本文選題：生長豬 + 模擬小腸液　； 參考：《中國農(nóng)業(yè)科學(xué)院》2015年碩士論文

【摘要】：本試驗(yàn)首先研究生長豬空腸液中消化酶的特性、空腸液消化酶的純化與模擬腸液的制備方法,在此基礎(chǔ)上探討了基于單胃動物仿生消化系統(tǒng)(SDS-Ⅱ)和模擬小腸液的仿生消化法測定日糧消化能和可消化氨基酸含量的可行性。從5個(gè)方面開展工作:試驗(yàn)一旨在研究不同生長階段及日糧條件下豬空腸液消化酶的活性,為模擬小腸液的制備參數(shù)提供參考。試驗(yàn)采用2×2兩因素完全隨機(jī)設(shè)計(jì),其中日糧設(shè)兩個(gè)水平,即玉米-豆粕型日糧和玉米-玉米干酒糟(DDGS)-米糠粕型日糧,豬生長階段設(shè)兩個(gè)水平,即生長期(60-100天)和育肥期(100-140天),測定空腸液中淀粉酶、胰蛋白酶、糜蛋白酶、脂肪酶的活性、總蛋白含量和p H。結(jié)果表明:日糧類型與生長階段對生長豬空腸液中各消化酶體積比活性及p H均無顯著影響(P0.05),豬空腸液單位體積主要消化酶平均活性和p H分別為:淀粉酶189.79 U/ml、胰蛋白酶62.10U/ml、糜蛋白酶7.36 U/ml、脂肪酶4.91U/ml和p H 6.44。試驗(yàn)二旨在探討空腸消化液中主要消化酶的活性與日糧養(yǎng)分消化率間的相關(guān)性,為模擬腸液制備中消化酶活性的誤差控制標(biāo)準(zhǔn)提供參考。玉米-豆粕型日糧和玉米-DDGS-米糠粕型日糧分別設(shè)16個(gè)重復(fù),每個(gè)重復(fù)1頭豬,測定日糧及糞樣的干物質(zhì)、鹽酸不溶灰分(AIA)、粗蛋白、總能及腸液中主要消化酶的活性,然后對每種日糧下的養(yǎng)分消化率與主要消化酶的活性進(jìn)行簡單相關(guān)分析和典型相關(guān)分析。結(jié)果表明:1)兩種日糧下,生長豬個(gè)體間4種主要消化酶活性的變異系數(shù)均在20%以上,而日糧干物質(zhì)、粗蛋白、能量消化率的變異系數(shù)均在1.5%以內(nèi);2)豬空腸液中淀粉酶、胰蛋白酶和糜蛋白酶活性與日糧養(yǎng)分消化率無顯著相關(guān)性(P0.05),而脂肪酶的活性與日糧養(yǎng)分消化率有一定的相關(guān)性,但對養(yǎng)分消化率變異的貢獻(xiàn)是有限的。試驗(yàn)三分為兩部分進(jìn)行:首先探討豬空腸液中消化酶的純化方法,然后比較以空腸液純化粉劑制備的模擬小腸液與以試劑酶制備的模擬小腸液在消化能力上的差異。采用2×2兩因素完全隨機(jī)設(shè)計(jì),即日糧和豬生長階段各設(shè)兩個(gè)水平。采用冰乙醇沉淀-凍干純化空腸液中的消化酶,測定純化過程中消化酶的活性損失和腸液純化粉劑的比活性。然后,根據(jù)四個(gè)處理下的空腸液中消化酶的活性,以試劑酶、處理組內(nèi)腸液純化粉劑+試劑酶、所有處理腸液混合液純化粉劑+試劑酶制備模擬小腸液分別測定日糧的水解物能值(EHGE),并與生物學(xué)法測值進(jìn)行相關(guān)性分析。結(jié)果表明:1)乙醇沉淀-凍干獲得的腸液消化酶活性的回收率均在60%以上,日糧類型和生長階段對純化粉劑的消化酶活性無顯著影響;2)乙醇沉淀過程中消化酶的活性損失顯著的高于凍干過程(P0.01);3)以腸液純化粉劑制備的模擬小腸液測定日糧的EHGE均顯著高于以試劑酶制備模擬腸液的相應(yīng)測值(P0.05),且與生物學(xué)法測值的相關(guān)性更高。以上結(jié)果表明,乙醇-沉淀凍干法提純的腸液消化酶粉劑可以更好地模擬小腸液。試驗(yàn)四旨在研究基于胃蛋白酶-模擬豬腸液-Viscozyme復(fù)合酶的仿生消化法估測豬棉籽粕日糧的消化能值。分別采用腸液純化粉劑和試劑酶模擬豬小腸液測定11種脫酚棉籽蛋白和19種棉籽粕日糧的酶水解物能值,并與生物學(xué)法測值進(jìn)行簡單相關(guān)分析。結(jié)果表明:1)采用腸液純化粉劑模擬小腸液測定脫酚棉籽蛋白的表觀酶水解物能值(AEHGE)和真酶水解物能值(TEHGE)與試劑酶測值間均具有很高的相關(guān)性(P0.05,r=0.96);2)采用腸液純化粉劑比試劑酶制備的模擬小腸液測定的棉籽粕日糧的酶水解物能值與生物學(xué)法測值的相關(guān)性更高(r=0.94 VS 0.87)。這表明基于豬源性消化酶粉劑制備的模擬小腸液在棉籽粕酶水解物能值的測定上優(yōu)于試劑酶。試驗(yàn)五研究基于胃蛋白酶-腸液純化粉劑兩階段仿生消化法測定棉籽粕可消化氨基酸的含量。以仿生消化法分別測定10種不同來源棉籽粕的可消化氨基酸含量,并與生物學(xué)法測值進(jìn)行相關(guān)性分析。結(jié)果表明:仿生消化法測定的棉籽粕除蛋氨酸外的其他氨基酸的可消化含量均與體內(nèi)法測值顯著相關(guān)(P0.05),這表明豬源性消化酶粉劑制備的模擬小腸液適用于棉籽粕可消化氨基酸含量的測定。
[Abstract]:In this experiment, we first studied the characteristics of digestive enzymes in the empty intestines of growing pigs, the purification of digestive enzymes in the jejunum and the preparation of simulated intestinal liquid. On the basis of this, the feasibility of the bionic digestion method based on the bionic digestive system (SDS- II) and the simulated intestinal fluid for the determination of the digestible energy and the content of digestible amino acids in the diet were discussed in 5 aspects. The experiment is to study the activity of digestive enzyme of pig jejunum in different growth stages and diets and provide reference for simulating the preparation parameters of small intestinal fluid. The experiment adopts 2 x 2 two factors completely random design, of which there are two levels of diet, namely corn soybean meal type diet and corn corn dry distiller's grains (DDGS) rice bran meal type diet. At the growth stage, there were two levels, namely, the growth period (60-100 days) and the fattening period (100-140 days). The activity of amylase, trypsin, chymotrypsin and lipase in the jejunum fluid, total protein content and P H. results showed that there was no significant effect on the volume ratio activity of digestive enzymes and P H in the growth stage and the growth stage (P0.05). The average activity of the main digestive enzyme and P H in the unit volume of the jejunum were: amylase 189.79 U/ml, trypsin 62.10U/ml, chymotrypsin 7.36 U/ml, lipase 4.91U/ml and P H 6.44. test two to explore the correlation between the activity of the main digestive enzymes in the jejunum digestive juice and the nutrient digestibility in the diet, to simulate digestive enzyme activity in the preparation of intestinal liquid The diet of corn soybean meal and corn -DDGS- rice bran meal had 16 repetitions, each repeated 1 pigs, measured the dry matter of the diet and feces, the insoluble ash (AIA), the crude protein, the total energy and the activity of the main digestive enzymes in the intestinal juice, and then the nutrient digestibility and the main elimination of each diet. The activity of the enzyme was analyzed by simple correlation analysis and canonical correlation analysis. The results showed that: 1) under two diets, the coefficient of variation of 4 main digestive enzyme activities among 4 growing pigs was more than 20%, while the coefficient of variation of dry matter, crude protein and energy digestibility were within 1.5%; 2) amylase, trypsin and chymotrypsin in pig's empty intestines. There was no significant correlation between sex and dietary nutrient digestibility (P0.05), but the activity of lipase had a certain correlation with the nutrient digestibility of the diet, but the contribution to the variation of nutrient digestibility was limited. The experiment was divided into two parts. First, the purification method of digestive enzymes in the pig jejunum solution was discussed, and then the preparation of the purified powder was prepared by the jejunum solution. The difference between the simulated small intestinal fluid and the simulated intestinal fluid prepared by the reagent enzyme was different. The 2 x 2 two factors were designed completely, and there were two levels in the diet and the growth stage of the pig. The digestive enzymes in the jejunum solution were purified by the ice ethanol precipitation and freeze drying. The loss of the activity of the elimination enzyme in the purification process and the ratio of the purified powder in the intestinal liquid were measured. Then, according to the activity of digestive enzymes in the four treated jejunum fluid, the reagent enzyme, the purified powder + reagent enzyme in the intestinal liquid of the treatment group, the hydrolysate energy value (EHGE) of the simulated small intestine liquid were prepared by the purified powder and reagent of the intestinal liquid mixture, and the correlation analysis was carried out with the biological method. The results showed that: 1) The recovery of the activity of digestive enzyme obtained by ethanol precipitation and freeze drying was above 60%, and the diet type and growth stage had no significant effect on the digestive enzyme activity of the purified powder; 2) the activity loss of digestive enzymes in the process of ethanol precipitation was significantly higher than that of the freeze drying process (P0.01); 3) the EH of the simulated intestinal fluid for the determination of the diet prepared by the purified powder of intestinal fluid GE was significantly higher than the corresponding measured value of the simulated intestinal liquid (P0.05) with the reagent enzyme, and the correlation was higher with the biological method. The above results showed that the enteric digestive enzyme powder purified by the ethanol precipitation freeze dry method could better simulate the small intestinal fluid. Four the experiment was designed to study the bionics based on the gastric protease mimic pig intestinal fluid -Viscozyme complex enzyme. Digestion method was used to estimate the digestibility of pig cottonseed meal diet. The enzyme hydrolysate values of 11 dephenolates and 19 kinds of cottonseed meal were measured by intestinal liquid purification powder and reagent enzyme simulated pig small intestine solution. The results showed that 1) used intestinal liquid to simulate small intestine liquid to determine dephenolol. The apparent enzyme hydrolysate value (AEHGE) and the value of the true enzyme hydrolysate (TEHGE) and the measured value of the reagents had a high correlation (P0.05, r=0.96); 2) the correlation between the hydrolysate of the hydrolysate of the cottonseed meal measured by the intestinal solution and the measured values of the cottonseed meal was higher than that of the reagents (r=0.94 VS 0.). 87). This shows that the simulated intestinal fluid based on porcine digestive enzyme powder is superior to the reagent enzyme in the determination of the enzyme hydrolysate of cottonseed meal. In test five, the content of digestible amino acids in cottonseed meal was determined by the two stage bionic digestion method based on pepsin - intestinal liquid purification powder. 10 different sources of cottonseed were determined by bionic digestion method. The results showed that the digestible content of other amino acids in the cottonseed meal, except methionine, was significantly correlated with the body method (P0.05), which showed that the simulated intestinal fluid prepared by the porcine digestible powder was suitable for the digestibility of cottonseed meal. Determination of the content of amino acids.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S828

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