本文關(guān)鍵詞： 綿羊瘤胃 支鏈氨基酸 組合效應(yīng) 消化代謝 微生物多樣性　出處：《新疆農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：本論文通過3個(gè)試驗(yàn)研究了不同支鏈氨基酸組合對(duì)瘤胃發(fā)酵參數(shù)及微生物種群區(qū)系的影響。明確了在不同精粗飼料比條件下BCAA(Val、Leu和Ile)的最佳適宜添加比例與組合效應(yīng),闡明了BCAA對(duì)結(jié)構(gòu)性碳水化合物在綿羊瘤胃消化代謝和整體消化的影響;揭示BCAA組合對(duì)瘤胃微生物多樣性和纖維降解菌生長的影響。試驗(yàn)一:本試驗(yàn)利用體外發(fā)酵培養(yǎng)技術(shù)和正交試驗(yàn)設(shè)計(jì),在飼料精粗比20:80、40:60和60:40的飼喂條件下,研究額外添加BCAA的16種組合對(duì)綿羊瘤胃微生物體外發(fā)酵的影響。結(jié)果表明:底物精粗飼料比20:80時(shí),異丁酸和戊酸濃度隨著Val水平的增加而呈顯著升高(P0.05);異戊酸濃度隨著Ile添加水平的增加而增大。底物精粗飼料比40:60時(shí),異丁酸濃度隨著Val添加水平的增加而升高,戊酸濃度隨著Leu、Ile添加水平的升高而有不同程度的升高,當(dāng)Val添加水平為0.67 mmol/L時(shí),異戊酸濃度最低。底物精粗飼料比60:40時(shí),異丁酸主要受Val的影響,隨著Val水平的增加異丁酸濃度顯著增加;異戊酸濃度受Leu和Ile的影響,戊酸濃度受Val和Ile的影響。3種不同發(fā)酵底物中添加BCAA組合對(duì)pH、NDF降解和乙酸/丙酸均無顯著影響。試驗(yàn)二:在試驗(yàn)一的基礎(chǔ)上,飼料精粗比20:80的飼喂水平條件下采用4*4拉丁方試驗(yàn)設(shè)計(jì),研究Val、Leu、Ile添加水平0.67、0、0.67 mmol/L(組合1),0.67、1.33、2.00 mmol/L(組合2),0.67、2、1.33 mmol/L(組合3)對(duì)綿羊瘤胃發(fā)酵各參數(shù)及營養(yǎng)物質(zhì)消化代謝的影響。結(jié)果表明:3種組合BCAA的添加對(duì)乙酸、丙酸、丁酸、TVFA、pH及其他營養(yǎng)物質(zhì)差異均不顯著(P0.05)。組合2的添加使異丁酸、異戊酸產(chǎn)量最大,顯著高于對(duì)照組(P0.05),使P保留率、NH3-N濃度在各個(gè)采樣時(shí)間均高于其他組。組合3的添加使NDF、TVFA日均濃度最大,顯著高于對(duì)照組(P0.05)。試驗(yàn)三:在試驗(yàn)二的基礎(chǔ)上,利用PCR-DGGE技術(shù)、高通量測(cè)序技術(shù)分析添加BCAA組合對(duì)瘤胃微生物區(qū)系的影響,結(jié)果表明:添加BCAA組合改變了瘤胃微生物的種類、數(shù)量及群落結(jié)構(gòu)的構(gòu)成,對(duì)細(xì)菌中Victivallis、Bacteroides、Butyrivibrio,真菌中unclassified Eurotiales sp.的豐度影響差異顯著,3種不同組合BCAA的添加均有提高綿羊Succiniclasticum豐度的趨勢(shì)。當(dāng)Val、Leu、Ile添加水平為0.67、2.00、1.33 mmol/L時(shí),丁酸弧菌屬(Butyrivibrio)的豐度提高了34.88%(P0.05)。綜上所述:在飼料精粗比20:80的飼喂水平下,額外添加Val、Leu、Ile的濃度為0.67、2、1.33 mmol/L時(shí),在體外使DM、NDF降解率達(dá)到最大;在體內(nèi),通過提高丁酸弧菌屬、降低食物谷菌屬、擬桿菌屬的相對(duì)豐度改變了瘤胃微生物的多樣性。
[Abstract]:Three experiments were conducted to study the effects of different branched amino acid combinations on rumen fermentation parameters and microbial population floras. BCAA(Val was determined under different feed ratios. The optimum proportion and combination effect of Leu and Ile were used to elucidate the effects of BCAA on the metabolism of structural carbohydrates in rumen digestion and overall digestion of sheep. The effects of BCAA combination on rumen microbial diversity and cellulolytic bacteria growth were revealed. Experiment 1: in this experiment, the in vitro fermentation culture technique and orthogonal design were used to determine the feed concentrate / diameter ratio of 20: 80. Under the feeding conditions of 40: 60 and 60: 40, the effects of 16 additional combinations of BCAA on rumen microbial fermentation in vitro in sheep were studied. The results showed that the crude feed ratio of substrate concentrate was 20: 80. The concentrations of isobutyric acid and valeric acid increased significantly with the increase of Val level. The concentration of isovaleric acid increased with the increase of Ile addition level, and the concentration of isobutyric acid increased with the increase of Val addition level, while the concentration of valeric acid increased with Leu at the ratio of 40: 60. When the level of Val was 0.67 mmol/L, the concentration of isovaleric acid was the lowest, and the crude feed ratio of substrate concentrate was 60: 40. Isobutyric acid was mainly affected by Val, and the concentration of isobutyric acid increased significantly with the increase of Val level. The concentration of isovaleric acid was affected by Leu and Ile, and the concentration of valeric acid was affected by Val and Ile. 3 kinds of different fermentative substrates were mixed with BCAA on pH. NDF degradation and acetic acid / propionic acid were not significantly affected. Test 2: on the basis of experiment 1, the Val was studied under the feed level of 20: 80 feed concentrate and feed ratio of 20: 80. Leulein Ile added 0.67 mmol / L (0.67 mmol / L) (combination 1, 0.671.33, 2.00 mmol / L). The effects of 1.33 mmol / L (combination 3) on the parameters of rumen fermentation and nutrient digestion and metabolism in sheep were studied. The results showed that the addition of BCAA to the three combinations had effect on acetic acid, propionic acid and butyrate. The difference of pH and other nutrients was not significant (P 0.05). The yield of isobutyric acid and isovaleric acid was the highest in combination 2, which was significantly higher than that in control group (P 0.05), and the retention rate of P was increased. The concentration of NH3-N in each sampling time was higher than that in other groups, and the daily average concentration of NH3-N was the highest with the addition of combination 3. Test 3: on the basis of experiment 2, the effects of adding BCAA combination on rumen microbial flora were analyzed by PCR-DGGE technique and high throughput sequencing technique. The results showed that the addition of BCAA changed the species, quantity and community structure of rumen microorganisms. The abundance of unclassified Eurotiales sp. was significantly different from that of Butyrivibrio. The Succiniclasticum abundance of sheep was increased with the addition of three different combinations of BCAA, when the addition level of BCAA was 0.67g / 2.00. 1.33 mmol/L. The abundance of Vibrio butyricum (Butyrivibrio) increased 34.88% P0.050.In conclusion, Val was added to the feed at the feed level of 20: 80. When the concentration of Leufen Ile was 0.67 mmol/L, the degradation rate of DM-NDF reached the maximum in vitro. In vivo, the relative abundance of Bacteroides changed the diversity of rumen microorganisms by increasing the genus Vibrio butyrate and reducing the relative abundance of the genus Glutazia.
【學(xué)位授予單位】：新疆農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S826.5

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