不同生理状态乳牛卵巢卵母细胞基因的差异表达

为了研究不同生理状态下的卵巢卵母细胞基因的表达情况,采集乳牛的卵巢,分离了卵母细胞,以单个卵母细胞的mRNA作为模板,用设计的随机引物和锚定引物,采用一步法RT-PCR扩增了卵母细胞基因。经聚丙烯酰胺凝胶电泳检测,发现有功能黄体和无黄体卵巢卵母细胞基因的电泳条带之间无明显差异,对一条明显差异条带进行回收、纯化,将纯化的PCR产物连接到T载体,阳性克隆经鉴定后,进行测序和同源性比较。结果显示,与体外培养的牛早期胚胎的一条序列具有很高的同源性。     

新疆地区奶牛子宫内膜炎病原菌的分离鉴定及其对抗菌药的敏感性试验

通过细菌形态学和细菌生化特性,分离和鉴定了新疆地区患子宫内膜炎的21头奶牛子宫分泌物中的病原菌分别为蜡样芽孢杆菌(24.5%)、非溶血性链球菌(18.4%)、大肠杆菌(14.3%)、溶血性链球菌(10.2%)、表皮葡萄球菌(10.2%)、金黄色葡萄球菌(10.2%)、胎儿弯曲菌(10.2%)和不动杆菌等.其中革兰氏阳性菌检出率较高(73.5%).子宫内膜炎病原菌以单一菌株感染为主(47.6%).采用Kirby-Barer法测定了鉴定菌株对13种抗微生物药的体外药物敏感性.链球菌、大肠杆菌和葡萄球菌对氨苄西林不敏感,而对卡那霉素和庆大霉素敏感性较高;大肠杆菌对青霉素和氨苄西林全部耐药(100%),对四环素和多西环素耐药率较高(均为83.3%);溶血性链球菌对青霉素和四环素全部耐药,对链霉素耐药率为80%;相对而言,表皮葡萄球菌和金黄色葡萄球菌的耐药率较低;氟喹诺酮类对各种分离菌株比较敏感.     

Variation in selected blood plasma constituents during the post-partum and breeding periods in dairy cows

Concentrations of insulin-like growth factor-I, albumin, beta hydroxybutyrate, glucose and urea were measured in 12 plasma samples collected over 9 weeks from mid-August from each of 40 cows in four seasonal herds which calved from mid-July. These herds comprised Jersey or Holstein-Friesian animals, with each breed grazed at two stocking rates. Daily dry matter intakes (kg dry matter/cow/day), which were estimated from pasture assessments before and after grazing, ranged from 4% (low stocked Jersey herd) to 38% (high stocked Friesian herd) below recommended intakes for each breed of cow in peak lactatipn. Stocking rate was associated with a 4% reduction in liveweight for cows in the high stocked herds and a 20% reduction in daily milk production (as a proportion of the low stocked herds' liveweight and milk production).

Eight cows produced at least one blood sample with a beta hydroxybutyrate concentration of more than 1.0 mmol/l. There were 42% of samples with glucose concentrations greater than 4.1 mmol/l. These two concentrations were the maximum values classified as normal for these two metabolites by the Ruakura Animal Health Laboratory. No concentrations of the other metabolites were outside their respective normal ranges. Breed, age and individual animals usually had greater effects on the average metabolite concentrations than did stocking rate.

The greatest effect was associated with sample day, especially for insulin-like growth factor-I and urea. A significant breed x stocking rate x sample day interaction in urea concentrations indicated that changes in the feeding value of the grazed pastures available to each herd were sufficient to produce separate fluctuating patterns in the daily average concentrations.

These results showed that frequent sampling on a herd basis would be necessary to identify the significance of factors which may be contributing to sample day variation in the concentration of metabolites in the plasma of pasture-fed cows.     

Follicular cysts in dairy cows

Follicular cysts are the most critical reproductive disorder in dairy cows and disturb the normal ovarian cycle, resulting in a prolonged interval from calving to conception. Therefore, this condition causes significant economic losses to the dairy industry. Two direct causal factors for cysts are suggested in this review; ovulation disorder and the delay of regression. Ovulation disorder has been accepted to be a main etiology of cystic follicle. This seems to be caused by the deficiency of positive feedback of estrogen to the hypothalamus, leading to the lack of luteinizing hormone surge. On the other hand, if a large anovulatory follicle is regressed immediately after the failure of ovulation, its follicle does not continue to grow, resulting in no cystic follicle being formed. Therefore, it is proposed that another cause of a cystic follicle is the delay (or absence) of the degeneration system of the follicle. This review will introduce these two causes separately, referring to recent advances in understanding the follicular cyst in dairy cows.     

Relationship among blood indicators of lipomobilization and hepatic function during early lactation in high-yielding dairy cows

Blood indicators are used as a tool to diagnose metabolic disorders. The present work was conducted to study the relationships among blood indicators of lipomobilization and hepatic function in high-yielding dairy cows. Two groups of Holstein cows were studied: 27 early lactation cows and 14 mid lactation cows from four different herds with similar husbandry characteristics in Galicia, Spain. Blood samples were obtained to measure beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), triglycerides (TG), and the activity of aspartate transaminase (AST) and gamma-glutamyl transferase. Cows in early lactation had higher levels of BHB and NEFA than mid lactation cows. High lipomobilization (NEFA > 400 µmol/L) was detected in 67% and 7% of early lactation and mid lactation cows, respectively, while subclinical ketosis (BHB > 1.2 mmol/L) was detected in 41% and 28% of the early lactation and lactation cows, respectively. TG concentrations were low in all cows suffering subclinical ketosis and in 61% of the cows with high lipomobilization. During early lactation, 30% of cows suffered hepatic lipidosis as detected by levels of AST. Compromised hepatic function was observed in early lactation cows as shown by lower concentrations of glucose, total protein, and urea.     

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.     

我国奶牛普通病的危害特点与防治对策

我国奶牛的普通病发病呈现逐年升高的趋势,尤其是内科病和产科病,大多数疾病呈隐性经过,因缺乏高效诊断技术这些疾病对我国奶牛业的健康发展危害较大,笔者着重分析了奶牛乳房炎、不孕症、营养代谢病、肢蹄病、胎衣不下、犊牛腹泻和真胃移位等常见疾病的发病情况、所造成的经济损失以及防治中存在的问题,并从流行病学调查、诊断技术、发病机理、防治药物、疾病监测体系、综合防控技术和奶牛标准化养殖等方面提出了奶牛普通病的防治对策。     

乳牛产后血浆瘦素动态与亚临床酮病的相关分析

对某奶牛场16头乳牛产前10d至产后56d血样的瘦素、酮体、葡萄糖、脂肪水平及其动态特征和相关性进行了检测。结果表明，血浆瘦素、血酮、血脂含量在不同乳牛和不同时间点之间均有显著差异。酮体在产后14～49d出现高峰，瘦素和血脂从产后第0d到产后第56d内逐渐升高。瘦素水平与血脂水平呈极显著正相关，血酮与血糖呈极显著负相关。亚临床酮病组乳牛的瘦素和血脂水平分别极显著或显著低于血酮正常组，其瘦素、血酮和血糖在试验期内的波动频率明显减少，跨度变长，瘦素在试验期内仅出现一个两端接近0、波峰不超过1．5ng／mL的波。证实，产后乳牛瘦素、血酮、血糖含量的低频率波动变化和产后8周内血浆瘦素跨时持久的单峰动态变化与乳牛亚临床酮病的发生有关。     

盐藻粉在奶牛生产中的应用

盐藻(Dunaliella salina)是一种单细胞无细胞壁的真核生物,主要用于提炼天然β-胡萝卜素和多糖。盐藻粉是盐藻干燥后获得的粉状颗粒,具有抗氧化、抗病毒、抗肿瘤和提高免疫力等生物学活性。目前,盐藻粉在人类和啮齿类动物上的研究较多,而其在奶牛生产中的应用很少。本文从瘤胃发酵、氧化应激调控、免疫功能、营养素利用和生产性能5个方面综述了盐藻粉对奶牛代谢的影响和潜在应用价值,以期为盐藻粉在奶牛生产中的应用提供参考依据。     

2005年上海地区乳牛球虫感染的季节动态

为了解上海地区乳牛球虫感染的季节变化情况，对上海地区3个牧场乳牛抽样直肠采集粪便，检查了718头乳牛粪样。结果，查出球虫阳性牛269头，平均感染率为37．46％，其中1月龄以内牛的感染率为33．89％，1～12月龄牛的感染率为42．33％，12月龄以上牛的感染率为25．95％。平均感染率最高的4月份为44．44％，最低的8月份为28．57％。3个牧场球虫阳性牛的感染强度（OPG值）为0～169000个，平均OPG值为9477个，其中1月龄以内牛的OPG值为8270个，1～12月龄牛的OPG值为4318个，12月龄以上牛的OPG值为145个。调查发现了6种球虫，分别是牛艾美球虫（Eimeria boris）、椭圆艾美球虫（E．ellipsoidalis）、邱氏艾美球虫（E．zurnii）、怀俄明艾美球虫（E．wyomingensis）、柱状艾美球虫（E．cylindrica）、亚球形艾美球虫（E．subspherica）。结果表明，2005年上海地区乳牛球虫感染率无明显季节差异，12月龄内乳牛的球虫感染率与感染强度均明显高于12月龄以上乳牛，乳牛球虫的优势虫种为牛艾美球虫、椭圆艾美球虫、邱氏艾美球虫。     

益生菌对奶牛亚急性瘤胃酸中毒的保护作用

亚急性瘤胃酸中毒对奶牛的生产性能有很大影响,使用益生菌来稳定奶牛分娩后-产奶过渡期瘤胃pH可以减轻这种代谢紊乱的症状。因此,本试验选择了体重(741±55)kg,产奶期在(212±19.5)d的奶牛4头,试验采用4×4拉丁方设计,共4种日粮:对照组饲喂基础日粮,处理1组为日粮添加0.5 g/d米曲菌,处理2组为日粮添加2.5 g/d米曲菌,处理3组为日粮添加2 g/d粪肠球菌和酿酒酵母菌的混合物。每组经过3 w的适应期、4 d酸中毒期以及3 d恢复期。结果显示:瘤胃最大pH范围在5.6～6.0,占比最高,与适应期和中毒期的结果相似。中毒期间,其他pH范围表现为显著差异(P <0.05)。中毒期,处理3组较对照组显著提高了瘤胃pH(P <0.05),同时处理3组瘤胃pH在5.6～6.0的占比也显著高于对照组(P <0.05)。日粮添加2.5 g/d米曲菌显著降低了产奶量(P <0.05)。奶牛遭受瘤胃酸中毒后,随着时间的推移,丙酸、丁酸和戊酸含量显著升高(P <0.05),而乙酸、异丁酸和异戊酸含量显著降低(P <0.05)。除了处理3组外,其他组瘤胃乳酸含量随时间的增加而升高(P <0.05)。饲喂12 d后,处理3组较对照组显著提高了乳酸含量(P <0.05)。瘤胃白球菌和大肠杆菌含量在饲喂2 h后显著升高(P <0.05),之后在6 h达到稳定。结论 :粪肠球菌和酿酒酵母菌复合物可以缓解奶牛亚急性瘤胃酸中毒的症状,米曲菌可以调控瘤胃pH,但高剂量米曲菌添加水平对瘤胃pH无显著影响。     

日粮添加碳酸氢钠对荷斯坦奶牛生产性能的影响

选择体重、胎次、泌乳期、乳脂率和产奶量基本一致的健康无病的中国荷斯坦奶牛30头,随机分成3组,分别为试验组Ⅰ,Ⅱ和对照组。试验组Ⅰ,Ⅱ奶牛日粮每日每头分别添加130、150g碳酸氢钠,对照组奶牛日粮每日每头分别添加110g碳酸氢钠,试验期28d。结果表明,试验组Ⅰ干物质采食量、粪便pH比对照组高1.41%、0.20(P0.05),非脂乳固体比对照组低0.4(P0.05);试验组Ⅱ干物质采食量、产奶量、4%标准乳、粪便pH分别比对照组高2.04%、2.01%、2.68%、0.42(P0.05),乳脂率高0.13%(P0.05),乳蛋白、非乳脂固体分别低0.36%、0.51%(P0.05)。另外,碳酸氢钠的增加使得牛乳中体细胞数显著降低,且与乳脂肪呈显著负相关(P0.05),而与乳蛋白质呈显著正相关(P0.05)。由此可知,在每头奶牛日粮中添加150g碳酸氢钠的范围内,随着日粮中碳酸氢钠的增加,能显著改善荷斯坦奶牛的生产性能,有效预防奶牛瘤胃酸中毒。     

影响奶牛生产性能及经济效益的因素

在规模化牛场的生产过程中,品种、营养、牛群结构、饲养管理、疾病防治等因素对奶牛生产及经济效益有很大影响.而选择高产品种、调整营养水平、建立合理的牛群结构、采取适宜的饲养管理和疾病防治措施,不仅能够提高奶牛的生产性能、繁殖性能,还能降低饲养成本、提高经济效益.文章对影响奶牛生产性能及经济效益的因素进行论述,为促进我国规模...     

Effect of rumen‐protected choline and methionine on physiological and metabolic disorders and reproductive indices of dairy cows

The objective of this study was to investigate the effect of feeding different levels of ruminally protected methionine and choline on the incidence of physiological and metabolic disorders, production, and some of the reproductive indices of Holstein dairy cows. Forty Holstein dairy cows in their first and second lactation were used from 4‐week pre‐partum through 20‐week post‐partum and randomly assigned to receive one of the following treatments: 18 g/day of rumen‐protected methionine (RPM), 60 g/day of rumen‐protected choline (RPC), 18 g/day of RPM + 60 g/day of RPC, and neither supplement (control). The treatments significantly affected services per conception and open days of lactating dairy cows (p < 0.05), but did not affect significantly on days to first oestrus and number of pregnant cows. RPM + RPC‐fed cows had the lowest open days, days to first oestrus and services per conception compared with other groups. The effect of treatments was significant on the incidence of metabolic and physiological problems except for foot/leg problems. Cows fed RPM+RPC had the lowest health problems compared with other groups (p < 0.05). Results indicate that the supplementation of RPM and RPC can improve reproductive performance and health status of dairy cows.     

Natural antibodies related to metabolic and mammary health in dairy cows

Natural antibodies (NAb) are defined as antibodies that circulate in normal healthy individuals under the absence of deliberate antigenic stimulation. Two types of NAb are distinguished: NAb towards exogenous antigens and NAb towards autoantigens (N(A)Ab). The objectives of the current study were threefold. First, we studied the relation between metabolic health and concentrations of NAb binding keyhole limpet hemocyanin (KLH) or lipopolysaccharide (LPS) in milk and plasma of dairy cows in early lactation. Second, we determined the presence of N(A)Ab binding transferrin, myosin and thyroglobulin in bovine milk. Third, we studied the relation between N(A)Ab in bovine milk and mammary health. For the first objective, dairy cows were either fed a control (C) (n = 8) or a diet where 2 kg of concentrates were replaced by an iso-energetic concentrate containing marine algae (ALG) from week −3 till 8 postpartum (experiment 1). Plasma and milk samples were analyzed weekly for NAb binding either KLH or LPS. Plasma was analyzed for glucose, non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA). For the second and third objective, milk samples were collected weekly from 96 dairy cows from week 2 till 9 postpartum and analyzed for milk composition and N(A)Ab binding myosin, transferrin and thyroglobulin (experiment 2). For both datasets, N(A)Ab titers are expressed as ²log values of the highest dilution giving a positive reaction. Data are expressed as means ± SEM. Repeated observations were analyzed in a mixed model. In experiment 1, no diet effect (P > 0.05) was observed on NAb binding LPS in plasma or milk, NAb binding KLH in milk was greater (P = 0.05) for cows fed the control diet. Concentration of NAb binding KLH and LPS in plasma was negatively related to plasma NEFA concentration (P < 0.05). In experiment 2, NAb binding myosin (5.66 ± 0.06), thyroglobulin (4.85 ± 0.06), and transferrin (5.76 ± 0.07) were identified in milk. Clinical mastitis incidence (9%) tended to be positively related to concentration of NAb binding myosin (P = 0.06) and negatively related to Nab binding transferrin (P = 0.08). In conclusion, NAb binding KLH and LPS in plasma and milk are related to metabolic health, as indicated by plasma NEFA concentration. Furthermore, this study demonstrates the presence of N(A)Ab in bovine milk and shows trends for a relation between N(A)Ab binding auto-antigens and mastitis. Future studies should confirm these trends and shed light on the predictive value of N(A)Ab in bovine milk for mammary health.     

夏季奶牛血液中热休克蛋白的表达量

选择40头体质健康的中国荷斯坦奶牛,根据胎次和泌乳天数,按照随机区组试验设计分为Ⅰ组（30kg/d）、Ⅱ组（30～35kg/d）、Ⅲ组（35～40kg/d）和Ⅳ组（40kg/d）。热应激前、热应激前期、热应激中期、热应激后期和热应激后分别于尾静脉采血,用ELISA试剂盒测定热休克蛋白（HSP）27,70,90的表达量。结果显示,Ⅳ组HSP27表达量最高,Ⅱ组表达量最低,Ⅳ组、Ⅲ组和Ⅰ组均显著高于Ⅱ组（P〈0.05）。HSP70表达量各组间没有明显差异,但随产奶量呈线性增加（P〈0.05）。HSP90的表达量,Ⅳ组和Ⅲ组明显高于Ⅱ组（P〈0.05）。HSP27的表达量热应激后差异较大;HSP70的表达量各组整个过程差异较大;HSP90的表达量在热应激前、热应激前期和热应激后差异较大。总之,在热应激过程,高产奶牛血清中热应激蛋白的表达量较高,HSP70表达量随产奶量呈线性增加,而不同热应激蛋白的变化规律差异较大。     

Activity of hepatic but not skeletal muscle carnitine palmitoyltransferase enzyme is depressed by intravenous glucose infusions in lactating dairy cows*

A positive energy balance in dairy cows pre‐partum may decrease hepatic carnitine palmitoyltransferase (CPT) enzyme activity, which might contribute to disturbances of lipid metabolism post‐partum. The purpose of this study was to investigate whether skeletal muscle CPT activity can also be downregulated during positive energy balance. Mid‐lactating dairy cows were maintained on intravenous infusion of either saline (control) or glucose solutions that increased linearly over 24 days, remained at the 24‐day level until day 28 and were suspended thereafter. Liver and skeletal muscle biopsies, as well as four diurnal blood samples, were taken on days 0, 8, 16, 24, and 32, representing infusion levels equivalent to 0%, 10%, 20%, 30% and 0% of the net energy for lactation (NE_L) requirement respectively. Glucose infusion increased serum insulin concentrations on day 16 and 24 while plasma glucose levels were increased at only a single time point on day 24. Serum beta‐hydroxybutyric acid concentrations decreased between day 8 and 24; whereas changes in non‐esterified fatty acids were mostly insignificant. Total lipid contents of liver and skeletal muscle were not affected by treatment. Hepatic CPT activity decreased with glucose infusion (by 35% on day 24) and remained decreased on day 32. Hepatic expression levels of CPT‐1A and CPT‐2 mRNA were not significantly altered but tended to reflect the changes in enzyme activity. In contrast to the liver, no effect of glucose infusion was observed on skeletal muscle CPT activity. We conclude that suppression of CPT activity by positive energy balance appears to be specific for the liver in mid‐lactating dairy cows.     

酮病对泌乳早期乳牛体内代谢和内分泌的影响

随机选取某奶牛场10头患临床酮病乳牛和10头同期健康对照组乳牛，检测了2组乳牛血液10项指标，阐明了酮病对泌乳早期乳牛体内代谢和内分泌的影响。结果显示，酮病乳牛血糖浓度极显著降低（P〈O．01），血浆NEFA和BHBA的浓度明显增高（P〈O．01）；酮病乳牛血浆Ins、LP、NPY、E2的浓度和Ins／Gn比值均明显降低（P〈O．05），而血浆Gn浓度未明显升高，P4未明显降低（P〉0．05）。表明，酮病乳牛体内某些激素协调作用紊乱会妨碍酮病乳牛能量负平衡的缓解，并将对产后生殖机能产生不良影响。     

Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

The present study was undertaken to develop a novel, practical and simple procedure for enzyme‐linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti‐estrone antibody and horseradish peroxidase‐labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra‐assay and inter‐assay coefficients of variation were 5.3–8.1% and 13.4–15.7%, respectively. The recovery rate was 78–102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.     

Loser cows in Danish dairy herds: definition, prevalence and consequences

During the last few years, many Danish dairy farmers have expressed increasing concerns regarding a group of cows, which we have chosen to term ‘loser cows’. Until now, a loser cow has not been described scientifically. We defined a loser cow on the basis of a clinical examination of the cow. A total of 15,151 clinical examinations were made on 6,451 individual cows from 39 randomly selected, large Danish dairy herds with loose-housing systems using a clinical protocol. Scores for the clinical signs lameness, body condition, hock lesions, other cutaneous lesions, vaginal discharge, condition of hair coat and general condition were converted into a loser cow score. Cows with a loser cow score of 8 or more were classified as loser cows. The overall prevalence of loser cows was 2.15%, 4.50% and 2.98% during the first, second and third round of herd visits, respectively.

The associations between the loser cow state and milk production, mortality, morbidity, culling and workload for the farmer were evaluated using data from herd visits and from the Danish Cattle Database and a number of different statistical techniques. It was concluded that the loser cow state has significant negative consequences for both the farmer and the cow. On average, loser cows yielded 0.61 to 2.24 kg energy corrected milk less per day than non-loser cows depending on parity. Hazard ratio for death or euthanasia was 5.69 for loser cows compared to non-loser cows. Incidence rate ratio for disease treatments was 0.69 for non-loser cows compared to loser cows. Loser cows were often culled in an ‘unfavourable’ way and generally caused extra workload for the farmer.

A simplified version of the loser cow score was evaluated and is recommended for future research and use in practice.