Salmonella Enteritidis 분리주에서의 선발된 불활화 백신균주의 방어효과 및 면역원성

강정무,원호근,김은희,노윤희,최환원,한태욱,Kang, Zheng-Wu,Won, Ho-Keun,Kim, Eun-Hee,Noh, Yun-Hee,Choi, Hwan-Won,Hahn, Tae-Wook 대한수의학회 2011 大韓獸醫學會誌 Vol.51 No.1

Salmonella Enteritidis (SE) has been a major causative agent of food-borne human disease due to consumption of contaminated eggs and poultry meat. To prevent SE infection in poultry, and therefore minimize human infections, vaccination with either killed or live SE vaccine is suggested. We evaluated a newly developed killed bacterin using a representative SE isolate in Korea. Among pool of SE isolates, two highly virulent isolates (the one isolate from chicken, the other from human) were selected by measuring mortality in mouse and chickens administered. The chickens were injected intramuscularly with killed vaccine and were challenged with highly virulent SE strain 3 week after vaccination. The recovered colony count (cfu/g) of spleen and cecal content in the vaccinated groups was reduced compared with those of the unvaccinated control group. The antibody level in the vaccinated groups was higher at 3 week post vaccination. These results indicate that vaccination with killed vaccine was effective in preventing the infection of virulent SE. Further study for a large number of layers should be needed for the effect of egg production, SE shedding in feces, persistence of antibody level.

Infectious bursal disease virus 국내분리주 및 백신주의 VP2 gene의 비교분석

김길동,강정무,김선중,권혁무,한태욱,Jin, Ji-Dong,Kang, Zheng-Wu,Kim, Sun-Joong,Kwon, Hyuk-Moo,Hahn, Tae-Wook 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.3

The VP2 full gene of Korean infectious bursal disease virus(IBDV) strain, SH/92, three attenuated vaccine strains, Bur706, Bursine-2 and CEV/AC strains, were amplified by reverse transcriptase-polymerase chain reaction and sequenced and compared with published VP2 gene sequences of IBDVs. The VP2 nucleotide sequence similarity between SH/92 and three vaccine stains was 95.6~96.5% whereas the nucleic acid similarity among three vaccine strains was 97.5~98.5%. The amino acid sequence similarity of VP2 of SH/92 compared with three vaccine strains was between 94.4 and 97.6% while the amino acid similarity among three vaccine strains was between 97.4 and 98.4%. The amino acid similarity between SH/92 and classical virulent strain, 52/70 and STC strain was 96.4 and 96.5%, respectively. The serine-rich heptapeptide was conserved in CEVAC and Bursine-2 as well as SH/92 but not in Bur706. The phylogenetic tree developed from amino acid sequences showed that SH/92 was categorized with vv IBDVs(HK46, OKYM, KKI, UPM94/273, SH95) in one branch while three vaccine strains were catagorized with STC strain in the other branch.

닭 Mycoplasma gallisepticum 6/85 생균 백신의 효능 평가

윤희준,강정무,김길동,신은경,정용운,정지혜,한태욱,Yoon, Hee-Jun,Kang, Zheng-Wu,Jin, Ji-Dong,Shin, Eun-Kyung,Jeong, Yong-Hoon,Jeong, Ji-Hye,Hahn, Tae-Wook 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.3

Mycoplasma gallisepticum (MG) continues to persist in many commercial layer farms in Korea,resulting in losses in egg production. Bacterins and live attenuated vaccines have been used for the prevention of losses caused by MG. One of these attenuated vaccines, MG 6/85 vaccine has been reported to be safe and efficacious in layers. However, MG 6/85 vaccine has not been evaluated for its safety and its efficacy in any commercial layer in Korea. Six-week-old specific pathogen-free (SPF) chickens were vaccinated with MG 6/85 vaccine by aerosol and were challenged with virulent MG R strain at 4 weeks after vaccination. The vaccinated group was able to resist challenge into the air sacs because the vaccinated group showed much less air sac lesion compared with the unvaccinated group. Each of two commercial layer farms was divided into vaccinated and unvaccinated groups. For each vaccinated gorup, MG 6/85 vaccine were sprayed at 17 week old on farm A and at 15 weeks old on farm B. Hen-day egg production, Hen-housed eggs, egg weight, mortality were evaluated until 50 week after vaccination.Compared with the unvaccinated group in each farm, the vaccinated group showed higher average egg production and egg weight, and higher hen-housed number. Results of this study are in agreement with other previous reports which demonstrated that MG 6/85 vaccine favorable effect on performance in commercial layers.

Salmonella Enteritidis 및 Salmonella Typhimurium을 함유한 이가 불활화백신인 Salenvac-T의 방어효과 및 임상연구

조영재,강정무,강경수,정승환,윤희준,서승원,한태욱,Cho, Youngjae,Kang, Zheng-Wu,Kang, Kyung-Soo,Jeong, Seunghwan,Yoon, Hee-Jun,Suh, Seungwon,Hahn, Tae-Wook 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.1

Commercial bivalent killed Salmonella vaccine Salenvac-T has been used in several countries in order to prevent salmonellosis with Salmonella enterica serovars Enteritidis (SE) and Typhimurium (ST) in poultry. However, this vaccine has not been used in poultry farms in South Korea. In this study, we evaluated the efficacy of Salenvac-T vaccine to protect against the challenge of virulent SE and ST, and the effect of the vaccine on egg production and mortality in layer hens. The colonization of liver, spleen and cecum with challenged SE and ST was reduced in vaccinated chickens compared with that of unvaccinated control group. The twice vaccination with Salenvac-T induced elevated antibody responses against both SE and ST detected by enzyme-linked immunosorbent assay (ELISA). The higher average hen-day production was observed in the vaccinated layer hens than in the unvaccinated layer hens without significance. The average mortality was lower in the vaccinated layer hens during the experiment period. The antibody responses to both SE and ST were persistently detected in the vaccinated layers. In summary, vaccination with Salenvac-T reduces colonization of internal organs and induces good antibody responses, thereby results in higher performance and lower egg contamination with SE and ST in layer hens.

Salmonella Gallinarum 세포외막단백질의 프로테옴 분석 및 닭에서의 방어능 효과

선지선,조영재,장주현,강정무,한장혁,한태욱,Sun, Jisun,Cho, Youngjae,Jang, Joo-Hyun,Kang, Zheng-Wu,Han, Jang-Hyuk,Hahn, Tae-Wook 한국축산식품학회 2013 한국축산식품학회지 Vol.33 No.2

Salmonella Gallinarum (SG) is known as an important pathogen that causes fowl typhoid in chickens. To investigate SG outer-membrane proteins (OMPs) as a vaccine candidate, we used proteomic mapping and database analysis techniques with extracted OMPs. Also, extracted OMPs were evaluated in several aspects to their safety, immune response in their host and protective effects. Our research has established a proteomic map and database of immunogenic SG-OMPs used as inactive vaccine against salmonellosis in chickens. A total of 22 spots were detected by 2-dimensional gel electrophoresis and immunogenic protein analysis. Eight spots were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight-Mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprinting (PMF) and categorized into four different types of proteins. Among these proteins, OmpA is considered to be an immunogenic protein and involved in the hosts' immune system. To estimate the minimum safety dose in chickens, 35 brown layers were immunized with various concentrations of OMPs, respectively. Consequently, all chickens immunized with more than a $50{\mu}g$ dose were protected against challenges. Moreover, intramuscular administration of OMPs to chickens was more effective compared to subcutaneous administration. These results suggest that the adjuvanted SG-OMP vaccine not only induces both the humoral and cellular immune response in the host but also highly protects the hosts' exposed to virulent SG with $50{\mu}g$ OMPs extracted by our method.

Salmonella enterica serovars Enteritidis의 온도감수성 변이주 및 폴리인산키나아제 변이주의 제작과 방어효과

김기주,박소연,조영재,곽정연,강정무,김은희,최환원,원호근,노윤희,한태욱,Kim, Kiju,Park, Soyeon,Cho, Youngjae,Kwak, Jeong-Yeon,Kang, Zheng-Wu,Kim, Eun-Hee,Choi, Hwan-Won,Won, Ho-Keun,Noh, Yun-Hee,Hahn, Tae-Wook 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.4

This study was focusing on evaluating the protection of polyphosphate kinase (ppk) deleted and/or temperature-sensitive (ts) Salmonella Enteritidis (SE) as an attenuated vaccine in chickens. We constructed SEppk, SEts and SEppk::ts mutants and screened those mutants by growth capability in vitro, protection study in mice model and antibody response in chickens. Among the mutants, SEppk::ts-3 was selected because it showed higher growth capability, good protection against highly virulent SE in mice model, and good antibody response in chickens. SEppk::ts-3 also showed good protection against highly virulent SE isolate because it decreased colonization of virulent SE challenge strain in spleen, liver and cecum compared with the non-vaccinated control. The SEppk::ts-3 mutant showed cross-protection against S. Gallinarum (SG) challenge although the its cross-protection rate was a little lower than that of SG9R, a commercial vaccine against SG infection. To use for live attenuated vaccine in chickens, it should further be characterized.

닭에서 Mycoplasma gallisepticum과 M. synoviae의 항체양성률 조사

장석현 ( Seok Hyun Jang ),강정무 ( Zheng Wu Kang ),정찬이 ( Chan Eee Jung ),윤종웅 ( Jong Ung Yoon ),한태욱 ( Tae Wook Hahn ) 한국가축위생학회 2010 韓國家畜衛生學會誌 Vol.33 No.1

Mycoplasma gallisepticum (MG) is major cause of chronic respiratory disease in chickens. M. synoviae (MS) most frequently occurs a subclinical upper respiratory infection but may result in airsacculitis and synovitis in chickens and turkeys. Both mycoplasmas induce economic losses by triggering chronic respiratory signs, airsacculitis and decreased egg production. For prevention of the infections, live attenuated andinactivated vaccines are commercially used for prevention of MG but not MS in Korea. Serum plate agglutination (SPA) and enzyme-linked immunosorbent assay (ELISA) have been commonly used for serological diagnosis for MG and MS. Recently, it is believed that MS spread in chickens is very seriously in Korea and respiratory infection with MS causes substantial loss in poultry farms. In this study, we investigated the serological prevalence of MG and MS in unvaccinated chickens between 2008 and 2009. The overall seroprevalence of MG was 24% of 2,094 for individual chickens and 24% of 189 farms. The overall seroprevalence of MS was 36% in 2,095 chickens and 39% in 198 farms. The results show that seropositive ratio of MS is higher than MG. The geographical prevalence of MG has been estimated in following sequence; Gangwon, Jeolla, Gyeonggi, Gyeongsang, and Chungcheong. The geographical prevalence of MS has been estimated as follows; Gangwon, Gyeonggi, Gyeongsang, Chungcheong, and Jeolla. Seasonal seroprevalencewas also examined, and it found that seroprevalence in spring, fall and winter was higher than that in summer in MG, but not in MS. No significant difference was shown in seroprevalence according to breed. Future study about pathogenicity of MS isolates would be needed and economical losses by MS outbreaks should be analyzed. Moreover, we compared sero-positivity obtained with SPA and ELISA. The kappa value of MG between SPA and ELISA was 0.8061 and the kappa value of MS between SPA and ELISA was 0.7649.

도축돈에서 분리된 살모넬라의 혈청형 및 유전형

최원종 ( Won Zong Choi ),정지헌 ( Ji Hun Jung ),원호근 ( Ho Keun Won ),강정무 ( Zheng Wu Kang ),한태욱 ( Tae Wook Hahn ) 한국가축위생학회 2008 韓國家畜衛生學會誌 Vol.31 No.1

Salmonella infections cause the disease in pigs but also some zoonotic Salmonella sero-types can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detec-tion of invA gene using Salmonella specific polymerase chain reaction(PCR), the epidemio-logical characteristics related to Salmonella strains cultured from pig samples in Gang-won areas using serotyping, random amplified polymorphic DNA(RAPD) and pulsed-field gel electrophoresis(PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine(TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioM?rieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23(52.3%) were S Eingedi, 10(22.7%) S Schwarzengrund, 9(20.5%) S Typhimurium, and 2(4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distin-guishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

비구조 단백질 유전자 primer를 사용한 RT-PCR에 의한 인플루엔자 A형 바이러스의 검출

문형선 ( Hyeong Sun Moon ),배윤영 ( Yoon Yeong Bae ),김길동 ( Ji Dong Jin ),강정무 ( Zheng Wu Kang ),한태욱 ( Tae Wook Hahn ) 한국가축위생학회 2009 韓國家畜衛生學會誌 Vol.32 No.2

Influeza type A virus have been worldwide problematic in animals as well as in humans. In this study, the use of reverse-transcriptase polymerase chain reaction (RT-PCR) was described for detecting influenza virus type A. The primer of RT-PCR was designed from an nonstructural (NS) gene of Influenza A virus. By RT-PCR, a product with the size of 189 bp was detected only when influenza virus type A was used as template. No products could be detected with Influenza virus type B as well as other respiratory pathogens. The detection limit of the RT-PCR was up to 100.3TCID50 which is comparable to the sensitivity of cell culture method. The RT-PCR could detect the influenza A virus from nasal turbinates of the ferrets infected with influenza virus type A not type B.