延边黄牛骨骼肌特异性钙蛋白酶基因多态性及其与肉质性状关系的研究

Calpain3(CAPN3)在肌肉组织中特异表达,其多态位点与畜禽诸多生产性状相关。本研究采用PCR-SSCP方法,设计2对基因特异性引物CAPN3-1、CAPN3-3,对96头延边黄牛CAPN3基因进行了SNPs检测,并将其与部分肉质性状进行了方差分析。试验结果显示,分别在CAPN3基因54982bp处发现A→G突变,56094bp处发现G→A突变。方差分析结果表明,2个位点中各个基因型与肉质嫩度、蒸煮损失均无显著相关性,CAPN3-3片段BB基因型仅在第3天时的pH显著高于AA纯合型个体(P0.05)。2个位点与肉质色度方差分析结果显示,2个位点中各个基因型黄色度呈显著相关(P0.05),与肉色中其他色度无相关性。CAPN3-1位点处,BB基因型较AA、AB基因型具有更高的黄色度值,此规律并不随时间的延续而改变,而在CAPN3-3位点处则无此规律。     

自动进排料密闭式固态发酵罐的设计

为解决当前固态发酵劳动强度大,生产效率低的问题,设计内置纵向排列带扇形缺口的浅盘组和升降旋转控制刮板的中心轴来实现自动进排料和翻料的固态发酵罐,可实现自动化生产,原位灭菌,提高生产效率。     

In vivo characterization of inflammatory biomarkers in swine and the impact of flunixin meglumine administration

Non-steroidal anti-inflammatory drugs (NSAID) are a family of chemicals that function to reduce pain, fever, and inflammation, and they are commonly used in people and animals for this purpose. Currently there are no NSAIDs approved for the management of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess efficacy. A previous in vitro study examining biomarkers of inflammation identified fourteen genes that were significantly altered in response to Escherichia coli lipopolysaccharide (LPS)-induced inflammation. In the present study, five of those fourteen genes were tested in vivo to determine if the same effects observed in vitro were also observed in vivo. Plasma levels of prostaglandin E(2) (PGE(2)), an essential mediator of fever and inflammation, were also determined. Two groups of swine were stimulated with LPS with the second group also treated with flunixin meglumine. Blood was collected at 0, 1, 3, 6, 8, 24, and 48h post LPS-stimulation. The RNA was extracted from the blood and quantitative real-time-PCR (qRT-PCR) was utilized to determine the expression patterns of CD1, CD4, serum amyloid A2 (SAA2), Caspase 1, and monocyte chemoattractant protein 1 (MCP-1). The LPS-stimulated animals demonstrated a statistically significant alteration in expression of SAA2 and CD1 at 3h post-stimulation. Flunixin meglumine treated animals' demonstrated reduced expression of CD1 in comparison to the LPS-stimulated swine at 24 and 48h post LPS-stimulation. Flunixin meglumine treated animals exhibited reduced expression of SAA2 at 48h post-stimulation compared to LPS-stimulated swine. Swine treated with LPS demonstrated statistically significant increases in plasma PGE(2) at 1h post-stimulation. Swine treated with flunixin meglumine had no increase in plasma PGE(2) levels at any time. These results demonstrate that PGE(2) production, along with two out of five genes (SAA2 and CD1) have the potential to serve as early biomarkers of inflammation as well as indicators of NSAID efficacy.     

牛初乳粉片混合工艺的优化研究

为提高牛初乳粉片的混合工艺的混合均匀度,采用响应面试验设计优化混合工艺参数。在单因素试验的基础上,确定采用投料方式Ⅲ,以混合时间、装载系数为主要影响因素,以混合均匀度为响应值,采用响应面法对混合工艺参数进行优化。结果表明:牛初乳粉片混合工艺最优参数为:混合时间15 min,装载系数35%,混合均匀度为94.68%,与模型预测的理论值96.52%均比较接近。采用响应面分析优化得到的牛初乳粉片混合工艺参数具有可行性和可靠性,可用于实际操作。     

重组质粒pACYC184-hok/sok的构建及稳定性研究

以低拷贝质粒pACYC184为载体,将hok/sok基因插入到载体BamH Ⅰ位点,构建重组质粒pACYC184-hok/sok稳定系统,同时构建hok/sok基因缺失突变重组质粒pACYC184-hok/sok(M).通过对构建的重组菌进行连续传代和对不同代次的重组质粒进行SphⅠ酶切,分析hok/sok基因的引入对宿主细胞生长的影响.结果表明,重组质粒pACYC184-hok/sok在无抗生素筛选压力下连续传代,传至第135代时质粒稳定率仍是100%,且传代前后的质粒SphⅠ酶切图谱没有发生变化,DNA序列测定表明,插入的hok/sok基因没有发生任何突变.突变重组质粒pACYC184-hok/sok(M)传代前后的质粒SphⅠ酶切结果虽没有发生变化,但其相应的重组菌传至第15代,质粒几乎完全丢失.生长曲线的测定结果表明,与舍pACYC184重组菌生长曲线相似,pACYC184-hok/sok重组菌生长较快,而pACYC184-hok/sok(M)重组菌生长缓慢.以上结果表明,含hok/sok稳定系统,其稳定性明显高于无hok/sok稳定系统的质粒pACYC184以及突变质粒pACYC084-hok/sok(M).重组质粒pAqCYC184-hok/sok具有良好的结构稳定性和分离稳定性.     

羽毛角蛋白降解的研究进展

羽毛角蛋白是一种硬性蛋白,在动物饲料、复合材料、食品包装和医药制品等领域展现出良好应用前景。但因羽毛角蛋白难溶解和降解的特殊性,一般方法很难获得可溶性角蛋白,酶法相对于传统物理和化学降解法具有反应条件温和、环境污染小和降解产物稳定等优点,是具有很高实用价值的羽毛降解途径。文中重点介绍角蛋白酶对羽毛的降解作用,并概述物理、化学方法的降解原理以及角蛋白的综合利用。     

新城疫病毒实时荧光RT-PCR检测方法的建立与应用

为满足新城疫病毒高通量快速检测的需要,针对新城疫病毒M基因序列,通过基因比对分析保守区域,经同源性分析后,设计合成多条引物和探针并对其筛选,建立了一种能够快速检测新城疫病毒的实时荧光RTPCR检测方法,并对该方法的特异性、灵敏性进行了评估。结果显示,该方法检测耗时短、特异性好,检测下限达10~(-4) ng/μL。利用该方法,对480份临床采集的各类家禽咽拭子样品进行检测,共检测出25份阳性,与常规RT-PCR检测方法结果一致,κ值为1(P 0.001)。结果表明,该方法快速、准确,可用于新城疫病毒的快速检测。     

乳酸乳球菌乳脂亚种IMAU60064增殖培养基的优化

以分离自西藏那曲县罗玛镇传统发酵酸牦牛奶中的乳酸乳球菌乳脂亚种（Lactococcus lactis subsp．Cremoris）IMAU60064为试验菌株,研究其最佳的培养条件。对碳源、氮源、缓冲盐等培养基成分及培养条件进行优化,并采用响应面法对优选的碳源、氮源和缓冲盐类的组成含量进行优化,得到IMAU60064的增殖培养基为：葡萄糖23g／L、大豆蛋白胨11g／L、牛肉膏11g／L、胰蛋白胨5g／L、NaAC1．8g／L、K2HP041．2g／L、柠檬酸钠1．2g／L、MgSO4·7H200．4g／L、MnSO4·5H2O54mg／L、L-半胱氨酸盐酸盐0．5g/L、吐温80为1g／L。Lactococcus lactis subsp．cremorisIMAU60064在此增殖培养基中经30℃,14h培养活菌数可达到3．26×10^8cFu／mL,比在MRS中（6．54×10^7CFU／mL）提高近5倍。     

灌丛草地放牧山羊的牧草适口性与嗜食性及山羊采食率研究

通过对放牧山羊的牧草适口性，嗜食性和采食率及其相互关系的分析，表明山羊放牧对灌丛草地的植物适口性、嗜食性比率和采食率均以灌木嫩枝叶，中禾草和苔草属植物较优。季节性规律为：春季山羊喜哺食灌木嫩枝叶，灌木嗜食性比率较高，采食率也较高，夏季山羊喜草本植物，草本植物的嗜食性比率高于灌木，秋季山羊较喜啃食灌木枝叶，嗜食性比率灌木高于草本植物，采食率低于春季，灌木采食率高于草本植物。