马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

关中驴对6株动物病毒抗原的免疫应答试验

以关中驴为试验动物 ,用猪瘟病毒(HCV )、犬细小病毒 (CPV )、兔瘟病毒(RHDV)、鸡新城疫病毒 (NDV)、伪狂犬病毒(PRV)等病毒进行免疫 ,检测所产生的抗体效价 ,并与猪、鸡、兔、犬、及猫等动物进行比较。试验结果表明 ,关中驴与对照动物产生的抗体效价一致 ,证明了关中驴可替代其它动物用于生物制品研究     

Assessment of the accuracy of testicular dysfunction detection in male donkey (Equus asinus) with the aid of colour-spectral Doppler in relation to plasma testosterone and serum nitric oxide levels

This study aimed to determine the usefulness of colour and pulsed Doppler modes for the accurate diagnosis of donkeys suffering from subfertility to determine whether testicular vascularity assessment could be an indicator for sperm functionality. The study sample was composed of 10 male donkeys with normospermia (control group) and 10 donkeys with hypospermia. Animals underwent scrotal circumference measurement, testicular Doppler examination, seminal evaluation, blood sampling and hormonal assay. Semen volume and concentration were significantly (p ≤ .05) lower in the subfertile group (30.25 ± 1.22 ml and 89.44 ± 2.55 × 10⁶/ml) as compared with the control group (82.76 ± 1.65 ml and 452.78 ± 1.25 × 10⁶/ml), and total sperm/ejaculation was significantly (p ≤ .05) higher in the normal donkeys (28.30 ± 2.32 × 10⁹/total ejaculated) as compared with the subfertile group. Intratesticular coloured area showed a marked decline in the hypospermic males. There was no significant difference between the two groups in testosterone level, although the normal group showed an increase in nitric oxide metabolites. Both Doppler indices of the three branches of the testicular artery were elevated significantly (p ≤ .05) in abnormal donkeys, whereas Doppler peak systolic and end-diastolic velocities were increased in the normal group. Male donkeys with subfertility demonstrated lower arterial vascularity parameters in the form of intratesticular coloured area and blood flow rate; therefore, the most optimal parameters for differentiating subfertile hypospermic from normospermic donkeys were found to be the two Doppler indices, velocities parameters, testicular blood flow rate and nitric oxide levels.     

两种运输车对驴运输应激的影响

本试验旨在研究改造运输车对缓解驴运输应激的效果。选取28头健康德州公驴,随机分为2组,从北京密云县运输至聊城市东阿县东阿黑毛驴研究所,路程约500 km,历时18 h。试验使用9.6 m长单层货车为运输车,对试验组(T)运输车进行改造,即加装顶层和双侧篷布、底部加装草帘和中部加装隔栏,对照组(C)不作处理。运输前后测定驴血液生化和激素指标,统计体重变化和发病驴数量。结果表明,与运输前相比,普通车组驴在落地当天检测到血清皮质醇(COR)、白蛋白(ALB)水平均显著升高(P<0.05),总蛋白(TP)、谷草转氨酶(AST)、热休克蛋白-90(HSP90)和肌酸激酶(CK)指标极显著升高(P<0.01),钙(Ca)浓度显著降低(P<0.05)。改造车组驴血液指标TP、AST和CK的变化范围低于普通车组。两组驴落地后血糖(GLU)水平均比运输前升高,但差异均不显著(P<0.05)。与运输前相比,改造车组驴落地后的体重减少低于普通车组;改造车组驴在落地后第10天平均体重接近运输前,而普通车组驴的平均体重在落地后第20天才基本恢复至运输前水平,即使用改造运输车辆可以尽早恢复体重。普通车组驴发病率为21.4%,改造车组为7.1%;两组均无死亡。综合上述结果,改造运输车可有效降低驴的运输应激,有利于提高动物运输福利,减少经济损失;本试验结果可为动物运输应激提供参考。     

德州驴朊病毒基因克隆与序列分析

根据已报道正常哺乳动物朊蛋白(PrP)基因序列设计引物,采用PCR法扩增了德州驴的PrP基因,将其克隆到T-Vector。序列测定及分析表明所克隆的德州驴PrP基因片段为767 bp,该基因内无内含子,包含了驴PRNP完整编码区序列,编码256个氨基酸的前体蛋白,推测其分子量约34 ku。通过对德州驴朊病毒核苷酸和氨基酸序列与其他7种动物的比较,我们发现有3个区域变异较大,分别与种属屏障和潜伏期有关。经推测驴的PrP基因型是ARQ型,对羊痒病中度敏感,但是至今没有马属动物感染羊痒病的报道。这为TSE种间传播的突破提供了基础数据。     

广灵驴分子系谱的建立及群体遗传结构分析

旨在对广灵县优种驴场保种群体进行调查的基础上构建分子系谱,并对其种群的遗传结构进行分析。本研究采集保种群成年、体况良好的广灵驴（体重350~400 kg）颈静脉血10 mL（n=107）,其中公驴13份,母驴94份,抗凝处理后提取全血DNA。采用12个微卫星标记进行荧光PCR扩增后,用ABI3730测序仪进行分型。分型结果采用Cervus 2.0和Pedigraph 2.4软件构建分子系谱,同时采用STRUCTURE2.3和Fstat软件计算群体遗传参数,采用R语言的hclust函数绘制7头公驴及其后代的系统发生NJ树（邻接树）。结果,对107头种驴进行了分子系谱构建,找到了30头子代的父亲和7头子代的母亲,系谱可靠性>90%;微卫星标记的平均观测杂合度（H_Obs）和多态信息含量（PIC）分别为0.676 5和0.593 9,标记遗传多样性较高;NJ树对7个公驴家系进行了聚类;群体分化系数（F_ST）为0.184,群体平均近交系数（F_IT）为0.033,群体内近交系数（F_IS）为-0.238,且群体处于哈代-温伯格平衡状态,存在很弱的近交。本研究建立了广灵驴保种群可靠性较高的分子系谱并对其遗传结构进行了分析,证明该群体遗传多态性较高,群体近交系数较低,处于较好的保种状态,具有较大的品种资源开发潜力。     

Ultrasound-guided retrobulbar nerve block in donkey cadavers

ObjectiveTo develop and describe an acoustic window to access and inject the retrobulbar region in donkey cadavers.Study designProspective cadaveric experimental study.AnimalsA total of 14 donkey cadaver heads.MethodsUltrasound landmarks for retrobulbar injection were identified by dissection of two cadavers. Ultrasound-guided retrobulbar injection of 3, 6 and 9 mL of contrast–lidocaine mixture (1:1) were evaluated using computed tomography (CT) in 12 cadavers for needle position and spread of injectate into the extraocular muscle cone.ResultsReal-time visualization of needle, extraocular muscle cone and optic nerve enabled successful intraconal instillation of injectate, as documented on CT images.Conclusions and clinical relevanceUltrasound-guided retrobulbar injections in cadaver donkeys provided adequate infiltration of the intraconal space.