目录
摘要Ⅰ
关键词Ⅰ
AbstractⅡ
引言
引言1
1材料与方法4
1.1实验动物4
1.2实验日粮4
1.3饲养管理5
1.4样品采集5
1.5主要实验仪器及试剂5
1.5.1实验主要仪器5
1.5.2实验主要试剂5
1.6瘤胃pH值的测定 5
1.7瘤胃液中SCFA浓度的测定 5
1.8实时荧光定量PCR检测SCFA吸收相关基因mRNA的表达6
1.8.1瘤胃上皮总RNA的提取6
1.8.2RNA的质量检测 6
1.8.3反转录7
1.8.4设计引物7
1.8.5Real time qPCR的产物检验7
1.8.6Real time qPCR扩增体系8
1.9数据分析8
2结果与分析9
2．1日粮精料水平对湖羊瘤胃pH和SCFA浓度的影响 9
2．2日粮精料水平对瘤胃上皮SCFA吸收相关基因mRNA表达的影响 9
3讨论11
4结论 15
参考文献 16
致谢 18
湖羊瘤胃吸收转运功能对高精料日粮的适应
摘 要
本实验研究通过渐进式增加湖羊日粮中精料水平的饲养实验、瘤胃pH和短链脂肪酸（Shortchain fatty acids, SCFAs）浓度测定及实时荧光定量PCR的方法探究湖羊瘤胃吸收转运功能对渐进式增加精料日粮的适应性。
本文研究内容和结果如下：
选用12头体况良好的湖羊，适应期饲喂10%精料+90%粗料全混合日粮。适应期后随机分为高精组（HC）、低精料组(LC)，每组6头，其中HC组装有瘤胃瘘管。实验期间，LC组全程10%精料+90%粗料全混合日粮饲喂，LC组每周增加20%精料，将精料比例从10%上升为70%。结果表明，湖羊瘤胃24小时pH值正常；饲喂高精料使乙酸、丙酸和丁酸及总SCFA浓度显著增加；基因检测发现，HC组与LC组相比，Na+/H *51今日免费论文网|www.51jrft.com +Q: ￥351916072￥ 
+交换体1(NHE1)、钠钾ATP酶（Na+/K+ATPase）、质子泵（Hv ATPase）、假定阴离子转运载体1(PAT1)和单羧酸转运蛋白(MCT1，MCT4)表达量增加，且差异显著（P<0.05）；NHE2、NHE3和阴离子交换载体2（AE2）在LC组表达量较高，差异不显著；LC组腺瘤下调蛋白(DRA)的表达量显著高于HC组（P<0.05）。
综上所述，SCFA转运载体的mRNA表达量升高，表示HC组瘤胃吸收SCFA的功能活跃。也从侧面证明渐进式增加精料的饲喂方法能够让湖羊瘤胃处于健康状态。24小时瘤胃pH曲线证明，HC组没有出现亚急性瘤胃酸中毒（Subacute ruminal acidosis, SARA），即湖羊已经适应了70%的高精料饲喂。
ADAPTATION OF RUMEN ABSORPTION AND TRANSPORT FUNCTION TO HIGHCONCENTRATE DIET IN HU SHEEP
ABSTRACT
In this study, the adaptability of rumen absorption and transport function of Hu Sheep to the progressive increase of concentrate diet was explored through feeding experiments of progressive increase of concentrate level in Hu Sheep diet, rumen pH and Shortchain fatty acids(SCFAs) concentration determination, and realtime fluorescence quantitative PCR.
The research contents and results of this paper are as follows:
12 Hu Sheep with healthy body condition were selected and fed with 10% concentrate +90% coarse feed at the adaptation stage.After the adaptation period, they were randomly divided into the highprecision group and the lowprecision control group, with 6 sheep in each group, among which 6 sheep in the highconcentration group were equipped with permanent fistulas.During the experiment, the lowconcentration control group was fed a full mixed diet of 10% concentrate +90% coarse feed, while the highconcentration group increased the diet concentrate from 10% to 70% by increasing the amount of concentrate by 20% per week.The results showed that the pH in rumen of Hu Sheep decreased as the proportion of dietary concentrate increased.The contents of acetic acid, propionic acid, butyric acid and total SCFA increased significantly.Gene testing showed that the expression levels of Na+/H+ exchanger1 (NHE1), Na+/K+ ATPase, vacuolated hydrogen ion ATPase (Hv ATPase), anion transporter 1(PAT1) and monocarboxylic acid transporter 1(MCT1, MCT4) were increased in the highconcentrate diet group compared with the lowconcentrate diet group, and the differences were significant (P <0.05).The expression levels of NHE2, NHE3 and anion exchange vector 2 (AE2) were higher in the lowconcentrate diet group, but the difference was not significant.The expression level of adenoma downregulated protein (DRA) in the lowconcentrate diet group was significantly higher than that in the highconcentrate diet group (P<0.05).

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