甘肃地区牛源金黄色葡萄球菌分子鉴定及RAPD分型

本研究目的是分离鉴定引起甘肃地区奶牛乳房炎的金黄色葡萄球菌,掌握其基因型情况。利用16S、23SrRNA保守序列PCR扩增对乳房炎奶样中的金黄色葡萄球菌进行鉴定,并进行RAPD基因分型。结果表明,310份奶样中共分离出金黄色葡萄球菌100株,RAPD结果显示这100株金黄色葡萄球菌均可得到清晰的RAPD指纹图谱,扩增产物在2～7条带之间,具有多种带型组成。通过聚类分析100株菌产生11个基因型,其中Ⅰ型4株,Ⅱ型4株,Ⅲ型10株,Ⅳ型13株,Ⅴ型7株,Ⅵ型24株,Ⅶ型16株,Ⅷ型6株,Ⅸ型4株,Ⅹ型10株,Ⅺ型2株。Ⅵ型为该地区的流行优势菌群,不同牛场各基因型菌株分布有明显差异。本研究说明牛场的环境与养殖条件对病原菌流行传播有明显的影响,这一结果对地区性奶牛乳房炎的防治提供了可靠的理论依据。     

牛体外受精卵的二步法培养体系的研究

以CR1aa为基础培养液,采用二步法对牛体外受精卵进行体外培养,完善牛体外受精卵的培养体系.实验一:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa 3 mg/mLBSA培养,后4 d换为CR1aa 50 mL/L FBS.处理组的囊胚率显著高于对照组,但卵裂率和囊胚孵化率无显著差异.实验二:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa培养,后4 d换为CR1aa 50 mL/L FBS.处理组的卵裂率显著高于对照组,但囊胚率和囊胚孵化率差异不显著.实验三:对照组连续7 d均为CR1aa 50 mL/L FBS 0.1mmol/L GSH培养,处理组前3 d为CR1aa 0.1 mmol/L GSH培养,后4 d换为CR1aa 50 mL/L FBS 0.1 mmol/L GSH.处理组的卵裂率显著高于对照组,囊胚率极显著高于对照组,但囊胚孵化率差异不显著.结果表明,GSH与二步法培养系统结合相对于传统的一步法培养系统更适于牛体外受精卵的体外培养.     

Treatment of rupture of suspensory ligament and superficial flexor tendon in a bull

Rupture of the suspensory ligament at the insertions on the proximal sesamoid bones, and of the superficial flexor tendon of the left fore limb, occurred in an adult Angus bull as a result of fighting. There was severe hyperextension of the metacarpophalangeal (MCP) joint with the dewclaws almost touching the ground. Radiographs revealed severe hyper-extension of the MCP joint with the sesamoid bones aligned directly distal to the metacarpus. Initially, a full length fiberglass cast was applied with the limb partially flexed within the cast and the heels elevated. The cast was replaced twice. The cast was removed after 136 days and the bull was bearing full weight on the limb. Prolonged immobilisation of the limb produced new bone in the area (a normal response in cattle) to cause ankylosis of the traumatized MCP joint and partial ankylosis of the carpus. The bull was being used for pasture breeding one year after the injury.     

Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.     

DIAGNOSTIC ULTRASOUND IMAGING OF SOFT TISSUES IN THE BOVINE DISTAL LIMB

In this study sonographic images of healthy flexor tendons, digital flexor tendon sheaths, metacarpophalangeal and metatarsophalangeal joints, and proximal and distal interphalangeal joints of the bovine limb were made. Then, synovial cavities in cadavers were filled with the necessary amount of fluid to make the lumina visible sonographically. After injecting 30–40 ml of saline solution into the digital flexor tendon sheath and into the metacarpophalangeal and metatarsophalangeal joints, or 8 ml in the proximal interphalangeal joint and 10 ml in the distal interphalangeal joint, the pouches of these synovial cavities were clearly demarcated. Afterwards, the sonographic image of synovial cavities distended by various inflammatory content were described. When the sonographic findings were compared to the findings in clinical patients, centesis and intraoperative procedures, there appeared to be a relationship as to the extent and location of the disorders as well as to the nature of the synovial effusion. Finally, we identified which planes on diseased bovine distal limbs were appropriate for obtaining optimal sonographic images.     

Developmental Competence of Frozen-thawed Blastocysts from Fair-quality Bovine Embryos Cultured with β-Mercaptoethanol

Two-hundred-and-thirty-one fair-quality embryos at the compacted morula stage collected from 89 superovulated cows were cultured in TCM199 or Brinster's BMOC-3 medium with or without 100 microM beta-mercaptoethanol (beta-ME). After 24 h culture, a total of 142 fair-quality embryos developed to the blastocyst stage, of which 106 were subsequently frozen with 1.8 M ethylene glycol. The mean cell number and development rates of frozen-thawed blastocysts from the fair-quality embryos cultured in TCM199 containing beta-ME were higher than those of the fair-quality embryos directly frozen without culture. The pregnancy rates obtained with frozen blastocysts from fair-quality embryos tended to be lower than those of non-cultured fresh fair-quality embryos and cultured fresh blastocysts. These results indicate that the inclusion of beta-ME in pre-freezing culture media improve the development of frozen-thawed blastocysts from fair-quality embryos, but not the pregnancy rate.     

牛卵泡卵母细胞的体外成熟和冷冻保存

在简化牛卵母细胞体外成熟的基础上,观察了保护剂、平衡温度、预平衡方法、解冻方法以及冷冻方法对牛卵泡卵母细胞冷冻的影响。结果表明:在体外成熟培养液中添加26.2 mmol/L的NaHCO3和对屠宰场卵巢进行选择更能促进牛卵母细胞的体外成熟;无论是程序冷冻还是玻璃化冷冻,乙二醇(EG)与甘油(GLY)相比更能促进牛卵泡卵母细胞的冷冻效果;在程序冷冻中,冷冻液中添加0.1 mol/L蔗糖比添加0.3mol/L的蔗糖更能促进牛卵泡卵母细胞的冷冻;在玻璃化冷冻中,37℃和25℃的平衡温度比4℃更适合牛卵泡卵母细胞的冷冻;在10%、5%、1%的预平衡浓度之间,5%的预平衡浓度即可达到预平衡的效果;在解冻时,多步脱除保护剂更能保护冷冻的卵母细胞;比较了几种最小样本量(minimum size sample,MSS)玻璃化冷冻方法,解冻成熟培养后的成熟率,拉细毛细玻璃管冷冻法为(41.67±3.19)%,极显著高于未拉细毛细玻璃管冷冻法(glassmicropipette,GMP)的(30.19±1.93)%和固体表面冷冻法(solid surface vitrification,SSV)的(28.33±2.89)%(P<0.01)。     

牛初乳酶解物对离体犊牛小肠上皮细胞增殖和吸收功能的影响

初乳经胃蛋白酶水解处理90分钟后所产生的水解物对离体犊牛小肠上皮细胞有明显的促增殖作用(P<0.01) ,而初乳经胰蛋白酶处理则不表现促增殖活性。初乳酶解物对促进细胞吸收葡萄糖的作用随着酶解处理时间的延长而逐渐增强 ,初乳经胰蛋白酶水解处理90分钟或经胃蛋白酶水解处理150分钟时的产物对细胞吸收葡萄糖的促进作用分别达到最强(P<0.01)。试验结果表明 ,初乳酶解物即小分子蛋白质(肽)具有刺激离体小肠上皮细胞增殖和功能发育的活性。     

Intraocular pressure in normal dairy cattle

Intraocular pressure (IOP) was measured in normal dairy cows by applanation tonometry. In the first study of 15 Holstein and 17 Jersey cows the mean IOP by Mackay-Marg tonometry was 27.5 ± 4.8 mmHg (range 16–39 mmHg); no significant differences ( P < 0.92) were observed between the Holstein and Jersey breeds. In the second study of 15 Holstein and 12 Jersey cows, the mean IOPs by Mackay-Marg and TonoPen-XL tonometry were 28.2 ± 4.6 mmHg (range 19–39 mmHg) and 26.9 ± 6.7 mmHg (range 16–42 mmHg), respectively. Comparisons of the Mackay-Marg and TonoPen tonometers indicated no significant differences ( P < 0.16). The mean and range of IOP in normal dairy cows within 2 SD (95% of the population) is 27 mmHg with a range of 16–36 mmHg.