【摘要】：本論文利用變性梯度凝膠電泳(DGGE)技術(shù)和實時熒光定量(q PCR)比較分析了飼喂不同中性洗滌纖維(NDF)水平日糧雜交水牛瘤胃細(xì)菌的多樣性和數(shù)量差異,試驗設(shè)計和結(jié)果如下:一、雜交水牛瘤胃微生物的多樣性:為了研究雜交水牛瘤胃內(nèi)與水牛耐粗飼相關(guān)的菌群,本試驗改變?nèi)占Z的營養(yǎng)組分,設(shè)計了3種不同日糧NDF水平(38%、44%、51%)。同時選擇3頭體況相近、健康、裝有永久性瘤胃瘺管的雜交水牛,采用3×3拉丁方試驗設(shè)計,試驗分三期,每一期飼喂21天,最后一天采集瘤胃內(nèi)容物,實驗室提取總DNA,利用DGGE分析瘤胃液相與固相細(xì)菌群落結(jié)構(gòu)。結(jié)果表明,(1)不同NDF水平日糧條件下,雜交水牛瘤胃固相與液相細(xì)菌群落區(qū)系都顯示出了一定的差異性,但差異性并不顯著。(2)在不同NDF水平日糧條件下,擬桿菌門與厚壁菌門均為瘤胃內(nèi)的優(yōu)勢菌群。(3)在雜交水牛瘤胃液相細(xì)菌區(qū)系中,當(dāng)NDF水平為44%時,瘤胃內(nèi)瘤胃球菌屬(Ruminococciis sp.)、解纖維熱酸菌(Acidothermus cellulolyticus)、解纖維梭菌(Clostridium cellulolyticum)與另兩組存在差異。在雜交水牛瘤胃固相細(xì)菌區(qū)系中,當(dāng)NDF水平為38%時,普雷沃氏菌科(Prevotellaceae)較另外兩組有差異。二、不同NDF水平日糧時雜交水牛瘤胃主要功能菌群的數(shù)量變化:通過Q-PCR的方法,分析了飼喂不同NDF水平日糧時雜交水牛瘤胃主要功能菌群的數(shù)量變化情況。結(jié)果表明:不同NDF水平日糧條件下,各組總普雷沃氏菌數(shù)量未產(chǎn)生顯著性差異,但普雷沃氏菌屬細(xì)菌數(shù)量在總細(xì)菌數(shù)量中占優(yōu)勢地位。反芻獸新月行單胞菌受到日糧NDF水平的影響,當(dāng)NDF水平為44%時,數(shù)量最多。纖維降解菌產(chǎn)琥珀酸絲狀桿菌、白色瘤胃球菌和黃色瘤胃球菌數(shù)量在各NDF水平下未產(chǎn)生顯著性差異,但高NDF水平組數(shù)量高于另外兩組,可能是由于日糧差異不夠顯著。不同NDF水平下脂解厭氧弧菌數(shù)量未受到影響。
[Abstract]:In this paper, denaturing gradient gel electrophoresis (DGGE) technique and real-time fluorescence quantitative (q PCR) (q PCR) were used to compare and analyze the diversity and quantity of rumen bacteria in buffalo fed with different neutral detergent fiber (NDF) levels. The results were as follows: 1. Diversity of rumen microbes in crossbred buffalo: in order to study the microflora in the rumen of crossbred buffalo, the nutrient components of diet were changed and three different dietary NDF levels (3844451%) were designed. At the same time, three crossbred buffaloes with similar body condition, healthy and permanent rumen fistula were selected. The experiment was designed by 3 脳 3 Latin square, and the experiment was divided into three stages, each of which was fed for 21 days, and the rumen contents were collected on the last day. Total DNA was extracted from the laboratory, and the structure of liquid and solid bacteria community in rumen was analyzed by DGGE. The results showed that: (1) under different NDF levels, the flora of rumen solid phase and liquid phase bacteria of crossbred buffalo showed some differences, but the differences were not significant. (2) under different NDF levels, the flora of bacteria was different. Bacteroides phylum and musculum thuringiensis were the dominant bacteria in rumen. (3) in the rumen liquid flora of hybrid buffalo, when the NDF level was 44, there was a difference between the two groups in the rumen bacteria, (Ruminococciis sp.), (Ruminococciis sp.), Clostridium cellulolyticum, (Acidothermus cellulolyticus), Clostridium fibrillolyticus, (Clostridium cellulolyticum). When the NDF level was 38, the (Prevotellaceae) of Prevoidae was different from that of the other two groups in the rumen solid phase flora of crossbred buffalo. Secondly, the quantity change of main functional flora in rumen of crossbred buffalo with different NDF level: the quantity change of main functional flora in rumen of crossbred buffalo fed with different NDF level was analyzed by Q-PCR method. The results showed that there was no significant difference in the number of Prevobacter in different diets with different NDF levels, but the number of Prevoia bacteria was dominant in the total number of bacteria. Ruminants were affected by dietary NDF level, and when NDF level was 444.The quantity was the most. There was no significant difference in the number of filamentous succinate bacteria, S. albicans and S. xanthoxylum produced by fibrinolytic bacteria, but the number of high NDF level group was higher than that of the other two groups, which may be due to the lack of significant difference in diet. The number of Vibrio acidolyticus at different NDF levels was not affected.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S823.5

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本文編號：2146288