含肽肝素渣水解物对早期断奶仔猪消化吸收与免疫功能影响的研究
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摘要
本研究分两个试验,分别研究了含肽肝素渣水解物的酶解工艺;探讨了含肽肝素渣水解物对早期断奶仔猪肠道组织结构、酶活性、有关血液生化指标、免疫器官指数、淋巴细胞数、血清猪瘟抗体OD值、腹泻指数以及生产性能的影响。
     试验一以肝素渣为原料,采用酶解法,按L_9(3~4)正交试验设计,确定了生产含肽肝素渣水解物的酶解工艺;分别对高压灭菌和煮沸1h后的原料进行细菌总数检查。结果表明,处理8的有效成分含量最高,其肽蛋白含量为41.92%,其中2-6肽和2-3肽的含量分别为12.64%和7.56%;原料经高压灭菌和煮沸1h后,细菌总数符合国家饲料卫生标准(GB13078-91)。
     试验二选择出生日龄相近(±2日龄)的商品代胜利白猪12窝,于21日龄断奶并从中选择72头,采用单因素随机区组试验设计,分成A、B、C、D、E、F6组,每组3个重复,每个重复4头猪。其中A组为对照组,不添加含肽肝素渣水解物(PIH)和血浆蛋白粉(SDPP),B、C、D组各添加1%、3%、5%PIH,E组添加2%SDPP,F组添加1%PIH与1%SDPP。各组日粮营养水平相同。试验期自断奶至49日龄止。于试验开始和第42、49日龄分别逐头称重并按重复组统计耗料量。在42日龄称重后,从每组中选择2头体重接近组平均体重的仔猪心脏采血,测定血液生化指标、淋巴细胞数和猪瘟抗体OD值,然后屠宰。取脾脏、下颌淋巴结和腹股沟淋巴结称重,计算免疫器官指数;取十二指肠食糜,测定淀粉酶、脂肪酶和胰蛋白酶活性;取十二指肠、空肠、回肠中部的肠管2cm,制作切片,观测肠绒毛高度和隐窝深度;试验期记录仔猪的腹泻情况,计算腹泻指数。试验结果表明:(1)消化酶活性。十二指肠食糜中淀粉酶活性除1%PIH组外,各试验组均显著高于对照组(p〈0.05),各试验组的脂肪酶活性与胰蛋白酶活性均显著高于对照组(p〈0.05),其中D组的淀粉酶和胰蛋白酶活性最高,3%PIH组的脂肪酶活性最高。2%SDPP组、1%SDPP+1%PIH组、3%PIH组以及5%PIH组之间以上三种消化酶活性的差异均不显著(p〉0.05);(2)小肠绒毛高度和隐窝深度。5%PIH组的小肠绒毛高度以及VH/CD值显著高于其它组(p〈0.05),十二指肠的隐窝深度各试验组显著低于对照组(p〈0.05),D组显著低于1%PIH组(p〈0.05);(3)免疫器官指数。各试验组的免疫器官指数均显著高于对照组(p〈0.05),且随着PIH添加量的增大有增加的趋势,而以5%PIH组的免疫器官指数最高。2%SDPP组、1%PIH+1%SDPP组以及3%PIH组之间的免疫器官指数的差异不显著(p〉0.05);(4)血液生化指标。5%PIH组和2%SDPP组的血铁和血钙浓度显著高于对照组(p〈0.05),而各试验组之间差异不显著(P〉0.05)。各试验组血清尿素氮含量显著低于对照组(p〈0.05),其中以5%PIH组的血清尿素氮含量最低,但与3%PIH组、2%SDPP组和1%PIH+1%SDPP组之间差异不显著
    
     蔡元丽:含肽肝素渣水解物对早期断奶仔猪消化吸收与免疫功能影响的研究
     (p>0.05),2%SDPP组和 10PIH+l%SDPP组与添加 PIH的各组之间的差异也不显
    著(p>0.05)。各试验组血清哈G含量显著高于对照组(p<0.05),2%SDPP组、
    loPIH+l0SDPP组以及3%PIH组之间差异不显著(p>0.05):(5)血液淋巴细胞数。
    除1%PIH组外的各试验组的血液淋巴细胞数差异不显著,但显著高于对照组
    (<0刀5),随着NH添加量的增大血液淋巴细胞数有增加的趋势;(6)血清猪瘟抗体
    OD值。50PIH组的血清抗体OD值显著高于对照组,且随PIH添加量的增大有增
    加的趋势,2%SDP组、1%NH+1%SDP组以及3%NH组之间差异不显著(P0.05);
     (7)腹泻指数。各试验组的腹泻指数显著低于对照组O<0刀5),而添加5%nH组腹
    泻指数最低,2%SDpp组、l%pIH+l%SDpP组以及3%pIH组之间差异不显著
     (p>O刀5);(8)生产性能。除1%nH组外的其他试验组牡、叩日龄的体重显著高
    于对照组(p<0.05),且随nH添加量的增加有增加的趋势,3OPIH组、5OPIH组、
    2%S*叩组和1%NN十1%**P组之间差异不显著(p>o刀5)。刀-42和刀*9日龄平
    均日增重各试验组显著高于对照组(p<0。05),其中以5%wN组最高,2%*0P组、
    l%PIH-l%SDPP组以及3%PIH组之间差异不显著(p>0.05)。30PIH组与5%PIH
    组的引-42日龄、ZI-49日龄的料重比显著低于对照组(p<0刀5),其中以5%PIH组的
    料重比最低,3%PIH组、5%PIH组、2%SDPP组和1%PIH+l%SDPP组之间的料重
    比差异不显著(p>0刀5)。结果分析表明,含肽肝素渣水解物能提高十二指肠消化酶
    活性,促进小肠的发育,增强仔猪的免疫功能,促进矿物元素和蛋白质在体内的沉
    积,有效地减轻了早期断奶仔猪的腹泻从而提高了仔猪的生产性能;含肽肝素渣水
    解物在早期断奶仔猪日粮中的适宜添加水平为3W5%:含肽肝素渣水解物可以代替
    2%血浆蛋白份,但与血浆蛋白粉无明显的协同作用。
The hydrolyzing conditions to produce porcine intestine hydrolysate (PIH) from a residue of heparin extraction were screened in the first part at this study. The safety of the residue was proved by microbiological examination. The Hydrolysate produced under the most suitable conditions contained 42% of peptides and proteins. The content of oligopeptides with less than six amino acid residues and oligopeptides with less than three amino acid residues are 12.64% and 7.56% respectively.
    In the second part of this study, 72 piglets weaned at 21-day old were randomly allocated to 6 treatments. For each treatment, there were 3 replicates and there were 4 piglets fof each replicate. The diets of these treatments had a same nutrient level, but contained 1%PIH, 3%PIH, 5%PIH, 2%spray dried plasma protein (SDPP), 1%PIH-H%SDPP respectively. Neither PIH nor SDPP was supplemented in the control group. The experiment studied when the piglets were 21-day old and finished at 49-day old. The piglets were weighted at 42 and 49-day old and feed intake was recorded. At 42-day old, 2 piglets from each treatment were slaughtered. Blood samples were analyzed for serum biochemical characteristics, number of lymphocytes and OD value of swine fever antibody. Immune organ indexes were calculated from the weight of spleen, lower jaw lymphnodes and inguen lympnnodes. Amylase, lipase and trypsin activities in duodenum chyme were measured. Tissue samples of duodenum jejunum and ileum were used to measured the villi height and crypts depth of small intestine. Cases of diarrhea were recorded during the experiment and diarrhea indexes were calculated.
    The result indicated that 1. With the exception of chyme amylase activity of pigtets receiving 1%PIH treatment, chyme amylase, lipase and trpsin activities of piglets receiving other treatments were significantly higher than piglets from the control (P<0.05). The highest amylase and trypsin activities were observed in pigets recerving 5%PIH treatment while the highest lipase activities was observed in piglets receiving 3%PIH treatment. There was no significant difference of enzyme activities between the treatment groups(P>0.05). 2. The villi height and VH/CD in the small intestines were higher in
    
    
    
    5%PIH group than other groups (p<0.05). Pigs fed the PIH or SDPP diets showed lower crypts depth than control group. 3. The immune organs indexes were significantly higher in PIH or SDPP groups than control group (p<0.05). The pigs fed 5%PIH diet had the highest immune organs index. The difference of immune organs indexes among 2%SDPP, 1%SDPP+1%PIH and 3%PIH groups was not significant (p>0.05). 4. Pigs fed 5%PIH and 2%SDPP diets showed higher serum iron and calcium concentrations compared to the control group (p<0.05). The blood urea nitrogen (BUN)concentration was lower in SDPP or PIH groups than control group (p<0.05). The BUN concentration of 5%PIH was the lowest. Pigs fed 5%PIH and 2%SDPP diets had higher immunoglobulin G (IgG) concentrations than control group (p<0.05). 5. Pigs fed experimental diets except fed 1%PIH diet showed more number of lymphocyte than pigs fed control diet (p<0.05). With the increase of of PIH level, the number of lymphocyte increased. 6. The OD value of swine fever antibody was higher in 5%PIH group than control group (p<0.05), but there was no significant difference among pigs fed 2%SDPP, 1%PIH+1%SDPP, 3%PIH diets(p>0.05). 7. Pigs fed SDPP or PIH diets had lower diarrhea indexes compared with pigs fed control diet (p<0.05). And the diarrhea index of 5%PIH group was the lowest. 8. At the 42 and 49-day old, the liveweighe of pigs fed SDPP or PIH diets except 1%PIH were higher than pigs fed control diet (p<0.05). The average daily gain from 21 to 42 day old and from 21 to 49-day old was higher in pigs fed SDPP or PIH diets than pigs fed control diet (p<0.05), with the pigs fed 5%PIH diet having the highest daily gain. Pigs fed 3%PIH or 5%PIH diet showed lower feed/gain ratio from the age of 21 to 42 days and from 21 to 49 days compared with pigs fed control diets. Pigs fed
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