• 한국동물위생학회지
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• 제42권4호
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• pp.237-243
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• 2019

Avian reovirus (ARV) is the pathogenic agent of tenosynovitis and malabsorption syndrome in broiler, which has caused significant economical losses due to poor feeder efficiency and stunting. In order to determine the prevalence of ARV infection in poultry farms in Jeonbuk province, Korea, we performed a surveillance study by testing 179 cecal samples from 131 broiler farms for virus detection, and 1,181 serum samples from 33 broiler farms (n=292) and 22 broiler breeder farms (n=1,525) for antibody detection in the province. Virological examination using RT-PCR showed that ARV were detected in 26.0% of the tested farms (34/131),with the highest positive rates in broilers of 6 days old or more in summer season. In serological test using ELISA, broiler and broiler breeder farms examined were all ARV antibody positive. In broiler, the positive rate and antibody titers showed a tendency to decrease with age in contrast to those of broiler breeders. Our results indicate that ARV is ubiquitous in broilers and broiler breeders in the province.

• 한국식물병리학회지
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• 제1권1호
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• pp.61-66
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• 1985

벼 흰빛잎마름병균주 Q7472와 Q7502의 항혈청을 사용하여 우리나라에서 수집한 벼 흰빛잎마름병균의 혈청형을 한천gel내 확산법에 의해 분류한 결과, 71균주중 65균주가 혈정형 A형, 5균주가 B-I, 1균주가 B-II형에 속하였다. 혈청형A에 속하는 균주는 $0.1-1\%$의 $CaCl_2$용액에서 자기의집이 거의 일어나지 않았으나 혈청형 B-I과 B-II에 속하는 균주는 심한 자기의집이 일어나 혈청형과 자기의집 사이에는 밀접한 관계가 있었다. 흰빛잎마름병균은 혈청학적 방법에 의해 쉽게 검색될 뿐만 아니라 한천gel내 확산법에 의해 벼 세균성줄무늬병균인 X.campestris pv. olyzicola의 부생세균인 E. herbicola와도 쉽게 구별되었다. 흰빛잎마름병균 검출은 병반을 NaCl용액에 넣은 후에 착압하거나 $100^{\circ}C$에서 1시간 열처리 후 착압한 즙액을 한천gel내 확산법에 의해 항혈청과 반응시킴으로써 가능하였으며, 병반이 적을때는 PSA배지에서 배양한 후 사용하면 효과적이었다.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제10권1호
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• pp.141-146
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• 1997

Postmortem examination was conducted on 350 (three hundred and fifty) chickens. Related samples (Liver, heart, ovary, spleen, bone-marrow, and caecal junction) were collected. The appropriate materials from the samples were cultured into different media. A total 40(forty) isolates of salmonella pullorum and S. gallinarum were identified and preserved. Characterization of the isolates were done by cultural, morphological, biochemical, and serological tests. Salmonella pullorum antigen was prepared from the local isolate, standardized and tested. This antigen was used in the field for the detection of pullorum or fowl typhoid infection or carrier birds. The antigen consisted of suspension of Salmonella pullorum in 0.50 percent sodium chloride plus 1.5 percent sodium sulfate and inactivated with 1% formalin U.S.P. and standardized with McFarland scale iv or by pour plate method containing 800 million organisms per milliliter and stained by the addition of alcoholic crystal violet. Sterility, safety and potency were tested and found as good as other international antigens. The antigen was found to retain its quality for six months when preserved at room temperatures. The test was made by mixing one drop of the antigen with a drop of blood or a drop of serum, on a glass plate or white tile. The locally produced antigen was as good as antigens from Japan, Hungary, Holland and India. A serological study was conducted with the locally prepared antigen in different farms, and the incidence was 0-4% in government farms, 5-10% in commercial imported breeds and 0-3% in cross breed local farms respectively.

• 대한수의학회지
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• 제47권4호
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• pp.417-423
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• 2007

Serological surveillance programs in animal populations are becoming increasingly important to estimate prevalence of a specific disease and subsequently to document disease-free status in a region or a country. For these purposes, the programs need to be based on both theoretical and economical aspects from the designing phase. From Aujeszky's disease (AD)-eradication program point of view, group of animals (aggregates, herds) not individual animal is the more important sampling unit of concern. In this study the authors therefore attempted to compute an appropriate sample size tailored to a current surveillance program against AD, assuming that the goal of this program is either herd-level prevalence estimation or documentation of AD-freedom. For prevalence estimation, assuming a finite population with imperfect sensitivity (Se) and specificity (Sp) of ELISA kit for AD diagnosis, the number of herds present, expected herd prevalence, and desired accuracy for a certain level of confidence, sample size was estimated at herd-level in the first stage and individual animal-level in the second stage. A two-stage sampling design was used to calculate a sample size to indicate AD-freedom. In this instance, the computation was based on the possible detection of a predetermined prevalence at a certain herd-level Se and Sp. This study indicated that the sample size varied with predetermined confidence, tolerance, Se and Sp at herd- and animal-level, and within- and among-herd prevalence. In general, smaller sample size was required to estimate AD prevalence than to document of AD-freedom. Compared to individual-based samples, two-stage sampling strategy requires a larger sample size to show disease-freedom. Statistical considerations including herd-level test characteristics when designing surveillance program also are further discussed.

• 한국동물위생학회지
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• 제23권4호
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• pp.349-360
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• 2000

An epidemiological survey was conducted to investigate fowl typhoid outbreaks in Kyungnam province of Korea. The causative agent, salmonella gallinarum was isolated from 68 chicken samples of tentatively diagnosed fowl typhoid cases occurred during the period from January 1996 to September 1999. Comparative studies were also carried out to evaluate the diagnostic methods for detection of S gallinam The results obtained were as follows; 1. Of the 68 cases of tentatively diagnosed fowl typhoid, 56 (82%) cases were determined as fowl typhoid by biochemical test and pathological findings. The other 12 (18%) cases were determined as paratyphoid. 2. Fowl typhoid outbreaks occur continuously all seasons in the year, however the incidence was remarkably increased from May to September. 3. The frequency of incidence of fowl typhoid in terms of regional distribution was relatively high in egg-laying hens facilities, and the mode of transmission is likely to be either egg-to-egg or lateral transfer by wild birds or rats. 4. All of 18 isolates from 56 cases were identified as S gallinarum by biochemical and serological test. 5. Antimicrobial drug susceptibility test against 18 isolates showed that the isolates were highly susceptible to ASH, CZ, CF and GM (above 90%), whereas those strains were 100% resistant to EM, NA and PC. 6. S gallinarum rfbS gene was targeted to be amplified by PCR for comparative detection of S gallinarum in the experimentally infected chickens. The amplified 720bp DNA fragment, which is specific in D serogroup strains of S enterica subspecies was confirmed by agarose gel electrophoresis. 7. A comparison made between fecal culture and PCR-method revealed that later-method was relatively higher in detection rate than that of former method for S gallinarum. 8. Comparison of currently applied methods, rapid serum agglutination test (RST) and microplate agglutination test (MAT), with experimentally infected chickens were made to evaluate sensitivity of detection by neutralizing antibody titration. Both methods detected neutralizing antibodies from the challenged chickens of 5 day post infection. However, positive reactions were determined after 7 and 9 days post infection by MAT and RST, respectively.

• 대한수의학회지
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• 제43권3호
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• pp.471-476
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by ovine herpesvirus 2 (OvHV-2). OvHV-2 is a gamma herpesvirus, which induces frequent latent infection and often difficult to detect its antigens and even specific nucleic acids because of its low viral copies in the infected tissues. Histopathology, serology and polymerase chain reaction (PCR) were compared for the diagnosis of MCF using 10 bison infected with OvHV-2. Histopathological diagnosis was performed using the criteria which was based upon the pathognomic lesions. Serological diagnosis was conducted using its serum with competitive ELISA for the detection of antibodies of OvHV-2. Also, the nest PCR was performed with peripheral blood leukocytes for the detection of OvHV-2-specific DNAs. Primers 556 and 775 were used for the primary amplification, and primers 556 and 555 were used for the secondary amplification. As the results, positive cases were 6 by histopahology, 9 by serology and 10 by PCR. As comparing with other diagnostic methods, PCR was found to be more sensitive than histopathology and serology. The recent development of molecular diagnostic assays has provided powerful tools for investigating how viruses survive in nature. Development of PCR specific for viruses has dramatically improved the accuracy of diagnosis of viruses in clinically infected animals. Furthermore, amplification of viral genomic material by nest PCR represents the most sensitive method for the detection of viruses and might be detected successfully even though very low viral DNA copies. So, it could be used as the first choice for the detection of viral DNAs with low copies such as the status of latent infection. However, it has also some limitation of application like as false negative results by PCR inhibitors and false positive results by contamination. The results of this study suggest that the use of molecular biological methods like PCR may increase the accuracy for the diagnosis of infectious diseases. However, in diagnostic laboratory, it is recommended that PCR assay must be conducted with other diagnostic methods for more reliable diagnosis.

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1401-1409
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• 2015

The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10⁰ fg/µl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

• 대한수의학회지
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• 제38권4호
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• pp.818-822
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• 1998

The persistence of porcine epidemic diarrhea virus(PEDV) infection was demonstrated in 7 swine farms employing continuous pig flow management even after seasonal outbreaks. Clinically, sporadic postweaning diarrhea was a major concern in those farms. Subsequently circulatory antibody detection using serum neutralizing test made useful for confirmation of PEDV persistent infections. The persistence of PEDV in the premise might have induced recurrence over the period of time.

• 대한수의학회지
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• 제42권4호
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• pp.545-550
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• 2002

The raccoon dog (Nyctereutes procyonoides) may be infected by Dirofilaria immitis. However, there has been no report on dirofilarial infection in the raccoon dog in Korea. In this study, we report on D. immitis infection in two wild raccoon dogs captured in the Daejeon area. The two raccoon dogs were referred to the Veterinary Teaching Hospital, Chungnam National University for diagnosis of D. immitis infection. The modified Knott's test for the detection of blood D. immitis microfilariae was positive, and serological test (FASTest$^{(R)}$ HW Antigen ELISA kit, Diagnostik Mega Cor, Austria) for D. immitis was positive as well. Additionally, D. immitis microfilariae were differentiated from other microfilariae by using acid phrnphatase histochemical staining (Leucognost-SP$^{(R)}$kit, Diagncstica MERCK, Germany). The two raccoon dogs were necropsed and D. immitis infection was confirmed.