过瘤胃胆碱在围产期奶牛的应用研究进展

奶牛养殖过程中,围产期被认为是最重要的时期之一,在该时期奶牛从干奶状态转为泌乳状态,生理上所受的应激造成奶牛泌乳早期采食量降低,易发生能量负平衡.此外,奶牛生产性能和繁殖性能也会降低,并常常伴有一些围产期疾病的发生,如脂肪肝、酮症、乳房炎等.胆碱能影响奶牛的生产与繁殖性能,参与奶牛的脂肪代谢,可降低脂肪肝发生率,提高免疫性能.对围产期奶牛饲喂过瘤胃保护胆碱(rumen protected choline,RPC),可改善奶牛机体代谢,增加小肠胆碱供应,进而提高奶产量、改善乳成分、缓解脂肪肝、减少围产期疾病的发生.本文综述了RPC对围产期奶牛干物质采食量、生产性能、乳成分以及健康状况的影响,旨在为围产期奶牛的养殖提供理论参考依据.     

A polymorphism in the insulin-like growth factor 1 gene is associated with postpartum resumption of ovarian cyclicity in Holstein-Friesian cows under grazing conditions

Background

Insulin-like growth factor 1 (IGF-1) gene is considered as a promising candidate for the identification of polymorphisms affecting cattle performance. The objectives of the current study were to determine the association of the single nucleotide polymorphism (SNP) IGF-1/SnaBI with fertility, milk production and body condition traits in Holstein-Friesian dairy cows under grazing conditions.

Methods

Seventy multiparous cows from a commercial herd were genotyped for the SNP IGF-1/SnaBI. Fertility measures evaluated were: interval to commencement of luteal activity (CLA), calving to first service (CFS) and calving to conception (CC) intervals. Milk production and body condition score were also evaluated. The study period extended from 3 wk before calving to the fourth month of lactation.

Results and discussion

Frequencies of the SNP IGF-1/SnaBI alleles A and B were 0.59 and 0.41, respectively. Genotype frequencies were 0.31, 0.54 and 0.14 for AA, AB and BB, respectively. Cows with the AA genotype presented an early CLA and were more likely to resume ovarian cyclicity in the early postpartum than AB and BB ones. No effect of the SNP IGF-1/SnaBI genotype was evidenced on body condition change over the experimental period, suggesting that energy balance is not responsible for the outcome of postpartum ovarian resumption in this study. Traditional fertility measures were not affected by the SNP IGF-1/SnaBI.

Conclusion

To our knowledge this is the first report describing an association of the SNP IGF-1/SnaBI with an endocrine fertility measure like CLA in cattle. Results herein remark the important role of the IGF-1gene in the fertility of dairy cows on early lactation and make the SNP IGF-1/SnaBI an interesting candidate marker for genetic improvement of fertility in dairy cattle.     

Dry period management and optimization of post-partum reproductive management in dairy cattle

Dry period and early post-partum management are decisive factors for fertility in lactating dairy cows. Previous studies have shown that decreased dry matter intake (DMI) and increased non-esterified fatty acids (NEFA) negatively affect fertility and subsequent milk production. The traditional dry period decreases DMI prior to parturition, resulting in a decrease in energy intake. A negative energy balance increases NEFA concentration, and increased NEFA may impair the immune system, especially by decreasing neutrophil function prior to parturition. Earlier studies have shown that post-partum health disorders, including retained placenta and metritis, were correlated with periparturient neutrophil function. In addition, decreased DMI is also linked to a reduced body condition score (BCS) in dairy cows. These events in the periparturient period negatively affect fertility. Some manipulation, such as shortening the dry period, may be a solution to increased DMI in the periparturient period, preventing post-partum disorders and subsequent fertility issues. This article aims to explain the effects of shortening the dry period on reproduction and early post-partum treatments to improve fertility. In addition, timed artificial insemination protocols will be discussed for use during the post-partum period to improve fertility in dairy cows.     

Evaluation of candidate genotype of leptin gene associated with fertility and production traits in Hardhenu (Bos taurus × Bos indicus) cattle

The present study was conducted on Hardhenu cattle to screen genomic region of leptin gene with an objective to find the association of genotypes with fertility and production traits. The association analysis with traits under study was analysed by least squares analysis of variance by taking SNPs genotype as fixed effects in the statistical model. The genotypic frequencies with respect to targeted loci g.92450765 G > A indicated that AG (0.54) genotype was highest in Hardhenu cattle. Chi-squared tests showed that g.92450765G > A SNP meet with the Hardy–Weinberg equilibrium (p > .05).The association analysis revealed significant association of genotypes with total milk yield (TMY) and 305 days milk yield (MY) (p < .05). Service period (SP) and calving interval (CI) were also found significantly associated with genotypes (p < .05). Whereas, lactation length (LL), dry days (DD) and age at first calving (AFC) did not divulge any significant association with genotype. The AG and GG genotypes were associated with higher milk yields as compared to AA genotype, indicating that allele G was associated with superior milk performance. However, AA genotyped cattle found to be favourable with SP, CI and artificial insemination (AI) per conception compared to AG and GG genotyped cows. Chi-square analysis revealed that genetic variants of g.92450765 G > A SNP of leptin gene differ significantly with regard to reproductive disorders incidence (p < .05). The frequency of GG genotype (88.89%) in the affected animal group was very high followed by AG. The animals with GG genotype were found to be more susceptible to reproductive disorders as suggested by the higher odd ratio value (16.00) in logistic model. These observations and their differential association with the fertility and production traits can be utilized as an aid to selection for genetic improvement of antagonistic traits in dairy cows.     

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.     

中国荷斯坦牛PIK3CB基因多态性及其与繁殖和产奶性状的关联分析

【目的】 探究磷脂酰肌醇-4,5-二磷酸3-激酶催化亚单位β（PIK3CB）基因多态性及其与中国荷斯坦牛繁殖和产奶性状的关系。【方法】 通过混池测序对中国荷斯坦牛PIK3CB基因进行单核苷酸多态性（SNP）位点筛选,采用竞争性等位基因特异性PCR （KASP）技术在1 160头健康泌乳中国荷斯坦牛中进行SNP分型并进行群体遗传学分析,采用线性模型进行SNP与11个繁殖和产奶性状基于单位点和单倍型组合的关联分析。【结果】 在PIK3CB基因中共检测到了17个SNPs,筛选出7个SNPs用于后续分析。关联分析发现,7个SNPs与多个目标性状存在显著或极显著的关联（P<0.05;P<0.01）;位于外显子区域的g.130433743 A>G位点AA基因型个体和位于可变剪接区域的g.130448069 G>A位点GG基因型个体,其经产牛首末次配种间隔、产奶量、乳蛋白量和乳脂量最低,体细胞评分最高,上述基因型个体具有较短的首末次配种间隔,而产奶性能相对较差;g.130387717 G>A位点AA基因型个体,其初配日龄和青年牛首末次配种间隔最低,产奶量、乳蛋白量和乳脂量最高,该基因型个体的繁殖和产奶性能均较好,上述3个SNPs位点可作为中国荷斯坦牛繁殖和产奶性状的候选位点重点关注。单倍型分析发现,PIK3CB基因的g.130387717 G>A、g.130430832 A>－、g.130433743 A>G、g.130433982 C>T、g.130446073 C>T和g.130448069 G>A 6个SNPs紧密连锁形成一个单倍型块,且与多个目标性状存在显著或极显著关联（P<0.05;P<0.01）,其中H2H3和H2H4单倍型组合个体的繁殖和产奶性能较好,为优势单倍型组合。【结论】 中国荷斯坦牛PIK3CB基因存在丰富的遗传变异,其多态性与繁殖和产奶性状存在关联,g.130433743 A>G、g.130448069 G>A和g.130387717 G>A位点可作为潜在分子标记,为中国荷斯坦牛的平衡育种提供理论依据。     

围产期奶牛的生理特点及营养代谢特征研究进展

围产期(产前3周至产后3周)是奶牛内分泌和代谢发生巨变的时期,此时期奶牛经历了不同程度的能量负平衡,可影响其产后生产性能和对疾病的易感性.本文综合阐述了围产期奶牛的生理特点及营养代谢特征,为以后围产期奶牛的饲养管理及科学研究提供新的思路.     

Differences between primiparous and multiparous dairy cows in the inter-relationships between metabolic traits, milk yield and body condition score in the periparturient period

During the early postpartum period dairy cows mobilize fat and muscle to support lactation. This is associated with alterations in blood metabolite and hormone profiles which in turn influence milk yield and fertility. This study developed models to determine how metabolic traits, milk yield and body condition score were inter-related at different times in the periparturient period and to compare these relationships in primiparous (PP, n=188) and multiparous (MP, n=312) cows. Data from four previous studies which included information on blood metabolic parameters, parity, milk yield, body condition score and diet were collated into a single dataset. Coefficients of polynomial equations were calculated for each trait between -1 week pre-calving and week +7 postpartum using residual maximum likelihood modelling. The completed dataset was used in a multiple correlation model to determine how the best fit curves were related to each other over time. PP cows had higher concentrations of insulin-like growth factor-I and lower beta-hydroxybutyrate concentrations throughout, higher leptin concentrations pre-partum and both the peak in non-esterified fatty acids and the nadir in urea concentration occurred earlier after calving. These differences were associated with significantly lower milk production. Leptin concentrations fell at calving and were related to body condition score. Insulin was negatively correlated with yield in MP cows only. In MP cows the relationship between insulin-like growth factor-I and yield switched from negative to positive between weeks +4 and +7. Both beta-hydroxybutyrate and urea were positively related to yield in PP cows. In contrast, in MP cows beta-hydroxybutyrate was negatively correlated with yield and urea was strongly related to body condition score but not yield. These results suggest that there are differences in the control of tissue mobilization between PP and MP cows which may promote nutrient partitioning into growth as well as milk during the first lactation.     

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。     

正常泌乳奶牛的血清瘦蛋白、血糖和血酮浓度及其相关性

随机选取正常的黑白花泌乳牛20头,按乳酮阳性分为阴性组(11头)和阳性组(9头)。对它们泌乳量、泌乳天数、血糖、血酮、血清Leptin等指标进行统计分析。结果显示血清Leptin、血糖、酮体等指标水平处于正常的范围,5项指标彼此之间的相关性不明显。但是,乳酮阳性组奶牛与阴性组比较,泌乳量降低,血清酮体升高,血糖水平降低,血清Leptin水平略高,除血清Leptin外,其余都差异显著(<0.05)。结果表明,正常的泌乳奶血清瘦蛋白等5项指标之间无显著的相关性,可能与多种因素有关,尤其是能量代谢的适应性调节。     

荷斯坦奶牛STAT5A基因的SNPs检测及其与产奶性状的关联分析

根据GenBank公布的牛STAT5A基因的2个SNPs（AJ237937和AF079568）的引物序列,采用PCR-SS-CP方法对758头中国荷斯坦奶牛进行了STAT5A基因多态性检测,并将其与5个产奶性状进行了关联分析。在SNP1的9501碱基处发现A→G突变;SNP2中发现2个连锁点突变,即12 440位T→C和12 550位的CCT插入/缺失;方差分析结果表明：SNP1对乳蛋白率有显著影响（P〈0.05）;SNP2对产奶量有极显著影响（P〈0.01）,对乳脂量、乳蛋白量有显著影响（P〈0.05）;单倍型组合对产奶量和乳蛋白量有极显著影响（P〈0.01）,对乳脂量有显著影响（P〈0.05）。多重比较分析表明SNP2的AB基因型的产奶量、乳脂量和乳蛋白量的最小二乘均数极显著（P〈0.01）或显著（P〈0.05）高于基因型AA;而单倍型组合H1H2的产奶量、乳脂量和乳蛋白量的最小二乘均数显著（P〈0.05）或极显著（P〈0.01）高于其它4种单倍型组合;H3H4对产奶量和乳蛋白量的效应极显著高于其它4种单倍型组合（P〈0.01）。结果提示：STAT5A基因可以作为奶牛产奶性状的候选分子遗传标记。     

奶牛围产期保健添加剂的研究现状

围产期是奶牛饲养中的一个非常特殊阶段,对奶牛的健康和产后的生产性能都有很大的影响。添加剂在奶牛围产期保健的应用越来越广泛。文中通过对预防低血钙、增强免疫和缓解能量负平衡3个方面来对奶牛围产期保健添加剂的研究现状进行综述。     

Anthelmintic treatment of dairy cows and its effect on milk production.

The results of more than 80 experiments on gastrointestinal parasitism and the impact of anthelmintic treatment on milk production in dairy cattle were reviewed. Abattoir surveys of culled dairy cows, faecal egg counts in milking cows, and serological tests and worm counts in cull cows in milk production studies were collated to assess the level of parasitism in dairy herds. The studies were divided into four general categories: induced infections in previously uninfected cattle; naturally infected cattle treated in mid-lactation; naturally infected cattle treated one to three times during the dry period and/or just before or just after parturition; and naturally infected cattle treated repeatedly from early lactation or given strategic treatments throughout the year. In most studies, the milk production of anthelmintic-treated cattle was compared with that of untreated controls. The anthelmintics investigated included members of the organophosphate, benzimidazole, imidazothiazole and macrocyclic lactone groups. The number of experiments in which the medicated (or uninfected) group had a higher milk yield was compared with the number of experiments in which the control (or infected) group had a higher yield. Overall, the studies demonstrated that grazing dairy cattle are likely to be infected with gastrointestinal nematode parasites, usually Ostertagia ostertagi and Cooperia species. These infections may be present as inhibited larvae, and a periparturient or spring rise is associated with their emergence. There is, at present, no reliable means of determining whether a cow or a herd may be parasitised subclinically at a level sufficient to interfere with milk production. In 70 of 87 experiments (80 per cent) there was an increase in milk production (P < 0.001) after anthelmintic treatment, with a median increase of 0.63 kg/day. In each of the four trial categories, a majority of the studies showed that anthelmintic treatment increased milk production. The yield of milk fat by the medicated cows was greater than by the controls in 26 of the 35 experiments in which that variable was studied (P < 0.01).     

中国不同牛种DGAT1基因遗传多样性及经济性状关联分析研究进展

乙酰辅酯A∶二酰甘油酰基转移酶(DGAT1)是甘油三脂合成过程中唯一的关键酶,对动物机体脂肪代谢、沉积起重要作用,已成为研究奶牛泌乳性状和肉牛主要经济性状的重要候选基因之一。近年来,随着相关研究的不断深入,研究者普遍认为DGAT1基因对奶牛的产奶量、乳脂率、乳蛋白率、乳糖率、乳脂肪酸及空怀天数、输精次数、初配年龄等繁殖性状均有显著影响,且与肉牛体脂也存在相关性。作者主要介绍了DGAT1基因的遗传多态性及其与经济性状关联分析在中国各地区不同品种奶牛、肉牛、水牛和牦牛上的研究进展,指出该基因对奶牛的产奶性能及肉牛的肉质性状、生长性状具有重要的调控作用。但由于环境条件、品种及选育背景等因素影响,DGAT1基因遗传多态性对不同地区不同牛群体的经济性状影响不尽相同。因此,要将DGAT1基因实际应用在品种选育工作中还需明确该基因对研究群体的具体遗传效应,并结合其他候选基因进行综合分析。     

Adiponectin links adipose tissue function and monocyte inflammatory responses during bovine metabolic stress

The periparturient period of dairy cows is characterized by intense lipid mobilization from adipose tissue leading to increased plasma concentrations of nonesterified fatty acids (NEFA). High NEFA are a predisposing factor for inflammatory based diseases. A major component of these diseases is uncontrolled macrophage/monocyte inflammatory responses. Changes in the endocrine activity of adipose tissue during the periparturient period could impact macrophage function by modifying the secretion of adipokines including adiponectin. Currently, the effects of adiponectin on monocyte activation in dairy cattle are unknown. In humans and rodents, this adipokine regulates monocyte phenotype and alterations in its plasma levels are linked with the development of inflammatory diseases. The objectives of this study were to establish associations between plasma adiponectin expression dynamics and different markers of lipid mobilization during the periparturient period of dairy cows and to characterize the effects of adiponectin on the inflammatory response of bovine monocytes. Plasma adiponectin, NEFA, BHB, albumin, and subcutaneous and retroperitoneal fat depots depth were measured during the periparturient period of dairy cows. In vitro, bovine monocytes were cultured with adiponectin to assess changes in pro-inflammatory responses following LPS stimulation. Results from this study demonstrate that alterations in plasma adiponectin levels in periparturient cattle are inversely correlated with the concentrations of plasma NEFA, an important marker of lipid mobilization. Furthermore, adiponectin exposure significantly decreased monocyte expression of TNFα after LPS stimulation thus markedly reducing their inflammatory response. Reduced plasma adiponectin during the periparturient period could predispose dairy cows to the development of uncontrolled monocyte inflammatory responses.     

Polymorphisms in the bovine leptin promoter associated with serum leptin concentration, growth, feed intake, feeding behavior, and measures of carcass merit

Leptin is the hormone product of the obese gene synthesized and secreted predominantly by white adipocytes. It functions as a lipostatic signal regulating BW, food intake, energy expenditure, reproduction, and certain immune system functions. Although previous studies have identified polymorphisms in the coding regions of the leptin gene in cattle that show considerable associations with feed intake, milk quality and quantity, and carcass fatness, no such associations have been reported for the leptin promoter. The current study reports associations between SNP in the 5' untranslated promoter region of the bovine leptin gene with serum leptin concentration, growth, BW, feed intake, feeding behavior, and carcass merit in hybrid cattle (n = 150). The study showed that animals with the TT genotype of a less frequent cytosine/thymine (C/ T) substitution (UASMS2; frequency of thymine allele equals 0.21) detected at position 528 in the bovine leptin promoter (GenBank Accession No. AB070368) show 48 and 39% increases in serum leptin concentration (P < 0.001), 39 and 31% increases in backfat thickness (P < 0.001), and 13 and 9% increase in marbling score (P = 0.01), compared with CC or CT genotypes, respectively. Animals with the TT genotype also show significantly higher feed intake (P < 0.001), growth rate, metabolic BW (P < 0.05), and live weight at slaughter (P < 0.10). Animals with the GG genotype of a more frequent cytosine/guanine (C/G) substitution (UASMS3; frequency of G allele equals 0.59) at position 1759 in the bovine leptin promoter (GenBank Accession No. AB070368) also show higher feed intake (P = 0.001), growth rate (P < 0.10), and BW (P < 0.01). The thymine allele of UASMS2 and the guanine allele of UASMS3 were separately associated with higher feeding duration (P < 0.05). The two SNP show significant linkage disequilibrium and could also be relevant in predicting other characteristics, such as milk yield and quality in cattle. These results, however, represent the initial associations of the polymorphisms with these traits, and further efforts are required to validate these findings in other populations.     

Leptin receptor expression and its change in association with the normalization of EGF profile after seminal plasma treatment in repeat breeder dairy cows

Factors associated with high milk production levels have been linked to alterations in the endometrial epidermal growth factor (EGF) profile, a cause of reduced fertility in dairy cows. Therefore, we examined the leptin system that connects nutritional status and reproduction in dairy cattle related to reduced fertility in repeat breeder cows. Plasma leptin concentrations were measured in 18 heifers, 20 high-yielding control cows, and 26 repeat breeder cows, showing an altered EGF profile. Then, all repeat breeder cows were infused with seminal plasma (SP) into the vagina at the next estrus to normalize the EGF profile, while heifers and control cows were infused with vehicle alone. All animals were examined for EGF profiles. Eighteen repeat breeder cows, nine heifers, and nine control cows were also determined for leptin receptor (Ob-R) expression levels in the estrous cycle before and after the infusion. SP normalized the EGF profile in 53.8% of the repeat breeder cows. Leptin concentrations were similar in all groups, regardless of the treatment results for the EGF profile. In contrast, Ob-R levels in repeat breeder and control cows were similar and higher than those in heifers before SP treatment. Ob-R in repeat breeders showing a normal EGF profile after treatment decreased to an intermediate level between heifers and control cows and may provide a clue to take measures against repeat breeding in dairy cows.     

副乳头对奶牛生产性能的影响及其相关基因LGR5的遗传表达分析

试验旨在探究副乳头（SNT）对奶牛产奶性能的影响及乳腺发育相关基因LGR5与副乳头发生的关系。收集325头有副乳头奶牛及同年出生的737头无副乳头奶牛的产奶性能数据,比较有、无副乳头奶牛间的产奶性能差异;利用实时荧光定量PCR技术检测有、无副乳头奶牛LGR5基因的表达差异,通过直接测序法测定有、无副乳头奶牛的LGR5基因序列,寻找该基因上的遗传突变。结果表明,无副乳头奶牛的日产奶量、高峰奶量及泌乳持续力等产奶性能均优于有副乳头奶牛,但未达到统计学显著水平;LGR5基因在有副乳头奶牛中的表达量显著高于无副乳头奶牛（P<0.05）;在LGR5基因第1内含子中检测到3个SNPs：rs42849472、rs42849471和rs42849470,有副乳头奶牛中3个SNPs的AA基因型频率均极显著高于无副乳头奶牛（P<0.01）。结果提示,LGR5基因可能与奶牛副乳头发生有关,可作为有效的候选基因。     

中国荷斯坦牛PIN1基因对产奶性状的遗传效应分析

前期研究通过荷斯坦公牛全基因组重测序鉴定到17个奶牛产奶性状候选功能基因,其中,肽基脯氨酸顺反异构酶基因(PIN1)参与甘油三酯代谢、甘油磷脂代谢以及mTOR信号通路,且位于产奶量和乳蛋白量性状QTL区间.为进一步系统分析PIN1基因是否对奶牛产奶性状具有遗传效应,本实验基于40头公牛的基因组DNA混池,采用PCR产物...