马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建     

范斌  陈卓  刘建华  鲍子磊  胡月  贾钦瑞  王雪竹  冉多良 《中国畜牧兽医》2018,45(10):2681-2690

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。  相似文献   

MAGNETIC RESONANCE, ULTRASOUND AND HISTOPATHOLOGIC CORRELATION OF ACUTE AND HEALING EQUINE TENDON INJURIES     

Jeffrey R.  Crass  MD  Ronald L.  Genovese  VMD  James A.  Render  DVM  Errol M.  Bellon  MD 《Veterinary radiology & ultrasound》1992,33(4):206-216

Ultrasonography and MRI have become valuable tools for imaging of tendon injuries. The current study examines the histopathologic basis for the imaging abnormalities. Five injured equine forelimbs and two normal contralateral limbs were studied with high resolution real time ultrasound and MRI. Histologic sections were made and correlated with the diagnostic images. All lesions were readily seen by both modalities. Lesions characterized by hemorrhage, edma, and cellular infiltration were sonolucent on ultrasound and bright on MRI images. MRI returned to normal as fibrogenesis ensued. Ultrasound images remained abnormal until fibrillar reallgnment occured with completion of the healing process. High resolution real time ultrasound and MRI both accurately reflect the tissue abnormalities in acute tendon injuries. The injured horse does appear to be an effective model for correlative imaging studies of tendon injuries.  相似文献   

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   总被引：3，自引：0，他引：3  

薛飞  贾斌  朱远茂  杨建德  王晓钧  相文华  赵立平  吕晓玲  沈荣显 《中国预防兽医学报》2003,25(5):321-325

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。  相似文献   

马鹿假台鹿采精技术的研究     

武春田  国泰  赵裕方  赵列平  张贵学 《黑龙江动物繁殖》1994,(3)

对６头试验公鹿用假台鹿采精，采精前对特制的假阴道以药用凡士林为滑润剂，注水充气，温度为４０℃±１℃，以适宜的压力和滑润度，将其安装在人工制做的假台鹿内。拨赶公鹿于采精场诱其爬跨。结果，试验的６头公鹿中有４头能够采精，共２２头次，射精量平均每头次１．５４ｍｌ，精子密度为１８．６—３７亿／ｍｌ，精子活力在０．８以上。９３年每头次生产细管冻精１０６．２５支（４２５支／４头次），是同年电刺激采精法平均６４．４支／头次（９０２支／１４头次）的１．６５倍，此法既能采到质量高的精液，又比电采法省工、省时、省力、安全，是一种理想的马鹿采精方法。  相似文献   

Insemination with Stallion Semen Frozen in 0,5 ml Straws     

R. Thomassen 《Reproduction in domestic animals》1993,28(4):289-293

Contents: An insemination trial using frozen semen is described. The freezing procedure was slightly modified from the Hannover method. The insemination dose consisted of 7 medium straws containing approx 1 10⁹ spermatozoa. A total of 28 mares of the Norwegian Trotter breed were inseminated during the 1991 season. During oestrus the mares were examined at 12 hour intervals, and the insemination was carried out after detection of ovulation. The pregnancy rate was 43% after the first insemination, increased to 68% after second and further to 75% after the third and last insemination. The foaling rate was 61.5%.  相似文献   

MAGNETIC RESONANCE IMAGING FEATURES OF EQUINE NIGROPALLIDAL ENCEPHALOMALACIA     

Sean G.  Sanders  DVM  Russell L.  Tucker  DVM  Rodney S.  Bagley  DVM  Patrick R.  Gavin  DVM  PhD 《Veterinary radiology & ultrasound》2001,42(4):291-296

Magnetic resonance imaging (MR) was used to make a diagnosis of equine nigropallidal encephalomalacia in a horse. Equine nigropallidal encephalomalacia is a neurodegenerative disease that has many characteristics with Parkinson-like diseases in humans. Historically, horses were euthanized based on clinical signs and exposure to the toxic weed, yellow star thistle (Centaurea solstitialis). Previously, the disease has only been confirmed on necropsy. MR imaging can provide accurate and sensitive visualization of typical lesions seen in the brain of horses affected with equine nigropallidal encephalomalacia. Lesions were seen on T1-weighted, T2-weighted and proton density images. There was no contrast enhancement following Gd-DTPA administration. Lesions seen on MR were confirmed at necropsy. Using MR to confirm a diagnosis of equine nigropallidal encephalomalacia will prevent unnecessary suffering of horses and expense to owners that would otherwise incur, while further diagnostics are performed.  相似文献   

Deep Freezing of Boar Semen Packaged in Plastic Bags and Straws     

C.O. Bwanga  S. Einarsson  H. Rodriguez-Martinez 《Reproduction in domestic animals》1991,26(3):117-125

For practical reasons, a large volume (i.e. 5 ml) of frozen boar semen per insemination dose is desirable, but successful freezing has not been achieved, since optimal cooling rates have not yet been established. Post-thaw motility and the acrosome intep'ty of semen from four boars frozen with a programmable freezin machine, in mini-(0.25 ml), maxi-(5 ml) plastic straws and in 10 × 5 cm PVC- or Teflon FEP-plastic bags (0.35 – 0.12 mm thick, 5 ml) was studied. The freezing of the semen was monitored using thermocouples placed in the straws and the bags. The freezing curve started from +5°C, at a rate of −3°C/min, to – 6°C, it was held for 1 min at −6°C, and was followed by further drop to −100°C at a rate of −20°C/min, with subsequent storage in LN₂. The bags had a much shorter freezing point plnteau, compared to the maxi-straws. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. The freezing procedure did not cause major acrosomal damages, significantly more normal apical ridges being present in the bags and mini-straws than in the maxi-straws. This in vitro evaluation indicates that the freezing method employed is satisfactory for freezing large volumes of boar semen into plastic bags .  相似文献   

Artificial Insemination with Frozen Semen in Ewes at Different Times of the Breeding Season     

K. Andersen Berg  J. Aamdal 《Reproduction in domestic animals》1991,26(1):27-30

Totally 13575 ewes of two different breeds, Dala and Spel, were inseminated with semen, frozen in straws and thawed at 70°C for 8 sec. An insemination dose of 0.2 ml containing approx. 150 × 10⁶ spermatozoa with at least 45 to 50% progressive motility was imerted 5 to 12 mm into the cervix. The insemination was performed once between 12 and 30 h after the onset of heat. The NR rate of the Dala ewes increased significantly during the season. The NR rate of the ewes inseminated before 15. November was 44.3%, from 15. to 20. November 52.2%. from 20. to 25. November 55.3% and from 25. November and later 61.4%. The corresponding values for the ewes of the Spel breed were 57.3, 58.7, 61.5 and 71.0% respectively, and only the difference between the two last values was statistically significant. The difference between the fertility of the two breeds was significant within each of the periods .  相似文献   

白枕鹤精液冷冻技术的研究   总被引：1，自引：0，他引：1  

张玲．刘大军  宋艳珠  李选芳 《黑龙江动物繁殖》1994,(1)

白枕鹤（ＧＲＵＳＶ　ＩＰＩＯ）羽毛青灰，红面，体型健美优雅，具有很高的观赏价值，已被国家列为二级保护动物。对白枕鹤类的人工授精、孵化、育雏以及冷冻精液技术的研究，可促进人工饲养条件下鹤类的增殖和种群的繁衍。本研究采用Ｂｅｌｔｓｖｉｌｌｅ稀释液做为基础液，４—６％ＤＭＳＯ做为冷冻保护剂，按常规冷冻法冻精配种受胎率达４０％，此项研究填补了我国在鹤类精液冷冻技术方面的空白。  相似文献   

影响精液冷冻和人工授精效果的因素     

吴结革  茆达干 《动物医学进展》2003,24(5):48-50

通过对精液冷冻保存的细胞反应原理的阐述 ,指出冷冻保护剂甘油对精液保存具有利弊效应 ,提出精子膜脂组成的差异使得不同品种的精子对冷冻损伤的易感性不同。雌性生殖道解剖结构的品种差异 ,精子形态 ,精子运行机制的细微差异 ,人工授精时间及精子的运行能力 ,采精方式等因素对精液冷冻保存和人工授精的成功有决定性作用。研究精子质膜的生物学特性可解决低活力精子的问题 ,然而这并不能解决冷冻后精子质量的个体差异。对精细胞基因组的研究可以找出这些个体的遗传差异。因此 ,冷冻精子和精原细胞 (用于细胞外注射 )的差异已经成为完整基因组问题。  相似文献