능성어, Epinephelus septemfasciatus의 바이러스성 신경괴사증 바이러스의 병원성 연구

손상규,전세규,Sohn, Sang-Gyu,Chun, Seh-Kyu 한국어병학회 1999 한국어병학회지 Vol.12 No.2

The pathogenicity of the fish nodavirus causing viral nervous necrosis (VNN) of sevenband grouper, Epinephelus septemfasciatus was examined in sevenband grouper and other marine fish by intramuscular injection. Sevenband groupers of 27~104 g in body weight were highly susceptible to the fish nodavirus, but yellowtail (537 g in body weight), red seabream (207 g), rock bream (43 g), flounder (41 g), tiger puffer (27 g) and rockfish (94 g) of the sizes used to this experiment were not. The pathogenicity of the viral agent to the sevenband grouper was high without regard to fish sizes at rearing water temperature over $20^{\circ}C$, but not at $15^{\circ}C$. Therefore, susceptibility of sevenband grouper to the viral agent causing VNN was thought to be water temperature-dependent rather than fish size (age-dependent). Compared to the infectivity of the viral agent to sevenband grouper with artificial infection methods, fish were successfully affected by intramuscular, intraperitoneal, oral, dipping and cohabitation administrations although there were slight differences in mortalities among infection methods. And survival sevenband grouper after infection with the fish nodavirus was resistant to the reinfection for a long time. 능성어의 바이러스성 신경괴사증 바이러스(fish nodavirus)의 병원성을 능성어 및 다른 해산어류를 대상으로 인위감염법에 의해 조사한 결과, 능성어 바이러스성 신경괴사증 바이러스는 능성어에서는 소형어(27 g)부터 대형어(104 g)까지 제사를 일으켜 강한 병원성을 나타냈지만, 실험에 사용한 크기의 방어(537 g), 참돔(207 g), 돌돔(43 g), 넙치(41 g), 자주복(27 g) 및 조피볼락(94 g)에는 폐사를 일으키지 않아 병원성을 나타내지 않았다. 그리고 사육수온별 능성어에 대한 바이러스의 병원성은 수온 $15^{\circ}C$에서는 병원성을 나타내지 않았고, 수온 $20\sim28^{\circ}C$에서는 수온이 높을 수록 병원성을 강하게 나타냈다. 그리고 바이러스는 근육주사, 복강 주사 및 경구감염뿐만 아니라 침지감염과 혼합사육감염에 의해서도 쉽게 능성어에 감염이 되지만, 감염 후 생존한 능성어에 대해서는 바이러스를 재감염시켜도 폐사가 일어나지 않았다.

PT-PCR 法에 의한 Infectious Pancreatic Necrosis Virus의 早期診斷

강호성(Ho Sung Kang),공희정(Hee Jeong Kong),구현나(Hyun Na Koo),박정우(Jeong Woo Park),손상규(Sang Kyu Sohn),박명애(Myung Ae Park),김진우(Jin Woo Kim),김한도(Han Do Kim) 한국수산과학회 양식분과 1997 韓國養殖學會誌 Vol.10 No.2

BFT 기법을 이용한 양식장 배출수가 고추 생육 및페놀함량에 미치는 영향

김용현(Yong Hyun Kim):허무룡(Moo Ryong Huh):박세원(Se Won Park):이재호(Jae Ho Lee):이준규(Jun Kyu Lee):손상규(Sang Gyu Sohn):이영식(Young Sik Lee),최혜승(Hye Sung Choi) 인간식물환경학회 2015 인간식물환경학회 학술대회 Vol.2015 No.추계

잉어류 바이러스성전신괴사증바이러스 (VSNCV) 백신 투여에 대한 잉어의 면역반응

조미영,손상규,김이청,김진우,오명주,정성주,박수일 한국어병학회 2003 한국어병학회지 Vol.16 No.3

양식 잉어류에서 심각한 피해를 야기하고 있는 VSNCV에 대한 백신을 제작하며 잉어에 투여한 후 특이적 및 비특이적 면역반응을 조사하였다. 백신은 포르마린처리백신 (FKV)과 가온처리백신 (HKV)를 제작하여 건강한 잉어에 0.2㎖씩 복강주사 하였으며, 2주 후 동일한 방법으로 boost처리하였다. 백신 투여 후 비특이적 면역반응 중 혈청 라이소자인 활성과 대식세포의 chemiluminescent (CL) reponses은 1차 주사 후 및 boost 처리 후 2일째부터 7일째까지 유의적인 증가를 나타내었으며, 이후 대조구 수준으로 감소하였다. ELISA법으로 항체가를 조사한 결과 대조구를 제외한 FKV와 HKV 투여구에서 주사 후 2주 째부터 boost처리 후 6주 째까지 유의적으로 증가하였으며, 그 중에서도 boost 처리 후 2주 째에 VSNCV로 공격 실험한 결과 FKV 투여구에서는 20%의 누적폐사율을 나타내어 방어력이 인정 되었으나, HKV투여구에서는 70%의 누적폐사율이 나타나 방어효과가 거의 없는 것으로 나타났다. VSNC is a viral disease causing significant economic losses in cultured carp Ciprinus carpio L. in Korea. Carps were immunized with prepared vaccines against VSNCV and examined specific and nonspecific immune responses. Carps were injected by 0.2㎖ of formalin-killed vaccine(FKV), heat-killed vaccine(HKV) or E-MEM, respectively and dealt with boost with same way two weeks later. The lysozyme activity of serum and chemiluminescent reponses of head-kidney leucocytes showed increased responses during 2-7 days post-first injection(pfi) and post-boost(pb) in the vaccinated fish, and then decreased to the level of control. As measured by ELISA, vaccinated groups showed a significant increase in VSNCV-specific serum antibodies between 2 weeks pfi and 6 weeks pb with a peak at 2 weeks pb. Results of the virus challenge showed that the fish vaccinated with FKV have induced protective immunity, white HKV injection hardly provided protection.

틸라피아(Tilapia sp .) 에서 분리한 Flexibacter columnaris 의 성장

전세규,손상규 한국수산학회 1985 한국수산과학회지 Vol.18 No.4

지느러미 腐蝕과 體色變化를 나타내는 틸라피아 病魚로부터 分離한 病原菌의 性狀은 다음과 같다. 1. 病魚로부터 分離한 病原菌은 Flexibacter columnaris로 同定되었다. 2. 分離한 病原菌液으로 人爲感染시켰을 때 전형적인 columnaris病 症狀을 나타냈다. 3. 病原菌은 kanamycin, tetracycline, amikacin 等에서 높은 感受性을 나타냈으나, sulfa劑에서는 거의 感受性을 나타내지 않았다. 4. 2% 이상 食鹽이나 50%이상 海水를 含有한 cytophaga 培地에서 病原菌은 자라지 못했다. 5. 증류수 대신 50% 이상 海水를 含有한 Cytophaga 培地에 病原菌을 接種시킨 후 1時問만에 約 10^6/㎖ 에서 10⁴/㎖로 菌數가 減少했는데 반하여, 5% 이상 食鹽을 含有한 cytophaga 培地에서는 接種 후 1時間만에 菌數가 約 10^5/㎖에서 10/㎖ 이하로 급격히 減少했다. The characteristics of a pathogen isolated from infected tilapia, which showed the external symptoms of fin erosion and body decoloration, were examined. The strain of isolated pathogen was identified as Flexibactor columnaris. The normal tilapia dipped into suspensions of the isolated pathogens has developed the infective symptoms. The strain of Flexibactor columnaris was sensitive to kanamycin, tetracycline, and amikacine, but resistant against sulfa drugs. This strain could not grow in the cytophaga media containing over 2% NaCl or over 50% sea water. In cytophaga broth containing more than 50% or sea water instead of distilled water, the number of Flexbacter columnaris decreased from about 10^6/㎖ to 10⁴/㎖ within one hour after inoculation. On the other hand, in cytophaga broth containing more than 5% of NaCl, the number rapidely reached less than 10/㎖ from about 10^5/㎖ within one hour after inoculation.

양식 능성어 , Epinephelus septemfasciatus 대량 폐사에 관한 연구

전세규,손상규,박명애,이생동 한국어병학회 1991 한국어병학회지 Vol.4 No.2

1. 自然感染 능성어는 외관적으로 먹이를 먹지 않고 體色이 옅은 밤색이나 짙은 밤색으로 변하며 몸이 휘어져 힘없이 가두리 바닥이나 수면에 누워서 斃死하였는데, 解剖를 해보면 腦出血과 脾臟 및 膽管이 膨大되어 있었다. 2. 自然感染魚 아가미에서 Trichodina sp. 寄生蟲이 다수 檢出되지만 大量 斃死의 直接 原因生物은 아니며, 또한 病魚의 臟器에서 優占的으로 分離되는 Vibrio屬 細菌은 病原性이 없는 海水常存 Vibrio 菌이었다. 3. RTG-2 및 CHSE-214 魚類 株化細胞에서 바이러스는 分離되지 않았지만, 病魚의 臟器磨碎濾過液으로 人爲感染시켰을 때 自然感梁魚와 類似한 症狀을 나타내면서 斃死하였다. 4. 自然感染魚 및 人爲感染魚의 肝 細胞質內 空胞部位에 크기가 45-60㎚ 정도되며 六角形 내지 多角形模樣을 한 virion들이 무수히 존재하였다. 5. 따라서 1990年과 1991年 高水溫期에 우리나라 南海岸 일원 해상가두리 양식장에서 사육중이던 능성어가 大量 斃死한 것은 바이러스性 疾病에 의한 것으로 생각된다. In the late summer of 1990 and 1991, mass mortality occured among cage-cultured grouper, Epinephelus septemfasciatus in south cost of Korea. The moribund fish didn't feed and became pale or dark chestnut colour and irregularly swimmed due to the loss of equilibrium, finally the diseased fish fell down side away on the bottom or the surface of cage showing the bent of body and died. The diseased fish showed the extensive hemorrahge in brain, the swelling of spleen and bile duct as the specific syptoms of internal organs. So the gill, skin and other organs of the diseased fish were examined for the presence of pathogenic parasites and bacteria. The parasitic Trichodina sp. were detected only from the gill lamella of the diseased fish, but these parasites seemed to be not a direct causative agents that induced the gross mortality of the cultured grouper. because these parasites were also observed in normal grouper, yellowtail, red seabream and rock bream co-cultured with the diseased grouper in same or near cages. In the viral examination, although isolation of the causative agent by the use of estabilshed cell Lines, RTG-2 and CHSE-214, was not succeed, the normal grouper inoculated intramuscularly with the filtered homogenate of the organs of the diseased fish showed the same external and internal signs with the naturally infected grouper. They died within a week. By using the naturally and the artificially infected fishes, electron microscopic observation revealed numerous hexagonal or polygonal particles in the cytoplasm of liver cells. Based on the these results, we suggest that the mass mortality of the cultured grouper would be occurred by the infection of a viral agent.

능성어, Epinephelus septemfasciatus 의 바이러스성 신경과시증 바이러스의 병원성 연구

전세규,손상규 한국어병학회 1999 한국어병학회지 Vol.12 No.2

능성어의 바이러스성 신경괴사증 바이러스(fish nodavirus)의 병원성을 능성어 및 다른 해산어류를 대상으로 인위감염법에 의해 조사한 결과, 능성어 바이러스성 신경괴사증 바이러스는 능성어에서는 소형어(27 g)부터 대형어(104 g)까지 제사를 일으켜 강한 병원성을 나타냈지만, 실험에 사용한 크기의 방어(537 g), 참돔(207 g), 돌돔(43 g), 넙치(41 g), 자주복(27 g) 및 조피볼락(94 g)에는 폐사를 일으키지 않아 병원성을 나타내지 않았다. 그리고 사육수온별 능성어에 대한 바이러스의 병원성은 수온 15℃에서는 병원성을 나타내지 않았고, 수온 20∼28℃에서는 수온이 높을 수록 병원성을 강하게 나타냈다. 그리고 바이러스는 근육주사, 복강 주사 및 경구감염뿐만 아니라 침지감염과 혼합사육감염에 의해서도 쉽게 능성어에 감염이 되지만, 감염 후 생존한 능성어에 대해서는 바이러스를 재감염시켜도 폐사가 일어나지 않았다. The pathogenicity of the fish nodavirus causing viral nervous necrosis (VNN) of sevenband grouper, Epinephelus septemfasciatus was examined in sevenband grouper and other marine fish by intramuscular injection. Sevenband groupers of 27∼104 g in body weight were highly susceptible to the fish nodavirus, but yellowtail (537 g in body weight), red seabream (207 g), rock bream (43 g), flounder (41 g), tiger puffer (27 g) and rockfish (94 g) of the sizes used to this experiment were not. The pathogenicity of the viral agent to the sevenband grouper was high without regard to fish sizes at rearing water temperature over 20℃, but not at 15℃. Therefore, susceptibility of sevenband grouper to the viral agent causing VNN was thought to be water temperature-dependent rather than fish size (age-dependent). Compared to the infectivity of the viral agent to sevenband grouper with artificial infection methods, fish were successfully affected by intramuscular, intraperitoneal, oral, dipping and cohabitation administrations although there were slight differences in mortalities among infection methods. And survival sevenband grouper after infection with the fish nodavirus was resistant to the reinfection for a long time.

양식 능성어로 부터 Fish Nodavirus 분리

오명주,전세규,손상규,박명애 한국어병학회 1998 한국어병학회지 Vol.11 No.2

1989년부터 남해안 일원 해상 가두리 양식장에서 사육중이던 능성어가 고수온기에 대량폐사하여 폐사율이 80%이었다. 발병한 병어는 채색흑화, 안구돌출, 평행감각 상실,회전운동과 몸통이 휘어지는 증상을 나타냈고, 조직학적으로는 뇌조직 및 안구망막의 신경세포가 공포 또는 괴사되었다. 병어로 부터 어류주화세포를 이용한 세포배양법으로 원인 바이러스는 분리되지 않았지만, 뇌 신경세포의 세포질에서 크기가 약 30 ㎚이며 외막(envelop)이 없는 다면체 모양의 바이러스 입자가 전가현미경적으로 관찰 되었다. 능성어로 부터 분리된 바이러스는 에테르나 열(50℃, 30분)처리에 의해 병원성이 실활되지 않았으나, 강산(pH 3)이나 강알카리(pH 11)에는 부분적으로 병원성이 실활되었다. 그리고 fish nodavirus인 Striped Jack Nervous Necrosis Virus(SJNNV)의 primer를 이용하여 인위감염 및 자연감염된 능성어를 대상으로 PCR을 한 결과 감염된 능성어의 뇌 및 안구조직에서 악 430 bp정도의 PCR 증폭산물이 검출되므로서 능성어를 폐사시키는 원인 바이러스는 SJNNV와 아주 유사한 nodavirus 였다. Since 1989, mass mortality has repeatly occurred in cage-cultured sevenband grouper, Epinephelus septemfasciatus along the southern coast of Korea in the summer season and usually reached over 80% within a few months. Diseased fish showed the clinical signs of anorexia, dark coloration, loss of eqilibrium, spinal swimming behaviour, vertebral deformity and inflation of swim bladder. Histopathologically, necrosis and/or vacuolation of the nerve cells in the brain and retina were observed. We previously reported that the causative agent was filtrable. The causative agent was not culturable in various fish cells; RTG-2, CHSE-214, BF-2, EPC and FHM. However, electron microscopic observation revealed unenveloped icosahedral viral particles with about 30 ㎚ in diameter in the cytoplasm of nerve cells of the brain. The characteristics of the virus was tested by an artificial infection with the filtrate of the homogenate of diseased fish. The pathogenicity of the virus was retained after treatment with ether or heat (50℃, 30 min) but partly lost by pH 3 or 11 treatment. These results suggest that the causative agent are similar to the fish nodavirus. In order to compare the causative agent with a fish nodavirus, Striped Jack Nervous Necrosis Virus (SJNNV), a polymerase chain reaction (PCR) was performed with primers specific to SJNNV. As a result, about 430 by PCR products were detected from the brain and the eye of both naturally and artificially infected sevenband grouper. All these results represent that the mass mortality in the cultured sevenband grouper is caused by the infection of a nodavirus similar to SJNNV and this is the first report of a fish nodavirus from the cultured sevenband grouper in Korea.

파장회전원판을 사용한 뱀장어 상육과 질병대책

김진우,전세규,손상규 한국수산학회 1985 한국수산과학회지 Vol.18 No.6

질병치료용으로 개발한 波狀回轉圓板을 순환여과 장치를 사용하여 199日만에 뱀장어를 3㎏에서 815.6㎏으로 성장시킬 수 있었다. 飼育水를 정화하기 위한 生物膜의 表面積이 1,127㎡였는데 波狀回轉圓板의 表面積이 800㎡였고, 水中에 沈積된 PVC 波板의 表面積은 327㎡였다. 이 장치는 汚物이 고이지 않으므로 Trichodina충과 Costia충의 치료가 용이하였다. The main purpose of this study is to evaluate a revolving plate-type biofilter system for mass culture of eels (Anguilla japonica) based on the experimental rearing for 199 days. Water quality, growth efficiency of fish and effects of fish disease control were critically evaluated. The experiment was conducted in two different units, each unit consisting of a cement tank containing 20㎥ of water. In unit A, a biofilter which includes 400 rotating undulated P. V. C. plates being 70㎝ in diameter which rotates at 6 rpm and also 400 undulated P. V. C. plates fixed in the settling chamber of an area of 66×62㎝. Water was continuously passed through the filter at a rate of 260ℓ/min., and supplemental water was added to the fish tank at a rate of 4㎥ a day. In unit B, the biofilter has 400 P. V. C. plates being 66×62㎝ each was installed in the settling tank. The results gained from the experimental rearing for 199 days from April 21, 1984 to November 5, 1984 are as follows. In the growth experiment, the weight of fish in unit A increased from 3.0㎏ to 815.6㎏, while in unit B, from 3.0㎏ to 416.0㎏. During the period of the experiment, in the both units the fish grew at an acceptable rate at the temperature at which they were held. Observing every aspect of eel culture, including growth rate, disease control and water quality, unit A appears to have adventages over unit B, which makes it particuraly attractive in intensive recirculating fish culture system. It was further observed that certain parasites such as Trichodina sp. and Costia sp. could easily be controled by appling 4ppm of KMNO₄.