本文選題：高精料日糧　切入點：綜合調(diào)控　出處：《南京農(nóng)業(yè)大學》2015年碩士論文　論文類型：學位論文

【摘要】：在現(xiàn)代奶牛的飼養(yǎng)過程中,為了滿足泌乳早期和中期的奶牛不斷增加的能量需求和提高奶產(chǎn)量,選擇高精料日糧飼喂的方式,但這種方式會引起亞急性瘤胃酸中毒等一系列代謝綜合征的發(fā)生。本試驗以泌乳中期荷斯坦奶牛為試驗動物,采用氣相色譜法、放射免疫分析法和分子生物學方法,研究高精料日糧對反芻動物瘤胃內(nèi)揮發(fā)性脂肪酸、發(fā)酵類型的影響,進一步研究其對瘤胃等消化道組織中GPR41和43相對表達量的影響。本試驗還用泌乳中期的奶山羊進行了綜合調(diào)控試驗,兩組試驗動物都飼喂精粗比6:4的高精料,將綜合調(diào)控組的精料進行制粒并且添加緩沖劑,來研究綜合調(diào)控法對高精料飼喂引起的機體代謝綜合征的緩解作用和對瘤胃等消化道組織GPR41和43相對表達量的影響,旨在為現(xiàn)代奶牛養(yǎng)殖業(yè)高精料飼喂方式提出改進的方法。試驗一:高精料日糧對反芻動物瘤胃內(nèi)揮發(fā)性脂肪酸和GPR41、43的影響:本試驗選用6頭泌乳中期的荷斯坦奶牛做高精料飼喂對反芻動物瘤胃內(nèi)揮發(fā)性脂肪酸、發(fā)酵類型和GPR41、43表達的影響的試驗。奶牛分成兩組,分別為高精料組(High concentrate,HC)(精粗比 6:4)和低精料組(High roughage,HR)(精粗比 4:6)。飼喂后通過瘤胃瘺管采集0-12 h瘤胃液,pH計檢測瘤胃液pH值,氣相色譜法分析瘤胃內(nèi)揮發(fā)性脂肪酸(VFA)。放射免疫法檢測外周血中炎性因子IL-1β、EL-6、TNFa的濃度。實時熒光定量PCR法檢測瘤胃和盲腸組織中GPR41、43的相對表達量。試驗結(jié)果為:高精料(精粗比6:4)飼喂奶牛會使瘤胃內(nèi)乙酸濃度極顯著降低;乙酸比例和乙丙比降低,丙酸比例升高,且差異顯著。飼喂日糧后高精料組瘤胃液pH值均值極顯著降低,pH值小于5.6和6.0的時間延長,且差異顯著。高精料組外周血中炎性因子IL-1β、IL-6、TNFa的濃度都高于低精料組,且IL-1β為差異極顯著,IL-6、TNFa為差異顯著。高精料組瘤胃組織GPR41、43的相對表達量顯著高于低精料組,盲腸組織GPR41、43的相對表達量極顯著高于低精料組。結(jié)論為:用高精料日糧飼喂奶牛改變了瘤胃的發(fā)酵類型。長期高精料飼喂使瘤胃pH均值降低,pH值低于5.6的時間超過3 h,外周血中炎性因子濃度升高,因此引起奶牛亞急性瘤胃酸中毒。高精料日糧可使瘤胃和盲腸組織中GPR41、43的表達量增加。試驗二:綜合調(diào)控對反芻動物瘤胃內(nèi)揮發(fā)性脂肪酸和GPR41和GPR43表達的影響:本實驗選用8頭泌乳中期奶山羊做綜合調(diào)控試驗研究綜合調(diào)控法能否緩解高精料日糧對反芻動物瘤胃內(nèi)揮發(fā)性脂肪酸產(chǎn)生的影響以及綜合調(diào)控法對GPR41、43表達的影響。將奶山羊分為兩組,分別為綜合調(diào)控組(Comprehensive regulation,CC)和高精料組(HC),在飼喂等重量的高精料條件下,將綜合調(diào)控組的精料進行制粒并且加入緩沖劑。飼喂后通過瘤胃瘺管采集0-10 h瘤胃液,pH計檢測瘤胃液pH值,氣相色譜法分析瘤胃內(nèi)揮發(fā)性脂肪酸(VFA)。放射免疫法檢測外周血中炎性因子IL-1β、IL-6、TNFa的濃度。實時熒光定量PCR法檢測瘤胃和盲腸等組織中GPR41、43的相對表達量。試驗結(jié)果為綜合調(diào)控組瘤胃內(nèi)乙酸和丁酸的含量極顯著高于高精料組,總VFA含量也極顯著地升高,乙酸比例和乙丙比顯著升高,丙酸比例顯著降低,顯著地調(diào)整了瘤胃發(fā)酵類型。經(jīng)過綜合調(diào)控pH值均值極顯著升高,pH值小于6.0的時間顯著縮短;炎性因子IL-1β、IL-6、TNFa的濃度低于高精料組,IL-1β為差異顯著,IL-6、TNFa為差異極顯著。奶山羊的瘤胃、十二指腸、盲腸和結(jié)腸組織都表達GPR41和43。調(diào)控組相比于高精料組,GPR41和43在瘤胃和十二指腸組織中表達升高,在盲腸和結(jié)腸中表達降低。結(jié)果表明,綜合調(diào)控法能夠顯著地調(diào)整瘤胃發(fā)酵類型,升高瘤胃內(nèi)環(huán)境pH值,減輕體內(nèi)炎癥反應(yīng),使動物在相對穩(wěn)定的瘤胃酸性環(huán)境下發(fā)酵產(chǎn)生大量的揮發(fā)性脂肪酸,從而有利于動物實現(xiàn)高產(chǎn),為現(xiàn)代牧業(yè)高精料飼喂方式提出改進的方法。
[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.

【學位授予單位】：南京農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S816

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