不同品种牛乳营养成分含量及脂肪酸含量和组成测定比较研究

[目的] 比较中国荷斯坦奶牛、荷斯坦奶牛和西门塔尔牛乳中常规营养成分含量、矿物质元素含量及脂肪酸含量和组成的差异。[方法] 选取中国荷斯坦奶牛、荷斯坦奶牛和西门塔尔牛各6头,每头奶牛收集200 mL生鲜乳样品,利用乳成分分析仪测定乳中常规营养成分含量,通过原子吸收分光光度计测定乳中矿物质元素含量,应用气相色谱仪测定乳中脂肪酸含量和组成。采用统计学方法对不同品种牛乳中的营养成分含量及脂肪酸含量和组成进行比较。[结果] 中国荷斯坦奶牛生鲜乳中乳脂含量显著（P<0.05）高于荷斯坦奶牛,乳蛋白含量显著（P<0.05）高于荷斯坦奶牛和西门塔尔牛;中国荷斯坦奶牛生鲜乳中乳钙含量显著（P<0.05）高于荷斯坦奶牛,乳锌含量显著（P<0.05）高于荷斯坦奶牛和西门塔尔牛。中国荷斯坦奶牛生鲜乳中豆蔻酸含量显著（P<0.05）低于荷斯坦奶牛,油酸和亚油酸含量显著（P<0.05）高于西门塔尔牛;中国荷斯坦奶牛生鲜乳中饱和脂肪酸含量显著（P<0.05）低于西门塔尔牛,不饱和脂肪酸含量显著（P<0.05）高于西门塔尔牛。[结论] 中国荷斯坦奶牛乳中部分营养成分和矿物质元素含量接近或优于荷斯坦奶牛和西门塔尔牛,脂肪酸含量和组成与荷斯坦奶牛相近,脂肪酸组成优于西门塔尔牛。     

奶牛SP1基因结构及对乳脂合成功能的初步分析

旨在探讨中国荷斯坦奶牛的特异性蛋白1（specificity protein,SP1）基因结构特征及其对奶牛乳脂合成的影响。根据NCBI已经公布的奶牛SP1基因序列（NM_001078027.1）,利用生物信息学分析其序列保守性、理化性质、蛋白亲水性、蛋白质结构及互作蛋白;采用PCR技术扩增并克隆SP1基因CDS序列。然后,选取6头健康的中国荷斯坦奶牛,分别取泌乳期和干奶期奶牛乳腺组织,运用实时荧光定量PCR和Western blot方法检测SP1基因在不同时期奶牛乳腺组织中的表达情况。分离并纯化泌乳期奶牛乳腺上皮细胞,通过SP1过表达及干扰检测其对乳脂合成的影响,分别进行3次独立试验。结果显示,SP1基因序列在不同物种间高度保守,与山羊相似度最高（98.94%）。奶牛SP1基因CDS区序列长2 361 bp,编码786个氨基酸,蛋白分子质量为80 902.17 u,理论等电点为6.94。平均疏水指数为-0.438,为不稳定的亲水性蛋白。SP1序列包含3个锌指结构,SP1蛋白二级结构以无规则卷曲（52.29%）为主。STRING蛋白互作分析结果显示,SP1与转录因子AP-1（JUN）、雌激素受体α（ERα）、MYC原癌基因蛋白（MYC）、TATA盒结合蛋白（TBP）等蛋白存在相互作用。实时荧光定量PCR和Western blot结果显示,SP1的mRNA和蛋白在泌乳期的表达量显著高于干奶期（P<0.01）。在奶牛乳腺上皮细胞中过表达SP1显著增加细胞甘油三酯的合成（P<0.01）,而干扰SP1的表达,甘油三酯合成显著降低（P<0.01）。以上结果提示,SP1正向调控乳脂合成,分析SP1基因结构和功能为深入研究SP1对泌乳奶牛乳脂合成调控机制提供理论依据。     

荷斯坦奶牛乳与牦牛乳的差异蛋白质组学分析

本试验利用蛋白质组学方法分析荷斯坦奶牛乳与牦牛乳乳清与乳脂肪球膜中的差异蛋白,比较功能差异,为开发不同品种牛乳提供帮助。通过离心法分离荷斯坦奶牛乳与牦牛乳的乳清与乳脂,分别提取蛋白,通过串联质谱标签（TMT）法进行蛋白质定性与定量分析,得到差异蛋白;对差异蛋白进行GO功能注释、KEGG代谢通路、蛋白质互作（PPI）等生物信息学分析。结果显示：牦牛乳的乳脂、乳蛋白、乳糖、总固形物和非乳脂固形物含量均显著高于荷斯坦奶牛乳（P<0.05）。在2种牛乳乳清中共鉴定到可信蛋白641种,在乳脂肪球膜中共鉴定到可信蛋白543种;在乳清中筛选出差异蛋白268种,其中牦牛乳比荷斯坦奶牛乳上调70种,下调198种;在乳脂肪球膜中筛选出差异蛋白267种,其中牦牛乳比荷斯坦奶牛乳上调68种,下调199种。GO功能注释及富集分析表明2种牛乳的乳清和乳脂肪球膜差异蛋白均主要富集在细胞外空间、小分子结合和免疫反应等方面。KEGG注释及富集分析表明2种牛乳的乳清和乳脂肪球膜差异蛋白主要富集在金黄色葡萄球菌感染、补体与凝血级联等通路。差异蛋白互作网络分析发现2种牛乳的乳清差异蛋白中甘油醛-3-磷酸脱氢酶（GAPD...     

Effect of Anionic Salts on Health and Production Performance of Holstein Cows in Peripartum

This study was to determine the effect of anionic salts on health and production performance of Holstein cows during the peripartum period. Twenty-four Holstein cows during the peripartum period were selected and divided into control group and experimental group. Twelve Holstein dairy cows in control group were fed diets with original concentrate,while twelve cows in experimental group were fed diets with concentrates adding anionic salts for the last 21 days before expected calving. All animals were ad libitum fed with the same diets after calving. The results showed that urine pH maintained 5.7 to 6.9,and blood pH was decreased significantly compared to control group (P<0.05). However,the contents of plasma Ca,glucose and alkaline phosphatase (ALP) activity in experimental group were significantly increased in 1 d after calving (P<0.05). What's more, the incidences of hypocalcemia,milk fever,retained placenta,displaced abomasum and mastitis were reduced in experimental group and the incidences of retained placenta was significantly decreased (P<0.05). No significant difference was detected in milk yield between two groups (P>0.05). In summary,supplementation of anionic salts concentrate could help to keep the health and plasma Ca homeostasis of Holstein cows.     

阴离子盐对围产期奶牛健康和生产性能的影响

试验旨在研究阴离子盐对围产期奶牛产后健康和生产性能的影响。试验选取24头围产前期经产奶牛分为对照组和试验组,对照组饲喂牧场原饲料、试验组于产前21 d饲喂含阴离子盐颗粒料,产犊后统一饲喂牧场原有日粮。结果显示,试验组奶牛尿液pH值维持在5.7~6.9;与对照组相比,试验组血液pH值显著降低（P<0.05）,血钙含量、血糖水平、碱性磷酸酶活性在产后1 d显著升高（P<0.05）;试验组低血钙、产乳热、胎衣不下、真胃移位和乳房炎等疾病的发生率降低,其中胎衣不下发病率显著降低（P<0.05）;但两组间产奶量无显著差异（P>0.05）。综上所述,饲喂阴离子盐颗粒料可提高奶牛的健康状况,维持血钙平衡。     

Definition of a non-alert downer cow syndrome and some case histories

The clinical, blood biochemical and autopsy findings on a cow affected by a 'non-alert downer cow syndrome' are described in detail and compared with brief clinical and biochemical details from six similar cases attended elsewhere. The significance of the biochemical observations is discussed in relation to the observations from many other clinical milk fever cases in a dairy practice in Victoria. This syndrome developed in 1.9 per cent of 584 cases of milk fever. The essential findings in the syndrome were that all cows had an initial clinical episode suggestive of milk fever but showed an unsatisfactory clinical response to calcium borogluconate therapy. After a day or two all cows became laterally recumbent, some exhibited expiratory moaning and all developed mucoid faeces which, in many cases contained spots of blood. All cases had significantly lower erythrocyte and plasma potassium concentrations than those in milk fever cases which responded to treatment.     

丙酸镁对泌乳早期奶牛体况、泌乳性能和代谢参数的影响

选用36头经产奶牛,根据泌乳期、上一泌乳期305 d产奶量和预产期,采用随机区组设计分为4组,对照组饲喂基础日粮,处理1,2和3组分别在基础日粮基础上添加丙酸镁50,100和150 g/d,研究丙酸镁对泌乳早期奶牛采食量、泌乳性能、血液代谢参数和尿酮浓度的影响。结果表明,添加丙酸镁对奶牛的采食量、乳脂率、乳蛋白率、乳糖率和乳干物质率无显著影响,添加丙酸镁100和150 g/d对产乳量、饲料转化效率、体况及代谢参数有改善,该二处理组产奶量、饲料转化效率、体况评分、能量平衡、血浆葡萄糖和胰岛素浓度均显著高于对照组(P<0.05),而血浆游离脂肪酸和β-羟丁酸浓度显著低于对照组(P<0.05),尿酮浓度(除100 g/d组产后7 d测定值与50 g/d组无显著差异外)显著低于对照组和50 g/d组(P<0.05)。根据试验结果,丙酸镁适宜添加量为100 g/d。     

采用四引物ARMS-PCR方法检测奶牛乳腺组织DGAT1基因单核苷酸多态性

试验旨在快速有效地检测泌乳奶牛的二酰甘油转酰基酶1（diacylgycerol acyltransferase 1,DGAT1）乳脂量性状的优势等位基因K232A单核苷酸多态性,确定DGAT1基因型进而确定泌乳性状,为中国荷斯坦奶牛分子标记辅助选择提供技术支持。选取6头泌乳初期（3头为高乳产量牛,3头为低乳产量牛）中国北方荷斯坦奶牛为研究对象,提取乳腺组织基因组DNA,分别设计1对外引物和1对内引物,建立一种四引物ARMS-PCR体系快速检测奶牛乳腺组织DGAT1基因单核苷酸多态性。结果发现,外引物扩增片段长度为512 bp,为PCR反应的阳性对照,基因型为232K扩增片段长度为369 bp,基因型为232A扩增片段长度为181 bp。PCR结果显示,6头牛的乳腺组织样本均由外部引物扩增出长度为512 bp的片段,泌乳期高乳产量奶牛和泌乳期低乳产量奶牛乳腺组织的特异性扩增片段长度均为181 bp。表明本研究选取的6头奶牛样本DGAT1基因K232A多态性均为232A型。提示该PCR鉴定方法能够快速有效地鉴定奶牛DGAT1基因型,可用于中国荷斯坦奶牛分子标记辅助选择。     

Effect of citrus pulp silage feeding on concentration of beta‐cryptoxanthin in plasma and milk of dairy cows

Citrus pulp is known to contain a functional molecule of beta‐cryptoxanthin which is one of the carotenoids showing anti‐oxidative capacity. Influences of citrus pulp silage feeding to dairy cows on beta‐cryptoxanthin concentration in plasma, other blood properties and milking performances were investigated. Four Holstein cows were fed total mixed ration (TMR) containing citrus pulp silage 20% dry matter (DM) for 2 weeks with free access to the TMR. Dry mater intake, milk production and milk components 2 weeks later were not altered compared with those of the control group without citrus pulp silage. Activities of aspartate aminotransferase, alanin aminotransferase and gamma‐glutamyltranspeptidase in plasma were not affected by feeding of citrus pulp silage. Concentrations of protein, albumin, sulfhydryl residue, ascorbic acid, thio‐barbituric acid reactive substance and urea nitrogen in plasma were also not altered by citrus pulp silage feeding. Concentration of beta‐cryptoxanthin in plasma was increased approximately 20‐fold compared with the control group (P < 0.05). Content of beta‐cryptxanthin in pooled milk fat fraction was also increased approximately three times compared with that of the control group. Feeding of TMR containing citrus pulp silage 15% DM for 30 days to eight dairy cows also increased plasma beta‐cryptoxanthin concentration 30‐fold compared with that before feeding.     

过瘤胃氨基酸高能复合物对奶牛产奶量及乳成分的影响

为研究不同过瘤胃氨基酸高能复合物对奶牛产奶量及乳成分的影响,试验选择胎次(3～5胎)、产奶量、产犊日期相近,体重(600±20)kg健康荷斯坦泌乳高峰期奶牛24头,随机分为4组,每组6头。1组为对照组,饲喂牛场常规日粮;2、3、4组为试验组,分别在常规日粮基础上添加过瘤胃氨基酸高能复合物Ⅰ、Ⅱ、Ⅲ500 g/(d.头)。正式期60 d。结果表明:奶牛日粮中添加过瘤胃氨基酸高能复合物后,均可明显提高奶牛产奶量、乳脂率、乳蛋白率及乳中总干物质含量。尤其是复合物Ⅲ,添加效果最为理想,平均日产奶量、乳脂率、乳蛋白率及总干物质含量分别比对照组提高21.19%(P<0.05)、21.31%(P<0.05)、9.72%(P<0.05)和10.34%(P<0.05),平均日校正乳产量提高35.70%(P<0.05)。     

Detection of Dairy Cow Mammary Tissue DGAT1 Gene Single Nucleotide Polymorphisms Using Tetra-primers ARMS-PCR Method

This study was aimed to detect the single nucleotide polymorphisms of diacylgycerol acyltransferase 1 (DGAT1) K232A, and determine the DGAT1 genotype and milk traits of dairy cow, which would provide a new technique for marker-assisted selection in China Holstein dairy cows. In the present study, six Northern China Holstein dairy cows (three were lactating cows with high quatity milk and three were lactating cows with low quatity milk) were used to detect mammary tissue DGAT1 gene K232A polymorphisms. Genome DNA was extracted from each cow, a pair of external primers and a pair of internal primers were designed to amplify DGAT1 gene. The results showed that PCR-amplified fragments were 512 bp (external band), 369 bp (232K allele) and 181 bp (232A allele), respectively. The exterenal band functions as the internal PCR-positive control. The tetra-primer ARMS-PCR amplifications yielded a 512 bp fragment and a 181 bp fragment, indicating that the six dairy cows were all homozygous 232A. The results indicated that the tetra-primers ARMS-PCR was a quick and convenient method to identify dairy cow DGAT1 gene K232A polymorphisms, which was suitable for marker-assisted selection in China Holstein dairy cows.     

苜蓿鲜草和青饲玉米对奶牛生产性能和乳品质的影响

探讨日粮中添加苜蓿鲜草和青饲玉米对奶牛生产性能及其乳成分的影响,本试验选用荷斯坦泌乳奶牛20头,随机分为2组,对照组饲喂秸秆和精料补充料;试验组饲喂苜蓿鲜草、青饲玉米和精料补充料日粮。试验结果表明,日粮中添加苜蓿鲜草和青饲玉米,可极显著提高奶牛的生产性能和乳脂率(P<0.01),显著提高干物质采食量和乳蛋白含量(P<0.05)。     

TMR中不同长度的苜蓿干草对泌乳奶牛生产性能的影响

试验旨在研究泌乳奶牛TMR中添加不同长度的苜蓿干草对奶牛采食及产奶性能的影响。采用3个3×3拉丁方试验研究了TMR中苜蓿干草长度分别为1、3、5 cm时对3头头胎及6头经产泌乳奶牛的采食及产奶性能的影响。结果表明:TMR中的苜蓿干草长度变化对奶牛的DM采食量无显著影响(P0.05);当苜蓿干草长度从1 cm增加到3 cm和5 cm时,奶牛的NDF、ADF及peNDF摄入量均显著增加(P0.05);3 cm和5 cm长度的苜蓿干草较1 cm的能显著提高奶牛4%FCM产奶量、产奶效率及乳脂率(P0.05)。结果提示,泌乳奶牛TMR中苜蓿干草以较长的5 cm为宜;较长的苜蓿干草不影响奶牛的干物质采食量,能增加其有效纤维的摄入量,提高其4%FCM产奶量、产奶效率及乳脂率。     

Rumen-protected choline administration to transition cows: effects on milk production and vitamin E status

We investigated the influence of rumen-protected choline (RPC) supplementation on milk production, lipid metabolism and vitamin E status in dairy cows receiving a silage-based diet. Twenty-six Italian Holstein multiparous cows were assigned by weight and average production in the previous lactation, to one of two groups: control (no RPC supplementation) and RPC (supplemented with 20 g/day rumen-protected choline chloride). Treatment began 14 days before expected calving and continued for 30 days after parturition. Choline administration significantly increased milk production during the first month of lactation and also the concentration (and total secretion) of choline in milk, but did not affect fat or protein concentrations in milk, or plasma levels of glucose, beta-hydroxybutyrate, cholesterol and non-esterified fatty acids (NEFA). However, around parturition, NEFA concentrations in plasma were lower in treated animals than in controls, suggesting improved lipid metabolism as a result of choline supplementation. Choline supplementation also increased alpha-tocopherol plasma concentrations, suggesting a novel aspect in dairy cows.     

中国荷斯坦奶牛CCR5基因克隆、生物信息学及表达分析

试验旨在克隆中国荷斯坦奶牛CC趋化因子受体5(C-C chemokine receptor type 5,CCR5)基因,对其进行生物信息学分析,并探究CCR5基因在奶牛炎性和健康组织中的表达水平。采用PCR技术扩增并克隆荷斯坦奶牛CCR5基因CDS区全长序列,连接pMD18-T载体并转化大肠杆菌DH5α感受态细胞,通过蓝白斑方法筛选阳性克隆并测序,对序列进行相似性比对及系统进化树构建;应用多种在线生物信息学软件对其编码蛋白进行分析,并利用实时荧光定量PCR方法检测CCR5基因在健康和炎症奶牛乳腺组织中的表达情况。结果显示,中国荷斯坦奶牛CCR5基因CDS区全长1 059 bp,编码352个氨基酸。相似性和遗传进化分析结果显示,奶牛CCR5基因与绵羊的遗传距离最近,高达96.0%,与鸡遗传关系最远,为61.0%,且在不同物种之间CCR5基因高度保守。中国荷斯坦奶牛CCR5蛋白分子质量为40.235 ku,理论等电点(pI)为9.30,为疏水性蛋白但不是分泌蛋白,主要存在于细胞质内;在CCR5蛋白二级结构中α-螺旋和无规则卷曲分别占51.14%和32.95%,三级结构模型预测结果与二级结构一致。实时荧光定量PCR结果显示,CCR5基因在健康奶牛乳腺组织中的表达量极显著低于炎性奶牛乳腺组织(P<0.01),提示其可能参与奶牛乳腺炎的发生过程。本试验结果为进一步研究奶牛CCR5蛋白的功能提供了理论依据,对探究奶牛CCR5基因在奶牛乳腺炎中的调控功能等具有重要意义。     

中国荷斯坦奶牛RPL23A、ACACB基因3个SNPs检测及其与产奶性状的关联分析

旨在开展奶牛群体高乳成分功能基因的验证与筛选,利用微流控芯片自主选育技术分析北京地区母牛群体高乳蛋白、高乳脂基因频率,同时分析基因多态性及其与产奶性状的相关性。本研究对北京地区8个大型奶牛场1 596头中国荷斯坦奶牛RPL23A、ACACB基因的多态性进行了检测,所有个体均为3胎以内的泌乳牛,收集每头牛所有测定日的产奶量、乳脂率、乳蛋白率等数据,同时对3个多态位点不同基因型与产奶性状进行了关联分析。RPL23A基因的SNP位点g.20146771G>A,在第一泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白量均达到极显著关联（P<0.01）,第二泌乳期,g.20146771G>A与5个产奶性状均呈极显著关联（P<0.01）。ACACB基因的SNP位点g.63962768C>T,在第一泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白量均呈极显著关联（P<0.01）,在第二泌乳期,与产奶量、乳脂量、乳脂率和乳蛋白率呈极显著关联（P<0.01）。结果表明,RPL23A、ACACB基因可以作为影响中国荷斯坦奶牛产奶性状的候选基因用于标记辅助选择,以上基因位点可能通过直接或间接的途径影响奶牛的乳脂或乳蛋白性状,对产奶性状起到重要调控作用。本研究为荷斯坦奶牛后续的标记辅助选择奠定了良好基础。     

Influence of race and crossbreeding on casein micelles size

Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production.     

A novel nutritional strategy to prevent milk fever and stimulate milk production in dairy cows

AIMS: To assess the efficacy of a feed supplement designed to reduce the dietary availability of calcium when fed during the last 2-3 weeks of pregnancy, on the incidence of postparturient hypocalcaemia (milk fever) and on milksolids (MS) production in pasture-fed dairy cows in early lactation. METHODS: In Experiment 1, late-pregnant Holstein/Friesian cows, 4-10 years old, grazing pasture, were either fed the supplement (1 kg/cow/day) for 2-4 weeks prior to calving (n=11) or no supplement (n=10). Plasma calcium concentrations were measured immediately before and after a 20 h fast intended to induce hypocalcaemia at the end of this period, 1-7 days before the cows calved. In Experiment 2, mixed-breed dairy cows, 3-10 years old in 7 commercial dairy herds were fed the same supplement (1 kg/cow/day) for 1-3 weeks immediately prior to calving (n=565) or no supplement (n=614), and incidences of clinical milk fever and MS production at 6-9 weeks post calving were compared between groups. RESULTS: In Experiment 1, plasma calcium concentrations were higher (p=0.01) in supplemented than in non-supplemented cows after fasting, indicating reduced susceptibility to fasting-induced hypocalcaemia. In Experiment 2, the incidence of milk fever was lower (6.4% vs 17.1%, p=0.001) and MS production was higher (p=0.01) in supplemented than in non-supplemented cows. CONCLUSIONS: These preliminary results demonstrate that feeding a supplement designed to reduce dietary calcium availability for 2-4 weeks immediately prior to calving reduced the susceptibility of cows to fasting-induced hypocalcaemia, reduced the incidence of clinical milk fever and increased MS production in early lactation. CLINICAL RELEVANCE: The concept and product reported here have potential to provide dairy farmers with a practical means to prevent hypocalcaemia and improve milk production during early lactation.     

Licking induced changes to the pattern of moxidectin milk elimination after topical treatment in dairy cows

Pour-on administration of the macrocyclic lactones anti-parasitic compounds in beef and dairy cattle is now worldwide accepted. However, the information available on their milk excretion pattern, after topical administration is rather limited. Additionally, the cattle licking behaviour has been proven to affect the kinetics of these anti-parasitic compounds. The purpose of this study was to investigate the influence of the natural licking behaviour on the plasma and milk disposition of moxidectin (MXD), topically administered (500 μg/kg) in lactating dairy cows. Ten lactating Holstein dairy cows (705 kg body weight) were allocated into two experimental groups ( n  = 5). The licking was prevented during 5 days postadministration in animals in group I, and the remaining cows (group II) were allowed to lick freely. MXD concentrations profiles were measured in plasma and milk over 15 days posttreatment. The licking restriction period caused marked changes in MXD disposition kinetics both in plasma and milk. Both plasma and milk MXD concentrations (partial AUC 0–5 days) were significantly lower ( P  < 0.05) in licking-restricted cows. After the 5-day of restriction period, the animals were allowed to lick freely, which permitted the oral ingestion of MXD, situation clearly reflected both in plasma profile and milk excretion pattern. Despite the enhanced MXD milk concentrations measured in free-licking cows, drug concentrations did not reach the maximum MXD residues limit.     

Analysis and Validation of the Differentially Expressed ENO1 in the Maternal Placenta of Cow with Retained Foetal Membrane

The paper aimed to analyze and validate the function of differentially expressed ENO1 in the cow with retained foetal membrane.In our research,we chose three healthy Holstein dairy cows and three Holstein dairy cows with retained foetal membrane of similar age,foetal times,weight and milk yield and divided them into two groups.The total protein of maternal placenta was extracted in the control and retained foetal membrane of cow.The differential expression of proteins were found out by the 2-DIGE,and the differential expression of ENO1 was validated by the Western blotting and Real-time quantitative PCR.The results showed that ENO1 was significant expression and large multiple in the two groups,and it was significant increased expression in the retained foetal membrane of cow after Western blotting and Real-time quantitative PCR (P=0.015<0.05;P=0.001<0.01).The ENO1 participated in the energy homeostasis,immune and fibrinolysis process which related to the retained foetal membrane of cow.It suggested that ENO1 was likely connected with the retained foetal membrane.