基于α-乳白蛋白基因序列的牛奶过敏原PCR检测

采用收集牛奶中脱落体细胞的方法提取牛奶DNA,针对牛奶过敏原蛋白α-乳白蛋白基因序列设计引物α-La-F/R,筛选适宜的实验条件,建立了检测牛奶过敏原的PCR方法.结果表明:PCR反应的最佳退火温度为56.5℃,引物浓度0.15μmol/L,循环数35,进一步实验表明该方法特异性良好,检测灵敏度可达到0.04ng DNA.将建立的PCR方法应用于10种食品的牛奶过敏原检测,实验结果与商品标示一致,表明该方法具有较高的准确性和可靠性,适用于实际商品中牛奶过敏原的检测.     

腐蹄病C型节瘤拟杆菌纤毛蛋白基因T程疫苗对家兔的免疫试验

将转化C型节瘤拟杆菌纤毛蛋白基因表达子粒的工程菌PAK/2pfsC在含羧苄青霉素营养肉汤培养基中培养,用MgCl2法在培养物上清中提取表达产物(纤毛蛋白).将表达的纤毛蛋白产物用弗氏完全佐剂乳化,制成疫苗.用疫苗免疫3只健康家兔,21 d后再次接种;定期采血,用对流免疫电泳和K凝集实验检测试验家兔的体液免疫应答及抗体水平.结果发现,免疫7 d即可产生相应抗体,21 d后抗体效价达到8 000×以上,而且高滴度抗体可维持6个月以上.试验表明,腐蹄病C型节瘤拟杆菌纤毛蛋白基因工程疫苗具有较好的免疫应答和免疫原性.     

土壤脲酶活性两种测定方法的比较

利用比色法和扩散法对金强河地区和甘肃农业大学校园内的土壤脲酶活性进行了分析。结果表明：比色法和扩散法测定的土壤脲酶活性平均值分别为（0．531±0．207）mg／g和（0．446±0．204）mg／g,两种测定方法的结果和变异系数差异均不显著（P〉0．05）。但从试验操作方面考虑,扩散法操作比较简单、所用药品种类少、时间短,是一种比较可行的土壤脲酶活性测定方法。     

预混料中维生素E的近红外光谱定量分析模型定标参考值选取

本研究旨在比较以检测值为定标参考值和以理论值为定标参考值2类近红外光谱(NIRS)定量分析模型的预测精准度差异。试验制备了225个维生素预混料样品,利用傅里叶变换近红外光谱仪采集样品在10 000～4 000 cm^-1内的NIRS,基于同一维生素预混料定标集,分别以检测值、理论值为定标参考值,借助偏最小二乘方法,筛选最优光谱预处理方式,构建维生素E的NIRS定量分析模型,并根据F-检验,采用同一独立验证集,评价NIRS定量分析模型的预测效果。结果表明：维生素E的NIRS定量分析模型A(Model-A),以检测值为定标参考值,最优光谱预处理方式为多元散射校正,定标集决定系数(R²)为0.992,最小交互验证标准误差(RMSECV)为4.50×10³IU/kg,相对分析误差(RPD)为11.2;维生素E的NIRS定量分析模型B(Model-B),以理论值为定标参考值,最优光谱预处理方式为变量标准化,定标集R²为0.991,RMSECV为4.94×10³IU/kg,RPD为10.6。...     

孵化厅设计的一般规则及其卫生防疫要求

1孵化厅设计的一般规则 孵化厅的设计内容主要包括两方面：其一是孵化厅平面建筑布局设计;其二是空气调节系统的设计。这两个方面并非截然分开,在设计中必须统筹兼顾,作为一个整体考虑。     

北大营示范场黑麦草引种与品比试验研究

利用1998年2－11月在云南省北大营示范场引种的10个黑麦草品种的观测资料，统计分析了在高、中、低3种施肥水平下黑麦草的累计产量、月平均产量及其生长速度的变化规律，研究了影响产量的主导因子及其监测模型。结果表明，在10个品种中，伯生（Bison Intermdiate）及安巴瑟（Embassy）具有良好的生产性能。黑麦草产量主要受气候、土壤肥力及品种等因素的影响，其中测草盘高度、降水量、蒸发量、日照时数和牧草生长天数等5个因子，可以较好地模拟、评估黑麦草产量的变化动态。     

Analysis of Rumen Microbial Diversity and Functional Prediction of Chinese Simmental Cattle Based on 16S rRNA High-throughput Sequencing Technology

To systematically explore the microbial diversity and function in the rumen of Chinese Simmental cattle,this experiment used 16S rRNA gene high-throughput sequencing technology to detect and analyze the rumen fluid samples of Simmental cattle (about 20 months old,the average weight was 550 kg),and its function was predicted by PICRUSt.The results showed that a total of 66 712 high-quality sequences were obtained from the Illumina Miseq sequencing platform,and 1 797 operational classification units (OTU) were obtained by cluster analysis.The taxonomic identification was divided into 13 phylums,20 classes,22 orders,33 families and 59 genuses;Firmicutes and Bacteroidetes were dominant bacteria,accounting for 62.46% and 22.45%,respectively;Based on the genus composition,[Eubacterium]_coprostanoligenes_group (9.16%),Ruminococcus_2 (7.90%),norank_f__Bacteroidales_RF16_group(6.23%),norank_f__Ruminococcaceae(4.79%),Rikenellaceae_RC9_gut_group(4.72%),norank_f_Bacteroidales_BS11_gut_group(4.49%),Ruminococcus_1(4.43%),etc.The PICRUSt function of the 16S rRNA genome predicted that the rumen flora function was mainly concentrated in carbohydrate transport and metabolism,and might contain a large amount of cellulose and lignin degrading enzyme genes.In summary,the high-throughput sequencing technology based on 16S rRNA gene fully revealed the diversity of rumen bacteria in Simmental cattle,and predicted that it was rich in protein decomposition and lignocellulose degrading enzymes.The basis of microbial cognition provided a reference for further research on rumen microbial functional genes closely related to some important nutritional and physiological functions.     

Pharmacokinetics of sanguinarine,chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half‐life (t_1/2) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (C_max) of DHSA (2.49 ± 1.4 μg/L) was higher that of SA (1.89 ± 0.8 μg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h^?1·kg^?1) with a t_1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.     

转基因动物制作技术及其基因载体研究进展

论文概述了家畜转基因动物的制作方法,初步分析了用于转基因技术的基因载体种类,深入探讨了不同载体的优缺点,并展望了基因载体的发展趋势.     

孕牛血清中早孕因子的分离纯化

采用DEAE－Sepharose(DEAE-琼脂糖凝胶）、CM－Sepharose(CM-琼脂糖凝胶）等离子交换层析、抗孕牛全血清IgG-Sepharose 4B亲和层析等技术，分离纯化妊娠4－5个月健康孕牛全血清中的早孕因子（EPF），用玫瑰花环抑制实验对各阶段产物EPF活性进行检测。结果表明，经层析过柱后的CM－ⅡA提取物有EPF活性。经SDS－PAGE银染显色证实它含有分子量为21、67、80KD的蛋白，实验结果支持21KD的多肽为主要EPF活性物成分。同时实验表明了亲和层析对进一步纯化EPF是可行的。