Illumina BovineSNP50 BeadChip Research Articles

Identification of genes associated with susceptibility to Mycobacterium avium ssp. paratuberculosis (Map) tissue infection in Holstein cattle using gene set enrichment analysis-SNP.

Multiple genome-wide association analyses have investigated susceptibility to bovine paratuberculosis, but few loci have been identified across independent cattle populations. A SNP-based gene set enrichment analysis (GSEA-SNP) allows expanded identification of genes with moderate effects on a trait through the enrichment of gene sets instead of identifying only few loci with large effects. Therefore, the objective of this study was to identify genes that were moderately associated with Mycobacterium avium ssp. paratuberculosis (Map) tissue infection using GSEA-SNP in Holstein cattle from the Pacific Northwest (PNW; n = 205) and from the PNW and Northeast (PNW+NE; n = 245) which were previously genotyped with the Illumina BovineSNP50 BeadChip. The GSEA-SNP utilized 4389 gene sets from five databases. For each annotated gene in the UMD3.1 assembly (n = 19,723), the most significant SNP within each gene and its surrounding region (10kb up- and downstream) was selected as a proxy for that gene. Any gene set with a normalized enrichment score > 2.5 was considered enriched. Thirteen gene sets (8 PNW GSEA-SNP; 5 PNW+NE) were enriched in these analyses and all have functions that relate to nuclear factor kappa beta. Nuclear factor kappa beta is critical to gut immune responses, implicated in host immune responses to other mycobacterial diseases, and has established roles in inflammation as well as cancer. Gene sets and genes moderately associated with Map infection could be used in genomic selection to allow producers to select for less susceptible cattle, lower the prevalence of the disease, and reduce economic losses.

Effect of two non-synonymous ecto-5\u2032-nucleotidase variants on the genetic architecture of inosine 5\u2032-monophosphate (IMP) and its degradation products in Japanese Black beef

BackgroundUmami is a Japanese term for the fifth basic taste and is an important sensory property of beef palatability. Inosine 5′-monophosphate (IMP) contributes to umami taste in beef. Thus, the overall change in concentration of IMP and its degradation products can potentially affect the beef palatability. In this study, we investigated the genetic architecture of IMP and its degradation products in Japanese Black beef. First, we performed genome-wide association study (GWAS), candidate gene analysis, and functional analysis to detect the causal variants that affect IMP, inosine, and hypoxanthine. Second, we evaluated the allele frequencies in the different breeds, the contribution of genetic variance, and the effect on other economical traits using the detected variants.ResultsA total of 574 Japanese Black cattle were genotyped using the Illumina BovineSNP50 BeadChip and were then used for GWAS. The results of GWAS showed that the genome-wide significant single nucleotide polymorphisms (SNPs) on BTA9 were detected for IMP, inosine, and hypoxanthine. The ecto-5′-nucleotidase (NT5E) gene, which encodes the enzyme NT5E for the extracellular degradation of IMP to inosine, was located near the significant region on BTA9. The results of candidate gene analysis and functional analysis showed that two non-synonymous SNPs (c.1318C > T and c.1475 T > A) in NT5E affected the amount of IMP and its degradation products in beef by regulating the enzymatic activity of NT5E. The Q haplotype showed a positive effect on IMP and a negative effect on the enzymatic activity of NT5E in IMP degradation. The two SNPs were under perfect linkage disequilibrium in five different breeds, and different haplotype frequencies were seen among breeds. The two SNPs contribute to about half of the total genetic variance in IMP, and the results of genetic relationship between IMP and its degradation products showed that NT5E affected the overall concentration balance of IMP and its degradation products. In addition, the SNPs in NT5E did not have an unfavorable effect on the other economical traits.ConclusionBased on all the above findings taken together, two non-synonymous SNPs in NT5E would be useful for improving IMP and its degradation products by marker-assisted selection in Japanese Black cattle.

Assessing the degree of stratification between closely related Holstein-Friesian populations

Genomic information is an important part of the routine evaluation of dairy cattle and provides the wide availability of animals genotyped using single nucleotide polymorphism (SNP) microarrays. We analyzed 2243 Polish and 2294 German Holstein-Friesian bulls genotyped using the Illumina BovineSNP50 BeadChip. For each bull, estimated breeding values (EBVs) calculated from national routine genetic evaluation were available for production traits and for somatic cell score (SCS). Separately for each population, we estimated SNP haplotypes, pairwise linkage disequilibrium (LD), and SNP effects. The SNP genetic covariance between both populations was estimated using a bivariate mixed model. The average LD was lower in the Polish than in the German population and, with increasing genomic distance, LD decays 1.7 times more rapidly in German than in Polish cattle. The comparison of SNP allele frequencies for base populations estimated separately using Polish and German data revealed a very good agreement. The comparison of genetic effects corresponding to various window lengths defined in bp emerged a systematic pattern: regardless of the length of the compared region, few significant differences were found for production traits, while many were observed for SCS. For each trait, the German population had much higher SNP variances than the Polish population and the genetic covariance estimates were all positive. Depending on traits’ inheritance mode, the additive genetic variation can be stored in many genes following the infinitesimal model (like for SCS) or distributed between genes with high effects and the polygenic “background” (like for production traits). Accounting for those differences has implications on the prospective international genomic evaluation.

Identification of additional loci associated with antibody response to Mycobacterium avium ssp. Paratuberculosis in cattle by GSEA-SNP analysis.

Mycobacterium avium subsp. paratuberculosis: (MAP) causes a contagious chronic infection results in Johne's disease in a wide range of animal species, including cattle. Several genome-wide association studies (GWAS) have been carried out to identify loci putatively associated with MAP susceptibility by testing each marker separately and identifying SNPs that show a significant association with the phenotype, while SNP with modest effects are usually ignored. The objective of this study was to identify modest-effect genes associated with MAP susceptibility using a pathway-based approach. The Illumina BovineSNP50 BeadChip was used to genotype 966 Holstein cows, 483 positive and 483 negative for antibody response to MAP, data were then analyzed using novel SNP-based Gene Set Enrichment Analysis (GSEA-SNP) and validated with Adaptive Rank Truncated Product methodology. An allele-based test was carried out to estimate the statistical association for each marker with the phenotype, subsequently SNPs were mapped to the closest genes, considering for each gene the single variant with the highest value within a window of 50kb, then pathway-statistics were tested using the GSEA-SNP method. The GO biological process "embryogenesis and morphogenesis" was most highly associated with antibody response to MAP. Within this pathway, five genes code for proteins which play a role in the immune defense relevant to response to bacterial infection. The immune response genes identified would not have been considered using a standard GWAS, thus demonstrating that the pathway approach can extend the interpretation of genome-wide association analyses and identify additional candidate genes for target traits.

Genome-wide association and pathway-based analysis using latent variables related to milk protein composition and cheesemaking traits in dairy cattle

The aim of this study was to perform genome-wide associations (GWAS) and gene-set enrichment analyses with protein composition and cheesemaking-related latent variables (factors; F) in a cohort of 1,011 Italian Brown Swiss cows. Factor analysis was applied to identify latent structures of 26 phenotypes related to bovine milk quantity and quality, protein fractions [αS1-, αS2-, β-, and κ-casein (CN), β-lactoglobulin, and α-lactalbumin (α-LA)], coagulation and curd firming at time t (CFt) measures, and cheese properties [cheese yield (%CY) and nutrients recovery in the curd] of individual cows. Ten orthogonal F were extracted, explaining 74% of the original variability. Factor 1%CY underlined the %CY characteristics, F2CFt was related to the CFt process parameters, F3Yield was considered as descriptor of milk and solids yield, whereas F4Cheese N underscored the presence of nitrogenous compounds (N) into the cheese. Four more F were related to the milk caseins (F5αS1-β-CN, F7β-κ-CN, F8αS2-CN, and F9αS1-CN-Ph) and 1 F was linked to the whey protein (F10α-LA); 1 F underlined the udder health status (F6Udder health). All cows were genotyped with the Illumina BovineSNP50 Bead Chip v.2 (Illumina Inc., San Diego, CA). Single marker regression GWAS were fitted. Gene-set enrichment analysis was run on GWAS results, using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway databases, to reveal ontologies or pathways associated with the F. All F but F3Yield showed significance in GWAS. Signals in 10 Bos taurus autosomes (BTA) were detected. High peaks on BTA6 (∼87 Mbp) were found for F6β-κ-CN, F5αS1-β-CN, and at the tail of BTA11 (∼104 Mbp) for F4Cheese N. Gene-set enrichment analyses showed significant results (false discovery rate at 5%) for F8αS2-CN, F1%CY, F4Cheese N, and F10α-LA. For F8αS2-CN, 33 Gene Ontology terms and 3 Kyoto Encyclopedia of Genes and Genomes categories were enriched, including terms related to ion transport and homeostasis, neuron function or part, and GnRH signaling pathway. Our results support the feasibility of factor analysis as a dimension reduction technique in genomic studies and evidenced a potential key role of αS2-CN in milk quality and composition.

Improving genetic evaluation using a multitrait single-step genomic model for ability to resume cycling after calving, measured by activity tags in Holstein cows

The objective of this study was to evaluate the improvement of the accuracy of estimated breeding values for ability to recycle after calving by using information of genomic markers and phenotypic information of correlated traits. The traits in this study were the interval from calving to first insemination (CFI), based on artificial insemination data, and the interval from calving to first high activity (CFHA), recorded from activity tags, which could better measure ability to recycle after caving. The phenotypic data set included 1,472,313 records from 820,218 cows for CFI, and 36,504 records from 25,733 cows for CFHA. The genomic information was available for 3,159 progeny-tested sires, which were genotyped using Illumina Bovine SNP50 BeadChip (Illumina, San Diego, CA). Heritability estimates were 0.06 for the interval from calving to first insemination and 0.14 for the interval from calving to first high activity, and the genetic correlation between both traits was strong (0.87). Breeding values were obtained using 4 models: conventional single-trait BLUP; conventional multitrait BLUP with pedigree-based relationship matrix; single-trait single-step genomic BLUP; and multitrait single-step genomic BLUP model with joint relationship matrix combining pedigree and genomic information. The results showed that reliabilities of estimated breeding values (EBV) from single-step genomic BLUP models were about 40% higher than those from conventional BLUP models for both traits. Furthermore, using a multitrait model doubled the reliability of breeding values for CFHA, whereas no gain was observed for CFI. The best model was the multitrait single-step genomic BLUP, which resulted in a reliability of EBV 0.19 for CFHA and 0.14 for CFI. The results indicate that even though a relatively small number of records for CFHA were available, with genomic information and using multitrait model, the reliability of EBV for CFHA is acceptable. Thus, it is feasible to include CFHA in Nordic Holstein breeding evaluations to improve fertility performance.

Genomic prediction and genome-wide association analysis of female longevity in a composite beef cattle breed.

Longevity is a highly important trait to the efficiency of beef cattle production. The objective of this study was to evaluate the genomic prediction of longevity and identify genomic regions associated with this trait. The data used in this study consisted of 547 Composite Gene Combination cows (1/2 Red Angus, 1/4 Charolais, 1/4 Tarentaise) born from 2002 to 2011 genotyped with Illumina BovineSNP50 BeadChip. Three models were used to assess genomic prediction: Bayes A, Bayes B and GBLUP using a genomic relationship matrix. To identify genomic regions associated with longevity 2 approaches were adopted: single marker genome wide association and Bayesian approach using GenSel software. The genomic prediction accuracy was low 0.28, 0.25, and 0.22 for Bayes A, Bayes B and GBLUP, respectively. The single-marker genome wide association study (GWAS)identified 5 loci with -value less than 0.05 after false discovery correction: UA-IFASA-7571 on chromosome 19 (58.03 Mb), ARS-BFGL-BAC-15059 on BTA 1 (28.8 Mb), ARS-BFGL-NGS-104159 on BTA3 (29.4 Mb), ARS-BFGL-NGS-32882 on BTA9 (104.07 Mb) and ARS-BFGL-NGS-32883 on BTA25 (33.77 Mb). The Bayesian GWAS yielded 4 genomic regions overlapping with the single marker GWAS results. The region with the highest percentage of genomic variance (3.73%) was detected on chromosome 19. Both GWAS approaches adopted in this study showed evidence for association with various chromosomal locations.

A genome-wide association study reveals a locus for bilateral iridal hypopigmentation in Holstein Friesian cattle

BackgroundEye pigmentation abnormalities in cattle are often related to albinism, Chediak-Higashi or Tietz like syndrome. However, mutations only affecting pigmentation of coat color and eye have also been described. Herein 18 Holstein Friesian cattle affected by bicolored and hypopigmented irises have been investigated.ResultsAffected animals did not reveal any ophthalmological or neurological abnormalities besides the specific iris color differences. Coat color of affected cattle did not differ from controls. Histological examination revealed a reduction of melanin pigment in the iridal anterior border layer and stroma in cases as cause of iris hypopigmentation. To analyze the genetics of the iris pigmentation differences, a genome-wide association study was performed using Illumina BovineSNP50 BeadChip genotypes of the 18 cases and 172 randomly chosen control animals. A significant association on bovine chromosome 8 (BTA8) was identified at position 60,990,733 with a -log10(p) = 9.17. Analysis of genotypic and allelic dependences between cases of iridal hypopigmentation and an additional set of 316 randomly selected Holstein Friesian cattle controls showed that allele A at position 60,990,733 on BTA8 (P = 4.0e–08, odds ratio = 6.3, 95% confidence interval 3.02–13.17) significantly increased the chance of iridal hypopigmentation.ConclusionsThe clinical appearance of the iridal hypopigmentation differed from previously reported cases of pigmentation abnormalities in syndromes like Chediak-Higashi or Tietz and seems to be mainly of cosmetic character. Iridal hypopigmentation is caused by a reduced content of melanin pigment in the anterior border layer and iridal stroma. A single genomic position on BTA8 was detected to be significantly associated with iridal hypopigmentation in examined cattle. To our knowledge this is the first report about this phenotype in Holstein Friesian cattle.

Genomic variation across cervid species in respect to the estimation of red deer diversity

AbstractThe aim of this study was to assess the genetic variations and relationships across evolutionary related cervid species in order to estimate the genetic diversity of the Red deer population that inhabits the forest area in the south-western part of Slovakia. The study was based on the application of cross-species SNP genotyping. The genomic data were obtained from a total of 86 individuals representing six genera (Axis,Dama,Cervus,Alces,Rangifer, andOdocoileus) using Illumina BovineSNP50 BeadChip. From 38.85% of successfully genotyped loci up to 1,532 SNPs showed polymorphism and were informative for subsequent analyses of the diversity and interspecific genetic relationships. Generally, a good level of observed heterozygosity was found across all species. The value of FIS(0.23±0.13) signalised the increase of a homozygous proportion within them. The application of molecular variance analysis to the hierarchical population structure showed that most of the variation was conserved within separate species (96%). The performed diversity analysis of Slovak Red deer population and comparative analysis of their phylogenic relationships among subspecies from genusCervusdid not identify a remarkable loss of genetic variability. Also, were not identified any degree of admixture that could be due to the historical background of deer farming in Slovakia or reintroduction and hybridisation by other species from genusCervus(C. canadensis, andC. nippon) which are the major risk of loss of autochthonous Red deer populations in many areas of Central Europe. The analysis of individual’s ancestry showed consistent results with patterns of evaluated group differentiations which means low migration rates among all species.

Identification of loci associated with susceptibility to subspecies () tissue infection in cattle.

Johne's disease is a contagious bacterial infection of cattle caused by ssp. (). A previous genome-wide association analysis (GWAA) in Holstein cattle identified QTL on BTA3 and BTA9 that were highly associated (P < 5 × 10) and on BTA1, BTA16, and BTA21 that were moderately associated (P < 5 × 10) with Map tissue infection. The objectives of this study were to validate previous GWAA results in Jersey cattle ( = 57), Holstein cattle from the Pacific Northwest (PNW, = 205) and a combined Holstein population from the PNW and the Northeast (PNW + NE, = 423), and also identify new loci associated with tissue infection. DNA was genotyped using the Illumina BovineSNP50 BeadChip, and the PNW + NE data was also imputed to whole genome sequence level using Run4 of the 1000 Bull Genomes project with Beagle v 4.1 and FImpute. Cases were ileocecal node positive and controls were negative for by quantitative PCR (qPCR). Individuals were removed for SNP call rate < 90%, and SNP were removed for genotype call rate < 90% or minor allele frequency < 1%. For the Jersey, PNW, and PNW + NE, GWAA were conducted using an allelic dosage model. For the PNW and the PNW + NE, an additional efficient mixed-model association eXpedited (EMMAX) analysis was performed using additive, dominance and recessive models. Seven QTL on BTA22 were identified in the Jersey population with the most significant ( = 4.45 × 10) located at 21.7 megabases (Mb). Six QTL were associated in the PNW and the PNW + NE analyses, including a QTL previously identified on BTA16 in the NE population. The most significant locus for the PNW was located on BTA21 at 61 Mb ( = 8.61 × 10) while the most significant locus for the PNW + NE was on BTA12 at 90 Mb ( = 2.33 × 10). No additional QTL were identified with the imputed GWAA. Putative positional candidate genes were identified within 50 kb 5' and 3' of each QTL. Two positional candidate genes were identified in Jersey cattle, 1 identified in the PNW and 8 in the PNW + NE populations. Many identified positional candidate genes are involved in signal transduction, have immunological functions, or have putative functional relevance in entry into host cells. This study supported 2 previously identified SNP within a QTL on BTA16 and identified 16 new QTL, including 2 found in the PNW and the PNW+NE, associated with tissue infection.

A COMPREHENSIVE CHARACTERIZATION OF GUELMOISE, A NATIVE CATTLE BREED FROM EASTERN ALGERIA

The present study was a global and a first characterization (Typology, biometric, biochemical and contribution on the genetic characterization) of the Guelmoise local cattle breed. A structured questionnaire surveys were carried out in 42 farms spread over the four wilayas of the north-eastern region of Algeria in order to characterize the local cattle population of Guelmoise and their mode of rearing. These surveys focused on the characteristics of the farms (socio-economic condition of the farmer, mode of breeding, reproduction and also to take samples of milk and blood). A total of 164 adult animals were selected for morphological description and body measurements (148 females and 16 males). The livestock system has an extensive nature and men are the main responsible for these farms. The height at withers (HaW), body length (BL), Heart Girth (HG), Spiral Tower (ST) and estimated weight (W) in Guelmoise cattle were 115 ± 0.07, 118 ± 0.08, 154 ± 0.09, 187 ± 0.11 cm and 258 , 12 ± 43.12 kg respectively in females and 171 ± 0.16, 176 ± 0.18, 187 ± 0.08, 225 ± 0.08 cm and 462 ± 59.67 kg respectively in males. Multivariate statistical analyses have allowed identifying four different classes were defined as follows morphometric measurements, weight and age in females as in males. Estimated age at first calving was 39.40 months while the calving interval varies between 14 to17 months and the reproductive life of the cows (13,7 years).The average daily production of milk 5,55 ± 1,67 liters in spring with a duration of lactation varies between 6 to 7 month. The physico-chemical characteristics of milk were focused on the Fat content (FC), Protein content (PC), Lactose (Lact), Dry extract (DE), Defatted dry extract (DDE) and freezing point depression (FPD) were respectively 2.85 ± 0.89%, 3.60 ± 0.39%, 4.82 ± 0.13%, 11.84 ± 0.90%, 9.13 ± 0.41% and 0.56 ± 0,01°C. Thegenotyping of 24 individuals of the Guelmoise population using the Illumina BovineSNP50 BeadChip, we found that the observed heterozygosity was globally equivalent to 0.30. The inbreeding measured by Fis was slightly positive (~0.04) indicating the absence of a phenomenon of selection. The data contribute to the feasibility of a conservation and selection programme for this breed and the results are useful for the implementation of a conservation strategy that should aim to conserve animals. The Guelmoise cattle can be used as an alternative genetic resource for production improvement programs.

Genome-wide mapping of 10 calving and fertility traits in Holstein dairy cattle with special regard to chromosome 18

Over the last decades, a dramatic decrease in reproductive performance has been observed in Holstein cattle and fertility problems have become the most common reason for a cow to leave the herd. The premature removal of animals with high breeding values results in both economic and breeding losses. For efficient future Holstein breeding, the identification of loci associated with low fertility is of major interest and thus constitutes the aim of this study. To reach this aim, a genome-wide combined linkage disequilibrium and linkage analysis (cLDLA) was conducted using data on the following 10 calving and fertility traits in the form of estimated breeding values: days from first service to conception of heifers and cows, nonreturn rate on d 56 of heifers and cows, days from calving to first insemination, days open, paternal and maternal calving ease, paternal and maternal stillbirth. The animal data set contained 2,527 daughter-proven Holstein bulls from Germany that were genotyped with Illumina's BovineSNP50 BeadChip (Illumina Inc., San Diego, CA). For the cLDLA, 41,635 sliding windows of 40 adjacent single nucleotide polymorphisms (SNP) were used. At each window midpoint, a variance component analysis was executed using ASReml. The underlying mixed linear model included random quantitative trait locus (QTL) and polygenic effects. We identified 50 genome-wide significant QTL. The most significant peak was detected for direct calving ease at 59,179,424 bp on chromosome 18 (BTA18). Next, a mixed-linear model association (MLMA) analysis was conducted. A comparison of the cLDLA and MLMA results with special regard to BTA18 showed that the genome-wide most significant SNP from the MLMA was associated with the same trait and located on the same chromosome at 57,589,121 bp (i.e., about 1.5 Mb apart from the cLDLA peak). The results of 5 different cLDLA and 2 MLMA models, which included the fixed effects of either SNP or haplotypes, suggested that the cLDLA method outperformed the MLMA in accuracy and precision. The haplotype-based cLDLA method allowed for a more precise mapping and the definition of ancestral and derived QTL alleles, both of which are essential for the detection of underlying quantitative trait nucleotides.

Genetic correlation and genome-wide association study (GWAS) of the length of productive life, days open, and 305-days milk yield in crossbred Holstein dairy cattle.

In this study, we estimated the genetic parameters and identified the putative quantitative trait loci (QTL) associated with the length of productive life (LPL), days open (DO), and 305-day milk yield for the first lactation (FM305) of crossbred Holstein dairy cattle. Data comprising 4,739 records collected between 1986 and 2004 were used to estimate the variance-covariance components using the multiple-trait animal linear mixed models based on the average information restricted maximum likelihood (AI-REML) algorithm. Thirty-six animals were genotyped using the Illumina BovineSNP50 Bead Chip [>50,000 single nucleotide polymorphisms (SNPs)] to identify the putative QTL in a genome-wide association study. The heritability of the production trait FM305 was 0.25 and that of the functional traits, LPL and DO, was low (0.10 and 0.06, respectively). The genetic correlation estimates demonstrated favorable negative correlations between LPL and DO (-0.02). However, we observed a favorable positive correlation between FM305 and LPL (0.43) and an unfavorable positive correlation between FM305 and DO (0.1). The GWAS results indicated that 23 QTLs on bovine chromosomes 1, 4, 5, 8, 15, 26, and X were associated with the traits of interest, and the putative QTL regions were identified within seven genes (SYT1, DOCK11, KLHL13, IL13RA1, PRKG1, GNA14, and LRRC4C). In conclusion, the heritability estimates of the LPL and DO were low. Therefore, the approach of multiple-trait selection indexes should be applied, and the QTL identified here should be considered for use in marker-assisted selection in the future.

Genome-wide association study for cheese yield and curd nutrient recovery in dairy cows

Cheese production and consumption are increasing in many countries worldwide. As a result, interest has increased in strategies for genetic selection of individuals for technological traits of milk related to cheese yield (CY) in dairy cattle breeding. However, little is known about the genetic background of a cow's ability to produce cheese. Recently, a relatively large panel (1,264 cows) of different measures of individual cow CY and milk nutrient and energy recoveries in the cheese (REC) became available. Genetic analyses showed considerable variation for CY and for aptitude to retain high proportions of fat, protein, and water in the coagulum. For the dairy industry, these characteristics are of major economic importance. Nevertheless, use of this knowledge in dairy breeding is hampered by high costs, intense labor requirement, and lack of appropriate technology. However, in the era of genomics, new possibilities are available for animal breeding and genetic improvement. For example, identification of genomic regions involved in cow CY might provide potential for marker-assisted selection. The objective of this study was to perform genome-wide association studies on different CY and REC measures. Milk and DNA samples from 1,152 Italian Brown Swiss cows were used. Three CY traits expressing the weight (wt) of fresh curd (%CYCURD), curd solids (%CYSOLIDS), and curd moisture (%CYWATER) as a percentage of weight of milk processed, and 4 REC (RECFAT, RECPROTEIN, RECSOLIDS, and RECENERGY, calculated as the % ratio between the nutrient in curd and the corresponding nutrient in processed milk) were analyzed. Animals were genotyped with the Illumina BovineSNP50 Bead Chip v.2. Single marker regressions were fitted using the GenABEL R package (genome-wide association using mixed model and regression-genomic control). In total, 103 significant associations (88 single nucleotide polymorphisms) were identified in 10 chromosomes (2, 6, 9, 11, 12, 14, 18, 19, 27, 28). For RECFAT and RECPROTEIN, high significance peaks were identified in Bos taurus autosome (BTA) 6 and BTA11, respectively. Marker ARS-BFGL-NGS-104610 (∼104.3 Mbp) was highly associated with RECPROTEIN and Hapmap52348-rs29024684 (∼87.4 Mbp), closely located to the casein genes on BTA6, with RECFAT. Genomic regions identified may enhance marker-assisted selection in bovine cheese breeding beyond the use of protein (casein) and fat contents, whereas new knowledge will help to unravel the genomic background of a cow's ability for cheese production.

Genome-wide linkage disequilibrium and past effective population size in three Korean cattle breeds.

The routine collection and use of genomic data are useful for effectively managing breeding programs for endangered populations. Linkage disequilibrium (LD) using high-density DNA markers has been widely used to determine population structures and predict the genomic regions that are associated with economic traits in beef cattle. The extent of LD also provides information about historical events, including past effective population size (Ne ), and it allows inferences on the genetic diversity of breeds. The objective of this study was to estimate the LD and Ne in three Korean cattle breeds that are genetically similar but have different coat colors (Brown, Brindle and Jeju Black Hanwoo). Brindle and Jeju Black are endangered breeds with small populations, whereas Brown Hanwoo is the main breeding population in Korea. DNA samples from these cattle breeds were genotyped using the Illumina BovineSNP50 Bead Chip. We examined 13 cattle breeds, including European taurines, African taurines and indicines, and hybrids to compare their LD values. Brown Hanwoo consistently had the lowest mean LD compared to Jeju Black, Brindle and the other 13 cattle breeds (0.13, 0.19, 0.21 and 0.15-0.22 respectively). The high LD values of Brindle and Jeju Black contributed to small Ne values (53 and 60 respectively), which were distinct from that of Brown Hanwoo (531) for 11 generations ago. The differences in LD and Ne for each breed reflect the breeding strategy applied. The Ne for these endangered cattle breeds remain low; thus, effort is needed to bring them back to a sustainable tract.

A genome-assisted MOET design for inbreeding prevention in the endangered Murnau-Werdenfelser cattle

Murnau-Werdenfelser cattle are an indigenous breed from Upper Bavaria in Germany. The population is endangered and is supported by the Bavarian government. The aim of the study was to select optimal unrelated mating partners for the Multiple Ovulation and Embryo Transfer (MOET) procedure and to present a new genomic technology to breeders. Samples from 143 animals of the Murnau-Werdenfelser breed were genotyped with the Illumina BovineSNP50 BeadChip using standard procedures. Genome-wide realized relationships in the form of an identity by descent (IBD) matrix among 143 individuals were estimated based on 50,864 SNP markers. In order to facilitate the discussion with breeders and to provide a basis for mating decisions, the individual DPSdistance matrix was presented in two-dimensional (2D) space. Based on the positions of both parents in the 2D plot, the potential positions of all offspring were derived. For each included donor dam, a separate 2D plot was prepared. Breeders accepted this graphical method well and used it smoothly for selection of the optimal MOET mates.

Estimation of local genetic ancestry in an admixed cattle population applying different methods

In current study, we estimated local genetic ancestry for Swiss-Fleckvieh dairy cattle population using two alternative approaches implemented in LAMP and MULTIMIX software. Swiss-Fleckvieh is a composite descending from Simmental and Red Holstein Friesian. Illumina BovineSNP50 Beadchip data for 485 animals were used. Genetic ancestries averaged across admixed animals indicated a high correlation of individual admixture at global levels (0.99), yet the correlation between genetic ancestries averaged across 7914 windows (5 SNPs each) was 0.55. The highest correlations of window-wise estimates were along chromosomes 2, 5, 12, 15, 21, 27 and 29 (≥0.80) and the lowest was −0.71 on chromosome 24. Based on LAMP results, a region on chromosome 13 (46.3–46.7 Mb) and three adjacent regions on chromosome 18 (18.8–19.2, 20.1–25.7 and 23.9–25.7 Mb) passed the threshold based on deviation from normal distribution of local ancestries assuming 1000 independent segments. Regarding MULTIMIX, another region on chromosome 22 (9.3–10.3 Mb) was significant. Although estimates from both programs are comparable at the global levels, there was considerable difference at the local levels.

Genome-wide association study for semen traits of the bulls in Chinese Holstein.

A genome-wide association study (GWAS) was performed to identify markers and candidate genes for five semen traits in the Holstein bull population in China. The analyzed datasetconsisted of records from 692 bulls from eight bull stations; each bull was genotyped using the Illumina BovineSNP50 BeadChip. Association tests between each trait and the 41188 informative high-quality SNPs were achieved with gapit software. In total, 19 suggestive significant SNPs, partly located within the reported QTL regions or within or close to the reported candidate genes, associated with five semen traits were detected. By combining our GWAS results with the biological functions of these genes, eight novel promising candidate genes, including ETNK1, PDE3A, PDGFRB, CSF1R, WT1, DSCAML1, SOD1 and RUNX2, were identified that potentially relate to semen traits. Our findings may provide a basis for further research on the genetic mechanism of semen traits and marker-assisted selection of such traits in Holstein bulls.

Identification of a haplotype associated with cholesterol deficiency and increased juvenile mortality in Holstein cattle.

Over the last decades, several genetic disorders have been discovered in cattle. However, the genetic background of disorders in calves is less reported. Recently, German cattle farmers reported on calves from specific matings with chronic diarrhea and retarded growth of unknown etiology. Affected calves did not respond to any medical treatment and died within the first months of life. These calves were underdeveloped in weight and showed progressive and severe emaciation despite of normal feed intake. Hallmark findings of the blood biochemical analysis were pronounced hypocholesterolemia and deficiency of fat-soluble vitamins. Results of the clinical and blood biochemical examination had striking similarities with findings reported in human hypobetalipoproteinemia. Postmortem examination revealed near-complete atrophy of the body fat reserves including the spinal canal and bone marrow. To identify the causal region, we performed a genome-wide association study with 9 affected and 21,077 control animals genotyped with the Illumina BovineSNP50 BeadChip (Illumina Inc., San Diego, CA), revealing a strong association signal on BTA 11. Subsequent autozygosity mapping identified a disease-associated haplotype encompassing 1.01 Mb. The segment of extended homozygosity contains 6 transcripts, among them the gene APOB, which is causal for cholesterol disorders in humans. However, results from multi-sample variant calling of 1 affected and 47 unaffected animals did not detect any putative causal mutation. The disease-associated haplotype has an important adverse effect on calf mortality in the homozygous state when comparing survival rates of risk matings vs. non-risk matings. Blood cholesterol values of animals are significantly associated with the carrier status indicating a codominant inheritance. The frequency of the haplotype in the current Holstein population was estimated to be 4.2%. This study describes the identification and phenotypic manifestation of a new Holstein haplotype characterized by pronounced hypocholesterolemia, chronic emaciation, growth retardation, and increased mortality in young cattle, denominated as cholesterol deficiency haplotype. Our genomic investigations and phenotypic examinations provide additional evidence for a mutation within the APOB gene causing cholesterol deficiency in Holstein cattle.

Whole-genome scan to detect quantitative trait loci associated with milk protein composition in 3 French dairy cattle breeds.

In the context of the PhénoFinLait project, a genome-wide analysis was performed to detect quantitative trait loci (QTL) that affect milk protein composition estimated using mid-infrared spectrometry in the Montbéliarde (MO), Normande (NO), and Holstein (HO) French dairy cattle breeds. The 6 main milk proteins (α-lactalbumin, β-lactoglobulin, and αS1-, αS2-, β-, and κ-caseins) expressed as grams per 100g of milk (% of milk) or as grams per 100g of protein (% of protein) were estimated in 848,068 test-day milk samples from 156,660 cows. Genotyping was performed for 2,773 MO, 2,673 NO, and 2,208 HO cows using the Illumina BovineSNP50 BeadChip (Illumina Inc., San Diego, CA). Individual test-day records were adjusted for environmental effects and then averaged per cow to define the phenotypes analyzed. Quantitative trait loci detection was performed within each breed using a linkage disequilibrium and linkage analysis approach. A total of 39 genomic regions distributed on 20 of the 29 Bos taurus autosomes (BTA) were significantly associated with milk protein composition at a genome-wide level of significance in at least 1 of the 3 breeds. The 9 most significant QTL were located on BTA2 (133 Mbp), BTA6 (38, 47, and 87 Mbp), BTA11 (103 Mbp), BTA14 (1.8 Mbp), BTA20 (32 and 58 Mbp), and BTA29 (8 Mbp). The BTA6 (87 Mbp), BTA11, and BTA20 (58 Mbp) QTL were found in all 3 breeds, and they had highly significant effects on κ-casein, β-lactoglobulin, and α-lactalbumin, expressed as a percentage of protein, respectively. Each of these QTL explained between 13% (BTA14) and 51% (BTA11) of the genetic variance of the trait. Many other QTL regions were also identified in at least one breed. They were located on 14 additional chromosomes (1, 3, 4, 5, 7, 15, 17, 19, 21, 22, 24, 25, 26, and 27), and they explained 2 to 8% of the genetic variance of 1 or more protein composition traits. Concordance analyses, performed between QTL status and sequence-derived polymorphisms from 13 bulls, revealed previously known causal polymorphisms in LGB (BTA11) and GHR (BTA20 at 32 Mbp) and excluded some other previously described mutations. These results constitute a first step in identifying causal mutations and using routinely collected mid-infrared predictions in future genomic selection programs to improve bovine milk protein composition.