氨基酸营养与肠道健康 PPT课件

蛋白质氨基酸营养与猪肠道健康谯仕彦2009-10-25重庆猪的肠道屏障及其功能氨基酸与猪的肠道健康日粮蛋白质水平与猪的肠道健康主要 内容 财务内部控制制度的内容财务内部控制制度的内容人员招聘与配置的内容项目成本控制的内容消防安全演练内容 一、猪的肠道屏障及其功能肠道屏障粘液层肠上皮层紧密连接层粘膜下固有层机械屏障生物屏障维持肠道微生态平衡降低肠道pH值激活肠道免疫系统活性促进营养素消化吸收肠道屏障上皮内淋巴细胞派尔集合淋巴小结肠上皮的识别功能M细胞肠道免疫球蛋白免疫屏障化学屏障胃酸胆汁各种消酶溶菌酶粘多糖常驻菌群微生态平衡注：研究较多的为肠上皮细胞屏障和免疫屏障猪肠道屏障功能微生物肠道屏障功能不全病原微生物入侵（细菌内毒素、炎症介质、细胞因子导致肠粘膜水肿）营养物质消化吸收障碍日粮抗原过敏反应肠粘膜缺血缺氧自由基损伤肠粘膜细胞小肠氨基酸代谢注：Asp，天冬氨酸；BCAA，支链氨基酸；BCKA，支链酮酸脱氢酶；CP，氨甲酰磷酸；Glu，谷氨酸；a-KG，a-酮酸；MTF，N5，N10－亚甲基四氢叶酸；OAA，草酰乙酸；Pyr，丙酮酸；TF，四氢乙酸Wu，1998抗氧化能量细胞修复信号分子粘液蛋白二、氨基酸与猪的肠道健康日粮真可消化苏氨酸对7-13kg猪生长性能的影响WangXu，QiaoShiyan,etal.2006,Anim.FeedSci.Technol.指标真可消化苏氨酸水平，％SEMP值0.370.751.11初始重,kg7.227.327.340.550.987末重,kg11.8013.0313.140.770.416ADG,g254318322170.021ADFI,g470477474120.920F/G,g1.861.501.500.060.001日粮真可消化苏氨酸对7-13kg猪蛋白质合成效率的影响指标真可消化苏氨酸水平，％SEMP值0.370.751.11肝脏99.9136.8144.010.450.020背最长肌5.09.85.21.180.017十二指肠粘膜13.386.851.86.410.012空肠粘膜75.5115.888.87.350.006回肠粘膜78.185.370.29.970.575十二指肠黏液58.579.649.17.550.047空肠黏液52.896.752.68.290.029回肠黏液55.977.975.25.770.032XuWang，ShiyanQiao，2007，J.nutr.日粮TID苏氨酸水平对大肠杆菌激发前后断奶仔猪空肠黏膜IgG含量的影响IgG:苏氨酸水平P>0.05E.coliP=0.06互作P=0.07IgG浓度日粮TID苏氨酸水平(g/kg)(µg/g)王旭,谯仕彦.2006日粮真可消化苏氨酸对21日龄断奶仔猪小肠黏液蛋白含量的影响WeiweiWang，ShiyanQiao,etal.,2009,J.Nutr.项目真可消化苏氨酸水平（%）SEM1P值0.370.740.891.11十二指肠0.313b0.466b0.806a0.396b0.0630.0004空肠0.196b0.429ab0.598a0.229b0.0830.0156回肠0.124b0.345a0.367a0.240ab0.0440.0086日粮真可消化苏氨酸对仔猪小肠黏液蛋白基因表达的影响WeiweiWang，ShiyanQiao,etal.,2009,J.Nutr.日粮真可消化苏氨酸对仔猪小肠黏液蛋白亚型的影响杯状细胞真可消化苏氨酸水平（%）SEM10.370.740.891.11十二指肠中性7.64.88.810.01.50酸性16.534.736.550.54.84含硫60.7ab70.7a66.0ab48.6b3.73空肠中性8.813.46.810.01.48酸性15.727.425.223.22.51含硫31.330.338.433.62.86回肠中性16.3b13.7b31.5a22.8ab3.29酸性19.2b30.7a28.0a25.2ab2.70含硫54.649.243.542.95.12WeiweiWang，ShiyanQiao,etal.,2009,J.Nutr.日粮真可消化苏氨酸对仔猪小肠黏膜上皮细胞凋亡的影响WeiweiWang，ShiyanQiao,etal.,2009,J.Nutr.小结日粮TID苏氨酸影响断奶仔猪肠黏膜和黏液蛋白质的合成日粮TID苏氨酸影响断奶仔猪肠黏液蛋白的基因表达，改变黏液蛋白的类型，从而影响小肠黏液屏障功能日粮TID苏氨酸影响小肠黏膜形态、黏膜上皮细胞通透性和细胞凋亡，从而影响小肠上皮细胞屏障功能2、精氨酸精氨酸作用的信号通路精氨酸FAK细胞信号PI-3KAP-1和cJun(丝氨酸激酶)(转录因子)血管再生血管舒张血流神经传递免疫反应线粒体功能蛋白激酶AmTOR蛋白质合成细胞增殖和迁移70kDaS6kinase(p70s6k)NO&多胺Rhoadsetal.(2004)Gut53:514-522.FAK,focaladhesionkinase.Rhoadsetal.(2008)JNutr(Inpress)日粮添加精氨酸对14日龄断奶仔猪小肠绒毛高度的影响日粮添加精氨酸对14日龄断奶仔猪小肠隐窝深度的影响AB断奶仔猪日粮中添加精氨酸对小肠绒毛形态的影响ZhenfengZhan,DefaLi，etal.,2008,J.Nutr.十二指肠空肠回肠一氧化氮(μmol/g)L-ArgL-ArgA内皮素(pg/mg)十二指肠空肠回肠L-ArgL-ArgB日粮添加精氨酸对14日龄断奶仔猪小肠中NO浓度的影响日粮添加精氨酸对14日龄断奶仔猪小肠中ET-1浓度的影响断奶仔猪日粮中添加精氨酸对小肠NO和血管内皮素的影响ZhenfengZhan,DefaLi，etal.,2008,J.Nutr.小结精氨酸影响肠道重量、肠粘膜重量、绒毛高度、隐窝深度、粘膜蛋白含量、粘膜DNA、肠上皮细胞的增殖，影响肠道屏障功能日粮适宜添加精氨酸可以作为一种有效的促进小肠微血管损伤修复的手段3、谷氨酰胺谷氨酰胺在肠道的生理功能为氨基酸、蛋白质和核酸的合成提供氮源为肠道供能约占总量的70％以上是肠道粘膜代谢的必需营养物质Gln缺乏，可使肠粘膜萎缩、绒毛变稀变短，屏障功能下降谷氨酰胺减轻肠粘膜损伤的可能机制参与三羧酸循环，生成ATP供能参与粘膜细胞核酸及蛋白质的合成代谢促进肠粘膜细胞的更新和再生保持肠粘膜细胞超微结构的完整性刺激机体产生高血糖素增加萎缩肠粘膜中谷氨酰胺酶活性Wuetal.1996.JNutr.126:2578-2584.十二指肠空肠谷氨酰胺添加比例谷氨酰胺添加比例绒毛高度或粘膜固有层深度断奶天数0%1%SE0%1%SE绒毛高度，µm0454339736536337270a358b32144234553447744734粘膜固有层深度，µm026919373053121923725617144234212230127916断奶仔猪日粮中添加谷氨酰胺对肠道绒毛高度和粘膜固有层深度的影响谷氨酰胺防止断奶仔猪空肠粘膜萎缩Domeneghini.2006.VetResComm.30:331–342.项目对照（n=16）0.5%谷氨酰胺组P值绒毛高度(V,µm)168.18±9.33207.63±9.330.0038隐窝深度(C,µm)109.97±13.73166.47±13.730.0049V/C值1.53±0.1031.25±0.1030.0215断奶仔猪日粮中添加0.5%的谷氨酰胺对小肠绒毛高度和隐窝深度的影响Domeneghini.2006.VetResComm.30:331–342.对照组（n=16）谷氨酰胺（n=16）P值肠上皮细胞（%）调亡（A）2.702.550.0109有丝分裂（M）39.1040.820.0159A/M值0.0690.0620.0023断奶仔猪日粮中添加0.5%的谷氨酰胺对小肠上皮细胞凋亡和有丝分裂的影响炎性介质TNF-α、IL-6、DAO、D乳酸增加，使肠粘膜通透性增强，导致肠道屏障功能降低断奶仔猪日粮中添加1%的谷氨酰胺对空肠谷胱甘肽和绒毛高度的影响WangJJetal.(2008)JNutr138:1025-1032项目对照组谷氨酰胺组GSSG/GSH,nmol/nmol0.087±0.0050.051±0.002*绒毛高度,µm269±7.5371±9.2*谷氨酰胺下调与凋亡有关的基因KLF-10-64%Laproteinhomolog-41%Peptidyl-prolylisomerase-37%MAPK-6-75%ICAM-1-38%WangJJetal.2008,JNutr138:1025-1032二、日粮蛋白质水平与猪的肠道健康试验 设计 领导形象设计圆作业设计ao工艺污水处理厂设计附属工程施工组织设计清扫机器人结构设计 与试验日粮日粮成分(%)1234小麦20.322.024.026.6玉米19.3321.7923.5323.79大麦20.322.024.026.6豆粕36.4530.2424.0217.66赖氨酸-0.20.40.8蛋氨酸0.020.080.140.02苏氨酸-0.090.180.27色氨酸--0.030.07异亮氨酸--0.010.02缬氨酸--0.090.09Bellego和Noblet,2002;猪起始体重:11kg试验日粮养分含量CP(%)22.420.418.416.9DE(MJ/kg)14.7514.5914.6314.50NE(MJ/kg)10.2810.3310.5210.52氨基酸(%)赖氨酸1.201.191.211.23蛋氨酸0.340.370.390.41苏氨酸0.820.810.800.81色氨酸0.270.240.230.24异亮氨酸0.950.860.750.75缬氨酸1.050.960.920.92组氨酸0.550.500.450.40Bellego和Noblet,2002日粮蛋白质水平对仔猪生产性能的影响指标日粮蛋白质水平(%)22.420.418.416.9ADG(g)642661690663ADFI(g)959103910611048Feed/gain1.501.581.541.58各种粪便所占比例(%)硬粪81.982.095.489.0软粪14.714.54.19.0液体状粪3.43.50.52.0排泄氮(g/d)10.79.46.85.1沉积氮(g/d)17.817.718.515.6饮水量(g/d)1941188718671645排尿量(g/d)757643625481Bellego和Noblet,2002降低日粮蛋白质水平对断奶仔猪生长性能和肠道健康的影响日粮CP水平(％)23.121.218.917.2DE(kcal/kg)3450345034503450SDLys,g/MJ0.930.930.930.93SDLys1.301.301.301.30SDThr0.880.820.810.81SDMet0.350.370.400.41SDTrp0.250.240.240.24Y.L.YaoandS.Y.Qiao,LivestockScience,2007结果：对生长性能和粪指数的影响日粮DE水平(%)23.121.218.917.2SEMP值始重(kg)6.806.756.756.720.030.935末重(kg)10.5210.6110.279.650.280.042ADG(g/d)266275252209170.027采食量(g/d)357370342307220.134饲料/增重1.351.341.361.490.040.036粪指数0.610.490.420.380.020.015血浆尿素氮(mg/L)139.394.480.460.55.8<0.01Y.L.YaoandS.Y.Qiao,LivestockScience,2007结果：肠道形态发育（单位um）日粮DE水平(%)23.121.218.917.2SEMP值十二指肠绒毛高度4544614574337.50.016隐窝深度1861791901804.50.717高度/深度2.452.572.412.410.080.575空肠绒毛高度4364294193938.80.017隐窝深度1971901921827.70.509高度/深度2.212.262.182.160.120.887回肠绒毛高度3993954003867.90.451隐窝深度1811761891755.80.194高度/深度2.202.242.192.211.00.437Y.L.YaoandS.Y.Qiao,LivestockScience,2007结果：粘膜蛋白和二糖酶活性日粮DE水平(%)23.121.218.917.2SEMP值粘膜蛋白含量(mg/g粘膜)55.956.554.251.83.10.078小肠二糖酶活性乳糖酶39.739.735.629.52.00.016蔗糖酶54.255.250.743.72.10.015麦芽糖酶85.387.086.985.72.40.753Y.L.YaoandS.Y.Qiao,LivestockScience,2007降低日粮蛋白质顺序补充氨基酸对断奶仔猪生长性能的影响日粮CP水平(％)21.0+Lys+Thr+Met17.0+Lys+Thr+Met+Trp+Ile+Val17.0+Lys+Thr+Met+Trp+Ile+Val+Gln17.0+Lys+Thr+Met+Trp+Ile+Val+Gln+ArgDE(kcal/kg)3450345034503450SDLys1.391.391.391.39SDThr0.960.960.950.96SDMet0.450.480.480.48SDTrp0.230.230.230.23SdIle0.800.940.940.94SDVal0.960.750.750.75SdArg1.361.051.041.34岳隆耀，谯仕彦，2008结果：对仔猪生长性能的影响（试验期7天）日粮CP水平(％)21.0+Lys+Thr+Met17.0+Lys+Thr+Met+Trp+Ile+Val17.0+Lys+Thr+Met+Trp+Ile+Val+Gln17.0+Lys+Thr+Met+Trp+Ile+Val+Gln+ArgP值始重(kg)6.76.76.66.70.93末重(kg)8.778.518.178.660.03ADG(g/d)2952592252810.01采食量(g/d)3814123404310.01饲料/增重1.291.591.511.540.02岳隆耀，谯仕彦，200840头杜×大×长仔猪,17±1d断奶,体重5.32±0.24kg，2个组，每组5圈，每圈5头处理1日粮CP水平，22.5%；处理2日粮CP水平17.6%CP试验期14天，第8天用5×1010cfu/mL的大肠杆菌口服感染，感染后1、3、7天屠宰日粮蛋白质水平对断奶仔猪生产性能和肠道健康的影响(Opapeju等,2009)Opapejuetal.,2009.J.Anim.Sci.日粮配方Item22.5%CP17.6%CPIngredient,%Corn53.2167.63Wheat14.015.00Soybeanmeal4.004.00Fishmeal6.006.00Wheypowder7.007.00Spray-driedbloodplasma2.002.00Casein7.90—Canolaoil3.643.34Limestone0.630.55Biofos0.510.83Vitaminandmineralpremix1.001.00Biolys—1.13DL-Met0.060.29L-Thr0.030.32L-Trp0.010.10L-Ile—0.37L-Val—0.44Calculatednutrientcomposition22.5%CP17.6%CPME,MJ/kg14.5114.51CP,%22.5017.64Lys,%1.471.47Met,%0.580.61Ile,%0.980.97Thr,%0.950.97Trp,%0.280.28Val,%1.231.21StandardizedilealdigestibleAA,%Lys1.351.35Met+Cys0.810.81Thr0.880.88Trp0.250.25Ile0.900.90Val1.121.12Ca,%0.800.80TotalP,%0.650.65饲料营养成分含量Item22.5%CP17.6%CPSEMInitialBW,kg5.345.300.24FinalBW,kg8.518.070.30ADG,gd1to7201a147b8.5d9to1033330421.81d12to1432234329.39Overall28626513.01G:F,g/gd1to71.00a0.82b0.02d9to101.201.110.10d12to141.081.060.07Overall1.061.040.05Fecalconsistencyscore24hafterchallenge0.680.320.1930hafterchallenge1.000.600.3048hafterchallenge1.40a0.50b0.2972hafterchallenge1.40a0.60b0.175dafterchallenge1.10a0.30b0.23Overall1.12a0.46b0.17结果：生产性能日粮蛋白质水平对回肠食糜中大肠杆菌K88数量的影响Taxonomiclevel,%22.5%CP17.6%CPSEMFhylumBacteroidetes1.52.50.50ClassunclassifiedBacteroidetes1.52.50.50厚臂菌门92.488.61.90杆菌纲11.613.91.15梭菌纲77.374.71.30梭菌目50.0a73.4b11.7FamilyLachnospiraceae18.2a43.0b12.4GenusRoseburia2.5a12.7b5.10OrderunclassifiedClostridia27.3a1.3b13.0ClassunclassifiedFirmicutes3.50.01.75PhylumLentisphaerae0.51.30.40ClassLentisphaerae0.51.30.40PhylumProteobacteria5.67.61.00ClassEpsilonproteobacteria02.51.25ClassDeltaproteobacteria0.52.51.00ClassGammaproteobacteria2.50.01.25ClassBetaproteobacteria2.01.30ClassunclassifiedProteobacteria0.51.30.40PhylumunclassifiedBacteria0.50.00.25日粮蛋白质水平对结肠菌群的影响Item22.5%CP17.6%CPSEMAmmoniaN,mg/L1dbeforechallenge237.85a168.97b20.683dafterchallenge293.99211.7771.287dafterchallenge414.97a182.65b73.89TotalVFA,mmol/L1dbeforechallenge17.2421.852.453dafterchallenge16.7515.321.477dafterchallenge17.8518.652.48日粮蛋白质水平对结肠氨浓度和总挥发性脂肪酸的影响Item22.5%CP17.6%CPSEMVillusheight,μm402.03405.5216.151dbeforechallenge330.16a458.52b26.383dafterchallenge412.92409.519.137dafterchallenge351.94a243.38a21.12Cryptdepth,μm1dbeforechallenge351.94a243.38a21.123dafterchallenge324.77312.1119.287dafterchallenge357.88299.4124.75Villusheight:cryptdepth1dbeforechallenge1.18a1.67b0.103dafterchallenge1.03a1.47b0.067dafterchallenge1.201.390.11日粮蛋白质水平对回肠绒毛发育的影响结论用大肠杆菌感染17日龄断奶仔猪17.6%CP添加合成氨基酸的日粮可取得和22.5%CP日粮相似的生产性能降低日粮蛋白质水平可减少结肠氨气的产生和蛋白质的发酵降低日粮蛋白质水平减少大肠和小肠中大肠杆菌的数量降低日粮蛋白质水平增加绒毛高度和隐窝深度的比,增加结肠中丁酸的产生Maximumquadratic=2.38R2=0.89日粮净能含量与育肥猪最小平均日增重百分比的折线和二次趋势图最小平均日增重百分比（%）日粮净能含量（Mcal/kg）Brokenpoint=2.36R2=0.84易学武、谯仕彦，2009Maximumquadratic=2.40R2=0.81日粮净能含量与育肥猪最小饲料转化率百分比的折线和二次趋势图最小饲料转化率百分比（%）日粮净能含量（Mcal/kg）Brokenpoint=2.36R2=0.80易学武、谯仕彦，2009日粮净能含量与育肥猪最小无脂瘦肉率百分比的折线和二次趋势图Maximumquadratic=2.42R2=0.86Brokenpoint=2.40R2=0.87最小无脂瘦肉率百分比（%）日粮净能含量（Mcal/kg）易学武、谯仕彦，2009仔猪采食不同蛋白水平日粮并补充氨基酸对氮平衡的影响岳隆耀、谯仕彦，2009不同赖氨酸水平的低蛋白日粮对生长猪氮排放的影响鲁宁、谯仕彦，2009低蛋白质日粮的环保意义指标日粮蛋白质降低1个百分点最大化效果(4-5个百分点)总氮排泄量-10%-50%空气中氨浓度-13%-60%饮水量-3%-28%排泄物总量-5%-30%Relendeau等，2000谢谢！