干奶期奶牛不同能量摄入对其肝脏胰岛素受体mRNA表达的影响

将健康围产期奶牛30头随机分为3组,预产前28 d分别饲喂奶牛标准日粮（能量摄入100%组）、标准日粮增加20%日粮（能量摄入120%组）和标准日粮减少20%日粮（能量摄入80%组）,产后各组奶牛均饲喂标准的产奶日粮,至产后56d结束,采用半定量RT-PCR方法,观测干奶期不同能量摄入水平对围产期健康奶牛肝脏胰岛素受体（IR）基因表达的影响.试验结果表明,干奶期低能饲喂奶牛,产后肝脏IR mRNA表达增加,提示低能饲喂有利于调节奶牛产后血糖平衡.     

干奶期不同能量摄入对围产期奶牛糖异生的影响

将围产期健康奶牛30头随机分为三组,分别于产前第28d开始饲喂标准日粮(能量摄入100%组)、标准日粮增加20%日粮(能量摄入120%组)和标准日粮减少20%日粮(能量摄入80%组),产后各组奶牛均饲喂标准泌乳日粮,至产后第56d结束,观察干奶期不同能量摄入水平对围产期健康奶牛糖异生的影响。结果表明,产前低能饲喂可以增加围产期奶牛血浆葡萄糖(GLU)浓度,肝脏丙酮酸羧化酶(PC)和磷酸烯醇式丙酮酸羧激酶(PEPCK)基因表达升高。说明干奶期低能饲喂奶牛,可以增强围产期奶牛的肝糖异生能力。     

干奶期不同能量摄入水平对奶牛产后能量负平衡的影响

选择围产期健康奶牛30头,随机分为三组,产前第28d开始,饲喂不同能量日粮,即对照组(C组)、高能量组(H组)和低能量组(L组);产后各组奶牛均饲喂标准泌乳日粮,至产后56d结束。研究干奶期不同能量摄入水平对产后奶牛能量负平衡的影响。结果表明:低能量组奶牛产后干物质摄入量显著高于其它两组、体重消耗较低,能量负平衡状态缓解。     

胰岛素、胰高血糖素对体外培养脂肪细胞胰高血糖素受体mRNA丰度的影响

为阐明胰岛素、胰高血糖素在奶牛脂肪代谢中的调控作用,应用RT-PCR法观察了胰岛素、胰高血糖素对体外培养脂肪细胞胰高血糖素受体(GLNR)mRNA丰度的影响。结果发现,随着培养液中胰岛素质量浓度的升高,GLNR mRNA表达逐渐增加(P<0.05);而随着培养液中胰高血糖素浓度的升高,GLNR mRNA表达逐渐降低(P<0.01)。本研究结果表明,胰岛素、胰高血糖素直接调控奶牛脂肪细胞胰高血糖素受体mRNA的表达。     

不同能量摄入水平对围产期奶牛肝MTP mRNA丰度的影响

将年龄、胎次相同，年产奶量大于7000kg的围产期荷斯坦奶牛30头随机分为3组，每组10头，低能量组（Ⅰ组）按中国奶牛营养标准2000年版减少20％日粮（能量摄入80％），高能量组（Ⅱ组）则按此标准增加20％日粮（能量摄入120％），对照组（Ⅲ组）按标准日粮（能量摄入100％）饲喂，试验从产前28d到产后56d结束，分娩后各组均按标准配制相同的泌乳日粮。分别于产前28、14d及产后1、14、28、56d采取肝活体组织。应用内对照RT—PCR方法检测肝脏组织MTPmRNA丰度。结果显示，各组奶牛肝MTPmRNA丰度均呈现先升高后降低的趋势。产后均高于产前，且产后1～28d差异显著（P〈0．01或P〈0．05），而后回降，至56d趋于平稳（P〉0．05）；Ⅲ组肝MTP mRNA丰度在产后1d达到峰值，而Ⅰ组和Ⅱ组在产后14d达到最大值（P〈0．01）；Ⅱ组产后1～28d显著高于I组。产后14d显著高于Ⅲ组（P〈0．01）；Ⅲ组产后1d和28d显著高于Ⅰ组（P〈0．01或P〈0．05）。这些结果表明，围产期奶牛能量摄入水平对肝MTP mRNA丰度有显著影响。     

不同能量摄入对围产期奶牛脂肪组织胰岛素受体mRNA丰度的影响

将年龄、胎次相同,年产奶量大于5000kg的围产期健康奶牛30头随机分为3组,每组10头。按照中国奶牛饲养标准(2000),低能组饲喂减少20%日粮(能量摄入80%组),对照组饲喂标准日粮(能量摄入100%组),高能组饲喂增加20%日粮(能量摄入120%组),试验从产前14天开始至产后28天结束,产后各组奶牛均饲喂标准日粮。采用半定量RTPCR方法检测了不同能量摄入对围产期奶牛脂肪组织胰岛素受体(InsR)mRNA丰度的影响。结果表明:各组脂肪InsRmRNA相对表达量从产前14天至产后28天均呈现先升高后降低的趋势,产后14天达到最大值,且产后高于产前;低能组产前InsRmRNA相对表达量低于高能组和正常组,产后1天至产后28天高于高能组和正常组,产后14天显著高于高能组(P<0.01)。可见,干乳期低能饲喂奶牛,产后脂肪InsRmRNA表达增加,提示产前适当降低能量摄入水平,有利于胰岛素抑制泌乳初期脂肪动员作用的发挥。     

干奶期不同能量摄入水平对产后奶牛生产性能的影响

将围产期健康奶牛30头随机分为3组．分别于产前28d开始饲喂NRC标准日粮（能量摄入100％组）、NRC标准增加20％日粮（能量摄入120％组）和NRC标准减少20％日粮（能量摄入80％组）．产后各组奶牛均饲喂标准日粮，至产后56d结束，观察干奶期不同能量摄入水平对产后奶牛生产性能的影响。结果表明，高能量组奶牛产后干物质采食量和产奶量均显著低于其他2组，并且体质量消耗明显。由此认为，干奶期能量摄入水平是影响和调节产后干物质摄入量、产乳量和能量平衡状态的重要因素。     

不同能量摄入水平对围产期奶牛肝载脂蛋白E mRNA丰度的影响

将年龄、胎次相同，年产奶量大于5000kg的围产期健康奶牛30头随机分为3组，每组10头。低能组饲喂中国奶牛饲养标准（2000）减少20％日粮（能量摄入80％组）、对照组饲喂中国奶牛饲养标准（2000）日粮（能量摄入100％组）和高能组饲喂中国奶牛饲养标准（2000）增加20％日粮（能量摄入120％组），试验从产前28d开始至产后56d结束，产后各组奶牛均饲喂标准日粮。采用内对照RT-PCR方法检测了不同能量摄入围产期奶牛肝组织载脂蛋白E（apolipoprotein E，apoE）mRNA丰度。结果表明：apoE mRNA相对表达量从产前28d至产后56d各组均呈现先升高后降低的趋势，各组apoE mRNA水平均在产后一天达到最大值（P〈0．01），且产后均高于产前（产后56d除外）；产前14d至产后28d低能组apoE mRNA相对表达量均高于其他两组（P〈0．05或P〈0．01），高能组在产后28d和56d高于对照组（P〈0．01）。可见，围产期奶牛能量摄入水平对肝组织载脂蛋白E mRNA丰度有显著影响。     

干乳期能量摄入水平对围产期奶牛肝载脂蛋白B100mRNA丰度的影响

将年龄、胎次相同，年产奶量大于5000kg的围产期健康奶牛30头随机分为3组，每组10头。Ⅰ组饲喂中国奶牛饲养标准（2000）减少20％日粮（能量摄入80％组）。Ⅱ组饲喂中国奶牛饲养标准（2000）日粮（能量摄入100％组），Ⅲ组饲喂中国奶牛饲养标准（2000）增加20％日粮（能量摄入120％组）。试验从产前28d开始至产后56d结束，产后各组奶牛均饲喂标准泌乳日粮。采用内对照RT—PCR方法，检测了干乳期摄入不同能量的围产期奶牛肝脏活体组织载脂蛋白B100（apoB100）mRNA丰度。结果表明。apoB100mRNA相对表达量，产前至产后Ⅰ、Ⅲ组均呈先升高后降低的趋势，Ⅲ组产后apoB100mRNA丰度降低了61．45％，Ⅰ组呈逐渐降低的趋势，各组均在产后28d达最低值（Ⅰ组，P〈0．05；Ⅱ、Ⅲ组，P〈0．01）。产前14d至产后1d，apoB100mRNA相对表达量Ⅲ组高于Ⅰ组，Ⅰ组高于Ⅱ组，组间差异极显著（P〈0．01）；产后28d。Ⅰ组高于Ⅱ、Ⅲ组（P〈0．05）。由此可见，围产期奶牛能量摄入水平对肝脏载脂蛋白B100mRNA丰度有显著影响。     

摄入不同能量的围产期乳牛肝低密度脂蛋白受体mRNA丰度的比较

将30头健康、经产、处于围产期的黑白花乳牛随机分为3组，每组10头。从产前28d开始，低能量组乳牛饲喂《中国奶牛饲养标准（2000）》减少20％日粮（能量摄入80％），对照组乳牛饲喂《标准》日粮（能量摄入100％），高能量组乳牛饲喂《标准》增加20％日粮（能量摄入120％），产后各组乳牛均饲喂标准日粮。至产后第56d结束试验；采用内对照RT-PCR方法检测摄入不同能量的围产期乳牛肝活体组织低密度脂蛋白受体（LDLR）mRNA丰度。结果，不同能量组乳牛肝LDLR mRNA丰度产前至产后均呈现先升高后降低的趋势。100％和120％能量组肝LDLR mRNA丰度在产后14d达最大值，且产后均高于产前（产后56d除外，P〈0．01或P〈0．05）；而80％能量组产后1d即达到最大值，产前14d至产后14d，LDLR mRNA相对表达量显著高于100％和120％能量组；产后28～56d，120％能量组显著高于80％和100％能量组（P〈0．01）。表明围产期乳牛能量摄入水平对肝LDLR mRNA丰度有显著影响。     

Characterization of ruminal dynamics in Holstein dairy cows during the periparturient period

We used four pregnant Holstein cows to delineate ruminal adaptations as cows transitioned from one lactation to the next. Cows were fed typical diets through far-off and close-up dry periods and lactation. We measured ruminal characteristics on day 72 (late lactation), 51 (far-off dry), 23 and 9 (close-up dry) prepartum and on days 6, 20, 34, 48, 62, 76 and 90 postpartum (early lactation). Measurements included: ruminal fill (weight of actual contents), ruminal capacity (volume of rumen when fully filled), digestibilities and ruminal passage rates. Ruminal capacity tended to increase linearly during early lactation but was stable during dry and transition periods. Both total and liquid fill decreased linearly during the dry period, increased across parturition, and increased linearly through early lactation. Dry matter fill decreased as cows were fed the close-up diet at day 23 prepartum then increased near parturition and continued to increase across early lactation. Solid passage rate was greatest when cows were fed the close-up diet, and decreased throughout the transition period. In lactation, solid passage rate responded quadratically with peak at day 48 followed by decreases through day 90 postpartum. Liquid passage increased linearly across the transition period. Total tract organic matter digestibilities increased linearly over the dry period with significant increases prior to or immediately after parturition, then they remained relatively stable over early lactation until they increased at day 90. Fibre digestibilities demonstrated quadratic responses over early lactation, being higher on day 6 and day 90 than at other times. Starch digestibilities decreased linearly across both the dry and transition periods with decreases in lactation until day 62 followed by increases until day 90. High producing lactating dairy cows go through a multitude of ruminal adaptations, in terms of digestion, passage, capacity and fill, as they transition from one lactation to the next.     

Heat influences on plasma insulin and glucagon in response to secretogogues in non-lactating dairy cows

Four non-lactating cows were offered a maintenance diet of hay wafer and a commercial concentrate. They were housed in a thermoneutral (TN; 20°C) and then a hot (30°C) environment in an artificial climate chamber. Glucose, arginine, butyrate, and insulin were administered through one jugular catheter, and from a catheter on the other side venous blood was collected. The peak increments in plasma insulin after the glucose and butyrate administrations were lower during heat exposure. The response of insulin after arginine injection was smaller in the hot compared with the thermoneutral environment; however, arginine injection resulted in a significantly higher secretion of glucagon in the hot environment. The response area of insulin after the insulin injection was smaller in the hot environment; however, insulin clearance rate was not changed. It is concluded that in non-lactating cows, insulin release is probably lower during heat exposure. With respect to plasma glucose during heat exposure, the lower basal values, lower concentrations after the end of the glucose infusion, and delayed recovery to basal values after the butyrate and insulin administrations observed, may indicate lower gluconeogenesis and glycogenolysis in the hot environment.     

Effects of amino acids infused into the vein on ghrelin‐induced GH,insulin and glucagon secretion in lactating cows

To investigate the effects of amino acids on ghrelin‐induced growth hormone (GH), insulin and glucagon secretion in lactating dairy cattle, six Holstein cows were randomly assigned to two infusion treatments in a cross‐over design. Mixture solution of amino acids (AMI) or saline (CON) was continuously infused into the left side jugular vein via catheter for 4 h. At 2 h after the start of infusion, synthetic bovine ghrelin was single injected into the right side jugular vein through the catheter. Ghrelin injection immediately increased plasma GH, glucose and non‐esterified fatty acids (P < 0.05) with no difference between both treatments. Additionally, plasma insulin and glucagon concentrations were increased by ghrelin injection in both treatments. The peak value of plasma insulin concentration was greater in AMI compared with CON (P < 0.05). Plasma glucagon concentration showed no difference in the peak value reached at 5 min between both treatments, and then the plasma levels in AMI compared with CON showed sustained higher values (P < 0.05). After plasma glucose concentration reached the peak, the decline was greater in AMI compared with CON (P < 0.05). These results showed that the increased plasma amino acids may enhance ghrelin action which in turn enhances insulin and glucagon secretions in lactating cows.     

Chromium supplementation and substitution of barley grain with corn: effects on metabolite and hormonal responses in periparturient dairy cows

Thirty-two multiparous Holstein cows were used to investigate the effects of chromium-l-methionine (Cr-Met) supplementation and dietary grain source on metabolic indices throughout the periparturient period. Cows were fed a total mixed ration with the concentrate portion based on ground barley (barley based diet, BBD)--or ground corn (corn-based diet, CBD) from 21 days before anticipated calving through 28 days after calving. The Cr-Met was supplemented at dosages of 0 or 0.08 mg of Cr/kg of metabolic body weight throughout the experiment. Thus, treatments were in a 2 (Cr-Met) × 2 (grain sources) factorial arrangement. Plasma glucose, non-esterified fatty acids, triglyceride, very low-density lipoprotein, blood urea nitrogen (BUN), insulin, glucagon, cortisol and insulin-like growth factor-I (IGF-I) concentrations were not affected by chromium supplementation on calving day (d 0). However, there was a trend for decreased β-hydroxybutyrate (BHBA) and increased cholesterol on d 0 in cows fed the Cr supplemented diet. On d 21 postpartum (wk 3 p.p.), plasma glucagon concentration tended to be greater in cows fed the Cr supplemented diet. However, other plasma metabolite and hormone measures were not affected by Cr supplementation in wk 3 p.p. There was no effect of grain source on d 0 plasma metabolic and endocrine measures. In wk 3 p.p., a significant difference was found only for plasma concentration of glucagon that was higher for cows fed the BBD compared with the CBD. There was an interaction of Cr-Met supplementation and grain source in wk 3 p.p. plasma concentration of BUN to decrease in CBD and increase in BBD, supplemented with Cr-Met. No Cr by grain interactions were observed for other plasma metabolic variables on d 0 and in wk 3 p.p. These results indicate that Cr-Met supplementation and substituting barley grain with corn throughout the transition period have only moderate effects on metabolic and endocrine parameters.     

Relationship between fat accumulation in the liver and energy intake, milk fat yield and blood metabolites in dairy cows

In the present study, 42 multiparous Holstein cows were used to investigate the relationship between fat accumulation in the liver and dry matter intake, milk yield and blood metabolites. Based on the percentage of fat in the liver cell at 2 weeks post‐parturition, the cows were classified into three groups. These groups were: (i) less than 10% of fat (normal group, n = 29); (ii) 10–20% of fat (mild group, n = 6); and (iii) more than 20% of fat (moderate group, n = 7). The bodyweight of the moderate group was high (771 kg) before calving. The sufficiency rates of total digestible nutrients (TDN) were remarkably decreased (approximately 65%) in early lactation. The milk fat yield and milk fat composition of the moderate group were higher (P < 0.05) than the other groups at 1 and 2 weeks post‐parturition. It was suggested that non‐esterified fatty acids (NEFA) mobilized from adipose tissues was directly used by the mammary gland for synthesis of milk fat. The percentage of bromsulfalein (BSP) retention of the moderate group was high (21.1%) at 30 min, and it showed that the BSP clearance function was significantly decreased. The concentrations of NEFA, β‐hydroxybutyric acid and glucose were appropriate indicators of energy status; however, aspartate aminotransferase, γ‐glutamyl transpeptidase and total bilirubin were not sensitive indicators of a moderate fatty liver. Thus, high‐yielding cows that calve in an overweight condition are more likely to develop excessive fat accumulation in the liver because of great mobilization from adipose tissues post‐parturition. In cows with a moderately fatty liver, a decrease in TDN sufficiency rates, an increase of milk fat yield and a reduction of liver function were observed in early lactation. The increase of serum NEFA and milk fat composition resulting from mobilization of adipose tissues helped to diagnose moderate fatty liver.     

Glucose transport and milk secretion during manipulated plasma insulin and glucose concentrations and during LPS‐induced mastitis in dairy cows

In dairy cows, glucose is essential as energy source and substrate for milk constituents. The objective of this study was to investigate effects of long‐term manipulated glucose and insulin concentrations in combination with a LPS‐induced mastitis on mRNA abundance of glucose transporters and factors involved in milk composition. Focusing on direct effects of insulin and glucose without influence of periparturient endocrine adaptations, 18 dairy cows (28 ± 6 weeks of lactation) were randomly assigned to one of three infusion treatments for 56 h (six animals each). Treatments included a hyperinsulinemic hypoglycaemic clamp (HypoG), a hyperinsulinemic euglycaemic clamp (EuG) and a control group (NaCl). After 48 h of infusions, an intramammary challenge with LPS from E. coli was performed and infusions continued for additional 8 h. Mammary gland biopsies were taken before, at 48 (before LPS challenge) and at 56 h (after LPS challenge) of infusion, and mRNA abundance of genes involved in mammary gland metabolism was measured by RT‐qPCR. During the 48 h of infusions, mRNA abundance of glucose transporters GLUT1, 3, 4, 8, 12, SGLT1, 2) was not affected in HypoG, while they were downregulated in EuG. The mRNA abundance of alpha‐lactalbumin, insulin‐induced gene 1, κ‐casein and acetyl‐CoA carboxylase was downregulated in HypoG, but not affected in EuG. Contrary during the intramammary LPS challenge, most of the glucose transporters were downregulated in NaCl and HypoG, but not in EuG. The mRNA abundance of glucose transporters in the mammary gland seems not to be affected by a shortage of glucose, while enzymes and milk constituents directly depending on glucose as a substrate are immediately downregulated. During LPS‐induced mastitis in combination with hypoglycaemia, mammary gland metabolism was more aligned to save glucose for the immune system compared to a situation without limited glucose availability during EuG.     

Comparison of cholesterol levels among lipoprotein fractions separated by anion-exchange high-performance liquid chromatography in periparturient Holstein–Friesian dairy cows

Changes in lipoprotein profiles occur in dairy cows during the periparturient period and in cows with transition cow disease. Here, the lipoprotein profiles of Holstein–Friesian dairy cows during the periparturient period were obtained by anion-exchange, high-performance liquid chromatography to evaluate the usefulness of lipoprotein profile evaluation during the periparturient period and in cows with fatty liver and milk fever. Lipoprotein levels (including total and high- (HDL-C) and low-density lipoprotein (LDL-C) cholesterol) in 10 healthy cows were low 4 weeks prepartum, with the lowest values at calving or within 1 week of calving; the values increased at 8 weeks postpartum. The lipoprotein levels were measured in 16 cows diagnosed with fatty liver (n=10) or milk fever (n=6) and compared to 10 healthy dairy cows. A significant difference was observed in HDL-C between healthy cows (at calving and 1 week postpartum), and the fatty liver and milk fever cows. Cows with fatty liver and milk fever had a lower mean HDL-C than the 10 healthy dairy cows at calving and 1 week postpartum. HDL-C might be a good indicator of energy balance for differentiating healthy cows from those with transition cow disease.