淬滅酶及沙蒿多糖對綿羊瘤胃微生態(tài)和營養(yǎng)代謝及免疫影響研究
發(fā)布時間：2019-04-29 06:30
【摘要】：本論文較為全面系統(tǒng)地開展了在不同NFC/NDF日糧中添加淬滅酶及沙蒿多糖對綿羊瘤胃發(fā)酵、瘤胃微生態(tài)和機體抗氧化能力及其免疫功能調(diào)控作用的研究。本研究選用9只健康并安裝有永久性瘤胃瘺管的鄂爾多斯細毛羊半同胞羯羊,按體重配對原則分為3組。其中A處理組飼喂NDF/NDF比為0.48日糧,B處理組NDF/NDF比0.76,C處理組NDF/NDF比1.53。首先利用體外產(chǎn)氣試驗進行添加劑量的篩選,然后采用自身對照試驗方法,開展三期動物試驗,第一期為對照期,第二期在基礎日糧上添加淬滅酶,第三期在基礎日糧基礎上添加沙蒿多,開展體內(nèi)添加效果評價。得到結果如下：1.通過體外產(chǎn)氣試驗篩選淬滅酶和沙蒿多糖的添加量。各處理淬滅酶添加量分別為A：1.0%、B：0.5%、C：1.0%,沙蒿多糖的添加量添加量分別為A：0.3%、B：0.5%、C：0.3%,測定了24小時累積產(chǎn)氣量及發(fā)酵完成后的pH值、VFA和NH3-N濃度。淬滅酶添加劑量以0.3%水平為宜；A組和C組日糧沙蒿多糖添加1.0%水平較好,B組選擇添加0.5%水平。2.對比各組結果pH值可以發(fā)現(xiàn),在晨飼前添加沙蒿多糖的高于對照組,在飼喂后逐漸轉(zhuǎn)變?yōu)榈陀趯φ战M,在多個時間點添加淬滅酶和沙蒿多糖提高了NH3-N濃度；從VFA濃度指標看出,添加淬滅酶及沙蒿多糖后,在前期主要是降低了乙酸和丙酸的濃度,然而在后期,添加淬滅酶和沙蒿多糖組卻較對照組提高乙酸丙酸等揮發(fā)性脂肪酸濃度的趨勢。3.通過16s rDNA高通量測定技術,對瘤胃內(nèi)細菌16S rRNA進行PCR擴增,得到了瘤胃內(nèi)細菌組成以及細菌群落多樣性變化情況。添加淬滅酶和沙蒿多糖后,降低了B日糧瘤胃內(nèi)細菌的shannon指數(shù)和simpson指數(shù)。通過特有OUT分析發(fā)現(xiàn),特有OUT順序為沙蒿多糖對照組淬滅酶組。通過細菌組成分析發(fā)現(xiàn),厚壁菌門和擬桿菌門是瘤胃中的優(yōu)勢菌群。通過樣品間顯著性差異分析(Metastats分析)發(fā)現(xiàn),添加淬滅酶增加了A日糧互養(yǎng)菌門細菌比例,降低了B日糧軟壁菌門的比例。添加沙蒿多糖增加了B日糧纖維桿菌門菌群比例。通過對種的對比發(fā)現(xiàn),添加淬滅酶減少了彎曲桿菌屬、紫單胞菌科細菌相關致病菌的比例,添加沙蒿多糖減少了Desulfo vibrionaceae屬、弧菌屬(Vibrio)、空腸彎曲菌細菌相關致病菌的比例。4.添加淬滅酶和沙蒿多糖提高了日糧的DM表觀消化率,添加沙蒿多糖同時提高了NDF的表觀消化率和日糧的氮代謝率,減少了糞N與進食N的比例,減少了B日糧條件下糞氮的排出。5.添加淬滅酶和沙蒿多糖提高了血清內(nèi)總抗氧化能力(T-AOC)、過氧化氫酶(CAT)的活性,降低了丙二醛(MDA)的含量,添加沙蒿多糖還提高了超氧化物歧化酶(T-SOD)的含量。均提高了血清內(nèi)總蛋白的含量,添加沙蒿多糖提高了血清中IgA的含量。6.添加淬滅酶減少了羊糞中大腸桿菌的數(shù)量,添加淬滅酶和沙蒿多糖均減少了羊糞中葡萄球菌的含量。
[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.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S826.5
本文編號：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.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S826.5
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