奶牛卵母细胞体外受精和胚胎培养研究

[目的]探讨适合于奶牛体外胚胎生产的最佳条件。[方法]以奶牛卵母细胞为试验材料,研究不同温度、运送时间对奶牛卵母细胞成熟的影响,并比较在相同培养条件下孤雌激活和体外受精对奶牛卵母细胞胚胎发育的影响。[结果]卵巢在37—42℃运输后的成熟率（11．3％）明显低于20—30℃、30-37℃的成熟率（79．3％,85．1％）,20～30℃、30～37℃组间差异不显著;运送时间6h后的卵母细胞成熟率（53．2％）明显低于4h内、4—6h的成熟率（85．6％,77．8％）,4h内、4～6h差异不显著;孤雌激活和体外受精胚胎在相同条件下的卵裂率（72．41％,68．18％）、8细胞率（45．24％,47．22％）、囊胚率（25．60％,18．89％）差异均不显著。[结论]卵巢在30℃左右4h内运回实验室最适宜,而孤雌激活和体外受精胚胎在相同条件下发育率差异不显著。     

A morphological assessment system for ‘show quality’ bovine livestock based on image analysis

Morphological assessment is one important parameter considered in conservation and improvement programs for bovine livestock. This assessment process consists of scoring an animal based on its morphology and is normally carried out by highly qualified staff. These animals are all of agreed ‘show quality’ and hence they are morphologically very similar.This paper presents a system designed to provide an assessment based on a lateral image of the cow. The system consists of two main parts: a feature extraction stage, to reduce the information on the cow in the image to a set of parameters, and a neural network stage to provide a score based on that set of parameters. For the image analysis section, a model of the animal is constructed by means of the point distribution model (PDM) technique. Later, that model is used in the searching process within each image, which is implemented using genetic algorithms (GAs). As a result of this stage, the vector of weights that describe the deviation of the given shape from the mean is obtained. This vector is used in the second stage, where a multilayer perceptron is trained to provide the desired assessment, using the scores given by experts for selected cows.The system has been tested with 138 images corresponding to 44 individuals of a special rustic breed, with very promising results, given that the information contained in only one view of the cow can not be considered complete.     

聚蔗糖水凝胶的制备及其在药物控制释放中的应用

[目的]研究聚蔗糖水凝胶的制备工艺,并考察制备工艺对溶胀度和牛血清蛋白（BSA）释放性能的影响。[方法]以环氧氯丙烷为交联剂,在碱性条件下制备聚蔗糖水凝胶,用BSA作为模型药物考察其释放性能。[结果]当聚蔗糖溶液浓度为16.7 g/L,交联剂用量为0.85 mol/L,反应温度为45℃,反应时间为5 h时,制得的聚蔗糖水凝胶弹性和硬度较好,其溶胀度在5.5～7.5。聚蔗糖水凝胶对BSA的释放性能表明,聚蔗糖水凝胶的交联程度决定BSA的释放速率,适当增加聚蔗糖水凝胶的交联程度有利于药物的缓慢释放。[结论]可以通过调控聚蔗糖水凝胶的溶胀度调节包埋在此水凝胶中BSA的释放速率。     

Disturbed bovine mitochondrial lipid metabolism: a review

In mammals, excess energy is stored primarily as triglycerides, which are mobilized when energy demands arise and cannot be covered by feed intake. This review mainly focuses on the role of long chain fatty acids in disturbed energy metabolism of the bovine species. Long chain fatty acids regulate energy metabolism as ligands of peroxisome proliferator-activated receptors. Carnitine acts as a carrier of fatty acyl groups as long-chain acyl-CoA derivatives do not penetrate the mitochondrial inner membrane. There are two different types of disorders in lipid metabolism which can occur in cattle, namely the hypoglycaemic-hypoinsulinaemic and the hyperglycaemic-hyperinsulinaemic type with the latter not always associated with ketosis. There is general agreement that fatty acid β-oxidation capability is limited in the liver of (ketotic) cows. In accord, supplemental L-carnitine decreased liver lipid accumulation in periparturient Holstein cows. Of note, around parturition concurrent oxidation of fatty acids in skeletal muscle is highly activated. Also peroxisomal β-oxidation in liver of dairy cows may be part of the hepatic adaptations to a negative energy balance (NEB) to break down fatty acids. An elevated blood concentration of nonesterified fatty acids is one of the indicators of NEB in cattle among others like increased β-hydroxy butyrate concentration, and decreased concentrations of glucose, insulin, and insulin-like growth factor-I. Assuming that liver carnitine concentrations might limit hepatic fatty acid oxidation capacity in dairy cows, further study of the role of acyl-CoA dehydrogenases and/or riboflavin in bovine ketosis is warranted.     

牛卵母细胞体外受精和培养技术

本文综述了牛卵母细胞体外受精和体外成熟技术的研究进展,包括精子的体外获能、卵母细胞成熟培养和体外受精、体外受精早期胚胎的培养等,并且对影响该技术的主要原因进行了分析。     

牛血清白蛋白对连香树PCR反应体系的优化

[目的]为了取得更好的PCR扩增效果,建立高效的PCR反应体系。[方法]首先通过6因素5水平的正交试验对连香树RAPD-PCR反应体系优化组合,然后不再改变该结果中的其他条件,而仅改变Taq酶的浓度及在反应体系中加入不同浓度的牛血清白蛋白（BSA）。[结果]在25组Taq酶和BSA浓度组合中,BSA改善连香树RAPD-PCR反应的最佳浓度为0.6μg/μl,Taq酶浓度可由1.0μg/μl减少至0.4μg/μl。最后优化得到的20.0μl连香树RAPD反应体系为：25mmol/LMg2＋2.0μl,dNTPs0.4μl,1U的TaqDNA酶1.0μl,10×Buffer缓冲液2.5μl,0.5OD引物0.8μl,约25ng模板0.6μl。[结论]BSA的适当加入减少了Taq酶的用量,在保证更好的试验效果前提下,节约了试验成本。     

利用耳组织、血液、牛奶样品检测牛病毒性腹泻病毒的比较

牛病毒性腹泻病以发热、口腔及消化道粘膜糜烂、腹泻为主要症状,可引起怀孕母牛流产或产畸型胎儿,给养牛业造成巨大的经济损失,该病已成为当前严重危害我国养牛业的传染病之一。由于大部分感染BVDV的牛不表现特征性的临床症状和病理变化,而且引起腹泻和消化道黏膜糜烂或溃疡的疾病很多,所以确诊需进行实验室检查。目前实验室检测BVDV的主要方法是利用美国IDEXX公司ELISA诊断试剂盒,通过BVDV抗原和抗体的检测区别牛群持续感染牛。用于检测BVDV的样品种类有许多种,绝大多数是采集血液(血清)或牛奶样品进行检测。本试验对奶牛进行耳组织采样,同时采集血液和牛奶,用三种样品在相同条件下同时检测BVDV抗原,用血清和牛奶样品检测抗体,比较样品的检测结果。结果表明,这三种样品检测结果具有一致性,且耳组织采样更宜于大面积筛查。     

Should the domestic buffalo (Bubalus bubalis) be considered in the epidemiology of Bovine Herpesvirus 1 infection?

Only limited information is available on the epidemiology and pathogenesis of Bovine Herpesvirus 1 (BoHV-1) in domestic buffalos. In this study, a virulent BoHV-1 field strain isolated from cattle was inoculated into buffaloes to evaluate their susceptibility to the virus and to investigate the establishment of viral latency through clinical, virological and serological investigations. Latency was also studied by attempting viral reactivation using pharmacological induction. Six of seven male, 5 months old buffaloes were intranasally inoculated with BoHV-1; the other animal was kept as negative control. The animals were clinically monitored during the post-infection (P.I.) and the post-pharmacological induction (P.P.) periods. During these periods, nasal and rectal swabs, and blood samples, with and without anticoagulant, were collected at 2–3 day intervals. On culling the animals, 206 days P.I., their trigeminal ganglia and tonsils were collected. No clinical signs referable to BoHV-1 were observed throughout the experimental period. However, seropositivity was detected in all infected animals within day 20 P.I., using BoHV-1 glycoprotein E and glycoprotein B competitive ELISAs (IDEXX) and virus neutralisation test. In real-time PCR (RT-PCR), five of these animals were positive, at least once, for nasal or rectal swabs, during the P.I. period. The sixth infected animal was found positive only in the trigeminal ganglia after culling. Ganglia were also positive for two other animals. Virus isolation in permissive cell-lines was successful for a part of the RT-PCR positive samples. The detected viruses were confirmed by genetic analysis as identical to the inoculated strain. No evidence of infection was observed in the negative control. This study represents the first experimental transmission of BoHV-1 in buffaloes, confirming their susceptibility to infection and their possible role as host/reservoirs of BoHV-1.