亚急性瘤胃酸中毒耐受性不同的奶牛瘤胃上皮形态及功能差异研究

本试验旨在探究SARA（亚急性瘤胃酸中毒）耐受性不同奶牛的瘤胃上皮形态及其功能差异。选取12头装有永久性瘤胃瘘管的泌乳中期荷斯坦奶牛,饲喂精粗比为4∶6的日粮,并根据瘤胃pH值的高低,分为SARA易感组（SUS,n=4）和SARA耐受组（TOL,n=4）。瘤胃上皮形态及功能分析结果显示,与TOL组比较,SUS组奶牛瘤胃上皮的棘突层和基底层厚度明显增厚（P<0.05）,SUS组奶牛瘤胃上皮组织中参与挥发性脂肪酸（VFA）吸收的PAT1、MCT4和DRA基因表达量较TOL组显著下调（P<0.05）,而H+转运载体NHE1、NHE2、NHE3和调节胞内pH的vH+ATPase和Na+/K+ATPase的表达量显著升高（P<0.05）;对参与调控瘤胃VFA代谢的基因定量结果表明,SUS组PDHA1和SREBP2的表达量显著高于TOL组（P<0.05）,而HMGCL-2的表达量显著降低（P<0.05）;此外,SUS组CDK2、CDK6和Cyclin D1、Bad及Caspase-9等参与瘤胃上皮细胞增殖与凋亡的基因表达量显著高于TOL组（P<0.05）。结果说...     

饲粮不同非纤维性碳水化合物/中性洗涤纤维对细毛羊氮代谢、瘤胃内环境及挥发性脂肪酸吸收的影响

本试验旨在研究饲粮不同非纤维性碳水化合物/中性洗涤纤维(NFC/NDF)对细毛羊氮代谢、瘤胃内环境及挥发性脂肪酸(VFA)吸收的影响。选用9只装有永久性瘤胃瘘管的内蒙古鄂尔多斯细毛羊半同胞羯羊,按体重随机分为3组,1组、2组和3组饲粮NFC/NDF分别为0.35、0.66和1.14,粗蛋白质含量均为13.50%。结果表明:1)饲粮NFC/NDF对摄入氮、粪氮、尿氮和沉积氮无显著影响(P>0.05),1组尿中尿素氮含量显著高于2组(P<0.05),极显著高于3组(P<0.01),并且2组显著高于3组(P<0.05)。2)采食1.5和3.0 h以后,1组氨态氮(NH_3-N)含量显著高于3组(P<0.05),采食4.5 h后,各组间NH_3-N含量无显著差异(P> 0.05)。pH均在6.0～6.8,1组pH显著高于3组(P <0. 05)。3)采食1. 5 h后,1组乙酸含量显著高于2组(P <0.05),极显著高于3组(P<0.01)。采食1.5、3.5和4.0 h后,1组和2组丙酸含量显著低于3组(P<0.05)。1组采食前及采食后丁酸含量均显著高于3组(P<0.05)。4) 2组和3组Na^+/H^+交换蛋白3(NHE3)和下调式腺瘤载体(DRA)基因表达量显著高于1组(P<0.05)。1组单羧酸转运蛋白4(MCT4)基因表达量显著高于2组和3组(P <0. 05)。而2组Na^+/H^+交换蛋白1(NHE1)和钠/钾ATP酶泵(Na^+/K^+ATPase)基因表达量显著高于1组和3组(P <0. 05)。综上所述,饲粮NFC/NDF为0.66时,对细毛羊瘤胃内环境和VFA吸收相关基因表达量有显著影响。     

胡麻饼代替豆粕对绵羊瘤胃代谢的影响

本文通过研究绵羊瘤胃发酵、消化酶活性及瘤胃上皮组织中挥发性脂肪酸(VFA)吸收相关基因表达量,探讨胡麻饼代替豆粕对绵羊瘤胃代谢的影响。试验选取5月龄、体重为(26.0±1.0)kg的杜泊×小尾寒羊杂交F1代公羔24只,随机分为4组[对照(CK)、Ⅰ、Ⅱ和Ⅲ组],每组6只羊,分别饲喂胡麻饼代替豆粕的饲粮,胡麻饼添加比例分别为0、6%、12%和18%。试验结束后,屠宰并取样,采集瘤胃液和瘤胃组织,研究瘤胃液发酵参数、消化酶活性及瘤胃上皮组织VFA吸收相关基因的表达。预试期10 d,正试期60 d。结果表明:1)Ⅰ和Ⅱ组的平均日增重(ADG)与CK组无显著差异(P0.05),但显著高于Ⅲ组(P0.05);Ⅱ组的料重比(F/G)显著低于CK组(P0.05)。2)与CK组相比,Ⅱ和Ⅲ组的羟甲基纤维素酶、β-葡萄糖苷酶和蛋白酶的活性显著提高(P0.05),Ⅲ组木聚糖酶和果胶酶活性显著提高(P0.05)。3)与CK组相比,胡麻饼代替豆粕对绵羊瘤胃液p H、丙酸和乳酸浓度、乙丙比无显著影响(P0.05),但Ⅱ组乙酸、丁酸和总挥发性脂肪酸(TVFA)的浓度显著提高(P0.05);Ⅰ、Ⅱ和Ⅲ组氨态氮(NH3-N)浓度均显著降低(P0.05),微生物蛋白(MCP)浓度显著提高(P0.05);4)与CK组相比,Ⅱ组单羧酸转运蛋白1(MCT1)和单羧酸转运蛋白4(MCT4)的mRNA相对表达量显著提高(P0.05),腺瘤下调蛋白(DRA)的mRNA相对表达量显著降低(P0.05);Ⅲ组Na+/H+交换蛋白(NHE3)的mRNA相对表达量显著提高(P0.05),MCT1、MCT4和DRA的mRNA相对表达量显著降低(P0.05);胡麻饼代替豆粕对阴离子交换蛋白2(AE2)的mRNA表达量无显著影响(P0.05)。综上,绵羊饲粮中一定比例的胡麻饼代替豆粕能够改善瘤胃代谢状况,在本试验条件下,饲粮中胡麻饼替代豆粕的适宜添加比例为12%。     

短链脂肪酸对奶牛瘤胃上皮细胞的炎性细胞因子和G-蛋白偶联受体41表达的影响

本试验旨在研究不同浓度短链脂肪酸(SCFA)对奶牛瘤胃上皮细胞的炎性细胞因子和G-蛋白偶联受体41(G PR41)表达的影响。试验分为4组,每组3个重复,分别用5、10、20和40 mmol/L的SCFA培养奶牛瘤胃上皮细胞24 h,收集细胞提取总RNA,通过实时荧光定量PCR(qRT-PCR)对炎性细胞因子、趋化因子和G PR41表达量进行测定。结果表明:添加20和40 mmol/L SCFA条件下,G PR41表达量显著高于添加5 mmol/L SCFA (P 0.05);与添加5和10 mmol/L SCFA相比,添加20和40 mmol/L SCFA显著加强了白细胞介素-1β(IL-1β)表达量(P0.05);与添加5 mmol/L SCFA相比,添加10、20和40 mmol/L SCFA显著加强肿瘤坏死因子-α(TNF-α)表达量(P 0. 05);此外,添加5、10和20 mmol/L SCFA之间的趋化因子20(CCL20)表达量差异不显著(P0.05),然而,添加40 mmol/L SCFA条件下,CCL20的表达量显著上调(P0.05);与添加5和10 mmol/L SCFA相比,添加20和40 mmol/L SCFA显著加强趋化因子2(CXCL2)和趋化因子8(CXCL8)表达量(P0.05);随着SCFA浓度逐渐上升,趋化因子3(CXCL3)表达量显著上调(P0.05)。综上所述,SCFA诱导GPR41表达,从而介导炎性细胞因子以及趋化因子的表达上调,介导瘤胃上皮保护性免疫反应。     

MCT1在犊牛消化道的分布以及SCFA对MCT1表达的影响

短链脂肪酸(SCFAs)是由纤维素性饮食在瘤胃内发酵产生的乙酸、丙酸、丁酸以及戊酸等组成的混合物。SCFA对维持反刍动物的能量平衡至关重要,目前对SCFA的吸收机制和影响因素至今尚不清楚,本试验主要研究一元羧酸转运蛋白(monocarboxylate/proton cotransporter isoform 1,MCT1)在犊牛消化道内的分布及SCFA对MCT1表达的影响。运用免疫印迹Western blot和实时荧光定量PCR检测MCT1在犊牛消化道内的分布和表达;在体外试验中,体外添加一定比例的SCFA(乙酸、丙酸和丁酸的混合物)处理犊牛原代瘤胃上皮细胞,检测SCFA对MCT1表达水平的影响。结果显示:MCT1在整个消化道内均有分布,并且在瘤胃内的表达水平显著高于其他部位(P<0.01);体外处理的瘤胃上皮细胞,正常组MCT1的表达水平显著高于(P<0.05)SARA组。结果表明:正常水平的SCFA能够促进MCT1的表达和转运能力,而过量的SCFA抑制MCT1的表达和转运能力。     

香芹酚和百里香酚对绵羊养分表观消化率、瘤胃发酵特性及纤维降解菌数量的影响

本试验旨在探讨香芹酚和百里香酚对绵羊养分表观消化率、瘤胃发酵特性及纤维降解菌数量的影响。选取9只健康装有永久瘤胃瘘管的小尾寒羊公羊,采用3×3拉丁方设计,预试期5 d,正试期15 d。对照组(CON)饲喂基础饲粮,试验1组(T₁组)在基础饲粮精料中添加香芹酚(2 g/d),试验2组(T₂组)在基础饲粮精料中添加百里香酚(2 g/d)。从正试期第10天开始,连续5 d采用全收粪法进行消化代谢试验,测定养分表观消化率;正试期最后1天在采食前(0 h)以及采食后1、3、5、7 h采集瘤胃液,测定发酵参数和纤维降解菌数量。结果表明:1)各组绵羊在采食后1 h瘤胃液氨态氮(NH 3-N)浓度均处于最高水平,采食后3 h瘤胃液pH均降到最低,而瘤胃液总挥发性脂肪酸(TVFA)浓度最高,采食后3、5 h瘤胃液微生物蛋白(MCP)浓度均处于较高水平,瘤胃液黄色瘤胃球菌、产琥珀酸瘤胃球菌数量在采食后3 h最高,白色瘤胃球菌数量在采食后1 h最高。2)从平均值看,与对照组相比,T₁组瘤胃液MCP、TVFA浓度显著升高(P<0.05),瘤胃液pH显著降低(P<0.05),干物质和粗蛋白质表观消化率显著升高(P<0.05),瘤胃液黄色瘤胃球菌、产琥珀酸瘤胃球菌数量显著升高(P<0.05),白色瘤胃球菌数量显著降低(P<0.05);T₂组瘤胃液MCP、TVFA浓度显著升高(P<0.05),各养分表观消化率无显著变化(P>0.05),瘤胃液白色瘤胃球菌数量显著降低(P<0.05)。综上所述,香芹酚和百里酚香均可影响绵羊瘤胃发酵特性及纤维降解菌数量,且香芹酚还可以显著提高绵羊的干物质和粗蛋白质表观消化率。     

饲粮中添加海南霉素对奶牛瘤胃发酵及甲烷产量的影响

本试验旨在研究海南霉素对荷斯坦奶牛瘤胃发酵模式及甲烷产量的影响。试验选用3头装有永久性瘤胃瘘管、体况相近的中国荷斯坦奶牛,采用3×3拉丁方试验设计,试验分为3期,每期15 d,试验设负对照组(不添加任何添加剂)、正对照组(添加10 mg/kg莫能菌素)和海南霉素组(添加7.2 mg/kg海南霉素)。结果表明:饲粮中添加海南霉素后,除在采食后2 h显著提高奶牛瘤胃内pH(P<0.05)外,其他时间均无显著影响(P>0.05);采食后0、2、8和10 h,海南霉素组氨态氮(NH3-N)浓度均显著低于负对照组(P<0.05),而采食后6 h,海南霉素有抑制NH3-N释放的趋势(P=0.06);采食后0、2、4和6 h,海南霉素组乙酸浓度及乙酸与丙酸的比值与负对照组相比均显著降低(P<0.05),丙酸浓度显著升高(P<0.05),总挥发性脂肪酸和丁酸浓度没有显著变化(P>0.05)。海南霉素显著抑制了瘤胃甲烷的产生(P=0.02),海南霉素组的奶牛甲烷呼出量为216.50 L/d,比负对照组降低了14.03%。由此得出结论:饲粮中添加海南霉素可以改变奶牛瘤胃的发酵类型,使其更趋向于丙酸型发酵,并显著降低动物的甲烷呼出量。     

高谷物日粮促进山羊瘤胃上皮单羧酸转运蛋白1及钠钾ATP酶mRNA的表达

本试验旨在研究高谷物日粮对山羊瘤胃上皮形态结构及单羧酸转运蛋白(monocarboxylate transporter, MCT)和钠钾ATP酶mRNA表达的影响。将10头装有永久性瘤胃瘘管的健康阉割公山羊随机分为饲喂全粗料日粮的对照组(Hay,0%谷物,n=5)和饲喂高谷物日粮的处理组(HG,65%谷物,n=5),试验期为7周。试验开始后,于每周晨饲后的0、2、3、4、6、8和12 h连续采集瘤胃液监测瘤胃pH值的变化,收集其中第0、3、6和12 h的瘤胃液待测挥发性脂肪酸(volatile fatty acid, VFA)浓度。试验的第50天,屠宰采集瘤胃上皮用于形态学及基因定量分析。研究结果显示:与全粗料组山羊相比,高谷物组山羊瘤胃pH值、乙酸浓度及乙丙比都显著下降(P<0.001),而瘤胃丙酸浓度、丁酸浓度及其他VFA浓度都显著升高(P<0.001);高谷物日粮组的瘤胃乳头长度显著高于对照组(P=0.001),瘤胃乳头宽度显著低于对照组(P<0.001),但是两组间的瘤胃乳头表面积并无显著差异;透射电镜结果显示,长期饲喂高谷物日粮导致瘤胃上皮细胞线粒体发生降解;实时定量PCR结果表明,与对照组相比,高谷物日粮显著升高了MCT1(P<0.001)和钠钾ATP酶(P=0.001)的mRNA表达量,显著降低了MCT4的mRNA表达量(P=0.041),但对MCT2的表达没有显著影响(P=0.305);进一步分析这些基因的mRNA表达量与pH值和VFA浓度之间的相关性,结果显示,MCT1和钠钾ATP酶的mRNA表达量与瘤胃pH值和乙酸浓度呈显著负相关,与总VFA、丙酸、丁酸的含量呈显著正相关,而MCT4的mRNA表达量与pH值呈显著正相关,与总VFA、丙酸、丁酸的含量呈显著负相关。以上结果提示:高精料引起的瘤胃pH值下降和VFA的变化可能与瘤胃上皮MCT和钠钾ATP酶表达量的变化相关。研究结果对深入认识高谷物饲喂引发的瘤胃功能紊乱具有重要意义。     

棕榈油和植物次生化合物对奶牛采食量、消化率及微生物种群的影响

本研究旨在确定添加棕榈油和豆荚对奶牛采食量、消化率、微生物蛋白合成和微生物种群的影响。试验选择4头泌乳早期、平均体重为(408±37)kg的奶牛,按照4×4拉丁方设计进行分组。分别饲喂4种日粮,对照组为基础日粮组,T1组为60 g/kg豆荚组,T2组为20 g/kg棕榈油组,T3组为60 g/kg豆荚组+20 g/kg棕榈油组。结果发现:对照组和棕榈油组粗蛋白质表观消化率较豆荚组显著提高10.26%和9.11%(P 0.05),而棕榈油+豆荚组中性洗涤纤维表观消化率较其他组显著提高3.29%、2.17%和2.97%(P 0.05)。奶牛采食4 h后,对照组瘤胃氨氮浓度较豆荚组显著提高35.11%(P 0.05),而对照组和棕榈油组氨氮浓度均值较其他两组分别提高12.64%、11.36%和12.07%、10.80%(P 0.05)。豆荚组尿囊素吸收量分别较其他组显著提高19.22%、11.94%和10.27%(P 0.05),同时微生物蛋白合成量最高(P 0.05)。豆荚+棕榈油组奶牛采食4 h后瘤胃纤维分解菌数量较其他组显著提高6.76%、8.22%和6.76%(P 0.05),而对照组和棕榈油组瘤胃蛋白水解菌数量较豆荚组显著提高21.87%和23.43%(P 0.05)。结论 :奶牛日粮添加棕榈油提高了纤维消化率和瘤胃纤维素分解菌的数量,而添加豆荚降低了粗蛋白质消化率、瘤胃氨氮浓度、瘤胃蛋白水解菌和原生动物数量,进而导致微生物蛋白合成量增加。     

不同蛋白质和碳水化合物饲料来源对牛瘤胃洗涤纤维降解和瘤胃液产物的定量研究

试验采用3×3拉丁方设计,3头安装永久性大口径瘤胃瘘管的青年母牛饲喂3种不同精饲料的日粮,3种精饲料分别为豆粕+玉米(日粮1),猪血粉+玉米(日粮2)和豆粕+小麦(日粮3)。精料日喂量按活牛体重的1%喂给,干玉米秸等量组成的日粮配给。试验为3个试验周期,每一周期为37d,其中适应期26d,在早晨采食后2、4、8、16和24h,分别5次实施瘤胃掏空,全量测定瘤胃物存留量。结果表明:日粮1在采食后2～8h瘤胃干物质、液体和NDF的存留量最低,与其他两种日粮相比差异显著(P<0.05);日粮1和日粮3瘤胃液氨氮存留量在采食后2～8h始终高于日粮2(P<0.05),但日粮3在采食4h后瘤胃液氨氮开始下降,而日粮1至采食后16h开始下降,日粮1瘤胃液氨氮数量采食后2～8h处于高峰状态,说明瘤胃微生物在瘤胃环境长时间一直保持稳定的降解能力;日粮1和日粮3在采食2～4h后处于较高的总挥发性脂肪酸(VFA)瘤胃存留量,日粮1在采食后8～16h瘤胃存留量降低,与日粮2和日粮3相比差异显著(P<0.05),日粮1对瘤胃VFA的快速移除,意味着瘤胃液的快速流出以及可能解释瘤胃固体物的快速移除。因此,日粮1和日粮3在牛瘤胃洗涤纤维降解和瘤胃液产物代谢模式发挥不同作用。     

牛瘤胃上皮细胞中挥发性脂肪酸吸收相关蛋白在白细胞介素10诱导下的差异表达研究

本研究旨在探讨炎症因子（白细胞介素10, IL-10）对牛瘤胃上皮细胞中挥发性脂肪酸（VFA）吸收相关蛋白基因表达的影响。试验采用随机区组设计,在瘤胃上皮细胞体外培养条件下,研究50 ng·mL^-1浓度的IL-10对牛瘤胃上皮细胞中核因子（NF-κB）、假定阴离子转运蛋白1（PAT1）、阴离子交换蛋白2（AE2）、单羧酸转运蛋白1（MCT1）、单羧酸转运蛋白4（MCT4）、钠氢离子交换蛋白1（NHE1）基因表达的影响。结果表明,在中性条件下,IL-10添加显著下调了NF-κB、PAT1、MCT1、NHE1基因的表达（P<0.05）,AE2基因的表达显著性上调（P<0.05）,而对MCT4基因的表达没有显著影响。在酸性条件下,IL-10添加组的NF-κB基因表达被显著性抑制（P<0.05）,而PAT1、MCT1、MCT4基因表达显著上调（P<0.05）。pH5.5组与pH7.2组比较,pH5.5组NF-κB基因表达显著性上调（P<0.05）。结果提示,在炎症条件下,白细胞介素10作为一种抗炎因子,可以缓解瘤胃上皮细胞炎症反应从而促进VFA吸收相关蛋白基因的表达。     

An energy-rich diet enhances expression of Na(+)/H(+) exchanger isoform 1 and 3 messenger RNA in rumen epithelium of goat

Rumen epithelial Na(+)/H(+) exchanger (NHE) catalyzes the exchange of extracellular Na(+) for intracellular H(+). Thus, it is of importance in the maintenance of Na and pH homeostasis of rumen epithelial cells. We have tested the hypothesis that an increase in energy and protein intake induces alterations of NHE isoform 1, 2, and 3 (NHE1, NHE1, and NHE3, respectively) mRNA abundance in the rumen epithelium of goats. Goats (n = 26) were randomly allocated to 2 experiments (n = 16 in Exp. 1, and n = 10 in Exp. 2) and fed either peanut straw ad libitum [PNS, n = 8 in Exp. 1, and n = 5 in Exp. 2; 600 kJ of ME/(kg(0.75)·d)] or PNS + concentrate [CF, n = 8 in Exp. 1, and n = 5 in Exp. 2; 1,000 kJ of ME/(kg(0.75)·d)] for 42 d. Concentrate (400 g/d) was given daily (0800 to 1700 h) in 4 equal portions at 3-h intervals. In Exp. 1, the goats were euthanized 2 h after the last portion of concentrate was fed, and in Exp. 2, the goats were euthanized after a fasting period of 16 h. In Exp. 1, goats in the CF treatment exhibited a greater ruminal short-chain fatty acid (SCFA) concentration (140.6 ± 1.30 mM) compared with those in the PNS treatment (114.3 ± 3.11 mM; P < 0.001), and pH decreased from 6.9 ± 0.09 to 5.9 ± 0.04 (P < 0.001). Correspondingly, the mRNA expression of NHE1 and NHE3 in the rumen epithelium was greater by 20% (P = 0.041) and 25% (P = 0.043) for goats in the CF treatment than for those in the PNS treatment. However, in Exp. 2, 16 h of fasting abolished differences in ruminal SCFA concentration, pH, and NHE mRNA expression between goats in the CF and PNS treatments. In both Exp. 1 and 2, a positive correlation was observed between ruminal SCFA concentration and expression of mRNA in NHE1 and NHE3, whereas expression was negatively correlated with ruminal pH. In in vitro studies with isolated rumen epithelial cells from goats fed dried grass, exposure to pH of 6.8 or to 20 mM SCFA increased (P < 0.01) NHE1 and NHE3 mRNA expression, as compared with exposure to pH of 7.4 or the absence of SCFA. A combination of reduced pH (6.8) and SCFA (20 mM) further enhanced (P < 0.05) NHE1 and NHE3 mRNA expression, indicating synergism between an increased concentration of SCFA and low pH (P < 0.05). Messenger RNA expression of NHE2 did not vary in vitro with pH (6.8) or SCFA (20 mM) or in vivo in Exp. 1 and 2. Thus, diet-dependent rumen epithelial NHE1 and NHE3 expression is probably related to ruminal SCFA concentration and pH, but that is not the case with NHE2.     

Post‐natal changes in MCT1 expression in the forestomach of calves

The monocarboxylate transporter 1 (MCT1) has been demonstrated to be involved in the transfer of short‐chain fatty acids (SCFA) and/or their intraepithelial metabolites from the rumen to the blood. As MCT1 plays a role in SCFA transfer, it is assumed that SCFA are the main substrates influencing its expression. However, there are hints that MCT1 may also be expressed during the early life of the animal when SCFA are not released in the forestomach. To figure out whether MCT1 expression in the forestomach is influenced independently of SCFA during that period, we studied post‐natal MCT1 expression immunohistochemically in the epithelia of omasum, atrium ruminis, saccus dorsalis ruminis, saccus ventralis ruminis and reticulum of calves born preterm and at term. The calves were nourished by colostrum or by milk‐based formula diet. MCT1 could be found in all the forestomach compartments tested, even in preterm calves. The protein was mainly oriented to the luminal side in the immature epithelium 24 h after birth. Orientation to the blood side of the cells developed during the first 4 days after birth. In the rumen epithelia (but not in the other forestomach compartments tested), orientation of MCT1 to the blood side of the cells was paralleled by an increase in the overall expression rate during the first 4 days after birth. As lactate levels were very high directly after birth, a lactate‐dependent substrate induction may have been the underlying mechanism. However, non‐specific changes due to general differential processes might also be the cause. Both early upregulation of MCT1 and high blood lactate levels may provide the epithelia with lactate as energy source.     

Ruminant Nutrition Symposium: Role of fermentation acid absorption in the regulation of ruminal pH

Highly fermentable diets are rapidly converted to organic acids [i.e., short-chain fatty acids (SCFA) and lactic acid] within the rumen. The resulting release of protons can constitute a challenge to the ruminal ecosystem and animal health. Health disturbances, resulting from acidogenic diets, are classified as subacute and acute acidosis based on the degree of ruminal pH depression. Although increased acid production is a nutritionally desired effect of increased concentrate feeding, the accumulation of protons in the rumen is not. Consequently, mechanisms of proton removal and their quantitative importance are of major interest. Saliva buffers (i.e., bicarbonate, phosphate) have long been identified as important mechanisms for ruminal proton removal. An even larger proportion of protons appears to be removed from the rumen by SCFA absorption across the ruminal epithelium, making efficiency of SCFA absorption a key determinant for the individual susceptibility to subacute ruminal acidosis. Proceeding initially from a model of exclusively diffusional absorption of fermentation acids, several protein-dependent mechanisms have been discovered over the last 2 decades. Although the molecular identity of these proteins is mostly uncertain, apical acetate absorption is mediated, to a major degree, via acetate-bicarbonate exchange in addition to another nitrate-sensitive, bicarbonate-independent transport mechanism and lipophilic diffusion. Propionate and butyrate also show partially bicarbonate-dependent transport modes. Basolateral efflux of SCFA and their metabolites has to be mediated primarily by proteins and probably involves the monocarboxylate transporter (MCT1) and anion channels. Although the ruminal epithelium removes a large fraction of protons from the rumen, it also recycles protons to the rumen via apical sodium-proton exchanger, NHE. The latter is stimulated by ruminal SCFA absorption and salivary Na(+) secretion and protects epithelial integrity. Finally, SCFA absorption also accelerates urea transport into the rumen, which via ammonium recycling, may remove protons from rumen to the blood. Ammonium absorption into the blood is also stimulated by luminal SCFA. It is suggested that the interacting transport processes for SCFA, urea, and ammonia represent evolutionary adaptations of ruminants to actively coordinate energy fermentation, protein assimilation, and pH regulation in the rumen.     

Differences in monocarboxylic acid transporter type 1 expression in rumen epithelium of newborn calves due to age and milk or milk replacer feeding

The aim of the present study was to investigate whether besides age and solid feed intake, monocarboxylic acid transporter type 1 (MCT1) expression in the rumen epithelium of calves is affected by liquid feed type [whole milk (WM) or milk replacer (MR)]. Thirty bull calves at the mean age of 5 days were randomly allocated to five experimental groups (six calves/group). Six calves were slaughtered immediately after allocation to the trial (5 days of life), eighteen calves were fed MR and slaughtered at week intervals (on 12, 19, 26 days of life respectively), and six calves were fed WM and slaughtered at the 26 days of life. MCT1 protein abundance and the MCT1 mRNA level were investigated in the dorsal and ventral sack of the rumen. Solid feed intake and short‐chain fatty acids (SCFA) concentration in the rumen fluid increased linearly with calves' age. The amount of the MCT1 protein and mRNA in the dorsal sac of rumen as well as the amount of MCT1 protein in the cranial ventral sac of rumen also increased linearly with calves' age. Calves fed WM had greater solid feed intake in the last week of the study as compared to calves fed MR, but SCFA concentration in the rumen fluid was not different. MCT1 mRNA expression in the cranial dorsal sac of rumen and protein MCT1 expression in both dorsal and ventral cranial sack of the rumen were higher in calves fed WM as compared to calves fed MR. This study confirmed age‐dependent changes of MCT1 expression in the rumen epithelium of newborn calves and showed that its expression might be affected by liquid feed type.     

高精料诱导的SARA对泌乳期山羊血液和瘤胃液中皮质醇的影响

选择7只装有永久性瘤胃瘘管的泌乳中期山羊,2×2拉丁方设计,分别饲喂精粗比为6∶4和4∶6的饲料,通过饲喂精粗比6∶4饲料建立泌乳期山羊瘤胃亚急性酸中毒(subacute ruminal acidosis, SARA)模型,研究SARA时山羊血液和瘤胃液中皮质醇浓度、肝脏皮质醇受体(glucocorticoid receptor, GR)mRNA表达及其他相关指标的变化。结果表明,精粗比6∶4日粮饲喂2周后成功诱导SARA状态(SARA组),采食后瘤胃液pH值低于5.8持续时间约6 h,而精粗比为4∶6组山羊瘤胃液pH均高于6.0 (对照组)。高精料日粮处理对瘤胃pH和乳酸以及瘤胃液和血浆内的脂多糖、皮质醇浓度有显著性影响(P<0.05),采食前SARA组山羊瘤胃液中乳酸浓度显著高于对照组(P<0.05),采食后0~4 h乳酸含量下降,6~10 h时逐渐增加并极显著高于对照组(P<0.01);2组间瘤胃液中脂多糖浓度无显著性差异(P>0.05),SARA组血浆脂多糖浓度在采食前和采食后6 h均显著高于对照组(P<0.05);采食前SARA组山羊血液中皮质醇浓度高于对照组(P=0.05),但采食后6 h两组间无显著差异(P>0.05); 2组山羊瘤胃液中皮质醇浓度在采食后2,4和6 h分别显著高于对照组(P<0.05),但在采食前和采食后10 h无显著差异(P>0.05)。Real-time PCR结果显示,SARA组山羊肝脏中GR mRNA表达显著下调(P<0.05)。本研究结果显示,泌乳期山羊发生SARA时糖皮质激素水平升高,负反馈下调肝脏GR的表达水平,提示SARA时机体处于应激状态,可能引起肝脏的物质代谢和营养物质重分配的改变。     

Diurnal Patterns of Ruminal Concentrations and Portal Appearance Rates of Short-chain Fatty Acids in Sheep Fed a Hay or a Concentrate/Straw Diet in Two Meals Daily

Abstract

Three rumen fistulated and catheterized sheep were meal-fed and used to study ruminal and arterial concentrations of short-chain fatty acids (SCFA) as well as portal appearance rates of SCFA and irreversible loss rate (ILR) of acetate in 24 h periods on a hay and a concentrate/straw diet, respectively.

Ruminal and arterial concentrations as well as portal appearance rates of SCFA and ILR of acetate were significantly affected by the intake of feed. Generally, the highest concentrations and appearance rates were obtained 2 h after feeding. The portal recovery of arterial acetate was not affected by feeding or diet. The 24 h means were 0.68 ± 0.01 and 0.67 ± 0.01 on the hay and the concentrate/straw diet, respectively. Partial correlation coefficients corrected for the effects of time, sheep, and diet were calculated for the relationships evaluated. The portal appearance rate of acetate (r = 0.52, P <0.001) and the portal net appearance rate of propionate (r = 0.68, P <0.001) were linearly related to the ruminal concentrations of the two SCFA. The logarithm of the portal net appearance rate of butyrate seemed to be linearly related to the logarithm of the ruminal concentration of undissociated butyric acid (r = 0.70, P <0.001) when the effect of time was omitted from the model. The portal appearance rate of acetate (r = 0.22, P <0.05) and the portal net appearance rate of propionate (r = 0.63, P <0.001) as well as butyrate (r = 0.55, P <0.001) were linearly related to the arterial concentration of the respective SCFA. The results show that within animal and diet the ruminal as well as arterial concentrations are good predictors of SCFA portal appearance rates in sheep fed roughage at maintenance. Ruminal and arterial concentrations of SCFA seem less reliable predictors of portal appearance rates of SCFA between diets and sheep. The portal appearance of SCFA, 5.6 ± 0.5 and 7.0 ± 0.3 mol d^?1, accounted for 44 ± 4 and 43 ± 2% of the calculated metabolizable energy intake on the hay and the concentrate/straw diets, respectively.