella abortus (BCSP31) revealed the prevalence of Brucella to be 26.5% (129/487) in blood samples and 21% (98/465) in tissue samples while, 16S rRNA PCR detected Brucella DNA in 6.8% (33/487) and 2.6% (12/465) in blood and tissue samples, respectively. Of PCR-positive samples, only 6.2% (30/487) of blood samples and 2.4% (11/465) of tissue samples were found to be positive by both BCSP31 and 16S rRNA PCRs. However, Brucella strains were isolated by blood culture from only two out of 487 blood samples (0.4%). This characterization and identification of pathogenic Brucella isolates is the first to clearly indicate that the organisms were Brucella abortus biovar 1.

• 한국동물위생학회지
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• 제35권4호
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• pp.269-273
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• 2012

A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

• 동의생리병리학회지
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• 제23권3호
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• pp.645-661
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• 2009

The water extract of Dohongsamul-Tang(DHSMT) has been traditionally used to stroke and brain injuries in Oriental Medicine. The present study was designed to investigate the effects of DHSMT on the gene expression profile of cerebral infarction by cDNA microarray in photothrombotic ischemia mouse model. Photothrombotic ischemia was induced in stereotactically held male BALB/c mice using rose bengal and cold light. MRI was performed 24 hours after inducing photothrombosis using 1.5 T MRI and 47 mm surface coil to obtain T2-weighted, and contrast-enhanced images. After MRI test, animal was sacrificed and the brain sections were stained for hematoxylin and eosin and immunohistochemistry. MRI and histological analysis revealed that lesion of thrombotic ischemia was well induced in the cortex with the evidence of biological courses of infarction. The target area of thrombotic infarction was 1 mm anterior to bregma and 3 mm lateral to midline with 2 mm in diameter, which were decreased by administration of DHSMT. To assess gene expression pattern of cerebral infarction, mRNA was isolated and reacted with microarray chip(Agilant's DNA Microarray 44K). Scatter and MA plot analysis were performed to clustering of each functional genes. M value [M=log2(R/G), A={log2(R ${\times}$ G)}/2] was between -0.5 and +0.5 with 40% difference. After pretreatment with DHSMT, the expression levels of mRNA of many genes involved in various signaling pathway such as apoptosis, cell cycle, cell proliferation, response to oxidative stress, immune response, angiogenesis, and inflammatory cytokine were markedly inhibited in photothrombotic ischemia lesion compared to the control group. These results suggest that DHSMT prevent ischemic death of brain on photothrombotic ischemia model of mice through modulation of gene expression at the transcriptional level.