淬灭酶及沙蒿多糖对绵羊瘤胃微生态和营养代谢及免疫影响研究
发布时间：2019-04-29 06:30
【摘要】：本论文较为全面系统地开展了在不同NFC/NDF日粮中添加淬灭酶及沙蒿多糖对绵羊瘤胃发酵、瘤胃微生态和机体抗氧化能力及其免疫功能调控作用的研究。本研究选用9只健康并安装有永久性瘤胃瘘管的鄂尔多斯细毛羊半同胞羯羊,按体重配对原则分为3组。其中A处理组饲喂NDF/NDF比为0.48日粮,B处理组NDF/NDF比0.76,C处理组NDF/NDF比1.53。首先利用体外产气试验进行添加剂量的筛选,然后采用自身对照试验方法,开展三期动物试验,第一期为对照期,第二期在基础日粮上添加淬灭酶,第三期在基础日粮基础上添加沙蒿多,开展体内添加效果评价。得到结果如下：1.通过体外产气试验筛选淬灭酶和沙蒿多糖的添加量。各处理淬灭酶添加量分别为A：1.0%、B：0.5%、C：1.0%,沙蒿多糖的添加量添加量分别为A：0.3%、B：0.5%、C：0.3%,测定了24小时累积产气量及发酵完成后的pH值、VFA和NH3-N浓度。淬灭酶添加剂量以0.3%水平为宜；A组和C组日粮沙蒿多糖添加1.0%水平较好,B组选择添加0.5%水平。2.对比各组结果pH值可以发现,在晨饲前添加沙蒿多糖的高于对照组,在饲喂后逐渐转变为低于对照组,在多个时间点添加淬灭酶和沙蒿多糖提高了NH3-N浓度；从VFA浓度指标看出,添加淬灭酶及沙蒿多糖后,在前期主要是降低了乙酸和丙酸的浓度,然而在后期,添加淬灭酶和沙蒿多糖组却较对照组提高乙酸丙酸等挥发性脂肪酸浓度的趋势。3.通过16s rDNA高通量测定技术,对瘤胃内细菌16S rRNA进行PCR扩增,得到了瘤胃内细菌组成以及细菌群落多样性变化情况。添加淬灭酶和沙蒿多糖后,降低了B日粮瘤胃内细菌的shannon指数和simpson指数。通过特有OUT分析发现,特有OUT顺序为沙蒿多糖对照组淬灭酶组。通过细菌组成分析发现,厚壁菌门和拟杆菌门是瘤胃中的优势菌群。通过样品间显著性差异分析(Metastats分析)发现,添加淬灭酶增加了A日粮互养菌门细菌比例,降低了B日粮软壁菌门的比例。添加沙蒿多糖增加了B日粮纤维杆菌门菌群比例。通过对种的对比发现,添加淬灭酶减少了弯曲杆菌属、紫单胞菌科细菌相关致病菌的比例,添加沙蒿多糖减少了Desulfo vibrionaceae属、弧菌属(Vibrio)、空肠弯曲菌细菌相关致病菌的比例。4.添加淬灭酶和沙蒿多糖提高了日粮的DM表观消化率,添加沙蒿多糖同时提高了NDF的表观消化率和日粮的氮代谢率,减少了粪N与进食N的比例,减少了B日粮条件下粪氮的排出。5.添加淬灭酶和沙蒿多糖提高了血清内总抗氧化能力(T-AOC)、过氧化氢酶(CAT)的活性,降低了丙二醛(MDA)的含量,添加沙蒿多糖还提高了超氧化物歧化酶(T-SOD)的含量。均提高了血清内总蛋白的含量,添加沙蒿多糖提高了血清中IgA的含量。6.添加淬灭酶减少了羊粪中大肠杆菌的数量,添加淬灭酶和沙蒿多糖均减少了羊粪中葡萄球菌的含量。
[Abstract]:In this paper, the effects of adding quenching enzymes and Artemisia annua polysaccharides to different NFC/NDF diets on rumen fermentation, rumen microecology, anti-oxidation capacity and immune function of sheep were studied systematically. Nine healthy half-sib Capricorn sheep with permanent rumen fistula were selected and divided into three groups according to the principle of body weight matching. The NDF/NDF ratio of A group was 0.48 days, the NDF/NDF ratio of B group was 0.76, and the NDF/NDF ratio of C treatment group was 1.53. First, the dosages were screened by the in vitro gas production test, and then the three stages of animal experiments were carried out with the method of self-control test. The first phase was the control period, the second phase was to add quenching enzymes to the basic diet. In the third stage, more Artemisia annua was added on the basis of basic diet, and the effect of in vivo addition was evaluated. The results are as follows: 1. The addition of quenching enzyme and polysaccharide of Artemisia annua was screened by gas production test in vitro. The contents of quenching enzyme were 1.0%, 0.5%, 1.0%, 0.3%, 0.5%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3% and 0.3%, respectively. The 24-hour cumulative gas production, pH value, VFA and NH3-N concentration after fermentation were determined. The optimal dosage of quenching enzyme was 0.3%, the dietary polysaccharide of Artemisia annua was better in group A and C, and 0.5% in group B, and 0.5% in group A and C, respectively. Compared with the pH value of each group, the addition of Artemisia annua polysaccharide before morning feeding was higher than that of the control group, and gradually changed to lower than that of the control group after feeding. The concentration of NH3-N was increased by adding quenching enzyme and polysaccharide of Artemisia annua at several time points. According to the VFA concentration index, the concentration of acetic acid and propionic acid decreased mainly in the early stage after adding quenching enzyme and polysaccharide of Artemisia annua, however, in the later stage, the concentration of acetic acid and propionic acid was decreased. The trend of increasing the concentration of volatile fatty acids such as acetic acid and propionic acid in the treatment group with quenching enzyme and Artemisia annua polysaccharide was higher than that in the control group. The bacterial composition and the diversity of bacterial community in rumen were obtained by PCR amplification of bacterial 16s rRNA by 16s rDNA high throughput assay. The shannon index and simpson index of bacteria in rumen of B diet were decreased by adding quenching enzyme and polysaccharide of Artemisia annua. Specific OUT analysis showed that the specific OUT sequence was the quenching enzyme group of Artemisia annua polysaccharide control group. Through the analysis of bacterial composition, it was found that thick-walled bacteria and Bacteroides were the dominant bacteria in rumen. The significant difference between samples (Metastats analysis) showed that the addition of quenching enzyme increased the proportion of bacteria in A diet and decreased the proportion of soft wall bacteria in B diet. The addition of Artemisia annua polysaccharide increased the proportion of fibrobacilli flora in B diet. Through the comparison of species, it was found that the addition of quenching enzyme reduced the proportion of bacteria related to Campylobacter, Vibrio and Artemisia polysaccharide decreased the proportion of bacteria related to Campylobacter, Vibrio (Vibrio), and Artemisia annua polysaccharide. Percentage of bacteria-related pathogens in Campylobacter jejuni. 4. The addition of quenching enzyme and polysaccharide of Artemisia annua increased the apparent digestibility of DM, increased the apparent digestibility of NDF and nitrogen metabolism, and decreased the ratio of fecal N to dietary N. Reduced the excretion of fecal nitrogen under B diet. 5. The addition of quenching enzyme and polysaccharide of Artemisia annua increased serum total antioxidant capacity (T-AOC), catalase (CAT) activity, and decreased the content of malondialdehyde (MDA). The addition of Artemisia annua polysaccharide also increased the content of superoxide dismutase (T-SOD). The content of total protein in serum was increased, and the content of IgA in serum was increased by adding polysaccharide of Artemisia annua. The amount of Escherichia coli in sheep dung was decreased by adding quenching enzyme and polysaccharide from Artemisia annua decreased the content of staphylococci in sheep dung.
【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S826.5
本文编号：2468018
[Abstract]:In this paper, the effects of adding quenching enzymes and Artemisia annua polysaccharides to different NFC/NDF diets on rumen fermentation, rumen microecology, anti-oxidation capacity and immune function of sheep were studied systematically. Nine healthy half-sib Capricorn sheep with permanent rumen fistula were selected and divided into three groups according to the principle of body weight matching. The NDF/NDF ratio of A group was 0.48 days, the NDF/NDF ratio of B group was 0.76, and the NDF/NDF ratio of C treatment group was 1.53. First, the dosages were screened by the in vitro gas production test, and then the three stages of animal experiments were carried out with the method of self-control test. The first phase was the control period, the second phase was to add quenching enzymes to the basic diet. In the third stage, more Artemisia annua was added on the basis of basic diet, and the effect of in vivo addition was evaluated. The results are as follows: 1. The addition of quenching enzyme and polysaccharide of Artemisia annua was screened by gas production test in vitro. The contents of quenching enzyme were 1.0%, 0.5%, 1.0%, 0.3%, 0.5%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3%, 0.3% and 0.3%, respectively. The 24-hour cumulative gas production, pH value, VFA and NH3-N concentration after fermentation were determined. The optimal dosage of quenching enzyme was 0.3%, the dietary polysaccharide of Artemisia annua was better in group A and C, and 0.5% in group B, and 0.5% in group A and C, respectively. Compared with the pH value of each group, the addition of Artemisia annua polysaccharide before morning feeding was higher than that of the control group, and gradually changed to lower than that of the control group after feeding. The concentration of NH3-N was increased by adding quenching enzyme and polysaccharide of Artemisia annua at several time points. According to the VFA concentration index, the concentration of acetic acid and propionic acid decreased mainly in the early stage after adding quenching enzyme and polysaccharide of Artemisia annua, however, in the later stage, the concentration of acetic acid and propionic acid was decreased. The trend of increasing the concentration of volatile fatty acids such as acetic acid and propionic acid in the treatment group with quenching enzyme and Artemisia annua polysaccharide was higher than that in the control group. The bacterial composition and the diversity of bacterial community in rumen were obtained by PCR amplification of bacterial 16s rRNA by 16s rDNA high throughput assay. The shannon index and simpson index of bacteria in rumen of B diet were decreased by adding quenching enzyme and polysaccharide of Artemisia annua. Specific OUT analysis showed that the specific OUT sequence was the quenching enzyme group of Artemisia annua polysaccharide control group. Through the analysis of bacterial composition, it was found that thick-walled bacteria and Bacteroides were the dominant bacteria in rumen. The significant difference between samples (Metastats analysis) showed that the addition of quenching enzyme increased the proportion of bacteria in A diet and decreased the proportion of soft wall bacteria in B diet. The addition of Artemisia annua polysaccharide increased the proportion of fibrobacilli flora in B diet. Through the comparison of species, it was found that the addition of quenching enzyme reduced the proportion of bacteria related to Campylobacter, Vibrio and Artemisia polysaccharide decreased the proportion of bacteria related to Campylobacter, Vibrio (Vibrio), and Artemisia annua polysaccharide. Percentage of bacteria-related pathogens in Campylobacter jejuni. 4. The addition of quenching enzyme and polysaccharide of Artemisia annua increased the apparent digestibility of DM, increased the apparent digestibility of NDF and nitrogen metabolism, and decreased the ratio of fecal N to dietary N. Reduced the excretion of fecal nitrogen under B diet. 5. The addition of quenching enzyme and polysaccharide of Artemisia annua increased serum total antioxidant capacity (T-AOC), catalase (CAT) activity, and decreased the content of malondialdehyde (MDA). The addition of Artemisia annua polysaccharide also increased the content of superoxide dismutase (T-SOD). The content of total protein in serum was increased, and the content of IgA in serum was increased by adding polysaccharide of Artemisia annua. The amount of Escherichia coli in sheep dung was decreased by adding quenching enzyme and polysaccharide from Artemisia annua decreased the content of staphylococci in sheep dung.
【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S826.5
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