Searching for DNA markers for milk production and composition on chromosome 6 in sheep

Several milk protein polymorphisms are potential tools for selection in dairy ruminants. However, research results for dairy sheep are not as conclusive as those for goats or cattle and are often controversial. The main objective of this study was to find and later use molecular genetic markers in selection to improve milk production and milk composition in Awassi ewes. Chromosome 6 was chosen because several studies have reported the presence of significant quantitative trait loci (QTL) affecting milk production traits on ovine and bovine chromosome 6. Altogether, genotypes for 13 microsatellite loci were determined for 258 ewes, which were purebred Awassi or Awassi-Merino crosses. Phenotypic data were lactation yield of milk, milk fat, protein and lactose (kg), average milk protein and fat percentage and average somatic cell count. Five out of the 13 microsatellites showed significant association with at least one of the examined traits.     

Evaluation of association between polymorphism within the thyroglobulin gene and milk production traits in dairy cattle

In dairy cattle, many studies have reported quantitative trait loci (QTL) on the centromeric end of chromosome 14 that affect milk production traits. One of the candidate genes in this QTL region – thyroglobulin (TG) – was previously found to be significantly associated with marbling in beef cattle. Thus, based on QTL studies in dairy cattle and because of possible effects of this gene on fat metabolism, we investigated the association of TG with milk yield and composition in Holstein dairy cattle. A total of 1279 bulls from the Cooperative Dairy DNA Repository Holstein population were genotyped for a single nucleotide polymorphism in TG used previously in beef cattle studies. Analysis of 29 sire families showed no significant association between TG variants and milk production traits. Within‐sire family analysis suggests that TG is neither the responsible gene nor a genetic marker in association with milk production traits.     

An independent confirmation of a quantitative trait locus for milk yield and composition traits on bovine chromosome 26

Several reports have demonstrated that bovine chromosome 26 (BTA26) harbours significant or suggestive quantitative trait loci (QTL) for milk production and composition traits in dairy cattle. Our previous study showed that a C/T substitution in the bovine TCF7L2 gene on BTA26 was significantly linked to QTL for protein yield (PY) in a Canadian dairy cattle population. Actually, this polymorphism was one of the markers derived from a genome‐wide screening of QTL for milk PY using an amplified fragment length polymorphism technique combined with a DNA pooling strategy. In the present study, 990 Holstein bulls with complete genotype and phenotype data from 14 sire families were analysed to confirm, if the QTL effects exist in other populations. Statistical analysis revealed that this marker was significantly associated with PY, protein percentage, milk yield and fat yield (FY) (p < 0.001) in the US Holstein population. These results indicate that this QTL region has a pleiotrophic effect on different milk traits and is portable in different populations.     

奶牛生物技术育种研究进展

奶业已成为我国农业战略产业的重要组成部分,与发达国家相比,我国奶牛在良种率、产奶量、乳品质等方面仍存在较大差距。面对日益增长的消费量和品质需求,加快奶牛育种显得尤为迫切。本文便针对我国奶业当前发展现状分析了我国奶牛产业发展存在的问题及利用生物技术进行育种的必要性和迫切性。从分子标记辅助选择、QTL 定位、全基因组关联分析、全基因组选择、转基因以及胚胎工程等方面阐述了近些年国内外奶牛生物技术育种研究现状和进展,讨论了目前奶牛生物技术育种过程中存在的问题,以期对我国奶业发展有所裨益。     

Consequences of selection for improving production traits on the frequency of deleterious alleles for fitness

In this study the effect of artificial selection on BLUP EBV for production traits on the allele frequencies of a pleiotropic QTL affecting both production and disease susceptibility was investigated. Stochastic simulations were used to model artificial selection on a production trait that is controlled, in part, by a biallelic QTL that also controls susceptibility to disease. The QTL allele increasing production also increased susceptibility to disease. Different modes of action and proportions of variation accounted for by the QTL were assessed for the production trait. The main results indicated that alleles that confer susceptibility to the disease could be maintained in the population over a long period, depending on the mode of action of the QTL. In addition, the results of the study indicate that, under various conditions, it is possible to find pleiotropic QTL that control 2 traits despite these traits appearing to be uncorrelated. Therefore, in practice, an estimate of the genetic correlation between 2 traits may be misleading when the presence of such a QTL exists. The results of this study have implications for breeding programs. For example, if a pleiotropic QTL exists that favors heterozygotes for a production trait, it would be very difficult to remove disease susceptibility alleles via traditional selection methods.     

A whole genome scan to map QTL for milk production traits and somatic cell score in Canadian Holstein bulls

The detection and mapping of genetic markers linked to quantitative trait loci (QTL) can be utilized to enhance genetic improvement of livestock populations. With the completion of the bovine genome sequence assembly, single nucleotide polymorphisms (SNP) assays spanning the whole bovine genome and research work on large scale identification, validation and analysis of genotypic variation in cattle has become possible. The objective of the present study was to perform a whole genome scan to identify and map QTL affecting milk production traits and somatic cell scores using linkage disequilibrium (LD) regression and 1536 SNP markers. Three and 18 SNP were found to be associated with only milk yield (MY) at a genome and chromosome wise significance (p < 0.05) level respectively. Among the 21 significant SNP, 16 were in a region reported to have QTL for MY in other dairy cattle populations and while the rest five were new QTL finding. Four SNP out of 21 are significant for the milk production traits (MY, fat yield, protein yield (PY), and milk contents) in the present study. Six and nine SNP were associated with PY at a genome and chromosome wise significant (p < 0.05) level respectively. Three and 17 SNP were found to be associated with FY at a genome and chromosome wise significant (p < 0.05) level. Five and seven SNP were mapped with somatic cell score at a genome and chromosome wise significant (p < 0.05) level respectively. The results of this study have revealed QTL for MY, PY, protein percentage, FY, fat percentage, somatic cell score and persistency of milk in the Canadian dairy cattle population. The chromosome regions identified in this study should be further investigated to potentially identify the causative mutations underlying the QTL.     

Genetics of resistance to mastitis in dairy cattle

Genetic variability of mastitis resistance is well established in dairy cattle. Many studies focused on polygenic variation of the trait, by estimating heritabilities and genetic correlation among phenotypic traits related to mastitis such as somatic cell counts and clinical cases. The role of Major Histocompatibility Complex in the susceptibility or resistance to intrammamary infection is also well documented. Finally, development from molecular genome mapping led to accumulating information of quantitative trait loci (QTL) related to mastitis resistance and better understanding of the genetic determinism of the trait. From economic and genetic analyses, and according to welfare and food safety considerations and to breeders and consumer's concern, there is more and more evidence that mastitis should be included in breeding objective of dairy cattle breeds. Many countries have implemented selection for mastitis resistance based on linear decrease of somatic cell counts. Given biological questioning, potential unfavourable consequences for very low cell counts cows are regularly investigated. Improvement of selection accuracy for mastitis resistance is ongoing and includes: advances in modelling, optimal combination of mastitis related traits and associated predictors, such as udder morphology, definition of global breeding objective including production and functional traits, and inclusion of molecular information that is now available from QTL experiments.     

Effects of DGAT1 variants on milk production traits in German cattle breeds

Various QTL mapping experiments led to the detection of a QTL in the centromeric region of cattle chromosome 14 that had a major effect on the fat content of milk. Recently, the gene encoding diacylglycerol O-acyltransferase (DGAT1) was proposed to be a positional and functional candidate for this trait. This study investigated the effects of a nonconservative lysine to alanine (K232A) substitution in DGAT1, which very likely represents the causal mutation, on milk production traits. Existing granddaughter designs for Fleckvieh and German Holstein, the two major dairy/dual-purpose breeds in Germany, were used to estimate allele frequencies and gene substitution effects for milk, fat, and protein yield, as well as fat and protein content. A restriction fragment length polymorphism assay was applied to diagnose the K232A substitution in DGAT1. Estimates of the allele frequencies for the lysine-encoding variant were based on maternally inherited alleles in sons and amounted to 0.072 for Fleckvieh and 0.548 for German Holstein. Effects of DGAT1 variants on content traits were pronounced; estimates of the gene substitution effect for the lysine-encoding variant were 0.35 and 0.28% for fat content and 0.10 and 0.06% for protein content in Fleckvieh and German Holstein, respectively. Conversely, negative effects of the lysine variant of -242 to -180 kg for Fleckvieh and -260 to -320 kg for German Holstein were revealed for milk yield from first to third lactation, resulting in enhanced fat yield of 7.5 to 14.8 kg in Fleckvieh and 7.6 to 10.7 kg in German Holstein. For protein yield, however, mainly negative effects of -3.6 to 0.2 kg in Fleckvieh and -4.8 to -5.2 kg in German Holstein were observed. Pearson correlations between residuals of milk yield and content traits were decreased when omitting DGAT1 effects in the analysis, thereby indicating that DGAT1 contributes to negative correlations between these traits. Molecular tests allow for the direct selection among variants; however, the benefits of the alternative alleles depend on economic weights given to the different milk production traits in the breeding goal.     

Nutrient Prioritization in Dairy Cows Early Postpartum: Mismatch Between Metabolism and Fertility?

For several decades, researchers worldwide report a decrease in fertility in high-yielding dairy cows, most probably based on conflicting metabolic and reproductive needs. The dairy herd manager's success at improving milk production has been accompanied by a negative trend for the most visible reproductive parameters such as calving intervals, number of days open and number of inseminations needed per pregnancy. In parallel, many research groups studied the metabolic and endocrine factors that influence follicular growth and the developmental competence of oocytes and embryos. In the past, herd managers and reproductive biologists each tried to tackle the same problems with limited consultation. More recently, the situation has improved significantly and theriogenologists, nutritionists and veterinarians now conduct research in multidisciplinary teams. This review paper starts in a general way by discussing nutrient prioritization towards the udder to guarantee milk production and by describing interactions between the somatotropic and gonadotropic axis. It then focuses on the consequences of the negative energy balance on follicular growth and environment, oocyte and embryo quality, not only by summarizing the currently accepted hypotheses but also based on clear scientific evidence at the follicular level. All this, with one question in mind: is there a mismatch between metabolism and fertility and what can the dairy manager learn from research to tackle the problem of reduced fertility?     

Estimation of genetic parameters for heat stress,including dominance gene effects,on milk yield in Thai Holstein dairy cattle

Heat stress in tropical regions is a major cause that strongly negatively affects to milk production in dairy cattle. Genetic selection for dairy heat tolerance is powerful technique to improve genetic performance. Therefore, the current study aimed to estimate genetic parameters and investigate the threshold point of heat stress for milk yield. Data included 52 701 test‐day milk yield records for the first parity from 6247 Thai Holstein dairy cattle, covering the period 1990 to 2007. The random regression test day model with EM‐REML was used to estimate variance components, genetic parameters and milk production loss. A decline in milk production was found when temperature and humidity index (THI) exceeded a threshold of 74, also it was associated with the high percentage of Holstein genetics. All variance component estimates increased with THI. The estimate of heritability of test‐day milk yield was 0.231. Dominance variance as a proportion to additive variance (0.035) indicated that non‐additive effects might not be of concern for milk genetics studies in Thai Holstein cattle. Correlations between genetic and permanent environmental effects, for regular conditions and due to heat stress, were ? 0.223 and ? 0.521, respectively. The heritability and genetic correlations from this study show that simultaneous selection for milk production and heat tolerance is possible.     

Theoretical efficiency of multiple-trait quantitative trait loci-assisted selection

The effectiveness of five selection methods for genetic improvement of net merit comprising trait 1 of low heritability (h² = 0.1) and trait 2 of high heritability (h² = 0.4) was examined: (i) two‐trait quantitative trait loci (QTL)‐assisted selection; (ii) partial QTL‐assisted selection based on trait 1; (iii) partial QTL‐assisted selection based on trait 2; (iv) QTL‐only selection; and (v) conventional selection index without QTL information. These selection methods were compared under 72 scenarios with different combinations of the relative economic weights, the genetic correlations between traits, the ratio of QTL variance to total genetic variance of the trait, and the ratio of genetic variances between traits. The results suggest that the detection of QTL for multiple‐trait QTL‐assisted selection is more important when the index traits are negatively correlated than when they are positively correlated. In contrast to literature reports that single‐trait marker‐assisted selection (MAS) is the most efficient for low heritability traits, this study found that the identified QTL of the low heritability trait contributed negligibly to total response in net merit. This is because multiple‐trait QTL‐assisted selection is designed to maximize total net merit rather than the genetic response of the individual index trait as in the case of single‐trait MAS. Therefore, it is not economical to identify the QTL of the low heritability traits for the improvement of total net merit. The efficient, cost‐effective selection strategy is to identify the QTL of the moderate or high heritability traits of the QTL‐assisted selection index to facilitate total economic returns. Detection of the QTL of the low h² traits for the QTL‐assisted index selection is justified when the low h² traits have high negative genetic correlation with the other index traits and/or when both economic weights and genetic variances of the low h² traits are larger as compared to the other index traits of higher h². This study deals with theoretical efficiency of QTL‐assisted selection, but the same principle applies to SNP‐based genomic selection when the proportion of the genetic variance ‘explained by the identified QTLs’ in this study is replaced by ‘explained by SNPs’.     

奶牛分子育种技术研究进展

分子育种技术是在DNA水平上直接进行选择,通过对随机分布于整个基因组的分子标记的多态性进行比较,更加全面地了解研究对象的多态性,可以获得更大的信息量,选种的准确性也更高,进而提高奶牛生产效率和经济效益。作者从奶牛分子育种技术的角度入手,阐述了数量性状基因的定位、分子标记辅助选择、转基因育种以及全基因组关联分析这几方面的研究难点、存在问题和研究进展,指出分子育种技术在奶牛育种及改良过程中具有非常重要的作用,进一步研究并应用分子育种技术,不仅可以加速我国奶牛核心群的全面建设,而且对培育种公牛也是非常必要的手段。     

The Research Progress of Molecular Technology for Dairy Cattle Breeding

Molecular breeding technology is direct selection at the DNA level by randomly distributed molecular markers throughout the genome polymorphism comparison,it is helpful to carry out more comprehensive study of polymorphism on investigated subjects,meanwhile,more information can be got,and the accuracy of selection will be higher,thereby,the milk production efficiency and economic benefits can be improved.From the perspective of the dairy cattle molecular breeding technology,the author expounds the research challenges,problems and research progress on several aspects,such as localization of QTL,MAS,transgenic breeding and GWAS,points out that molecular breeding techniques have a very important role in dairy cattle breeding and improvement process.Further research and application of molecular breeding technology not only can accelerate the development of dairy cows in the core group of comprehensive construction,but also is very necessary to cultivate a bull.     

Triennial Lactation Symposium: Effects of stress on postpartum reproduction in dairy cows

The objective was to review the effects of production stressors on reproductive performance of dairy cows. It has been well documented that genetic selection for milk yield over the last 50 yr has been associated with reduced fertility. In addition to negative associations between yield and conception rate, there is also an association between milk production and expression of behavioral estrus. Stress caused by production diseases in high-yielding dairy cows also contributes to the problems of poor fertility. Lameness results in reduced intensity of estrus and can contribute to ovulation failure, which is largely due to reduced pre-ovulatory estradiol secretion and failure of the LH surge. Mastitis has been associated with prolonged intervals to dominant follicle selection, and in animals with uterine infection the dominant follicle grows slower and produces less estradiol. In a recent study, we identified that milk yield was associated with an increased incidence of uterine infection, which is known to contribute to reduced fertility and prolonged calving-to-conception intervals. The incidence of uterine disease was 73% in high-yielding, compared with 45% in low-yielding cows. As well as effects at the ovary, various models of stress have also been shown to perturb endocrine secretion in the hypothalamus and anterior pituitary. In conclusion, the adverse effects on fertility associated with genetic selection for yield in dairy cows is, in part, associated with increased incidences of production disease-induced stress but is also associated with high milk yield.     

Identification of genomic regions related to age at first calving and first calving interval in water buffalo using single-step GBLUP

In Brazil, water buffaloes have been used to produce milk for mozzarella cheese production. Consequently, the main selection criterion applied for the buffalo genetic improvement is the estimated mozzarella yield as a function of milk, fat and protein production. However, given the importance of reproductive traits in production systems, this study aimed to use techniques for identifying genomic regions that affect the age at first calving (AFC) and first calving interval (FCI) in buffalo cows and to select candidate genes for the identification of QTL and gene expression studies. The single-step GBLUP method was used for the identification of genomic regions. Windows of 1 Mb containing single-nucleotide polymorphisms were constructed and the 10 windows that explained the greatest proportion of genetic variance were considered candidate regions for each trait. Genes present into the selected windows were identified using the UOA_WB_1 assembly as the reference, and their ontology was defined with the Panther tool. Candidate regions for both traits were identified on BBU 3, 12, 21 and 22; for AFC, candidates were detected on BBU 6, 7, 8, 9 and 15 and for first calving interval on BBU 4, 14 and 19. This study identified regions with great contribution to the additive genetic variance of age at first calving and first calving interval in the population of buffalo cows studied. The ROCK2, PMVK, ADCY2, MAP2K6, BMP10 and GFPT1 genes are main candidates for reproductive traits in water dairy buffaloes, and these results may have future applications in animal breeding programs or in gene expression studies of the species.     

Mapping of quantitative trait loci for lactation persistency traits in German Holstein dairy cattle

A whole genome scan to map quantitative trait loci (QTL) for persistency of milk yield (PMY), persistency of fat yield (PFY), persistency of protein yield (PPY) and persistency of milk energy yield (PEY) was performed in a granddaughter design in the German Holstein dairy cattle population. The analysis included 16 paternal half‐sib families with a total of 872 bulls. The analysis was carried out for the first lactation and for the first three lactations combined using univariate weighted multimarker regression. Controlling the false discovery rate across traits and data sets at a level of 0.15 and treating the four persistency traits as different traits revealed 27 significant QTL. A total of 12 chromosomes showed significant QTL effects on a chromosomewise basis. The DGAT1 effect was highly significant for PPY and protein yield. A haplotype analysis using results of previous studies of the same design revealed a co‐segregation of various persistency QTL and QTL affecting health traits like dystocia and stillbirth and functional traits like non‐return rate 90 and somatic cell score.     

全基因组选择及其在奶牛育种中应用进展

全基因组选择是指基于基因组育种值（GEBV）的选择方法,指通过检测覆盖全基因组的分子标记,利用基因组水平的遗传信息对个体进行遗传评估,以期获得更高的育种值估计准确度。由于可显著缩短世代间隔,全基因组选择作为一项育种新技术在奶牛育种中具有广阔的应用前景,目前已经成为各国的研究热点。不同国家的试验结果表明,奶牛育种中基于GEBV的遗传评估可靠性在20%~67%之间,如果代替常规后裔测定体系,可节省92%的育种成本。本文综述了全基因组选择的基本原理及其在各国奶牛育种中的应用现状和所面临的问题。     

奶山羊产奶性状相关基因的筛选与功能研究进展

奶山羊作为乳用品种的山羊,因其具有奶质优良、产奶性能稳定、乳汁营养丰富等特点,逐渐成为畜牧业发展的一大亮点。经过多年的选种选育,我国现已拥有了多个地方奶山羊品种。随着分子育种技术的发展,对于奶山羊产奶性状相关基因的研究越来越多。对奶山羊产奶性状相关基因的研究进展进行综述,以期为奶山羊优秀基因资源的研究、保护和利用提供理论依据。     

A multiple-phenotype imputation procedure as a method for prediction of cheese-making efficiency in Spanish Assaf sheep

Sheep milk is mainly intended to manufacture a wide variety of high-quality cheeses. The ovine cheese industry would benefit from an improvement, through genetic selection, of traits related to the milk coagulation properties (MCPs) and cheese yield-related traits, broadly denoted as “cheese-making traits.” Considering that routine measurements of these traits needed for genetic selection are expensive and time-consuming, this study aimed to evaluate the accuracy of a cheese-making phenotype imputation method based on the information from official milk control records combined with the pH of the milk. For this study, we analyzed records of milk production traits, milk composition traits, and measurements of cheese-making traits available from a total of 1,145 dairy ewes of the Spanish Assaf sheep breed. Cheese-making traits included five related to the MCPs and two cheese yield-related traits. The milk and cheese-making phenotypes were adjusted for significant effects based on a general linear model. The adjusted phenotypes were used to define a multiple-phenotype imputation procedure for the cheese-making traits based on multivariate normality and Markov chain Monte Carlo sampling. Five of the seven cheese-making traits considered in this study achieved a prediction accuracy of 0.60 computed as the correlation between the adjusted phenotypes and the imputed phenotypes. Particularly the logarithm of curd-firming time since rennet addition (logK₂₀) (0.68), which has been previously suggested as a potential candidate trait to improve the cheese ability in this breed, and the logarithm of the ratio between the rennet clotting time and the curd firmness at 60 min (logRCT/A60) (0.65), which has been defined by other studies as an indicator trait of milk coagulation efficiency. This study represents a first step toward the possible use of the phenotype imputation of cheese-making traits to develop a practical methodology for the dairy sheep industry to impute cheese-making traits only based on the analysis of a milk sample without the need of pedigree information. This information could be also used in future planning of specific breeding programs considering the importance of the cheese-making efficiency in dairy sheep and highlights the potential of phenotype imputation to leverage sample size on expensive, hard-to-measure phenotypes.     

Genomic selection

Genomic selection is a form of marker-assisted selection in which genetic markers covering the whole genome are used so that all quantitative trait loci (QTL) are in linkage disequilibrium with at least one marker. This approach has become feasible thanks to the large number of single nucleotide polymorphisms (SNP) discovered by genome sequencing and new methods to efficiently genotype large number of SNP. Simulation results and limited experimental results suggest that breeding values can be predicted with high accuracy using genetic markers alone but more validation is required especially in samples of the population different from that in which the effect of the markers was estimated. The ideal method to estimate the breeding value from genomic data is to calculate the conditional mean of the breeding value given the genotype of the animal at each QTL. This conditional mean can only be calculated by using a prior distribution of QTL effects so this should be part of the research carried out to implement genomic selection. In practice, this method of estimating breeding values is approximated by using the marker genotypes instead of the QTL genotypes but the ideal method is likely to be approached more closely as more sequence and SNP data is obtained. Implementation of genomic selection is likely to have major implications for genetic evaluation systems and for genetic improvement programmes generally and these are discussed.