传染性法氏囊病病毒（IBDV）人工感染SPF鸡的病理学观察

取３２０只３０～４０Ｈ龄ＳＰＦ鸡，经滴鼻和点眼接种传染性法氏囊病病毒（ＩＢＤＶ），接种后２４～７２ｈ出现死亡，病死率５１．２５％，死亡鸡剖检变化主要为法氏囊肿胀、出血，甚至呈紫葡萄样，脾脏肿大、出血，骨骼肌出血，腺胃肌胃交界处出血。病理组织学变化，呈现以法氏囊淋巴滤泡内髓质淋巴细胞坏死为主的特征性病变，并可在巨噬细胞浆内发现病毒包涵体。电镜观察，在法氏囊内淋巴细胞、异染性细胞、巨噬细胞浆内，见大量晶格状排列的病毒粒子和包涵体，表明ＩＢＤＶ首先损害法氏囊淋巴滤泡髓质内未成熟的Ｂ淋巴细胞，病毒在淋巴细胞内以包涵体方式增殖。     

套式反转录—聚合酶链反应检测恙螨及鼠肺内肾综合征出血热病毒RNA

选择覆盖肾综合征出血热病毒（ＨＦＲＳＶ）膜蛋白（ＭＰ）基因的保守区核苷酸序列合成２对引物,采用异硫氰酸胍一步抽提ＲＮＡ,建立了ＲＴ－ＰＣＲ检测鼠体恙螨及游离恙螨体内ＨＦＲＳＶ－ＲＮＡ的方法,扩增产物经凝胶电泳及斑点印迹杂交证实具有特异性。结果显示,ＨＦＲＳＶ抗原阳性鼠体恙螨５０只组、１０只组、游离螨５０只组,ＨＦＲＳＶ抗原阳性鼠肺１０００ｍｇ、５００ｍｇ组经ＲＴ－ＰＣＲ检测为阳性;ＨＦＲＳＶ抗原阳性鼠体恙螨５只组,ＨＦＲＳＶ抗原阴性鼠体恙螨５０只和１０只组,游离螨１０只和５只组,鼠肺ＨＦＲＳＶ抗原阳性１００ｍｇ组均未见有明显扩增带。进一步用套式反转录－聚合酶链反应（ＮｅｓｔｅｄＲＴ－ＰＣＲ）检测,在ＲＴ－ＰＣＲ未测出ＨＦＲＳＶ－ＲＮＡ的各组中均检测有ＨＦＲＳＶ－ＲＮＡ。结果表明ＮｅｓｔｅｄＲＴ－ＰＣＲ具有高特异、高敏感的特点,可用于检测恙螨体内微量ＨＦＲＳＶ－ＲＮＡ,为确认恙螨作为ＨＦＲＳＶ的传播媒介提供了分子生物学证据。     

全混合颗粒料中添加不同类型蛋氨酸对湖羊生长性能及血清生化指标的影响

本试验旨在研究在湖羊全混合颗粒料中添加不同类型蛋氨酸(蛋氨酸钙盐与蛋氨酸羟基类似物异丙酯)对湖羊生长性能及血清生化指标、抗氧化指标和激素含量的影响.选取体重为(19.99±2.04)kg的健康湖羊72只,随机分为3组:对照组、试验Ⅰ组(在基础饲粮中添加1.5 kg/t蛋氨酸钙盐)和试验Ⅱ组(在基础饲粮中添加1.5 kg...     

羊布鲁菌16MOmp25真核表达载体的构建

利用聚合酶链式反应（PCR）,以羊布鲁菌16M基因组为模板克隆Omp25基因,设计含有EcoRI和xhoI酶切位点的引物序列,定向克隆到真核表达载体pCMV-HA上,构建真核表达重组质粒pCMV-HA～Omp25。经测序及双酶切验证正确,证明成功构建了pCMV-HA—Omp25真核表达质粒,为表达纯化外膜蛋白Omp25,研究其功能奠定基础。     

家蚕病原性微孢子虫孢子表面蛋白的选择性分离与总蛋白比较分析

选择性分离了Ｎｏｓｅｍａｂｏｍｂｙｃｉｓ孢子表面蛋白和总蛋白，采用ＳＤＳ－ＰＡＧＥ检测发现，孢子表面蛋白由分子量不同的３种亚基（３２０００、２６０００、２４５００Ｄａ）组成，其总蛋白至少包括２５个条带。同时将从养蚕生产中收集到的两种微粒子ＭＡ－１和ＭＤ－１以及从野外昆虫收集到的微孢子虫Ｐｈａ－Ｍ和Ｈａ－Ｍ与Ｎ．ｂｏｍｂｙｃｉｓ进行了比较，发现不同微孢子虫间蛋白组成有差异。     

高职兽医专业实践教学体系的构建

对高职兽医专业实践性教学体系中存在的问题及模型构建的目的进行了介绍.以期使学生的动手能力和解决生产实践中存在问题的能力以及组织生产能力得到提高,同时把加强实践教学环节作为提高教学质量的重要手段进行研究和探索,从而在强化实践教学环节中逐步建立起适合高职兽医专业的实践教学体系.     

The 16MΔvjbR as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection

Brucellosis brings great economic burdens for developing countries. Live attenuated vaccines are the most efficient means for prevention and control of animal Brucellosis. However, the difficulties of differentiating of infection from vaccine immunization, which is essential for eradication programs, limit their applications. Therefore, the development of a vaccine that could differentiate infection from immunization will overcome the limitations and get extensive application. VjbR is a quorum sensing regulator involving in Brucella's intracellular survival. The vjbR∷Tn5 mutants have been proven effective against wild type strain challenge, implying its possibility of use in vaccine candidate development. To further evaluate this candidate gene, in the present study, the antigenicity of purified recombinant VjbR protein was analyzed. Antibodies to Brucella melitensis VjbR could be detected in sera from patients and animals with brucellosis but not in control ones, implying the potential use of this protein as a diagnostic antigen. Then a vjbR mutant of B. melitensis 16M was constructed by replacing the vjbR with kanamycin gene. The mutant showed reduced survival in macrophage and mice. Vaccination of BALB/c mice with 16MΔvjbR conferred significant protective immunity against B. melitensis strain 16M challenges, being equivalent to which induced by the license vaccine Rev.1. The vjbR deletion mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon and interleukin-10. The most importance is that, the use of vjbR mutants as vaccines in association with diagnostic tests based on the VjbR antigen would allow the serological differentiation between infected and vaccinated animals. These results suggest that 16MΔvjbR is an ideal live attenuated vaccine candidate against B. melitensis and deserves further evaluation for vaccine development.     

牛结核γ-干扰素ELISA检测方法在检疫工作中的应用

为评价牛γ-干扰素ELISA检测方法检测牛结核的效果及国产试剂盒的检测效果,本试验首先将国产试剂盒与Prionics试剂盒对42份相对阳性的样品和105份相对阴性的样品进行对比研究。然后对5个规模化牛场的3000头奶牛首先进行国产单纯结核菌素颈部皮内变态反应试验,3天后选取皮内变态反应阳性和可疑及部分阴性牛共418头,进行牛γ-干扰素试验。结果国产试剂盒对阳性和阴性样品的检测敏感性和特异性分别为95.2%和100%,与Priobics试剂盒的符合率为99.3%。表明国产试剂盒与进口试剂盒的检测能力一致,牛γ-干扰素检测方法准确可靠。5个牛场的3000头奶牛单纯结核菌素颈部皮内变态反应试验阳性为138头,可疑105头。γ-干扰素试验对418头奶牛的检测,其中阳性74头,与颈部皮内变态反应（可疑牛暂时视为阴性）的符合率为60.5%。     

酸奶及乳饮料中大肠杆菌变化趋势研究

[目的]为了探讨实际生产过程中酸奶和乳饮料感染大肠杆菌（E.coli）后的变化情况及影响因素。[方法]将大肠杆菌标准菌株接种至酸奶、乳饮料及培养基中,设置不同的储藏温度、pH,定期测定储藏过程中大肠杆菌含量变化情况。[结果]随着储藏时间延长,不同储藏温度（4℃和25℃）,不同pH(4.2和3.6)的酸奶和乳饮料中大肠杆菌含量均减少,相对于储藏于4℃的产品,储藏于25℃的产品中菌含量降低更快;当产品pH为3.6时,菌含量减少速率高于pH为4.2的产品。将大肠杆菌置于不同pH的培养基中培养,结果表明,当pH<3.8时,大肠杆菌的活性将会受到抑制,证明了pH对大肠杆菌的影响。[结论]本研究为实际生产加工过程中产品冷藏及脱冷条件下大肠杆菌的检验检测和产品质量控制提供理论依据和理论指导。