세망내피증 바이러스 항체검출을 위한 ELISA 표준화

성환우,이수정,Sung, Haan Woo,Lee, Su Jeong 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.4

Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

깔짚 교체 및 재사용 육계농장 분리 대장균의 항생제 내성 양상

성환우,최강석,권혁무,이영주,Sung, Haan-Woo,Choi, Kang-Seuk,Kwon, Hyuk-Moo,Lee, Young-Ju 대한수의학회 2017 大韓獸醫學會誌 Vol.57 No.3

The isolation rate of Escherichia (E.) coli in poultry litter was investigated at 44 broiler farms, 20 that used fresh litter and 24 that used recycled litter. The patterns of resistance to antibiotics of the E. coli isolates were compared. In litter sampled before the rearing period, the isolation rate of E. coli was higher at farms that used fresh litter; E. coli was present in the litter in 94.5% (35 out of 37 flocks tested) of the farms that used fresh litter vs. 51.2% (21 out of 41 flocks) of the farms that used recycled litter. The susceptibility of the 93 isolates of E. coli to 13 antibiotics was studied. Before the rearing period, E. coli isolates from the farms that recycled litter showed higher resistance rates than isolates from farms that replaced litter with fresh litter. Comparing the antibiotic resistance patterns of isolates from litter sampled before and at the end of the rearing period, the antibiotic resistance rates at the end of the rearing period increased dramatically compared with rates before the rearing period.

국내 분리 세망내피증 바이러스의 면역억제능

성환우,김선중,Seong, Hwan-woo,Kim, Sun-jung 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.4

Humoral and cellular immune responses are depressed in chickens infected with reticuloendotheliosis virus(REV). The extent of depression is influenced by the age of infection and strain of virus. This study was conducted for investigation of immunosuppressive effects of a Korean isolate of REV. Chickens infected with REV-HI, a Korean isolate, at 1 day old were severely suppressed in the vaccinal immunity against Newcastle disease, infectious bronchitis and infectious bursal disease. But these immunosuppressive effects were not observed in chickens infected with the virus at 2 weeks of age, or contact infected by growing in-contact with inoculated chickens from one day old. The clinical signs following infectious laryngotracheitis(ILT) vaccination in chickens infected with REV-HI at 1 day old were more severe than those of uninfected chickens, and some of REV-infected chickens(21.4%) were died after the vaccination. Mortality following virulent ILT virus infection was increased in REV-HI infected chickens. Effects of REV infection at one day old to susceptibilities to subsequent Chicken anemia agent (CAA) infection were also studied. Chickens were infected with REV-HI at 1 day old and subsequently inoculated CAA at 1, 7, 14 and 28 days old, respectively. Mortalities of the chickens infected with REV-HI and subsequent CAA infection were 100, 100, 40 and 0%, respectively, whereas 23, 8, 0 and 0% of chickens infected with only CAA were died, respectively. These above all results suggest that a Korean isolate of REV may be highly immunosuppressive.

세망내피증 바이러스의 조기검출과 국내 분리주의 면역억제성 및 종양원성

성환우 한국가금학회 1995 한국가금학회 정기총회 및 학술발표회 Vol.12 No.-

중합효소연쇄반응을 이용한 닭 종양성 질병의 감별진단에 관한 연구

성환우,김선중,Seong, Hwan-woo,Kim, Sun-jung 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.1

The present study attempted to apply polymerase chain reaction (PCR) to develop a rapid differential diagnosis among Marek's disease, reticuloendotheliosis and avian leukosis. The primers chosen to detect Marek's disease virus (MDV) flank the 132bp tandem direct repeat of the MDV genome. The primers selected for reticuloendotheliosis virus (REV) and avian leukosis virus (ALV) are based on proviral long terminal repeats of spleen necrosis virus and Rous-associated virus-2 genomes, respectively. The specific PCR products of MDV, REV and ALV were observed with each primer and the reaction was not cross-reacted among the viruses. MDV-specific DNA was also amplified from the MDV-induced lymphoma (MDCC-MSB1) but not from the REV-induced tumor and ALV-induced lymphoma (LSCC-1104B1). In addition, proviral DNA of REV from REV-induced tumor and proviral DNA of ALV from ALV-induced lymphoma were also amplified by REV-specific and ALV-specific PCRs, respectively. Therefore these three PCR methods may be used to rapidly differentiate among MDV, REV and ALV-associated tumors in diagnosis.

국내 분리주를 이용한 오리 바이러스성 간염 생백신주의 개발

성환우,김재홍,송창선,한명국,이윤정,모인필,김기석,Sung, Haan-woo,Kim, Jae-hong,Song, Chang-seon,Han, Myung-guk,Lee, Youn-jeong,Mo, In-pil,Kim, Ki-seuk 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.1

Duck viral hepatitis is an acutic, highly infectious viral disease of young ducklings. The most practical means for controlling duck viral hepatitis is the vaccination of ducklings or of a breeding stock. We attempted to develop a vaccine strain of duck hepatitis virus (DHV) using a Korean isolate by serial chicken embryo passages. The propagation of DHV in chicken embryos was carried 140 passages. After the $50^{th}$ passage, of which the virus was non-pathogenic for ducklings, approximately every $20^{th}$ passage of the virus was tested for vaccinal efficacy. Both the $70^{th}$ and $90^{th}$ passage of the virus gave good protection against challenge infection to a DHV-DRL reference strain(type 1) and a virulent Korean isolate. The $110^{th}$, $125^{th}$ and $140^{th}$ passage of the virus were less protective than the $70^{th}$ and $90^{th}$ passage, which means that more than $110^{th}$ passage may lead to over-attenuation of the virus. Ducklings vaccinated with the chicken-embryo-adapted virus by oral, intramuscular or eye drop administration showed earlier resistance to challenge infection from 3 to 7 days postvaccination. Of the above methods, ducklings vaccinated intramuscularly presented the most rapid resistance against challenge. The minimum immune dose of the chicken-embryo-adapted virus in ducklings was also studied. Ducklings inoculated with a dose of $10^{2.0}\;ELD_{50}$ and below were not fully protected against challenge with a virulent DHV, showing a protection rate of 67% to 73%, but ducklings inoculated with a dose of $10^{3.0}\;ELD_{50}$ and over were completely protected. The virus yield of the chicken-embryo-adapted DHV was examined at 24hrs and 48hrs of the incubation time in the allantoic fluid, embryo head and embryo minus head of the embryonating egg. In all three components, the titer of the virus was higher at 48 hours than that at 24 hours after incubation. And the titer of the virus was higher in the embryo minus head, embryo head and the allantoic fluid, in order. Field trials for evaluating the efficacy of the attenuated DHV as a live vaccine were done in duck farms with about 25% mortality of flocks resulting from duck viral hepatitis. After the use of the experimental vaccine, the mortality due to duck viral hepatitis was dramatically reduced in the farms. These results indicated that the attenuated DHV using a Korean isolate could be a good candidate as a live vaccine strain of DHV in Korea.

산란계에서의 칸디다증 국내 발생 사례

성환우,권혁무,한정희,모인필,Sung, Haan-Woo,Kwon, Hyuk-Moo,Han, Jeong-Hee,Mo, In-Pil 대한수의학회 2010 大韓獸醫學會誌 Vol.50 No.4

Candidiasis is a mycosis caused by the mycelial yeast of the Candida genus which is opportunistic pathogen of humans, animals, and birds. Under some conditions such as prolonged antibiotic therapy, overcrowding, and immunosuppression, the opportunistic Candida can cause disease. Chicken candidiasis is sporadically occurred and characterized by unsatisfactory growth, listlessness, roughness of feathers, and death. A case of 23 weeks old layer with history of increased mortality and anemia was submitted to our Lab. At necropsy, the characteristic lesions were observed in the crop and proventriculus. The whitish pseudomembrane, that are peeled easily, was found in the crop. Proventriculus was swollen and the mucosa was covered with hemorrhagic exudate. The histological changes of the affected crop are epithelial hyperplasia, hydropic degeneration, and mycelia formation. Smears made from the necrotic mucosal surfaces of the crop revealed the presence of large number of yeast cells and mycelia. Pure cultures of yeast colonies were obtained from the potato dextrose agar. The yeast cells were identified as Candida albicans by gene sequencing. To our knowledge, this is the first report of candidiasis in chickens with anemia in Korea.

오리 간염 바이러스의 분리와 국내 분리주의 약독화

성환우,김재홍,Sung, Haan-woo,Kim, Jae-hong 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.1

Duck viral hepatitis is an acute, highly infectious viral disease of young dacklings aged from two days to three weeks. The significant lesion associated with the disease was enlarged liver including necrotic foci and numerous hemorrhagic spots. We have isolated five strains of duck hepatitis virus (DHV) from field cases showing about 20% mortality with a sign of opisthotonos. When a-day-old ducklings were intramuscularly inoculated with one of the isolates, 92% of the birds were died within 5 days. We attempted to develop an attenuated strain of duck hepatitis virus (DHV) using one of the isolates by serial chicken embryo passages. The propagation of DHV in chicken embryos was carried 140 passages. The virus titer increased gradually from the $21^{st}$ through the $50^{th}$ passage, but there was no significant increase of virus titer in subsequent passages after then. Through the serial passages, the virulence of the virus for chicken embryos was gradually increased but decreased for ducklings. The pathogenicity of the virus for ducklings was preserved up to the $21^{st}$ passage but disappeared at the $50^{th}$passage. An attenuated Korean isolate which was passaged 140 times in chicken embryos gave good protection in ducklings against both challenge infection to a Korean virulent strain and to a DHV-DRL strain, a type 1 reference strain of DHV, which indicated that the Korean isolates could be classified as DHV type 1. And the above results suggest that an attenuated Korean isolate can be used for developing a live DHV vaccine.

학술 3 - 닭 전염성빈혈 예방대책

성환우,Seong, Hwan-U 대한수의사회 2012 대한수의사회지 Vol.48 No.7

닭 빈혈인자(CAA)감염증이란

성환우 대한양계협회 1991 월간 양계 Vol.23 No.5