丁酸钠对高精料日粮模式下奶山羊乳腺内组胺介导的炎症反应的影响

本试验旨在研究饲喂高精料日粮以及日粮中添加丁酸钠对奶山羊乳腺内组胺介导的炎症反应的影响。选用泌乳中期的18头萨能奶山羊,随机分成3组,每组各6头。对照组(LC)饲喂粗精比为6∶4的低精料日粮,即日粮包括60%粗料和40%精料;试验一组(HC)饲喂粗精比为4∶6的高精料日粮,即日粮包括40%粗料和60%精料;试验二组(BHC)饲喂粗精比为4∶6的高精料日粮,并在日粮中添加20 g丁酸钠。试验期为20周。试验开始前两周,所有试验动物手术安装瘤胃瘘管。试验第20周采集瘤胃液、外周血液和乳腺组织样品。结果显示:与LC组相比,HC组瘤胃液、外周血液、乳腺组织中的组胺含量显著升高,外周血液中的炎性因子IL-1β、IL-6和TNF-α显著升高,乳腺组织中的组胺受体基因表达显著升高,炎性因子在基因和蛋白表达水平上均显著升高。与HC组相比,BHC组瘤胃液、外周血液、乳腺组织中的组胺含量显著下降,外周血液中的炎性因子均显著降低,乳腺组织中的组胺受体基因表达显著降低,炎性因子的基因和蛋白表达显著下降。结果表明:泌乳奶山羊长期饲喂高精料日粮后,瘤胃产生的组胺经消化道吸收并转运到乳腺组织中,刺激炎症的产生;而通过添喂丁酸钠,组胺和炎性因子的产生显著降低,乳腺组织炎症反应减轻。     

SARA状态下瘤胃产生的LPS对奶牛乳腺组织细胞凋亡的影响

本试验采用12头泌乳中期的荷斯坦奶牛,随机分为两组,每组6只。其中一组饲喂低精料日粮,即日粮中含有40%的精料和60%的粗料,精粗比为4∶6,作为对照(LC)组;另外一组饲喂高精料日粮,精粗比为6∶4,作为试验(HC)组。试验周期为18周,试验开始前为所有奶牛安装瘤胃瘘管,试验后期采集瘤胃液,乳动脉、静脉血和乳腺组织样品,测定瘤胃液p H值和LPS含量、血浆中LPS含量、乳腺组织细胞中凋亡相关基因如caspase-3、caspase-8、caspase-9、bax和bcl-2等的表达、凋亡相关蛋白酶的活性和NF-κB通路中的蛋白表达。结果显示:长期高精料饲喂使奶牛瘤胃p H值均值降低,一天中p H值低于5.6的时间超过3 h;高精料组血浆和瘤胃液中LPS含量均极显著高于低精料组(P0.01);高精料组caspase-3的基因表达显著高于低精料组(P0.05),而bcl-2的基因表达显著低于低精料组(P0.05);高精料组caspase-3和caspase-8的酶活性相比于低精料组都显著升高(P0.01);高精料组NF-κB p65和磷酸化的NF-κB p65的蛋白表达量均显著高于低精料组(P0.05)。     

高精料对奶山羊盲肠中SCFAs-GPR41/43介导的炎症反应的影响

为了探讨高精料长期饲喂过程中,反刍动物肠道内的短链脂肪酸(SCFAs)在奶山羊盲肠中通过其特异性受体G蛋白偶联受体41/43(GPR41/43)对其下游的炎症信号通路是否有影响,选用12只萨能奶山羊随机分为2组,对照组(LC)饲喂低精料(精∶粗=4∶6,n=6),试验组(HC)饲喂高精料(精∶粗=6∶4,n=6)日粮,经10周饲喂试验,取盲肠组织样及盲肠内容物进行气相色谱短链脂肪酸含量、肠内容物pH、实时荧光定量(RT-q PCR)及蛋白印迹的检测。结果显示:相比于LC组,HC组乙酸(P0.01)、丙酸(P0.01)及丁酸(P0.05)的含量极显著或显著升高,且HC组pH值明显降低(P0.05);HC组GPR41/43、肿瘤坏死因子-α(TNF-α)及趋化因子20(CCL20)基因表达量相比于LC组极显著升高(P0.01),白细胞介素-1β(IL-1β)、IL-6、IL-8、IL-10及CCL5等基因表达量显著升高(P0.05);HC组GPR41/43蛋白表达量显著高于LC组(P0.05)。提示:高精料饲喂后,反刍动物盲肠组织中通过短链脂肪酸活化GPR41/43进而激活下游炎症信号通路诱导组织内炎症反应。     

饲喂高精料日粮对泌乳期山羊泌乳性能及抗氧化能力的影响

为了研究长期饲喂高精料日粮对泌乳期山羊泌乳性能和乳腺组织抗氧化能力的影响,实验选用12只健康经产泌乳中期关中奶山羊,随机分为两组,分别饲喂精粗比为35∶65(低精料组, LC, n=6)和65∶35(高精料组, HC, n=6)的日粮。实验期共10周,实验期间采集羊奶与血液;实验结束后采用静脉注射的方法对山羊进行安乐死,随后采集乳腺组织并迅速置于液氮中速冻后保存于-70 ℃待测。结果显示,HC组山羊乳产量显著高于LC组(P<0.01),同时乳中乳糖,乳蛋白的比例随着泌乳时间的增加而显著升高。与LC组山羊比较,HC组山羊乳腺组织中SREBP1和GLUT1 mRNA表达显著下降(P<0.05);磷酸化AKT1蛋白表达呈下降趋势(P=0.08);除此之外,HC组山羊乳腺组织的总抗氧化能力T-AOC(P=0.06)和GSH酶活均呈下降趋势(P=0.08)。这些结果表明,长期饲喂高精料日粮可降低乳腺组织的抗氧化能力,同时可降低乳中乳脂率以及乳腺组织中SREBP-1和P-AKT的表达。     

高精料日粮对奶山羊肝、脾炎性因子表达的影响

为探究饲喂高精料日粮对泌乳奶山羊肝、脾组织的影响及机制,将14只泌乳初期关中奶山羊随机分为对照组(6只)和试验组(8只),对照组饲喂普通(精粗比35∶65)日粮,试验组饲喂高精料(精粗比65∶35)日粮,试验期19周。于试验的0、1、4、8、12、15、16、18、19周每周采集所有羊乳汁测定乳脂率,6～18周每周日采集所有羊粪便测定pH,试验结束当日采集肝、脾组织,制作组织切片观察肝、脾组织形态变化;采用RT-qPCR检测肝、脾组织中炎性因子及TLR4通路关键蛋白的基因表达;采用Western blot技术检测TLR4通路关键蛋白的表达水平。结果显示,长期饲喂高精料日粮造成奶山羊乳脂率降低,试验的16、18和19周与对照组差异显著(P0.05);试验组奶山羊粪便pH均值从第9周开始下降,13～18周与对照组差异显著(P0.05);试验组奶山羊肝细胞颗粒变性或空泡变性,肝小叶中央静脉周围有炎性细胞浸润,脾组织无明显变化;肝组织中促炎因子IL-6和TNF-β以及TLR4与NF-κB2基因表达显著升高(P0.05);脾组织中促炎因子TNF-β和抗炎因子IL-10以及TLR4基因表达显著升高,NF-κB2基因表达显著降低(P0.05);肝组织中与TLR4通路相关的p38、JNK、ERK和p65这4种蛋白的表达变化不明显,脾组织p38、ERK和p65表达与对照组相比虽差异不显著,但都呈现下降趋势。综上所述,长期饲喂高精料日粮导致奶山羊乳脂率和粪便pH降低,通过上调NF-κB的基因表达诱导肝组织中IL-6和TNF-β基因表达升高,从而对肝造成炎性损伤,对脾组织影响不明显。     

高精料日粮对奶牛肝脏细胞凋亡的影响

选用12头安装瘤胃瘘管和肝、门静脉血管插管的泌乳中期荷斯坦奶牛,研究高精料日粮对反刍动物肝脏细胞凋亡的影响。试验随机分成2组:高精料组(HC)和低精料组(LC)。饲喂后通过瘤胃瘘管采集0~12 h瘤胃液,测定瘤胃p H值;生化分析法检测外周血中谷草转氨酶(GOT)、谷丙转氨酶(GPT)、乳酸脱氢酶(LDH)、白蛋白(ALB)和总蛋白(T-Pro)含量;检测肝、门静脉血液中脂多糖(LPS)的含量;采用RT-q PCR法检测肝脏中凋亡相关基因的表达;分光光度计法测定肝脏细胞中Caspase-3、Caspase-8的活性;Western blot法测定NF-κB的蛋白表达量。结果:20周后,高精料组瘤胃内p H低于5.6且每天持续3 h以上,显示奶牛发生SARA。与低精料组相比,高精料组门静脉LPS的浓度显著升高,肝功能受到影响,肝脏中促凋亡基因表达上调,抑凋亡基因表达下调,NF-κB的蛋白表达量升高,Caspase-3、Caspase-8的酶活性均表达上调,且Caspase-3有显著性的差异。结果表明,长期饲喂高精料,使消化道中的LPS由门静脉进入肝脏,引起肝脏的炎性损伤,增加肝细胞的细胞凋亡程度。     

复合缓冲剂对奶山羊乳成分、血浆生化指标及激素含量的影响

本试验旨在研究长期饲喂高精料饲粮下添加复合缓冲剂(碳酸氢钠20 g/d、氧化镁12 g/d、丁酸钠20 g/d)对奶山羊乳成分、血浆生化指标及激素含量的影响。选择8头安装门静脉及肝静脉瘘管处于泌乳中期的奶山羊随机分为2组,分别饲喂基础饲粮(高精料饲粮组,HG组)和基础饲粮+复合缓冲剂(高精料饲粮+复合缓冲剂组,BG组)。预试期为7 d,正试期为143 d。结果显示:与HG组相比,复合缓冲剂显著升高采食后010 h瘤胃液p H平均值(P0.05);复合缓冲剂显著降低肝静脉血浆中非酯化脂肪酸的含量(P0.05),对门静脉和肝静脉血浆中葡萄糖、β-羟丁酸含量无显著影响(P0.05),对肝脏组织中甘油三酯和总蛋白含量无显著影响(P0.05);复合缓冲剂显著降低门静脉血浆中胰岛素和胰高血糖素的含量(P0.05),对门静脉和肝静脉血浆中胰岛素样生长因子1和生长激素的含量无显著影响(P0.05);复合缓冲剂显著或极显著升高产奶量、乳脂率、乳蛋白率、乳非脂固形物率(P0.05或P0.01)。综合得出,在长期高精料饲粮饲喂条件下,由碳酸氢钠、氧化镁、丁酸钠组成的复合缓冲剂能通过影响奶山羊血浆生化指标及激素含量,影响体机营养物质代谢,最终有效提高产乳量与改善乳品质。     

奶牛亚急性瘤胃酸中毒对肝脏内组胺代谢及NF-κB炎性信号通路的影响

为研究奶牛亚急性瘤胃酸中毒(SARA)时血液内组胺浓度的变化及对奶牛炎性信号通路的影响,通过对健康奶牛饲喂高精料日粮,诱发亚急性瘤胃酸中毒,手术安装血管瘘管,采集门静脉和肝静脉血液样品,组胺ELISA试剂盒检测血液中组胺含量,并通过RT-PCR对在体肝脏中的NF-κB、环氧合酶-2(COX-2)、IL-6、TNF-α表达水平进行相对定量和绝对定量检测。结果表明,高精料组奶牛门静脉内组胺浓度较对照组均有所增加,高精料组组胺平均浓度(19.99±0.21)μg/L,较低精料组(18.12±0.35)μg/L差异显著(P<0.05);高精料奶牛的组胺平均清除率(57.23%)显著高于低精料组(50.28%),说明高精料组奶牛组胺在肝脏内代谢率高于低精料组;高精料组NF-κB、COX-2、IL-6和TNF-α的表达较低精料组均有升高,且NF-κB、COX-2、IL-6和TNF-α四个基因的表达有显著升高(P<0.05)。本试验证明了奶牛SARA中血液内组胺浓度的升高可诱导肝脏对其转化和降解的效率增加,且组胺含量的增高可诱导奶牛肝脏中NF-ΚB信号通路的打开,进而导致COX-2等炎性基因表达。     

高精料对泌乳奶山羊瘤胃上皮氧化应激和胆固醇代谢的影响

为了研究长期或短期饲喂高精料日粮对泌乳期奶山羊瘤胃上皮组织氧化应激和胆固醇代谢的影响,实验选用17只健康的经产泌乳中期关中奶山羊,随机分为3组:饲喂低精料组(对照组,LC,n=5);长期饲喂高精料组(HL, n=7),19周饲喂期;短期饲喂高精料组(HS,n=5),4周饲喂期。实验结束后采集瘤胃组织,用PBS反复清洗,于液氮中速冻后置于-80 ℃冰箱保存。结果显示,与对照组相比,HL和HS组山羊瘤胃上皮组织中细胞周期相关基因CDK2和CDK4 mRNA表达水平显著升高(P<0.05),且HS组p-ERK1/2蛋白表达显著升高(P<0.05),但GPR41和GPR43蛋白表达无显著变化(P>0.05);HL和HS组山羊瘤胃上皮组织促细胞凋亡基因Casepase9 mRNA表达显著升高(P<0.05),HS组抗凋亡Bcl-2/Bax mRNA表达比例呈下降趋势(0.050.05);HS组瘤胃上皮组织中胆固醇含量下降达显著水平(P<0.05),且HL组呈下降趋势(0.05+ ATPase mRNA表达显著升高(P<0.05),NHE2蛋白表达升高但未达显著差异水平(P>0.05)。与HL组相比,HS组山羊瘤胃上皮组织中总抗氧化能力T-AOC显著升高(P<0.05);且胆固醇含量显著下降(P<0.05)。以上结果说明,短期饲喂高精料日粮可加快泌乳期奶山羊瘤胃上皮组织的更新,提高组织总抗氧化能力;而长期饲喂高精料日粮未引起细胞增殖相关蛋白和总抗氧化能力的显著变化;饲喂高精料日粮可以加快瘤胃上皮组织对VFA的转运。此外,饲喂高精料日粮引起瘤胃上皮组织内胆固醇含量降低,胆固醇酰基转移酶基因表达的显著上调,提示其对瘤胃上皮组织更新的潜在影响。     

谷氨酰胺对饲喂高精料的奶山羊瘤胃上皮的保护效应研究

本研究旨在探讨过瘤胃谷氨酰胺对饲喂高精料的奶山羊瘤胃上皮屏障的保护效应。实验采用随机区组设计,将12头健康奶山羊随机分为高精料组(n=6)和高精料添加谷氨酰胺组(n=6),谷氨酰胺添加量为50 g/(d·头)。谷氨酰胺处理4周后,每组各选取4头山羊进行屠宰。HE染色、扫描和透射电镜法观察瘤胃上皮的组织形态学变化,实时荧光定量 PCR 法检测瘤胃上皮炎症因子及紧密连接蛋白mRNA的相对表达水平。结果表明,与对照组相比,添加谷氨酰胺显著降低了瘤胃液中丙酸浓度(P<0.01),有降低瘤胃乳酸、提高丁酸浓度的趋势,但对瘤胃液pH、乙酸、异丁酸、总挥发性脂肪酸浓度无显著影响(P>0.05);对组织形态的观察结果表明,添加过瘤胃谷氨酰胺可维持瘤胃复层扁平上皮各细胞层的完整性,保护角质层,减少细胞凋亡;并显著降低瘤胃上皮组织中TNF-α的mRNA相对表达量(P<0.05),显著提高occludin的mRNA相对表达量(P<0.05);但对IFN-γ、IL-1β、IL-6、IL-10、claudin-1、claudin-4和ZO-1 mRNA表达量无显著影响(P>0.05)。结果说明,高精料日粮下添加过瘤胃谷氨酰胺可降低瘤胃上皮的炎症反应,改善上皮紧密连接,从而维护瘤胃上皮屏障的正常功能。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.