6株牛传染性鼻气管炎病毒BICP4基因序列的同源性分析

牛传染性鼻气管炎病毒的立即早期启动基因IER4.2编码感染细胞蛋白BICP4基因。各种疱疹病毒的BICP4基因的Ⅰ，Ⅲ，Ⅴ区变异性较大，并为其特征区。为了比较分离自不同牛种，不同部位的5种病毒株BICP4蛋白I区基因的差异性，本文根据已发表的IBBV K22毒株重复序列IRs中BICP4的基因序列设计了一特异性引物，对各毒株进行PCR扩增，均得到约540bp的片段，将其克隆，测序并连同K22株进行同源性比较。结果表明，6株病毒的核苷酸序列及氨基酸序列同源性均在98%以上，说明这些毒株重复序列中的早期基因高度保守，在一定程度上也显示出IBRV的高度保守性。     

单克隆抗体ELISA检测犊牛粪便中冠状病毒抗原

用两株单克隆抗体混合后包被ＥＬＬＩＳＡ微孔板，加牛冠状病毒抗原，加经白陶土处理过的兔抗牛冠状病毒免血清，再加上辣根氧化物酶标记的羊抗兔ＩｇＧ抗体，加底物质显色间接ＥＬＩＳＡ检测牛冠状病毒抗原获得成功。用此方法检测细胞培养的牛冠状病毒上清液，其敏感度高出血凝试验（ＨＡ）８倍，从武汉市附近３个奶牛场采集犊牛腹泻粪便１０５份，用单抗ＥＬＩＳＡ检测牛冠状病毒抗原，共检出阳性３６份，并与血凝及血凝抑制试验，     

切割取样对胚胎发育的影响

体视显微镜下,采用徒手持金属刀片和自制的玻璃切割针分别对小鼠和牛胚胎进行切割取样,并对取样后的胚胎进行体外培养48h,其发育率分别为76.7%(46/60)和80%(16/20),两者差异不显著(P>0.05);奶牛新鲜胚胎切割取样后,胚胎移植妊娠率47.1%(8/17),比对照组妊娠率59.0%(23/39)差异显著(P<0.05)。冷冻-解冻胚胎切割取样后移植妊娠率42.9%(6/17)虽然也比对照组50%低,但是差异不显著(P>0.05)。     

奶牛乳腺炎防治的免疫学和药理学机制

奶牛乳腺炎是一种复杂的多因素疾病，给世界各国奶牛业造成了巨大的经济损失，而至今未能提出一个彻底解决的办法。本文综述了奶牛乳腺炎的致病因素、乳腺的免疫机制和乳腺炎治疗的药理学机制方面的研究进展，重点剖析了乳腺特殊的免疫体系和乳腺炎治疗的药理学特点，分析阐明了奶牛乳腺炎免疫和治疗失败的原因。     

牛腔前卵泡的简易机械分离方法

采用两种机械法分离牛腔前卵泡。方法一 (M -1) ,皮肤移植刀切割、剪碎、过滤镜下直接捡卵法。方法二 (M -2 ) ,皮质片剪碎、离心、悬浮、过滤镜下捡卵法。M -1平均每卵巢采卵数为 46 9个± 18 2个 ,M -2为 6 8 4个± 12 5个 ;处理时间M -1为 2 5 4min± 6 7min ,M -2为 46 3min± 16 8min ,两种方法采集腔前卵泡数量及处理时间均存在显著差异。平均每小时采集卵泡数分别为 99 5和 87 2个 ,M -1多于M -2。两种方法获得腔前卵泡大小分布规律也有明显差异 ,M -1采集腔前卵泡直径为 6 0～ 15 0 μm ,绝大部分是次级卵泡 ,而M -2获取腔前卵泡则偏小。总的看来 ,M -1处理时间短 ,回收效率高 ,方法简便 ,且分离卵泡直径较大 ,适于体外培养 ,是机械分离牛腔前卵泡的一种理想方法。     

奶牛乳房炎多联苗(B)的临床免疫试验

用奶牛乳房炎多联苗 (B)免疫家兔 ,14 d血清中金黄色葡萄球菌抗体效价可达 1∶ 8,2 8d抗体水平明显升高 ,可达 1∶ 32 ,注苗后 35 d攻毒 ,攻毒后 2 d保护率为 93.3% (14 /15 ) ,自然保护率为 2 0 % (1/5 ) ,10 d保护率为 6 6 .6 % (10 /15 ) ,自然保护率为 2 0 % (1/5 ) ;用多联苗 (B)臀部肌肉注射免疫泌乳牛 ,试验期间 4个月内 ,3ml剂量组临床型乳房炎月平均发病率为 11.0 9% ,其对照组为 2 0 .19% ,发病率降低 4 5 .2 9% ,差异极显著 (P<0 .0 1) ;5 m l剂量组月平均发病率为 13.0 1% ,其对照组为 31.36 % ,发病率降低 5 8.5 1% ,差异极显著 (P<0 .0 1) ;3ml剂量组与 5 m l剂量组之间发病率无明显差异 (P>0 .0 5 ) ;注苗后泌乳牛血清抗体水平可持续 4个月 ,且 30 d时效价水平最高。     

Comparative studies of strains of infectious bovine rhinotracheitis virus isolated from latently infected calves

Three strains (479 C, 778 TL, 982 LE) of infectious bovine rhinotracheitis (IBR) virus isolated from latently infected calves were compared with the prototype strain of IBR virus (LA strain) in studies which included restriction endonuclease analysis, experimental infection, and reciprocal cross protection tests in cattle. From the restriction endonuclease analysis it appeared that the 3 "latent" viruses were derived from the same isolate, and that it differed slightly from the LA strain. However, latency does not seem to have affected the pathogenicity or the immunogenicity of the virus. This is demonstrated by the identical clinical and virologic response of calves subjected to experimental infection with the various strains under study, and by the finding that when the LA strain and a "latent" strain (982 LE) were tested in cross protection tests in cattle, they proved to be mutually protective.     

Inflammation-related changes in cyclic AMP and cyclic GMP in bovine mastitis

Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) cocentrations in milk and plasma samples from healthy and mastitic cows were determined by radioi-immunoassay and compared with prostaglandins (PGE2, PGF2 and thromboxane B2 [TXB2]), phospholipids and other relevant parameters in milk and blood. The concentrations of cAMP were about five times higher in plasma (p<0.01) than in milk, whereas the cGMP concentration in milk was three times higher (p<0.01) than that in plasma in both healthy and diseased animals. In mastitic milk, the cAMP and cGMP concentrations were 19% and 65% and in blood plasma 13% and 84% higher respectively than in healthy animals. In milk, cyclic nucleotide concentrations correlated with the markedly elevated cell count and also with the prostaglandin concentration and pH. In blood, cAMP correlated positively with phospholipids and cGMP with reduced glutathione (GSH). These changes are considered to be important in the disease process and, in particular, the increase in cGMP deserves further study.     

Reduced protein kinase C activity and endogenous protein phosphorylation in ethionine-induced fatty liver in cows

Protein kinase C (PKC) activity was evaluated and the phosphorylation of its endogenous substrates was explored in fatty liver induced by administration of ethionine (an analogue of methionine) to cows in order to assess the relevance of PKC-dependent phosphorylation in the development of fatty liver. PKC activity was decreased in both the cytosolic and the total particulate fractions from fatty livers, compared to the corresponding fractions from control liver. The mode of activation by the PKC cofactors (1-oleoyl-2-acetyl-sn-glycerol, 12-O-tetradecanoylphorbol-13-acetate, phosphatidylserine and Ca²⁺) was similar in both control and fatty livers, suggesting a quantitative but not a qualitative change in PKC in fatty liver. At least three substrate proteins (34 kDa, 26 kDa and 19 kDa) were found in the cytosolic fraction and their phosphorylation was reduced in fatty liver. These results suggest that impairment of the signal transduction pathway mediated by PKC is involved in the pathogenesis of fatty liver in cows.Abbreviations ATP adenosine triphosphate - EGTA ethylene glycol bis(-aminoethylether)-N,N,N,N-tetraacetic acid - NEFA non-esterified fatty acid - OAG 1-oleoyl-2-acetyl-sn-glycerol - PKC protein kinase C - PS phosphatidylserine - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - TG triglyceride - TPA 12-O-tetradecanoylphorbol-13-acetate