高精料日粮对反刍动物瘤胃及盲肠组织中GPR41、43表达的影响
本文选题:高精料日粮 切入点:综合调控 出处:《南京农业大学》2015年硕士论文 论文类型:学位论文
【摘要】:在现代奶牛的饲养过程中,为了满足泌乳早期和中期的奶牛不断增加的能量需求和提高奶产量,选择高精料日粮饲喂的方式,但这种方式会引起亚急性瘤胃酸中毒等一系列代谢综合征的发生。本试验以泌乳中期荷斯坦奶牛为试验动物,采用气相色谱法、放射免疫分析法和分子生物学方法,研究高精料日粮对反刍动物瘤胃内挥发性脂肪酸、发酵类型的影响,进一步研究其对瘤胃等消化道组织中GPR41和43相对表达量的影响。本试验还用泌乳中期的奶山羊进行了综合调控试验,两组试验动物都饲喂精粗比6:4的高精料,将综合调控组的精料进行制粒并且添加缓冲剂,来研究综合调控法对高精料饲喂引起的机体代谢综合征的缓解作用和对瘤胃等消化道组织GPR41和43相对表达量的影响,旨在为现代奶牛养殖业高精料饲喂方式提出改进的方法。试验一:高精料日粮对反刍动物瘤胃内挥发性脂肪酸和GPR41、43的影响:本试验选用6头泌乳中期的荷斯坦奶牛做高精料饲喂对反刍动物瘤胃内挥发性脂肪酸、发酵类型和GPR41、43表达的影响的试验。奶牛分成两组,分别为高精料组(High concentrate,HC)(精粗比 6:4)和低精料组(High roughage,HR)(精粗比 4:6)。饲喂后通过瘤胃瘘管采集0-12 h瘤胃液,pH计检测瘤胃液pH值,气相色谱法分析瘤胃内挥发性脂肪酸(VFA)。放射免疫法检测外周血中炎性因子IL-1β、EL-6、TNFa的浓度。实时荧光定量PCR法检测瘤胃和盲肠组织中GPR41、43的相对表达量。试验结果为:高精料(精粗比6:4)饲喂奶牛会使瘤胃内乙酸浓度极显著降低;乙酸比例和乙丙比降低,丙酸比例升高,且差异显著。饲喂日粮后高精料组瘤胃液pH值均值极显著降低,pH值小于5.6和6.0的时间延长,且差异显著。高精料组外周血中炎性因子IL-1β、IL-6、TNFa的浓度都高于低精料组,且IL-1β为差异极显著,IL-6、TNFa为差异显著。高精料组瘤胃组织GPR41、43的相对表达量显著高于低精料组,盲肠组织GPR41、43的相对表达量极显著高于低精料组。结论为:用高精料日粮饲喂奶牛改变了瘤胃的发酵类型。长期高精料饲喂使瘤胃pH均值降低,pH值低于5.6的时间超过3 h,外周血中炎性因子浓度升高,因此引起奶牛亚急性瘤胃酸中毒。高精料日粮可使瘤胃和盲肠组织中GPR41、43的表达量增加。试验二:综合调控对反刍动物瘤胃内挥发性脂肪酸和GPR41和GPR43表达的影响:本实验选用8头泌乳中期奶山羊做综合调控试验研究综合调控法能否缓解高精料日粮对反刍动物瘤胃内挥发性脂肪酸产生的影响以及综合调控法对GPR41、43表达的影响。将奶山羊分为两组,分别为综合调控组(Comprehensive regulation,CC)和高精料组(HC),在饲喂等重量的高精料条件下,将综合调控组的精料进行制粒并且加入缓冲剂。饲喂后通过瘤胃瘘管采集0-10 h瘤胃液,pH计检测瘤胃液pH值,气相色谱法分析瘤胃内挥发性脂肪酸(VFA)。放射免疫法检测外周血中炎性因子IL-1β、IL-6、TNFa的浓度。实时荧光定量PCR法检测瘤胃和盲肠等组织中GPR41、43的相对表达量。试验结果为综合调控组瘤胃内乙酸和丁酸的含量极显著高于高精料组,总VFA含量也极显著地升高,乙酸比例和乙丙比显著升高,丙酸比例显著降低,显著地调整了瘤胃发酵类型。经过综合调控pH值均值极显著升高,pH值小于6.0的时间显著缩短;炎性因子IL-1β、IL-6、TNFa的浓度低于高精料组,IL-1β为差异显著,IL-6、TNFa为差异极显著。奶山羊的瘤胃、十二指肠、盲肠和结肠组织都表达GPR41和43。调控组相比于高精料组,GPR41和43在瘤胃和十二指肠组织中表达升高,在盲肠和结肠中表达降低。结果表明,综合调控法能够显著地调整瘤胃发酵类型,升高瘤胃内环境pH值,减轻体内炎症反应,使动物在相对稳定的瘤胃酸性环境下发酵产生大量的挥发性脂肪酸,从而有利于动物实现高产,为现代牧业高精料饲喂方式提出改进的方法。
[Abstract]:In the breeding process of modern dairy cows, in order to meet the early and mid lactation cows increased energy demand and increase milk yield, high concentrate diets fed, but in this way will cause subacute ruminal acidosis and a series of metabolic syndrome. In this experiment, the test for lactating Holstein cows animal, by gas chromatography, radioimmunoassay and molecular biological methods of high concentrate diets on ruminant animal rumen volatile fatty acids, influence the fermentation type, further study on the GPR41 of rumen and other tissues of digestive tract and 43 relative expression effect. This experiment has also carried out a comprehensive control test in mid lactation dairy goats, two groups of experimental animal feeding forage to concentrate ratio of high concentrate 6:4, will concentrate control group for granulation and add a buffer, to study the comprehensive control method of high precision Feeding caused by metabolic syndrome and effects on ruminal digestive tract tissue of GPR41 and 43 relative expression of influence, to put forward the improvement methods for the modern dairy industry high concentrate feeding mode. Experiment 1: high concentrate diets on the effects of volatile fatty acid and GPR41,43 in the rumen of ruminant animal experiment 6 lactating Holstein cows do high concentrate feeding of ruminant animal rumen volatile fatty acids, effects of fermentation type and GPR41,43 expression. The cows were divided into two groups, high concentrate group (High, concentrate, HC) (concentrate ratio 6:4) and low concentrate group (High roughage HR, (4:6) the ratio of concentrate to roughage). After feeding through the acquisition of 0-12 h rumen rumen, rumen pH value pH detection, analysis of rumen volatile fatty acids by gas chromatography (VFA). Radioimmunoassay in peripheral blood inflammatory factor IL-1 beta, EL-6, The concentration of TNFa. The relative expression of GPR41,43 detection of rumen and cecal tissues by real-time fluorescence quantitative PCR method. The test result is: high concentrate (coarse than 6:4) of dairy cows fed diets will make the rumen acetic acid concentration decreased significantly; the proportion of acetic acid and propylene ratio decreased, the proportion of propionic acid increased significantly. Feeding diets after high concentrate group rumen pH values significantly decreased, the pH value is less than 5.6 and 6 of the time, and the difference was significant. The high concentrate group in peripheral blood inflammatory cytokine of IL-1, IL-6, TNFa concentrations were higher than the low concentrate group, and IL-1 beta is extremely significant difference, IL-6. TNFa is significant difference. The relative expression of high concentrate group GPR41,43 rumen tissues were significantly higher than that in low concentrate group, the relative expression of cecal tissue GPR41,43 content was significantly higher than that in low concentrate group. Conclusion: the use of high concentrate diets of dairy cows fed diets change the type of rumen fermentation. The long-term high concentrate feeding rumen p The mean H decreased, pH value is lower than 5.6 of the time more than 3 h, elevated concentrations of inflammatory factors in peripheral blood, thus causing subacute ruminal acidosis in dairy cows. High concentrate diets can make the expression of GPR41,43 in rumen and cecal tissue increased. Experiment two: comprehensive regulation effect on the expression of volatile fatty acids and GPR41 and GPR43 in rumen of ruminant animal: in this experiment, 8 lactating dairy goat do experimental research on comprehensive regulation method of comprehensive regulation can alleviate the impact of high concentrate diet on ruminant animal rumen volatile fatty acids and the comprehensive control method of GPR41,43 expression. The goats were divided into two groups, respectively. Control group (Comprehensive regulation, CC) and high concentrate group (HC), the Fed's weight in high concentrate conditions, will concentrate control group for granulation and adding buffer. After feeding by rumen fistula collected 0-10 H Rumen fluid pH detection rumen pH value analysis of rumen volatile fatty acids by gas chromatography (VFA). Radioimmunoassay in peripheral blood inflammatory factor IL-1 beta, IL-6, the concentration of TNFa. The relative expression of GPR41,43 was detected by real time fluorescence quantitative PCR in rumen and cecal tissue in the test. The content of acetic acid and butyric acid in the rumen of comprehensive control group was significantly higher than that of high concentrate group, the total VFA content was also significantly increased, the proportion of acetic acid and ethylene were significantly higher than that of propionic acid ratio decreased significantly, significantly adjust the rumen fermentation type. After comprehensive regulation of the average pH value increased significantly, the pH value is less than 6 the time was significantly shortened; inflammatory cytokine of IL-1, IL-6, TNFa concentration is lower than the high concentrate group IL-1 beta is significant difference, IL-6, TNFa were extremely significant difference. The dairy goat rumen, duodenum, cecum and colon tissue expressed GPR41 and 43. compared to the control group High concentrate group, and 43 GPR41 in the rumen and duodenum tissue expression increased, decreased expression in the cecum and colon. The results show that the integrated control method can significantly adjust the type of rumen fermentation, rumen environment increased pH value, reduce the inflammation in the body, the animal in a relatively stable rumen fermentation to produce volatile acidic environment a large number of fatty acids, which is beneficial to the animal to achieve high yield, modern animal husbandry high concentrate feeding way proposes the improvement method.
【学位授予单位】:南京农业大学
【学位级别】:硕士
【学位授予年份】:2015
【分类号】:S816
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