E.stiedai感染兔免疫器官内球虫抗原的免疫组化定位

用免疫组化SABC法,对斯氏艾美耳球虫感染后兔肝脏、脾脏和肝门淋巴结中球虫抗原的分布进行了观察。结果表明,在肝脏窦状隙、胆管上皮细胞、肝门淋巴结和脾脏内可见到明显的呈棕黄色阳性反应区,表明在这些区域和部位有相应的E.stiedai孢子囊、裂殖子或子孢子抗原存在,且抗原数量以肝脏内最多,其次为脾脏,最少者为肝门淋巴结;显微镜下还可见到大量坏死的肝细胞、淋巴细胞以及朗罕氏巨细胞,说明多核巨细胞也参与了对球虫的吞噬和处理过程。     

斯氏艾美耳球虫子孢子的移行途径

关于斯氏艾美耳球虫Ｅｉｍｅｒｉａ ｓｔｉｄｅａｉ子孢子的移行途径争议尚大。作者借光镜和电镜互补的手段研究了子孢子从肠腔到肝内胆管上皮细胞的移行。所取材料为人工感染５－５０万卵囊的１月龄幼兔的血液，肠道，淋巴结，脾和肝，取材料时间为感染后２－７２ｈ至４－６ｄ。依据子孢子大量出现于肠系膜淋巴结和小量出现于脾脏而未见于其它部位淋巴结，以及肝内胆管周围营养血管中有大量子孢子出现等观察结果，提出了子孢子移行     

猪繁殖-呼吸综合征活疫苗对仔猪的安全性试验

本试验用猪繁殖-呼吸综合征（PRRS）活疫苗和国内分离的PRRS强毒CH—1a株接种PRRS阴性的断奶仔猪，分别在接种后的3、7、14d各剖杀1头，取各脏器分别做冰冻切片和病理切片观察。用间接免疫荧光法检测各脏器PRRS病毒的分布。结果表明，PRRS活疫苗在免疫初期，抗原主要分布在脾脏、淋巴结，其次是肾脏和肺脏，少见于肝脏和心脏，第14d时在脾、淋巴结和肾脏有一定量的抗原，而肺脏相比则数量很少，肝脏和心脏未检到PRRS病毒抗原的存在，表明接种PRRS活疫苗随着时间的推移抗原分布呈下降趋势。而强毒抗原分布以脾脏最多，依次是肾脏、肺脏、淋巴结、肝脏、心脏，接种后第14d仍能在各脏器检到PRRS病毒抗原。病理组织学检测结果表明，活疫苗产生以下颌淋巴结、脾脏增生为特征的免疫应答，组织损伤轻微，对肺的病变较少，且仔猪生长良好。强毒则引起以大面积的肺泡隔增宽为特点的间质性肺炎和微循环障碍的病理变化，淋巴小结、脾脏滤泡发生崩解与周围界限不清，个别淋巴细胞核浓缩，组织损伤严重。本试验表明弱毒疫苗对仔猪是安全的。     

家养野猪自然感染猪圆环病毒2型的病理学观察

经PCR检测南京某养殖场送检的表现为多系统衰竭综合征的家养野猪病原为猪圆环病毒2型(PCV2)。采用常规石蜡切片法和免疫组织化学法对患病仔猪的病理组织学变化特点及PCV2抗原在淋巴结内的分布进行了研究。结果显示,剖检病死仔猪病变主要表现为全身淋巴结肿大和间质性肺炎。组织病理学变化主要表现为淋巴结、脾和扁桃体等免疫器官淋巴滤泡萎缩,淋巴细胞数量减少,有大量巨噬细胞浸润及多核巨细胞;非淋巴组织病变主要为肺脏呈典型的间质性肺炎,肝脏局灶性坏死及淋巴细胞浸润;肺脏和肝脏病灶内也可见少量多核巨细胞。PCV2主要分布于淋巴组织内单核巨噬细胞、支气管黏膜上皮、肺泡壁Ⅱ型上皮细胞内,肾小球内也可见少量分布。     

猪弓浆虫病的诊治与虫体分离

江西某农场的猪种猪和肥猪几乎几时暴发以高热为特征的疫病，病理变化主要为淋巴结出血、肿大脾脏、肝脏肿大，肺炎、心脏出血。肺门淋巴结触片多见量弓浆虫滋养体，病猪血清的弓浆虫抗原，抗体均阳性。接种病料的小鼠于接种后２５天死亡，分离出一株弓浆虫，因此诊断猪场种场和肥猪暴发的疫病与弓浆虫病，并推测本次弓浆虫病的病原来自饲料中的鱼粉。     

猪弓形虫自然感染病例病理组织学观察

为探究猪弓形虫感染对猪器官组织学结构的影响,对信阳某猪场自然发病猪进行病理学诊断。处死病猪后采取肝脏组织用福尔马林固定,姬姆萨染色法进行染色。采集肺、肝、肾、淋巴结和心等组织,石蜡包埋,HE染色,光学显微镜下观察。在用姬姆萨染色的肝脏切片上能够观察到弓形虫的速殖子,分别在肺、肝、肾、脾脏、淋巴结和心脏等组织器官切片中发现弓形虫滋养体或假囊,表明该场疾病是由猪弓形虫引起的,各组织器官出现出血、炎性细胞浸润、嗜酸性粒细胞和中性粒细胞增多等病理变化。     

用特殊染色对兔肝球虫的不同形态及肝脏病变的观察

为了研究兔肝球虫的形态,并观察其引起的肝脏病变,试验对检出的患球虫病兔的肝脏进行了眼观检查和分类,并采集不同类型的肝球虫病变做石蜡切片,用普通染色和特殊染色处理后进行病理组织学观察。结果表明:眼观检查可将肝球虫的病变分为肿大型肝病变和局部萎缩(硬化)型肝病变两类,病理组织学检查可将肝病变分为萎缩性肝球虫病变和局部增生性肝球虫病变两类。组织切片通过特殊染色,不仅能鉴别不同的肝脏病变,而且可区分不同发育阶段的肝球虫。说明可以通过对肝球虫所致肝病变的观察和特殊染色对肝球虫不同发育阶段进行鉴别。     

不同品种地方鸡感染柔嫩艾美耳球虫后组织总超氧化物歧化酶活性的比较

选择30日龄无球虫感染的鹿苑鸡与油鸡各60只、大骨鸡与固始鸡各50只,分别随机分成感染组和对照组,感染组每只鸡经嗉囊接种柔嫩艾美耳球虫孢子化卵囊1×105个.感染后第2、4、6、8、12天分别扑杀感染组和对照组,采集心脏、肝脏、脾脏、肾脏和盲肠,测定组织中总超氧化物歧化酶(T-SOD)活性.结果显示,与对照组相比,T-SOD的活性在4个品种感染鸡的脾脏、肝脏、心脏和盲肠组织中下降,而在肾脏组织中增高;具体为,在鹿苑鸡的脾脏和大骨鸡的肝脏与心脏先增高后下降,而在鹿苑鸡的肝脏与盲肠、油鸡的肾脏和固始鸡的肓肠中未发生显著变化.研究表明,球虫感染引起的T-SOD活性变化在不同品种鸡和同一品种鸡的不同组织中是不一致的;4个品种鸡组织中T-SOD活性变化与其对柔嫩艾美耳球虫的敏感性程度之间关系不十分明晰,但球虫感染引起油鸡的盲肠组织内T-SOD活性持续下降.     

柔嫩艾美尔球虫裂殖子免疫原性的研究

本文评价了柔嫩艾美尔球虫第二代裂殖子的免疫原性。试验采用弗氏完全佐剂乳化的裂殖子抗原及从组织中分离得的活游离殖子，以不同剂量的不同时间接种健康小鸡，随后用同源球虫进行攻击。效果评价指标为：（１）比较各抗原组的鸡只死亡率及盲肠病变记分；（２）各抗原组鸡只的增重差异。结果表明，柔嫩艾美尔球虫第二代裂殖子抗原不能刺激鸡体产生保护性免疫力；具有活力的裂殖子接种于健康鸡盲肠能诱发部分免疫力，但较自然感染者为     

抗堆型艾美耳球虫子孢子单抗对阶段特异性抗原的识别

为了用针对堆型艾美耳球虫子孢子的5株单抗检测各代裂殖子的反应性,分别用甲醇、丙酮、戌二醛和自然干燥等4种固定方法处理裂殖子,观察其对染色结果的影响;并运用免疫荧光技术（IFA）,对单抗和裂殖子进行染色。结果显示,5株单抗中,单抗Easp-3H6和单抗Easp-5G10识别堆型艾美耳球虫裂殖子的抗原位于裂殖子的顶端,单抗5B4对各阶段裂殖子有较弱的反应,而单抗Easp-3G3和5G7不与堆型艾美耳球虫任何一代的裂殖子发生反应,表明鸡堆型艾美耳球虫的侵入阶段存在共有抗原,即种内各发育阶段的抗原。     

Trail antigen in Eimeria stiedai sporozoites associated with a thrombospondin-related motif and the entry of cultured cells

In order to examine the antigenic similarity and specificity of the trail antigen of Eimeria stiedai and Etp 100, a microneme protein of Eimeria tenella, monoclonal antibodies to the trail antigen of E. stiedai sporozoites were selected by an indirect immunofluorescent antibody method. The monoclonal antibody of one clone, 3D10, reacted with the anterior portion of non-fixed sporozoites. By immunoblotting, the monoclonal antibody was found to react with a 100 kDa antigen of E. stiedai sporozoites, and a 117 kDa antigen of E. tenella sporozoites and merozoites. It was also found to react with a recombinant protein with thrombospondin-/properdin-like motifs homologous to E. tenella microneme protein Etp 100. The monoclonal antibody significantly inhibited the penetration of E. stiedai sporozoites into cultured rabbit hepatobiliary epithelial cells. These results suggest that E. stiedai sporozoites have a trail antigen, located in the anterior region on the outer surface of the sporozoites, which has an epitope with thrombospondin-/properdin-like motifs similar to E. tenella microneme protein Etp 100. This protein may play an important functional role in the process of penetration of host cells.     

Detection and Localization of Aleutian Disease Virus and its Antigens in vivo by Immunoferritin Technique

Tissues from mink infected with aleutian disease virus were examined by the electron microscope for the presence of virus particles. Virus-like particles, measuring 22 nm in diameter, were observed in macrophages of spleen, mesenteric lymph node and in Kupffer cells in liver of mink ten to 13 days after infection. The virus-like particles were usually present in vacuoles inside the cytoplasm of macrophages and Kupffer cells and, occasionally, similar particles were observed inside the nucleus. Cells from uninfected mink did not contain such patricles. To correlate the existence of these virus-like particles with the presence of aleutian disease virus antigen in infected cells, tissues were processed for immunoferritin technique. It was found that aleutian disease virus antigen was present in vacuoles inside the cytoplasm of cells from the infected spleen, lymph node and liver, and that the location was similar to that of the 22 nm virus-like particles. In addition, some viral antigen was also detected as cytoplasmic granular material. The nuclei of some cells also contained aleutian disease virus antigen. The pattern of aleutian disease virus antigen was similar to the distribution of virus-like particles in cells of infected tissue. It is suggested that virus replication occurs inside the nucleus with subsequent accumulation of virus in the vacuoles of the cytoplasm.     

牛病毒性腹泻病毒在绵羊垂直感染中的抗原分布

将新疆分离的病毒性腹泻病毒（BVDV）野毒株SHN－98和标准毒株Oregon C-24分别接种无BVDV感染的羔羊和怀孕100天左右的母羊，建立BVDV在绵羊垂直感染的动物模型，应用病毒抗原定位检测的免疫组化染色技术，探求BVDV在实验性感染绵羊经胎盘感染胚胎、羔羊过程中，病毒在感染组织、靶器官、靶细胞中的分布规律。结果表明：BVDV通过母羊的胎盘感染胚胎。BVDV在感染妊娠母羊体内主要分布于心肌、肝、肺、脾、淋巴结、胃肠粘膜、脑神经、子宫粘膜、胎盘等器官组织，其中以胃肠粘膜、脾、淋巴结、脑神经、子宫粘膜、胎盘等器官组织中分布量较多；BVDV在垂直感染胚胎体内主要分布于胸腺、淋巴结、脑、肝、肾、心肌、脾、肺、胃肠粘膜、脐带等器官组织，其中以胸腺、胃肠粘膜、脑神经等器官组织中分布量较多；BVDV在垂直感染羔羊体内主要分布于心、肾、胃肠粘膜、脾、胸腺、淋巴结、大脑、小脑、海马、视丘、视神经、眼球脉络膜等器官组织，其中以胃肠粘膜、大小脑、用、视神经、淋巴结等器官组织中分布较多，在组织分布中，BVDV对上皮细胞、神经胶质细胞、淋巴细胞有较强的亲嗜性，SHN－98和OregonC-24在垂直传播中组织器官的分布无明显差异。     

免疫组化法检测布氏杆菌在人工感染牛体内的分布

将布氏杆菌分不同剂量皮下注射接种于健康牛,于接种后45 d剖杀,采集相关的器官组织制备组织切片,应用免疫组化方法检查细菌在牛体内的分布。结果显示,肝脏、脾脏、肾脏、淋巴结出现强阳性信号;在高剂量注射的牛体内,肺脏、颌下淋巴结出现阳性细胞;心脏和健康牛组织呈阴性反应。阳性信号主要位于感染细胞的胞质中,偶尔出现在细胞核内。     

Porcine postweaning multisystemic wasting syndrome in Korean pig: detection of porcine circovirus 2 infection by immunohistochemistry and polymerase chain reaction.

This report describes the first diagnosis of porcine circovirus (PCV) infection in weaned pigs with postweaning multisystemic wasting syndrome in Korea by immunohistochemistry and polymerase chain reaction. The most unique lesions were multifocal granulomatous inflammation affecting lymph nodes, liver, and spleen, characterized by infiltrates of epithelioid macrophages and multinucleated giant cells. Circoviral antigen was detected in formalin-fixed sections and was usually present in large, round, dendritic cells in the white pulp of spleen and remnants of follicles in lymph nodes. Lymphoid follicles in the tonsils also contained PCV antigen. A 530-bp DNA fragment of circovirus was successfully amplified from all tested lymph nodes, liver, and spleen.     

Expression of tissue factor in experimental bovine pneumonic pasteurellosis and endotoxemia

Tissue factor (TF), a cell surface-associated cofactor and activator of coagulation factor VII, has been implicated in the local and systemic activation of coagulation associated with sepsis. This study describes the pattern of TF expression in experimental bovine pneumonic pasteurellosis and endotoxemia. Immunohistochemical techniques were used to localize TF antigen in tissue sections. Tissue factor expression was not observed in tissues from control animals. In response to Pasteurella haemolytica challenge, TF was expressed within alveolar walls, by mononuclear inflammatory cells within alveoli, and in walls of arteries, arterioles, bronchi, and bronchioles. Tissue factor was not detected in unaffected lung, liver, spleen, lymph node or kidney tissue. Administration of Escherichia coli endotoxin intravenously resulted in tissue factor expression in lung, spleen, and lymph node tissue. Results of this study indicate that TF is expressed locally at sites of inflammation and systemically in endotoxemia. Therefore, TF may be involved in coagulation events associated with pneumonic pasteurellosis.     

Persorption of luminal antigenic molecule and its specific antibody via apoptotic epithelial cells of intestinal villi and Peyer's patches into peripheral blood in rats

The possibility of persorption of bovine serum albumin (BSA) molecules from mucous epithelial cells and its mechanism were investigated in rats orally pre-immunized by BSA for 14 consecutive days. In the small and large intestines, both the BSA antigen (BSA-Ag) and its specific antibody (SpAb) were absorbed by the epithelial cells at the late apoptotic stage (ApoEp), and were subsequently transcytosed by membranes of the small vesicles. The basal cytoplasms containing highly-concentrated BSA-Ag and SpAb were occasionally fragmented into small cytoplasmic droplets that were secreted into the lamina propria. In Peyer's patches, both BSA-Ag and SpAb were more actively absorbed and transcytosed toward the dome area by the ApoEp of the dome apex than by the M cells. BSA-Ag and SpAb were finally persorbed into the portal blood and lymph, but were never secreted into the bile. They were also engulfed by macrophage-like cells in the villous lamina propria, mesenteric lymph node and spleen, and by hepatocytes in the liver. These findings suggest that sensitized soluble luminal antigens are taken up by ApoEp in the small intestine and are finally persorbed into the peripheral blood. The uptake of luminal antigen might be mediated by its luminal SpAb.     

Comparative studies on the distribution and population of immunocompetent cells in bovine hemal node, lymph node and spleen

The distribution and population of immunocompetent cells in bovine hemal node, mesenteric lymph node and spleen were analyzed comparatively by immunohistochemistry and flow cytometry. Many CD8(+) cells, CD172a(+) cells and γδ T cells were found in the lymphatic cord along the sinus of the hemal node and the splenic red pulp. A few CD8(+) cells and γδ T cells were distributed diffusely in the paracortex and medullary cord of the mesenteric lymph node. Many germinal centers were recognized in the lymphatic regions such as the cortex and white pulp of these lymphoid organs. The populations of CD8(+) cells and γδ T cells in the hemal node and the spleen were higher than those of the mesenteric lymph node. In addition, the populations of CD21(+) cells and MHC class II(+) cells in the hemal node and the mesenteric lymph node were higher than those of the spleen. The results suggest that the hemal node has an important role in both cellular and humoral immunity as well as the lymph node and the spleen in cattle.     

Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax-embedded tissues

This study evaluates the cross-reactivity of seven anti-human and one anti-bovine antibodies in formalin-fixed, paraffin-embedded tissue samples of liver and mesenteric lymph nodes of 13 striped dolphins (Stenella coeruleoalba). Four antibodies (CD3, IgG, lysozyme and S100 protein) reacted with striped dolphin lymph nodes in a similar pattern to that observed in the species of origin. The anti-human MHC class II mAb reacted strongly with macrophages and dendritic-like cells of striped dolphins, whereas a small number of lymphocytes were labelled with this antibody. These antibodies were used to study the immunophenotype of the inflammatory infiltrated in non-specific chronic reactive hepatitis (eight cases) and chronic parasite cholangitis (two cases) and normal liver (three cases) of striped dolphins. Non-specific chronic reactive hepatitis was composed of inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells in portal spaces and hepatic sinusoids. Lymphonodular aggregates observed in chronic parasitic cholangitis showed a cellular distribution similar to that found in lymph node cortex, including the presence of S100+ and MHC class II+ dendritic-like cells in lymphoid follicles and interfollicular areas. This result suggests that those inflammatory infiltrates are highly organised to enhance antigen presentation to B and T cells.