【摘要】：奶牛機(jī)體的抗氧化能力與機(jī)體的健康水平息息相關(guān),處于泌乳早期的高產(chǎn)奶牛易發(fā)生氧化應(yīng)激,引起與氧化應(yīng)激相關(guān)的疾病。提升奶牛機(jī)體的營養(yǎng)水平及監(jiān)測(cè)機(jī)體的健康狀況,對(duì)減少疾病的發(fā)生和疾病監(jiān)測(cè)具有重要價(jià)值。葡萄籽原花青素(GSPE)屬于多酚化合物,具有很強(qiáng)的抗氧化特性,作為奶牛飼料添加物鮮有報(bào)道。本研究進(jìn)行三項(xiàng)試驗(yàn),試驗(yàn)一探究GSPE對(duì)泌乳早期奶牛血漿抗氧化能力和酮病等指標(biāo)的影響;試驗(yàn)二了解GSPE對(duì)牛奶抗氧化能力的影響,綜合評(píng)價(jià)其對(duì)奶牛機(jī)體的作用;試驗(yàn)三篩選評(píng)價(jià)奶牛機(jī)體抗氧化狀況的乳樣指標(biāo)。具體研究如下:試驗(yàn)一:葡萄籽原花青素對(duì)奶牛血漿抗氧化能力和酮病等指標(biāo)的影響試驗(yàn)選擇16頭無臨床疾病,泌乳天數(shù)、產(chǎn)奶量、體重、胎次、體況評(píng)分相近的泌乳早期奶牛,按完全隨機(jī)設(shè)計(jì)分為4組(G0、G20、G40、G80),每組隨機(jī)接受1個(gè)試驗(yàn)處理,即飼喂0、20、40和80mg/kgBW/day的GSPE。于 Oh(灌喂前)、3h、6h、12h、24h、48h、72h、96h、120h、144h 和 168h采集血樣,測(cè)定下列氧化/抗氧化指標(biāo):總抗氧化能力TAC(血漿鐵還原力法)、谷胱甘肽過氧化物酶GPX(DTNB法)、超氧化物歧化酶SOD(羥胺法)、丙二醛MDA(硫代巴比妥酸法),過氧化氫H2O2(鉬酸顯色法)。檢測(cè)酮病等指標(biāo):葡萄糖GLU(GOP-PAP法-雙劑型)、游離脂肪酸NEFA(可見光比色法)及酮體KET(改良水楊醛比色法)。結(jié)果顯示,與GO組相比,G20、G40、G80在血漿氧化抗氧化方面TAC、GPX、SOD、MDA和H2O2均未產(chǎn)生顯著影響(P0.05)。在代謝變化方面,G20、G40、G80組的GLU含量均值均高于GO組,在“試驗(yàn)時(shí)間×分組”效應(yīng)上,飼喂GSPE對(duì)血漿GLU產(chǎn)生極顯著的影響(P0.01),且G40組的效果優(yōu)于G20、G80;G40相對(duì)于GO組NEFA含量在“試驗(yàn)時(shí)間×分組”效應(yīng)上在生理濃度范圍內(nèi)顯著升高(P0.05);飼喂GSPE對(duì)酮體未產(chǎn)生顯著影響(P0.05)。結(jié)果表明,飼喂GSPE可提高泌乳早期奶牛機(jī)體的血漿葡萄糖水平,對(duì)于減少泌乳早期奶牛能量負(fù)平衡的發(fā)生具有積極作用,以40mg/kg BW/day的GSPE效果較好。試驗(yàn)二:葡萄籽原花青素對(duì)奶牛乳汁抗氧化能力和酮體含量的影響選擇48頭泌乳早期奶牛,按完全隨機(jī)設(shè)計(jì)分為4組:G0、G20、G40、G80。對(duì)照組(GO)飼喂原奶牛場(chǎng)的基礎(chǔ)日糧;G20、G40和G80組,分別在基礎(chǔ)日糧中添加20、40和80mg/kg BW/day的GSPE。試驗(yàn)期42 d,GSPE各劑量組每天上午灌喂一次GSPE溶液,GO組同時(shí)灌喂一次同等劑量的牛場(chǎng)飲用水,并于Od(灌喂前),7d,14d,21d,28d,35d,42d常規(guī)采集奶樣分離乳清,測(cè)定TAC、GPX、H2O2及KET指標(biāo)。結(jié)果顯示,G0、G20、G40和G80四組奶牛乳清TAC、GPX、H202在分組效應(yīng)上,G40組奶牛的乳清GPX活力極顯著的高于G0組(P0.01),且GSPE各劑量組在試驗(yàn)期間乳清GPX活力均高于G0組;乳清TAC、H2O2在GSPE各劑量組與G0組間差異不顯著(P0.05),但相對(duì)于G0、G20、G80組,G40組的乳清TAC較高,乳清H202含量較低。從奶牛乳汁抗氧化狀態(tài)變化上分析,40mgGSPE/kgBW/day的劑量,在改善奶牛乳汁抗氧化性能方面的效果更好。在代謝變化方面,酮體水平在GSPE各劑量組與G0組間均無顯著差異(P0.05)。研究表明,在泌乳早期奶牛日糧中添加GSPE對(duì)乳汁抗氧化性能方面無負(fù)面影響,且40mg/kg BW/day的GSPE可改善乳汁GPX活力。試驗(yàn)三:奶牛血漿與乳清中抗氧化及酮體指標(biāo)的含量及相關(guān)性分析本試驗(yàn)選取南寧市某奶牛場(chǎng)48頭泌乳早期健康奶牛,連續(xù)采樣7周,于每周末清晨空腹采集血樣和奶樣,并及時(shí)制備血漿和乳清,分別獲得336份樣品。測(cè)定TAC、GPX、H2O2、KET水平。結(jié)果顯示:乳清和血漿的TAC 含量分別為 6.89±1.96U/mL 和 4.97±1.99U/mL,GPX 分別為38.96±18.23U/mL 和 122.17±64.43U/mL,H2O2 分別為 7.78±2.97mmol/L 和16.80±5.31mmol/L,KET 分別為 8.54±2.56mg/dL 和 3.25±1.48mg/dL,其差異均達(dá)極顯著水平(P0.01)。乳清與血漿GPX呈極顯著正相關(guān)(r=0.552,P0.01)、TAC呈顯著正相關(guān)(r=0.113,P0.05)、H2O2呈顯著正相關(guān)(r=0.124,P0.05)。健康奶牛乳清與血漿KET無顯著相關(guān)性(r=0.058,P0.05),但乳清 KET 與血漿 TAC 顯著負(fù)相關(guān)(r=-0.122,P0.05),與血漿GPX極顯著負(fù)相關(guān)(r=-0.326,P0.01)。結(jié)果表明,乳清TAC、H2O2、KET可作為評(píng)估奶牛機(jī)體健康狀況的備選指標(biāo);乳清GPX可作為奶牛GPX普檢中的評(píng)價(jià)指標(biāo)。綜上所述:日糧中添加GSPE,對(duì)酮體水平?jīng)]有顯著影響,可以提高泌乳早期奶牛機(jī)體的血漿葡萄糖水平,改善乳汁GPX活力,其中以40mg/k BW/day的GSPE添加效果較好。檢測(cè)乳清GPX活力,可作為普查與監(jiān)控泌乳早期奶牛健康與GPX水平的參考方法。
[Abstract]:The antioxidant capacity of dairy cows is closely related to the health level of the body. High yield cows in early lactation are prone to oxidative stress and cause diseases related to oxidative stress. It is of great value to improve the nutrition level and monitor the health status of the body, and to reduce the occurrence of disease and disease monitoring. GSPE, which belongs to polyphenols, has strong antioxidant properties and is rarely reported as an additive for dairy cattle feed. Three experiments were conducted in this study to explore the effects of GSPE on plasma antioxidant capacity and ketosis in early lactating cows. Test two to understand the effect of GSPE on the antioxidant capacity of milk, and to evaluate its effect on dairy cows. Effect of the body; test three screening and evaluation of milk samples for the antioxidant status of dairy cows. Experiment 1: Experiment 1: Test on the effects of grape seed proanthocyanidins on the antioxidant capacity and ketosis of dairy cows, select 16 early lactating cows with no clinical diseases, lactation days, milk production, body weight, birth weight, and body condition. Complete random design was divided into 4 groups (G0, G20, G40, G80). Each group was randomized to receive 1 experimental treatments, namely, GSPE. in Oh (before feeding) and Oh (before feeding), 3h, 6h, 12h, 24h, etc., to determine the following oxidation / antioxygenation indexes: Total antioxidant capacity (plasma iron reduction force), glutathione peroxide Enzyme GPX (DTNB), superoxide dismutase SOD (hydroxylamine method), malondialdehyde MDA (thiobarbituric acid method), hydrogen peroxide H2O2 (molybdate chromogenic method). Detection of ketosis, such as glucose GLU (GOP-PAP method - double dosage form), free fatty acid NEFA (visible light colorimetric method) and ketone body KET (modified salicylaldehyde colorimetric method). The results show that G20, G40, and G40 are compared with GO group. TAC, GPX, SOD, MDA and H2O2 were not significantly affected in plasma oxidation resistance (P0.05). In the metabolic changes, the mean value of GLU content in G20, G40 and G80 groups was higher than that in the GO group. The amount of GSPE was significantly increased in the physiological concentration range (P0.05) in the "test time x group" effect (P0.05). The results showed that feeding GSPE could improve the plasma glucose level of the early milk cow body, and had a positive effect on reducing the energy negative balance of milk cows in early lactation, and 40mg/kg B The effect of GSPE in W/day was better. Experiment two: the effect of grape seed proanthocyanidins on the antioxidant capacity and the content of ketone body in dairy milk selected 48 early lactating cows and divided into 4 groups according to the complete random design: G0, G20, G40, G80. control group (GO) fed the basic diet of raw milk cattle; G20, G40 and G80 groups were added to basal diet to add 20,40 and 80mg/kg The GSPE. test period of /day was 42 d, and each dose group of GSPE was fed a GSPE solution every morning. The GO group was fed with the same dose of drinking water at the same time, and Od (before feeding), 7d, 14d, 21d, 28d, and 35d. In group G40, the GPX activity of dairy cow's whey was significantly higher than that of G0 group (P0.01), and the GPX activity of whey in each dose group of GSPE was higher than that of the G0 group during the experiment, and the difference between the whey TAC and H2O2 was not significant (P0.05) between the GSPE dosage groups and the G0 group (P0.05). The effect of 40mgGSPE/kgBW/day was better in improving the antioxidant activity of dairy milk. There was no significant difference in the level of ketone body between the GSPE dose group and the G0 group (P0.05) in the metabolic change (P0.05). The study showed that the addition of GSPE to the milk of early lactating dairy cows had no negative effect on the antioxidant activity of milk. Effect, and the GSPE of 40mg/kg BW/day can improve the activity of milk GPX. Test three: the content and correlation analysis of antioxidant and ketone body in dairy cow plasma and whey. The experiment selected 48 early lactating healthy cows in a dairy farm of Nanning city for 7 weeks, collecting blood samples and milk samples on the empty abdomen on the morning of every morning, and preparing plasma and milk in time. The levels of TAC, GPX, H2O2, and KET were measured in 336 samples respectively. The results showed that the TAC content in whey and plasma were 6.89 + 1.96U/mL and 4.97 + 1.99U/mL respectively, GPX was 38.96 + 18.23U/mL and 122.17 + 64.43U/mL respectively, and H2O2 was 7.78 + 2.97mmol/L and 16.80 + respectively, respectively, 8.54 and 3.25 + respectively, respectively. The difference was very significant (P0.01). There was a very significant positive correlation between whey and plasma GPX (r=0.552, P0.01), TAC showed significant positive correlation (r=0.113, P0.05), H2O2 showed significant positive correlation (r=0.124, P0.05). The plasma GPX was significantly negatively correlated (r=-0.326, P0.01). The results showed that whey TAC, H2O2, KET could be used as an alternative index for evaluating the health of dairy cows; whey GPX could be used as an evaluation index in dairy cow GPX examination. Sugar level can improve the activity of milk GPX, and the effect of 40mg/k BW/day GSPE is better. The detection of GPX activity in whey can be used as a reference method for screening and monitoring the health and GPX level of dairy cows in early lactation.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S858.23

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