本文選題：奶牛 + 熱應(yīng)激�。� 參考：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：本研究旨在研究熱應(yīng)激對(duì)泌乳奶牛泌乳活動(dòng)造成的影響,并對(duì)這些影響的機(jī)制進(jìn)行初步研究。研究包括以下三個(gè)方面。1、高溫氣候環(huán)境對(duì)泌乳奶牛機(jī)體代謝和泌乳性能的影響選取6頭相同泌乳期的荷斯坦奶牛,正常飼喂和飲水。實(shí)驗(yàn)期從6月29日至8月5日,共計(jì)35天。每日10:00和18:00記錄氣溫,統(tǒng)計(jì)產(chǎn)奶量,每周取全天奶樣檢測(cè)乳蛋白、乳脂肪和乳糖含量。于實(shí)驗(yàn)開始第1周和最后1周采集頸靜脈血液檢測(cè)相關(guān)生化指標(biāo)和血液中相關(guān)激素的含量。結(jié)果表明:隨著環(huán)境氣溫由26 ℃升高至38 ℃,實(shí)驗(yàn)?zāi)膛＞a(chǎn)奶量由15.2kg/d下降至10kg/d;乳蛋白產(chǎn)量由0.47kg/d下降至0.30 kg/d,乳脂肪產(chǎn)量由0.46kg/d下降至0.28kg/d,乳糖產(chǎn)量由0.71kg/d降至0.46kg/d。血液中 TP(94.18±0.35115.07±1.04g/L,p=0.049),球蛋白(69.58±0.2088.95±1.10g/L,p=0.045)和葡萄糖(2.88±0.043.54±0.11 mmol/L,p=0.03)含量均顯著升高;LDL-C(0.18±0.030.29±0.01mmol/L,p=0.004)和 AKT(4.17±0.3135.50±4.15 U/L,p=0.004)極顯著升高;HDL-C 顯著降低(3.23±0.082.20±0.04 mmol/L,p=0.033)。血液中 GH(1.19±0.031.54±0.06ng/mL,p=0.037)和 IGF-I(80.05±10.62-100.72 ± 21.78ng/mL,p=0.048)含量顯著升高;皮質(zhì)醇(288.21 ±49.73573.76 ±11.71ng/mL,p=0.009)含量極顯著升高,胰島素(22.48±2.5325.66±1.95IU/L,p=0.057)和胰高血糖素(597.18±58.11658.02±49.47pg/mL,p=0.06)濃度均有升高。結(jié)果提示:在中國(guó)南京夏季高溫高濕條件下,可誘發(fā)奶牛發(fā)生熱應(yīng)激,奶牛內(nèi)分泌和營(yíng)養(yǎng)代謝均發(fā)生變化,表現(xiàn)泌乳量減少,乳品質(zhì)降低。2、熱應(yīng)激時(shí)乳成分前體物重分配變化與乳品質(zhì)降低的關(guān)系研究通過探討血液中氨基酸和游離脂肪酸等"乳成分前體物"與乳脂肪、乳蛋白合成之間的關(guān)系,揭示熱應(yīng)激時(shí)泌乳奶牛乳品質(zhì)降低的可能機(jī)制。實(shí)驗(yàn)動(dòng)物飼養(yǎng)與處理同第三章。實(shí)驗(yàn)開始和結(jié)束時(shí)采集頸靜脈血液,RP-HPLC和ELISA發(fā)分別檢測(cè)血液中游離氨基酸和游離氨基酸含量。結(jié)果表明:隨著氣溫的升高(由26 ＞升高至最高38 ＞),乳脂肪、乳蛋白含量降低,實(shí)驗(yàn)?zāi)膛Ｑ褐锌侼EFA顯著升高(178.20±16.70272.42±20.53μg/mL,p=0.025);Glu、Asp、Gly 和 Val 等主要的生糖氨基酸(811.28±259.223254.05±666.79μmol/L,p=0.035)以及 Leu、Ile 等生酮氨基酸均升高或顯著升高(169.26±32.27502.76±105.50 μmol/L,p=0.046);總支鏈氨基酸(Val、lle、Leu)含量顯著增加(198.36±52.571474.85±355.83μmol/L,p=0.02870.05),在必需氨基酸中的所占比例由54.75%增至67.89%。結(jié)論:在夏季高溫奶牛熱應(yīng)激狀態(tài)下,乳脂肪、乳蛋白含量均降低,但血液中NEFA,游離氨基酸,特別是支鏈氨基酸含量顯著升高。提示熱應(yīng)激時(shí)血液中高水平的NEFA和游離氨基酸并未完全用于乳脂肪和乳蛋白的合成,還發(fā)揮了除此之外的供能、糖異生及免疫反應(yīng)等其他作用,而后者的作用優(yōu)先于參與乳脂肪、乳蛋白的合成。熱應(yīng)激時(shí)乳成分前體物的重分配變化可能是導(dǎo)致乳蛋白、乳脂肪下降的機(jī)制之一。3、熱應(yīng)激致奶牛乳成分前體物發(fā)生重分配機(jī)制初探在前兩章的實(shí)驗(yàn)中,我們發(fā)現(xiàn)熱應(yīng)激可以引起奶牛乳蛋白和乳脂肪的前體物發(fā)生重分配,并引起牛奶中乳蛋白和乳脂肪含量降低。為了探究乳蛋白和乳脂肪前體物重分配引起乳成分含量降低的機(jī)制,本部分實(shí)驗(yàn)對(duì)實(shí)驗(yàn)?zāi)膛＿M(jìn)行了肝臟樣品組織采集。采用Real-time PCR方法,對(duì)奶牛肝臟內(nèi)糖異生關(guān)鍵酶PC和PEPCK基因,肝臟組織中 GHR、IGFR,HSP70 及 Keap1-Nrf2 通路及其下游 HO1,GCLM,NQO1,GCLC 等蛋白進(jìn)行了 mRNA表達(dá)檢測(cè)。結(jié)果發(fā)現(xiàn):在熱應(yīng)激時(shí),奶牛肝臟內(nèi)PC(0.917±0.0081.08±0.025,p=0.045)和PEPCK(0.87±0.0091.29±0.132,p=0.032)表達(dá)水平顯著升高;GHR(1.05±0.121.39±0.02,p=0.007)表達(dá)水平極顯著上調(diào),IGFR(1.07±0.051.28±0.018,p=0.036)顯著上調(diào);肝臟中HSP70(0.35±0.1651.55±0.012,?=0.006)表達(dá)極顯著增高;Keap1(1.07±0.241.32±0.27,p=0.046)和Nrf2(1.59±0.181.73±0.19,p=0.049)基因表達(dá)顯著升高,其下游4個(gè)基因中,HOl(0.98±0.081.19±0.02,p=0.037)和NQO(0.92±0.081.43±0.02,p=0.026)表達(dá)顯著提高,GCLM(0.69±0.100.97±0.05,p=0.059)和GCLC(0.84±0.071.29±0.11,p=0.061)表達(dá)有升高,但差異不顯著。結(jié)果提示:熱應(yīng)激時(shí)肝臟中糖異生作用和抗應(yīng)激作用均增強(qiáng),血液中升高的乳蛋白、乳脂肪前體物在GH-IGF-I軸和Keap1-Nrf2-ARE信號(hào)通路的介導(dǎo)下更多地參與了糖異生過程和抵抗細(xì)胞的應(yīng)激損傷,過多消耗了乳成分合成的前體物,減少了乳蛋白和乳脂肪的合成。
[Abstract]:The purpose of this study was to study the effects of heat stress on lactation activities of lactating dairy cows and to study the mechanism of these effects. The study included the following three aspects: the following three aspects: the effects of high temperature climate on the metabolism and lactating performance of lactating dairy cows, 6 Holstein cows with the same lactation period, normal feeding and drinking water. From June 29th to August 5th, a total of 35 days. The temperature was recorded at 10:00 and 18:00 per day, milk production was measured, milk protein, milk fat and lactose content were taken every week. The biochemical indexes of the jugular blood test and the content of related hormones were collected at the first first and the last 1 weeks of the experiment. The results showed that the environmental temperature was 26 degrees centigrade. The milk yield of dairy cows decreased from 15.2kg/d to 10kg/d at 38 C, milk protein yield decreased from 0.47kg/d to 0.30 kg/d, milk fat yield decreased from 0.46kg/d to 0.28kg/d, lactose yield decreased from 0.71kg/d to TP (94.18 + 0.35115.07 + 1.04g/L, p=0.049), globulin and glucose. The content of (2.88 + 0.043.54 + 0.11 mmol/L, p=0.03) increased significantly, and LDL-C (0.18 + 0.030.29 + 0.01mmol/L, p=0.004) and AKT (4.17 + 0.3135.50 + 4.15 U/L, p=0.004) significantly increased; HDL-C significantly decreased (3.23 + 0.04, 0.04, 0.04) and 80.05 + The content of g/mL, p=0.048) increased significantly; the content of cortisol (288.21 + 49.73573.76 + 11.71ng/mL, p=0.009) increased remarkably. The concentration of insulin (22.48 + 2.5325.66 + 1.95IU/L, p=0.057) and glucagon (597.18 + 58.11658.02 + 49.47pg/mL, p=0.06) increased. The results suggested that the cows could induce cows under high temperature and high humidity in summer in Nanjing, China. During heat stress, the endocrine and nutritional metabolism of dairy cows varied, showing the decrease of milk secretion, the decrease of milk quality by.2, the relationship between the change of milk composition precursor redistribution and the decrease of milk quality during heat stress. The relationship between the amino acid and free fatty acids in the blood and milk fat and milk protein synthesis was explored. The possible mechanism of milk quality reduction in lactating cows during heat stress. Experimental animals were fed and treated in the same third chapters. The jugular blood was collected at the beginning and end of the experiment. The contents of free amino acids and free amino acids in the blood were detected by RP-HPLC and ELISA hair respectively. The results showed that with the increase of temperature (from 26 > to the highest 38), milk fat With the decrease of milk protein content, the total NEFA in the blood of the experimental cows increased significantly (178.20 + 16.70272.42 + 20.53 g/mL, p=0.025), and the main carbohydrate amino acids such as Glu, Asp, Gly and Val (811.28 + 259.223254.05 + 666.79 mu mol/L, p=0.035) and Leu, and so on were increased or significantly increased (169.26 + 105.50 105.50 mu. 6) the content of total branched chain amino acids (Val, LLE, Leu) increased significantly (198.36 + 52.571474.85 + 355.83 mu mol/L, p=0.02870.05), and the proportion of the essential amino acids increased from 54.75% to 67.89%. conclusion: in the heat stress state of high temperature dairy cows in summer, the milk fat and milk protein content decreased, but the blood NEFA, free amino acids, especially the branched amino groups. It is suggested that the high levels of NEFA and free amino acids in the blood are not completely used in the synthesis of milk fat and milk protein, but also play other roles, such as energy supply, sugar isogenesis, and immune response. The role of the latter is preceded by the participation of milk fat and the synthesis of milk protein. The redistribution changes may be one of the mechanisms leading to the decrease of milk protein and milk fat,.3, the mechanism of redistribution of milk ingredients precursors caused by heat stress in the first two chapters of the experiment, we found that heat stress can cause heavy distribution of milk protein and milk fat precursors, and cause milk protein and milk fat content in milk. In order to explore the mechanism of milk protein and milk fat precursor redistribution caused by the redistribution of milk ingredients, the experimental dairy cows were collected in this experiment. The Real-time PCR method was used to detect the key enzymes PC and PEPCK in the liver of dairy cows and the GHR, IGFR, HSP70 and Keap1-Nrf2 pathways in the liver tissue and their lower levels. The expression of HO1, GCLM, NQO1, GCLC and other proteins was detected by mRNA expression. The results showed that the expression level of PC (0.917 + 0.0081.08 + 0.025, p=0.045) and PEPCK (0.87 + 0.0091.29 + 0.132, p=0.032) in the liver of dairy cows increased significantly during heat stress, and the expression level of GHR (1.05 + 0.02, 0.02, 0.007) was significantly up-regulated. The expression of HSP70 (0.35 + 0.1651.55 + 0.012, =0.006) was significantly increased in the liver, and the expression of Keap1 (1.07 + 0.241.32 + 0.27, p=0.046) and Nrf2 (1.59 + 0.181.73 + 0.19, p=0.049) increased significantly, and the expression of HOl (0.98 + 0.081.19 + 0.02, p=0.037) and 0.92 + 0.02 (0.92) increased significantly. The expression of GCLC (+ 0.100.97 + 0.05, p=0.059) and GCLC (0.84 + 0.071.29 + 0.11, p=0.061) increased, but the difference was not significant. The results suggested that the glucose and isogenesis and anti stress in the liver were enhanced during heat stress. The milk protein, the milk fat precursor, and the milk fat precursor were more involved in the glucose differentiation under the GH-IGF-I axis and the Keap1-Nrf2-ARE signaling pathway. The process of production and resistance to stress damage of cells, too much consumed the precursors of milk composition, and reduced the synthesis of milk protein and milk fat.
【學(xué)位授予單位】：南京農(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)】：S823

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