高精料日粮对奶山羊乳腺组织NOD1炎性信号通路的影响

本试验旨在研究高精料日粮对奶山羊乳腺组织NOD1炎性信号通路的影响与调控。选取18头泌乳中期的健康萨能奶山羊,试验前分别安装瘤胃瘘管和乳静脉血管插管,随机分成3组:饲喂精粗比为4∶6的低精料日粮作为对照组(LC);饲喂精粗比为6∶4的高精料日粮作为处理组(HC);饲喂高精料日粮并添加丁酸钠作为调控组(BHC),试验时间为20周。试验结束后采集瘤胃液、乳静脉血和乳腺组织样品用于后续的瘤胃pH值、乳静脉血中炎性因子浓度、乳腺组织中炎症相关基因m RNA和蛋白水平的检测。结果显示:一天内HC组的瘤胃pH值小于5.8持续4 h且显著低于LC组,表明亚急性瘤胃酸中毒(SARA)诱导成功;饲喂4 h后BHC组瘤胃pH值显著高于HC组而与LC组差异不显著,表明丁酸钠能够缓解瘤胃pH的持续降低。HC组乳静脉血中炎性细胞因子IL-1β、IL-6和TNF-α的浓度显著高于LC组,而BHC组的炎性细胞因子溶度显著低于HC组并与LC组无显著差异;与LC组和BHC组相比,HC组奶山羊乳腺组织炎症相关基因NOD1、RIP2、NF-κB、IL-1β、IL-6的表达量显著上调;NOD1、NF-κBp65、NF-κBpp65的蛋白表达水平和m RNA水平变化一致。结果表明,长期饲喂高精料日粮能够激活奶山羊乳腺组织的NOD1-NF-κB炎性信号通路,诱导乳腺炎症应答,日粮中添加丁酸钠可以一定程度地缓解乳腺组织的炎症反应。     

高精料日粮对泌乳期奶山羊脂肪代谢的影响

为了探究长期饲喂高精料日粮对泌乳期奶山羊乳脂形成的影响机制,为科学饲养泌乳期的奶山羊提供理论基础。选用处于泌乳初期的14只关中奶山羊,随机分为2组,饲喂日粮精粗比为35∶65的作为对照组,65∶35的作为试验组,试验期为19周。结果表明,与对照组相比,饲喂高精料日粮的奶山羊血液谷丙转氨酶浓度显著升高(P0.05),甘油三酯有降低的趋势;试验组肝细胞呈肿胀变性或空泡样变性;与对照组相比,试验组奶山羊肝脏内脂类的分解和合成代谢相关基因表达均未发生显著变化。说明长期饲喂高精料日粮可导致血液中谷丙转氨酶升高,这可能与肝细胞受损有关,但脂类代谢的相关基因表达均没有发生显著变化。     

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（P<0.05）;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（P<0.05）;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（P<0.05）;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（P<0.05）;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。     

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。     

饲喂高精料日粮对奶牛泌乳及相关调节因子的影响

为了解高精料日粮对泌乳奶牛乳产量的影响,选择12只健康经产奶牛,分别饲喂精粗比为40∶60(对照组)和60∶40(高精料组)的日粮,试验期12周,每周监测乳产量。试验期间每周采集血液样品用于测定外周血生长激素(GH)和胰岛素样生长因子1(IGF-1)含量;试验结束,采集瘤胃液置-70℃保存待测,肝脏和乳腺组织于液氮速冻后置-70℃保存待测。结果表明,高精料日粮的饲喂显著降低了瘤胃pH(P0.05);奶牛外周血GH含量和IGF-1含量从第7周开始显著降低(P0.05);肝脏组织中生长激素受体(GHR)、蛋白质酪氨酸激酶2(JAK2)、信号传导与转录激活子5A(STAT5A)及信号传导与转录激活子5B(STAT5B)mRNA表达水平显著下调(P0.05);乳腺组织中,胰岛素样生长因子1受体(IGF-1R)mRNA表达水平显著下调(P0.05)。结果提示,长期饲喂高精料日粮会降低血液中GH和IGF-1含量,抑制GH-IGF-1轴的基因表达及关联的JAK2-STAT5信号途径的相关基因表达,从而减少了乳产量。     

高精料日粮对湖羊发情、孕酮水平及黄体PLINs基因表达的影响

高精料日粮会引起瘤胃代谢机能异常,引发机体内分泌紊乱。本试验旨在研究高精料日粮对湖羊发情、孕酮水平及黄体组织围脂滴蛋白(PLINs)家族成员mRNA表达的影响。试验选用5月龄雌性湖羊20只,分为低精料组(LC,精粗比2∶8,n=10)和高精料组(HC,精粗比6∶4,n=10)。于正试期第20 d进行同期发情处理,在正试期的第58天采集卵巢,分离黄体,采集血液检测孕酮(P_4)和脂多糖(LPS)的含量,统计平均日增重(ADG)及干物质采食量(DMI)。应用实时定量PCR检测黄体PLINs基因的表达量,并对LPS浓度、卵巢重、黄体数量、孕酮水平及PLINs基因表达量进行相关性分析。结果显示,高精料组ADG显著高于低精料组(P0.05),DMI显著低于低精料组(P0.05)。高精料组血清LPS浓度显著高于低精料组(P0.05)。高精料和低精料组母羊自然发情率分别为30%和70%(P=0.089)。2组母羊黄体期血清P_4浓度、卵巢重和黄体数量均无显著差异(P0.05)。PLINs家族成员mRNA在黄体组织中均有不同程度的表达,其中PLIN2 mRNA表达量最高,且低精料组PLIN2 mRNA表达量显著高于高精料组(P0.05),而PLIN3、PLIN4和PLIN5的mRNA表达量则显著低于高精料组(P0.05)。LPS浓度与PLIN2 mRNA表达量呈显著负相关(P0.05),与PLIN3 mRNA表达量呈显著正相关(P0.05)。PLIN3 mRNA表达量与PLIN4、PLIN5呈显著正相关(P0.05),与PLIN2呈显著负相关(P0.05)。结果表明,高精料日粮饲喂使湖羊血清LPS浓度升高,可能不利于母羊的发情。黄体组织PLINs基因在高精料组和低精料组之间的表达差异,提示PLINs基因表达可能受LPS及饲料中精粗比的影响,同时为进一步研究营养与繁殖的关系提供数据支撑。     

高精料日粮对山羊瘤胃和盲肠发酵及生物胺生成与吸收的影响研究

本实验旨在探究饲喂高精料日粮对山羊瘤胃和盲肠发酵、生物胺生成与吸收的影响研究。试验选用6头装有永久性瘤胃瘘管的雄性波杂山羊,随机分成低精料组与高精料组,实验期24 d。结果显示,与低精料组相比,饲喂高精料日粮的山羊瘤胃液中总挥发性脂肪酸、乙酸、丙酸、丁酸、戊酸、总支链脂肪酸、酪胺、腐胺、组胺、甲胺和色胺浓度显著性增加(P<0.05),而瘤胃pH和乙丙比显著降低(P<0.05);饲喂高精料日粮的山羊盲肠内容物中的乙酸、丙酸、总挥发性脂肪酸浓度和上述5种生物胺浓度显著增加(P<0.05);同时,瘤胃和盲肠静脉血中的上述5种生物胺的浓度显著提高(P<0.05)。相关性分析结果表明,瘤胃液中酪胺、腐胺、甲胺和色胺浓度与瘤胃静脉血所对应的生物胺浓度呈显著正相关(P<0.05),且盲肠内容物中酪胺、腐胺、甲胺和色胺与盲肠静脉血中对应的生物胺的浓度呈显著正相关(P<0.05)。结果说明,饲喂高精料日粮促进了山羊瘤胃和盲肠发酵,提高了瘤胃与盲肠内容物中生物胺浓度,暗示高精料日粮可能提高消化道上皮的通透性,促进消化道上皮对生物胺的吸收。     

赖氨酸添加水平对高精料日粮体外瘤胃发酵特性的影响

试验旨在探究不同赖氨酸（lysine, Lys）添加水平对高精料日粮体外瘤胃发酵特性的影响。试验设置5个组,分别添加0、0.05%、0.10%、0.20%和0.40%的Lys,每个组5个重复,其中0添加作为对照组,体外发酵48 h后收集发酵液用以检测瘤胃发酵参数,并采用多项指标综合指数（MFAEI）对添加效果进行综合评估。结果发现：(1)添加Lys对12、24、36、48 h产气量以及有机物消化率均无显著影响（P>0.05）,0.05%、0.10%、0.20%添加组的甲烷产量显著高于0.40%添加组（P<0.05）;(2)添加Lys显著提高了发酵液的pH(P<0.05),0.05%、0.10%、0.20%添加组的乙酸、丙酸、丁酸、戊酸和总挥发性脂肪酸的含量显著高于0.40%添加组（P<0.05）,0.10%、0.20%添加组的异戊酸含量显著高于0、0.40%添加组（P<0.05）;(3)MFAEI分析发现,添加0.20%的Lys对体外瘤胃发酵参数的改善效果最好。本试验结果表明,高精料日粮中添加0.20%的Lys可以改善瘤胃发酵。     

Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats

Background: To understand the impact of feeding a high-concentrate diet to mid-lactating goats for a long time on liver metabolism and inflammatory response, two dimensional polyacrylamide gel electrophoresis(2-DE) and real-time PCR method were employed to detect proteins differentially expressed in liver and their m RNAs expression in goats fed high concentrate diet(HC) or low concentrate diet(LC). Twelve lactating dairy goats were randomly assigned to either a HC diet group(65 % concentrate of dry matter; n = 6) or a LC diet group(35 % concentrate of dry matter; n = 6) for 10 wk.Results: Twenty differentially expressed spots(≥2.0-fold changes) in the hepatic tissues were excised and successfully identified using MALDI TOF/TOF. Of these, 8 proteins were up-regulated, while the rest 12 proteins were down-regulated in HC goats compared to LC. Differential expressed proteins including alpha enolase 1(ENO1), glutamate dehydrogenase 1(GLUD1), glutathione S-transferase A1(GSTA1), ATP synthase subunit 5β(ATP5β), superoxide dismutase [Cu-Zn](SOD1), cytochrom c oxidase subunit Via(COX6A1) and heat shock protein 60(HSP60) were further verified by real-time PCR and/or western blot at m RNA or protein expression level. Consistent with the 2-DE results, a significant decrease of β-actin protein expression and SOD enzyme activity was observed in liver of HC goats(P 0.05), while ENO1 protein expression was significantly up-regulated in HC compared to LC goats(P 0.05). However, western blot analysis did not show a significant difference of hepatic HSP60 protein between HC and LC group, which did not match the decrease of HSP60 content detected by 2-DE analysis. Real-time PCR showed that glutathione S-transferase P1(GSTP1) and SOD1 m RNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase(COX3) and ATPase 8(ATP8) m RNAs expression were markedly increased compared to LC(P 0.05). Gene Ontology(GO) analysis revealed that HC diet resulted in altered expression of proteins related to catalytic and mitochondrial metabolism in the liver, and may increase the stress response with up-regulating the expression of differentiation 14(CD14) cluster and serum amyloid A(SAA) as well as C-reactive protein(CRP) in the liver.Conclusions: These results suggest that feeding high concentrate diet to lactating goats for 10 wk leads to the activation of the inflammatory response, and decreases the anti-oxidant capacity, and subsequently impairs the mitochondrial function in the liver.     

Induction by endotoxin of the inflammatory response in the lactating and dry bovine mammary gland

Quarters of three lactating and three dry cows were infused with 10 micrograms endotoxin and the inflammatory response induced in the mammary gland was compared. The response of the dry glands was much less than that shown by the lactating glands. The swelling in the lactating quarters was severe after four hours and declined slowly; in the dry quarters the swelling was mild and transient. Antitrypsin levels, which reflect vascular permeability, were much greater in the lactating quarters, and the somatic cell counts were similarly much higher in the lactating quarters. One dry quarter gave no response other than a slight increase in antitrypsin after four hours.     

高精料对泌乳期山羊肝脏氨基酸分配与重分配及乳蛋白的影响

为了探讨饲喂不同精粗比日粮对肝脏氨基酸分配与重分配的影响及可能的机制,本研究选择6只安装了肝脏多血管瘘的健康泌乳奶山羊,分别饲喂精粗比为40∶60(对照组)和60∶40(高精料组)的饲料,2×2拉丁方设计,饲喂期16周。期间取乳样,测定乳蛋白含量,并通过肝门静脉、肝静脉和股动脉血管瘘取血, RP-HPLC法测定血浆中游离氨基酸含量。结果表明,高精料组山羊乳中乳蛋白含量显著低于低精料组(P<0.05);肝门静脉血液中各游离氨基酸含量均高于低精料组,而肝静脉和股动脉血液中各游离氨基酸含量均低于低精料组,即高精料组各游离氨基酸在肝脏中的消耗量均多于低精料组,通过股动脉进入乳腺组织的氨基酸含量也少于低精料组。本研究结果显示,长期饲喂高精料日粮,乳蛋白含量降低。其机制与肝脏中氨基酸分配与重分配发生改变,氨基酸在肝脏中消耗量增大,进入乳腺组织合成乳蛋白的前体物氨基酸减少有关。     

灌注葡萄糖对奶山羊乳腺营养摄取的影响

本试验研究了阴外动脉灌注葡萄糖对乳腺营养物质吸收和乳成分的影响。选用体重和产奶量相近的4只泌乳中期关中奶山羊,做阴外动脉和腹部皮下静脉手术安装血插管,采用4×4拉丁方设计,每天分别灌注葡萄糖0、12、24g和36g。结果表明:乳脂率随葡萄糖灌注量的增加而显著降低(P0.01)。灌注36g/d处理组乳脂产量显著低于其他处理组(P0.01);葡萄糖灌注提高了奶山羊的产奶量,24g/d处理组的产奶量显著高于对照组(P0.01);36g/d处理组乳腺对葡萄糖的摄取量和乳糖的产量都显著高于对照组(P0.01)。甘油三酯和非脂固形物的含量各组间差异不显著(P0.05),但葡萄糖灌注提高了乳蛋白产量,其中24g/d处理组乳蛋白产量最高,并显著高于对照组(P0.05)。葡萄糖的灌注显著降低了奶山羊干物质的采食量(P0.01)。葡萄糖的灌注对乳腺氨基酸的摄取影响不显著。葡萄糖的灌注使氨基酸合成乳蛋白的产出率提高1%～9%,葡萄糖合成乳糖的产出率提高0%～5%,有效改善了乳成分。     

Effects of dietary monensin and sodium propionate on net nutrient flux in steers fed a high-concentrate diet

Four Holstein steers (mean body weight, 211 +/- 20 kg) were utilized in a Latin-square design with a 2 X 2 factorial arrangement of treatments to investigate the effects of monensin (0 or 220 mg/d) and sodium propionate (0 or 450 g/d) on net nutrient flux. Steers were surgically prepared with hepatic portal and mesenteric venous catheters and an elevated carotid artery, after which they were adjusted to their basal diet (85% concentrate) and initial treatment over 19 d. Samples of arterial and portal venous blood were taken hourly over 3 h for the final 3 d of each 2-wk period. Portal blood flow was determined by primed continuous infusion of para-aminohippurate. No changes were seen in dry matter intake, portal blood flow, or net portal flux of any of the volatile fatty acids with the exception of butyrate flux, which decreased with monensin addition. Addition of monensin decreased net portal flux of ammonia, decreased recycling of urea, and tended to increase the net portal flux of glucose. Addition of sodium propionate increased the net portal flux of glucose and decreased the net portal flux of alpha-amino-N. These results are interpreted to suggest that changes in the products of ruminal fermentation may not be exactly translated into the products appearing in the portal circulation, and more information is needed to describe these relationships.     

Effect of nutrient intake on mammary gland growth in lactating sows

Sixty-one primiparous sows were used to determine the response of mammary gland growth to different energy and protein intakes during lactation. After birth, litter size was set to 9 or 10 pigs. Sows were slaughtered at selected times up to 30 d of lactation. Individual sows were fed one of four diets that were combinations of different amounts of energy and protein (3.0 Mcal ME and 8.0 g lysine/kg diet; 3.0 Mcal ME and 16.2 g lysine/kg diet; 3.5 Mcal ME and 6.4 g lysine/kg diet; or 3.5 Mcal ME and 13.0 g lysine/kg diet). Mammary glands were collected at slaughter and trimmed of skin and the extraneous fat pad. Each gland was weighed, cut in half to measure cross-sectional area, ground, and stored at -20 degrees C for chemical analysis. Frozen, ground tissue was used to determine dry matter, dry fat-free tissue (DFFT), total tissue protein, ash, and DNA content. Only glands known to have been suckled were included in this data set. Response surface regression was used for statistical analysis. The percentage of protein, fat, ash, and DNA in each suckled mammary gland was affected only by total energy intake (P<.05). The percentage of dry tissue and fat decreased as the total energy consumed during lactation increased, whereas the percentage of protein and DFFT increased as total energy intake increased. There were quadratic effects (P<.05) of both total energy and protein intake on wet weight, dry weight, protein amount, DFFT amount, and DNA amount of each suckled mammary gland during lactation. This study shows that mammary gland growth is affected by nutrient intake during lactation. The weight of suckled mammary glands and the amount of mammary tissue protein, DFFT, and total DNA were maximal on d 27.5 of lactation when sows had consumed an average of 16.9 Mcal of ME and 55 g of lysine per day during lactation. Provision of adequate amounts of nutrients to sows during lactation is important for achieving maximal growth of mammary glands and maximal milk production.     

Negative effects of long-term feeding of high-grain diets to lactating goats on milk fat production and composition by regulating gene expression and DNA methylation in the mammary gland

Background: It is well known that feeding a high concentrate(HC) diet to lactating ruminants likely induces subacute ruminal acidosis(SARA) and leads to a decrease in milk fat production. However, the effects of feeding a HC diet for long periods on milk fatty acids composition and the mechanism behind the decline of milk fat still remains poorly understood. The aim of this study was to investigate the impact of feeding a HC diet to lactating dairy goats on milk fat yield and fatty acids composition with an emphasis on the mechanisms underlying the milk fat depression. Seventeen mid-lactating dairy goats were randomly allocated to three groups. The control treatment was fed a low-concentrate diet(35% concentrate, n = 5, LC) and there were two high-concentrate treatments(65% concentrate, HC), one fed a high concentrate diet for a long period(19 wks, n = 7, HL); one fed a high concentrate diet for a short period of time(4 wk, n = 5, HS). Milk fat production and fatty acids profiles were measured. In order to investigate the mechanisms underlying the changes in milk fat production and composition,the gene expression involved in lipid metabolism and DNA methylation in the mammary gland were also analyzed.Results: Milk production was increased by feeding the HC diet in the HS and HL groups compared with the LC diet(P 0.01), while the percentage of milk fat was lower in the HL(P 0.05) but not in the HS group. The total amount of saturated fatty acids(SFA) in the milk was not changed by feeding the HC diet, whereas the levels of unsaturated fatty acids(UFA) and monounsaturated fatty acids(MUFA) were markedly decreased in the HL group compared with the LC group(P 0.05). Among these fatty acids, the concentrations of C15:0(P 0.01), C17:0(P 0.01), C17:1(P 0.01), C18:1 n-9 c(P 0.05), C18:3 n-3 r(P 0.01) and C20:0(P 0.01) were markedly lower in the HL group, and the concentrations of C20:0(P 0.05) and C18:3 n-3 r(P 0.01) were lower in the HS group compared with the LC group. However, the concentrations of C18:2 n-6 c(P 0.05) and C20:4 n-6(P 0.05) in the milk fat were higher in the HS group. Real-time PCR results showed that the m RNA expression of the genes involved in milk fat production in the mammary gland was generally decreased in the HL and HS groups compared with the LC group. Among these genes, ACSL1, ACSS1 2, ACACA, FAS, SCD, FADS2, and SREBP1 were downregulated in the mammary gland of the HL group(P 0.05), and the expressions of ACSS2, ACACA, and FADS2 m RNA were markedly decreased in the HS goats compared with the LC group(P 0.05). In contrast to the gene expression, the level of DNA methylation in the promoter regions of the ACACA and SCD genes was increased in the HL group compared with the LC group(P 0.05). The levels of ACSL1 protein expression and FAS enzyme activity were also decreased in the mammary gland of the HL compared with the LC group(P 0.05).Conclusions: Long-term feeding of a HC diet to lactating goats induced milk fat depression and FAs profile shift with lower MUFAs but higher SFAs. A general down-regulation of the gene expression involved in the milk fat production and a higher DNA methylation in the mammary gland may contribute to the decrease in milk fat production in goats fed a HC diet for long time periods.