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자연발생한 돼지 다발성 장막염 예로부터 Haemophilus parasuis의 분리와 면역조직화학적 진단
배유찬,강문일,황의경,손현주,최상호,Bae, You-chan,Kang, Mun-il,Hwang, Eui-kyong,Sohn, Hyun-joo,Choi, Sang-ho 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.4
From Jan. 1996 to Oct. 1997, 29 pigs with 40-70 days old showing dyspnea inappetite and polyserositis were collected and carried out necropsy, bacterial culture, histopathology, avidin biotin complex(ABC) stain, fluorescent antibody(FA) test, and electron microscopy. In the study, 4 strains from 3 pigs were isolated from meninges, pleura and synovial fluid and also were identified as Haemophilus parasuis serovar 5. Main histopathological lesions of 29 pigs with polyserositis were frequently composed of fibrinous peritonitis(27), pleurisy(22), interstitial pneumonia(21), fibrinous epicarditis(20), fibrinopurulent meningitis(8) and synovitis(4). By ABC stain, 11/29(37.9%) pigs with polyserositis were confirmed to have H parasuis serovar 5 in the cytoplasm of macrophages and neutrophils in cerebral meninges, epicardium, pleura surface of lung or serosa of spleen. ABC stain(20.8~40.0%) to detect H parasuis serovar 5 in tissues was more sensitive than bacterial culture(10.3%), but less sensitive than FA test(62.5%) using frozen tissues even though the result of 8 cases. By electron microscopy, a bacterium was also detected in the cytoplasm of macrophages in purulent exudate of cerebral meninges. Consequently, we confirmed that H parasuis serovar 5 has been involving to cause pigs with polyserositis and can be detected by FA and ABC stain as reliable diagnostic tools.
김재훈,황의경,배유찬,손현주,박중원,윤용덕,Kim, Jae-hoon,Hwang, Eui-kyung,Bae, You-chan,Son, Hyun-joo,Park, Jung-won,Yoon, Yong-dhuk 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.3
Both immunoelectron microscopy(IEM) and immunogold conjugate immunoelectron microscopy (IGC-IEM) techniques were developed for the detection of porcine epidemic diarrhea virus(PEDV) from the feces. Fecal samples were incubated sequentially with anti-PEDV monoclonal antibody(MoAb) and immunogold conjugated goat anti-mouse IgG+IgM. Then negatively stained, mounted on the formvar carbon-coated copper EM grids and observed by the transmission electron microscope. By the direct electron microscopy(DEM), coronavirus particles were observed from 17 cases of total 33 fecal samples of grower pigs and sows. The virons of coronavirus were moderately pleomorphic but mostly spherical, with a diameter ranged from 90 to 190nm. PED virus particles were identified from 15 cases of 17 DEM positive samples by the IEM and IGC-IEM techniques. Aggregates of PED virus coated with specific antibody were seen in fecal samples incubated with homologous anti-PED virus MoAb but not in control samples incubated with anti-TGE virus MoAb. Following incubation with immunogold-conjugated secondary antibody, the gold granules were usually distributed around and among the virus particles and soluble and viral particle-associated antigen. So, IEM and IGC-IEM techniques were proved a rapid and sensitive methods for detection and identification of PED virus from fecal and intestinal contents.