Bos indicus type of growth hormone receptor gene is retained in Japanese Black cattle

The growth hormone receptor (GHR) gene is responsible for growth and carcass traits, and polymorphisms associated with the variation of meat production are thought to occur in the liver‐specific promoter of the GHR gene in cattle. The aim of this study was to analyse the structure of the liver‐specific promoter of GHR in Japanese Black cattle, as the relationship between GHR polymorphism and meat production is poorly understood in this breed. Typically in European cattle, the LINE‐1 element, a family of retrotransposons, is inserted in the liver‐specific promoter. However, a short GHR promoter without the LINE‐1 sequence was found in the Japanese Black breed as in Bos indicus cattle. The frequency of the short allele was approximately 60%. In addition, 24 of 29 Holstein/Japanese Black crosses carried the short allele from their sire. The present result suggests that the short allele for GHR may be a candidate marker for improving meat production of Japanese Black cattle.     

Relationship of the bovine growth hormone gene to carcass traits in Japanese black cattle.

The bovine growth hormone gene (bGH) possesses three haplotypes, A, B and C, that differ by amino acid mutations at positions 127 and 172 in the fifth exon: (leucine 127, threonine 172), (valine 127, threonine 172) and (valine 127, methionine 172) respectively. The correlation between meat quality or carcass weight and these haplotypes was investigated in Japanese black cattle. Altogether, 940 bGH haplotypes were compared with respect to six carcass traits: carcass weight, longissimus muscle area, rib thickness, subcutaneous fat thickness, beef marbling score and beef colour. The frequency of the B haplotype was higher (0.421) than that of A (0.269) and C (0.311). High carcass weight and low beef marbling were associated with haplotype A (p < 0.05 and p < 0.01 respectively), whereas beef marbling was increased by haplotype C (p < 0.05). Estimated regression coefficient of the A haplotype substitution effect for carcass weight and beef marbling score were 5.55 (13.1% of the phenotypic SD) and -0.31 (17.0%) respectively. That of the C haplotype for beef marbling score was 0.20 (11.0%). The other traits showed no relationship to the haplotypes examined. The results of this investigation suggest that information pertaining to bGH polymorphisms in Japanese black cattle could be used to improve the selection of meat traits.     

Bovine growth hormone gene polymorphism affects stress response in Japanese Black cattle

We investigate the associations between growth hormone (GH) gene polymorphism and behavioral and physiological responses to stressors and learning ability in Japanese Black cattle. Flight distance test was conducted in the first experiment. Steers with haplotype C of GH gene polymorphism avoided human approaches at a significantly greater distance than ones without haplotype C (C: 1.9 ± 0.9, non‐C: 1.0 ± 0.2 m, P < 0.05). An open‐field test was conducted in the second experiment. Behavioral responses did not differ significantly between steers with and without haplotype C. Increases of heart rates to dropping of iron pipes was significantly higher in steers with haplotype C (C:161.7 ± 21.8, non‐C:130.7 ± 31.3%, P < 0.05). Despite basal serum concentrations not being different between steers with and without haplotype C, serum cortisol in blood sampling immediately after severe confinement in a race tended to be higher in steers with haplotype C (P = 0.1). The maze test was conducted as the third experiment. There was no difference in performance in the maze test between steers with and without haplotype C. It is concluded that genetic polymorphism of GH may affect stress responses through GH concentration in steers.     

Haplotype C of growth hormone (GH) gene in Japanese Black cattle: Structure of GH protein and a novel method for detection of the gene

From a series of studies on single nucleotide polymorphisms (SNPs) in the bovine growth hormone (GH) gene of Japanese Black cattle, the type‐C (127^Val and 172^Met) that is specific for this breed has been intensively focused upon because of the economic importance for carcass traits, such as intramuscular oleic acid contents. In the present study, we intended to analyze the 3‐D structure of GH of haplotype C, and developed a novel method to detect the type C gene. Three‐D analysis of the type C protein showed that the amino acid residues (127^Val and 172^Met), which are present in the third and fourth helixes, respectively, and are important for binding with GH receptors, are shifted to deeper positions in the molecule compared with that for type A (127^Leu and 172^Thr), implying the alteration of binding interaction with receptors. A novel, efficient and cost‐effective method (Dot‐blot‐SNP technique) for type C genotyping was successfully established, of which the basal method was a reported genotyping of SNPs for a large number of plants, reducing the cost to 10% or less of direct sequencing.     

IARS mutation causes prenatal death in Japanese Black cattle

Isoleucyl‐tRNA synthetase (IARS) c.235G > C (p.V79L) is a causative mutation for a recessive disease called IARS disorder in Japanese black cattle. The disease is involved in weak calf syndrome and is characterized by low birth weight, weakness and poor suckling. The gestation period is often slightly extended, implying that intrauterine growth is retarded. In a previous analysis of 2597 artificial insemination (AI) procedures, we suggested that the IARS mutation might contribute toward an increase in the incidence of prenatal death. In this study, we extended this analysis to better clarify the association between the IARS mutation and prenatal death. The IARS genotypes of 92 animals resulting from crosses between carrier (G/C) × G/C were 27 normal (G/G), 55 G/C and 10 affected animals (C/C) (expected numbers: 23, 46 and 23, respectively). Compared to the expected numbers, there were significantly fewer affected animals in this population (P < 0.05), suggesting that more than half of the affected embryos died prenatally. When the number of AI procedures examined was increased to 11 580, the frequency of re‐insemination after G/C × G/C insemination was significantly higher at 61‐140 days (P < 0.001). The findings suggested that the homozygous IARS mutation not only causes calf death, but also embryonic or fetal death.     

Identification of leptin gene polymorphisms associated with carcass traits and fatty acid composition in Japanese Black cattle

Previous studies have indicated that some leptin gene polymorphisms were associated with economically important traits in cattle breeds. However, polymorphisms in the leptin gene have not been reported thus far in Japanese Black cattle. Here, we aimed to identify the leptin gene polymorphisms which are associated with carcass traits and fatty acid composition in Japanese Black cattle. We sequenced the full‐length coding sequence of leptin gene for eight Japanese Black cattle. Sequence comparison revealed eight single nucleotide polymorphisms (SNPs). Three of these were predicted to cause amino acid substitutions: Y7F, R25C and A80V. Then, we genotyped these SNPs in two populations (JB1 with 560 animals and JB2 with 450 animals) and investigated the effects on the traits. Y7F in JB1 and A80V in JB2 were excluded from statistical analysis because the minor allele frequencies were low (< 0.1). Association analysis revealed that Y7F had a significant effect on the dressed carcass weight in JB2; R25C had a significant effect on C18:0 and C14:1 in JB1 and JB2, respectively; and A80V had a significant effect on C16:0, C16:1, C18:1, monounsaturated fatty acid and saturated fatty acid in JB1. The results suggested that these SNPs could be used as an effective marker for the improvement of Japanese Black cattle.     

Identification of the core rumen bacterial taxa and their population dynamics during the fattening period in Japanese Black cattle

The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period.     

Nucleotide sequence of myostatin gene and its developmental expression in skeletal muscles of Japanese Black cattle

Myostatin, a member of the transforming growth factor‐β superfamily, is a well known negative regulator of skeletal muscle growth. In the present study, the 6660 bp nucleotide sequence of the myostatin gene in Japanese Black cattle (JBC), including the entire coding region of 1128 bp, was determined. The amino acid sequence deduced from the nucleotide sequence of JBC was well conserved with its sequence of other cattle, although it was found that an Α→G transition at nucleotide position 641 results in the substitution of asparagine by serine at amino acid position 214. In order to examine the expression pattern of the myostatin gene in the skeletal muscles of JBC, its expression in three skeletal muscles, Semitendinosus (ST) muscle, Biceps femoris muscle and Longissimus lumborum muscle, of fetal and calf stages was analyzed by real time polymerase chain reaction. The highest level of the myostatin expression was observed in the fetal stage. In calf stages the highest expression was observed in ST muscle compared with the other two muscles. These results suggest that a higher expression of myostatin gene, especially in the fetal stage and in ST muscle during calf stages, is involved in the arrest in skeletal muscle growth and that its functional domains and genomic structure in JBC are well conserved with those in other mammals.     

Estimation of genetic parameters for growth and feed utilization traits in Japanese Black cattle

Performance test results of 3250 sire candidates were used to estimate the genetic parameters of growth and feed utilization traits in Japanese Black cattle. Growth traits analyzed were six body measurements at the end of the performance test and daily gain (DG) during the test. Feed utilization traits were intakes and conversions of concentrate, roughage, digestible crude protein and total digestible nutrient (TDN). Genetic (co)variance components were estimated by the restricted maximum likelihood procedure using an expectation maximization algorithm under the two‐trait animal model. Heritabilities for growth traits ranged from 0.40 to 0.70 and for feed utilization traits from 0.21 to 0.74. Genetic correlations of DG were positive with feed intake (0.15–0.77) and negative with feed conversions (?0.63 to ?0.30). These relationships indicate that the selection based on DG improves feed efficiency but it simultaneously increases feed intake. Feed conversions showed genetic correlations ranging from ?0.09 to 0.03 with total available energy consumption, TDN intake. Thus the results suggested that feed conversions were not efficient selection criteria to decrease TDN intake and to improve comprehensive feed utilization ability.     

A clinical case of a subepiglottic cyst in a Japanese Black calf

A 36-day-old Japanese Black calf exhibited wheezing associated with dyspnea from birth. Arterial blood gas analysis revealed a low oxygen partial pressure of 51 mmHg, low oxygen saturation of 83%, and high carbon dioxide partial pressure of 58.8 mmHg. Computed tomography, endoscopy, and ultrasonography showed cyst formation under the epiglottis. When the cyst was aspirated under ultrasonic guidance to secure the airway, 30 ml of viscous white turbid content was aspirated. The cyst shrank immediately after aspiration, but the wheezing and respiratory symptoms resumed 7 days after aspiration. Therefore, the cyst was surgically removed from the ventral side of the neck. No cyst remodeling was observed 30 days after surgical removal.     

Genetic parameters for gestation length and the relationship with birth weight and carcass traits in Japanese Black cattle

The objectives of this study were to estimate genetic parameters for gestation length (GL), including estimation of maternal effects, and to investigate the genetic relationships of GL with birth weight and carcass traits in a Japanese Black cattle population. The original data comprised 34 775 records of animals born from October 1999 to August 2003. Two different models were used to analyze the data for GL. The first model (M1) included direct genetic effect of the calf and maternal genetic effect as random effects. The second model (M2) treated GL as a trait of the dam and included direct genetic effects only. M1 was used in bi-variate analysis. The direct and maternal heritabilities for GL estimated from M1 were 0.53 and 0.14, respectively. This result shows that GL is moderately inherited and can be controlled genetically. The direct × maternal genetic correlation for GL was −0.73. Direct genetic correlations of GL with carcass traits were close to zero. However, genetic correlation of maternal GL with carcass weight was moderate (0.25).     

沃金黑牛GPR41基因多态性及其与不同阶段生长性状的相关性分析

[目的]研究旨在探讨GPR41基因多态性对沃金黑牛不同阶段生长性状的影响。[方法]试验共选取63头沃金黑牛(去势公牛)为研究对象,采用Sanger测序技术法检测单核苷酸多态性(SNP)位点,利用Haploview软件进行单倍型分析。利用“牛等家养动物群体遗传分析工具V1.0”评价沃金黑牛群体遗传结构,利用SPSS 19.0软件分析GPR41基因多态性与生长性状的相关性。[结果]结果发现,沃金黑牛GPR41基因外显子2存在2个突变位点,分别为S1:Chr18:46058902 bp处的G/T突变和S2:Chr18:46058908 bp处的C/T突变;S1、S2位点多态性与沃金黑牛不同生长阶段部分生长性状存在显著相关(P<0.05);2个SNPs位点共形成3种有效单倍型组合,单倍型组合H2H2(TT/CC)显著影响沃金黑牛体斜长与背膘厚。[结论]结果表明,GPR41基因SNP位点存在遗传多态性,可用于沃金黑牛(去势公牛)主要生长性状的基因标记参考、稳定与优化。     

Genetic parameters of postnatal mortality and birth weight in Japanese Black calves

Postnatal mortality (PM) of calves is an economically important trait, because a high mortality rate of calves decreases farm income and significantly increases production costs. In spite of the economic importance of PM, genetic studies on PM have been scarce for Japanese Black cattle. The objectives of this study were to estimate the direct and maternal heritability for PM using a Bayesian threshold model with Gibbs sampling in Japanese Black cattle. The original data consisted of 43 723 records of animals born from January 2005 to June 2006. Four binary traits of mortality were considered: D1‐14, D15‐60, D61‐180 and D1‐180 with numbers indicating the period of risk in days after birth. Direct heritabilities for PM ranged from 0.12 to 0.31 and maternal heritabilities for PM were lower than the direct heritabilities and ranged from 0.02 to 0.16. Direct genetic correlations between D1‐14 and D15‐60, between D15‐60 and D61‐180, and between D1‐14 and D61‐180 were 0.501, 0.689 and 0.062, respectively. Direct genetic correlations of birth weight with D1‐14, D15‐60, D61‐180 and D1‐180 were 0.089, 0.482, 0.434 and 0.394, respectively. These results indicated that different genes were responsible for early and late PM and early PM (D1‐14) can be genetically reduced without influencing birth weight.     

Performance of using opposing homozygotes for paternity testing in Japanese Black cattle

Genome-wide single nucleotide polymorphism (SNP) markers in Japanese Black cattle enable genomic prediction and verifying parent–offspring relationships. We assessed the performance of opposing homozygotes (OH) for paternity testing in Japanese Black cattle, using SNP genotype information of 50 sires and 3,420 fattened animals, 1,945 of which were fathered by the 50 genotyped sires. The number of OH was counted for each sire–progeny pair in 28,764 SNPs with minor allele frequencies of ≥0.05 in this population. Across all pairs of animals, the number of OH tended to increase as the pedigree-based coefficient of relationship decreased. With a threshold of 288 (1% of SNPs) for paternity testing, most sire–progeny pairs were detected as true relationships. The frequency of Mendelian inconsistencies was 2.4%, reflecting the high accuracy of pedigree information in Japanese Black cattle population. The results indicate the utility of OH for paternity testing in Japanese Black cattle.     

DNA polymorphism of insulin-like growth factor-binding protein-3 gene and its association with birth weight and body weight in cattle

Insulin‐like growth factor‐binding protein‐3 (IGFBP‐3) is a protein that binds the majority of insulin‐like growth factors in circulation for regulation of its action on growth and metabolism of the animals. Animals belonging to Hariana, Holstein‐Friesian (HF) and their crossbreds (HF × Hariana) were studied using polymerase chain reaction‐restriction fragment length polymorphism and nucleotide sequencing of the IGFBP‐3 gene. A 651‐bp fragment of the IGFBP‐3 gene spanning over a part of exon 2, complete intron 2, exon 3 and a part of intron 3 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in HF and crossbred cattle revealing polymorphism in both the populations. The frequency of AA, AB and BB genotypes was 0.65, 0.32 and 0.03 in crossbreds and 0.29, 0.65 and 0.06 in HF respectively. The allelic frequency of the A and B allele was 0.81 and 0.19 in crossbreds and 0.62 and 0.38 in HF cattle respectively. Only one restriction pattern (AA genotype) was observed in all the animals of Hariana breed of Bos indicus showing the absence of polymorphism. Nucleotide sequencing revealed a C → A mutation in the intron 2 region of the IGFBP‐3 gene as the cause of the polymorphism. Least squares analysis revealed a significant effect (p < 0.05) of genotypes on birth weight and body weight (weight at 12, 18 and 24 months of age) of the animals. Animals of AB genotype showed higher birth weight and body weight than the animals possessing AA genotype.     

Variance components due to direct genetic, maternal genetic and permanent environmental effect for growth and feed-efficiency traits in young male Japanese Black cattle

Variance components and genetic parameters were estimated using data recorded on 740 young male Japanese Black cattle during the period from 1971 to 2003. Traits studied were feed intake (FI), feed‐conversion ratio (FCR), residual feed intake (RFI), average daily gain (ADG), metabolic body weight (MWT) at the mid‐point of the test period and body weight (BWT) at the finish of the test (345 days). Data were analysed using three alternative animal models (direct, direct + maternal environmental, and direct + maternal genetic effects). Comparison of the log likelihood values has shown that the direct genetic effect was significant (p < 0.05) for all traits and that the maternal environmental effects were significant (p < 0.05) for MWT and BWT. The heritability estimates were 0.20 ± 0.12 for FI, 0.14 ± 0.10 for FCR, 0.33 ± 0.14 for RFI, 0.19 ± 0.12 for ADG, 0.30 ± 0.14 for MWT and 0.30 ± 0.13 for BWT. The maternal effects (maternal genetic and maternal environmental) were not important in feed‐efficiency traits. The genetic correlation between RFI and ADG was stronger than the corresponding correlation between FCR and ADG. These results provide evidence that RFI should be included for genetic improvement in feed efficiency in Japanese Black breeding programmes.     

Accuracy of imputation of single nucleotide polymorphism marker genotypes from low‐density panels in Japanese Black cattle

Using target and reference fattened steer populations, the performance of genotype imputation using lower‐density marker panels in Japanese Black cattle was evaluated. Population imputation was performed using BEAGLE software. Genotype information for approximately 40 000 single nucleotide polymorphism (SNP) markers by Illumina BovineSNP50 BeadChip was available, and imputation accuracy was assessed based on the average concordance rates of the genotypes, varying equally spaced SNP densities, and the number of individuals in the reference population. Two additional statistics were also calculated as indicators of imputation performance. The concordance rates tended to be lower for SNPs with greater minor allele frequencies, or those located near the ends of the chromosomes. Longer autosomes yielded greater imputation accuracies than shorter ones. When SNPs were selected based on linkage disequilibrium information, relative imputation accuracy was slightly improved. When 3000 and 10 000 equally spaced SNPs were used, the imputation accuracies were greater than 90% and approximately 97%, respectively. These results indicate that combining genotyping using a lower‐density SNP chip with genotype imputation based on a population of individuals genotyped using a higher‐density SNP chip is a cost‐effective and valid approach for genomic prediction.     

Interval mapping of growth quantitative trait loci in Japanese Black beef cattle using microsatellite DNA markers and half-sib regression analysis

A confirmatory scan for the regions of bovine chromosome 1 segregating the quantitative trait loci (QTL) influencing birthweight, weaning weight, yearling weight, and preweaning and postweaning average daily gains was performed by genotyping half‐sib progeny of four Japanese Black sires using microsatellite DNA markers. Data were analyzed by generating an F‐statistic every 1 cM on a linkage map by the regression of phenotype on the probabilities of inheriting an allele from the sire after adjusting for the fixed effects of sire, sex, parity and season of birth as well as age as a covariate. Permutation tests at chromosome‐wide significance thresholds were carried out over 10 000 iterations. A significant QTL for birthweight at 114 cM was detected in the sire 2 family. This identification of a birthweight QTL in Japanese Black cattle may be useful for the implementation of marker‐assisted selection.     

Allelic frequencies and association with carcass traits of six genes in local subpopulations of Japanese Black cattle

Marker‐assisted selection (MAS) is expected to accelerate the genetic improvement of Japanese Black cattle. However, verification of the effects of the genes for MAS in different subpopulations is required prior to the application of MAS. In this study, we investigated the allelic frequencies and genotypic effects for carcass traits of six genes, which can be used in MAS, in eight local subpopulations. These genes are SCD, FASN and SREBP1, which are associated with the fatty acid composition of meat, and NCAPG, MC1R and F11, which are associated with carcass weight, coat color and blood coagulation abnormality, respectively. The frequencies of desirable alleles of SCD and FASN were relatively high and that of NCAPG was relatively low, and NCAPG was significantly associated with several carcass traits, including carcass weight. The proportions of genotypic variance explained by NCAPG to phenotypic variance were 4.83 for carcass weight. We thus confirmed that NCAPG is a useful marker for selection of carcass traits in these subpopulations. In addition, we found that the desirable alleles of six genes showed no negative effects on carcass traits. Therefore, selection using these genes to improve target traits should not have negative impacts on carcass traits.     

Genetic diversity of growth hormone receptor gene in cattle

Growth hormone receptor (GHR) belongs to a member of the cytokine receptor superfamily. Polymorphism of presence or absence of an approximately 1.2 kbp LINE-1 element is observed in bovine GHR gene. The present study was carried out for estimating the genetic diversity and the origin of the LINE-1 element in 10 European, Southeastern Asian and East Asian cattle breeds or populations. Genotyping of the LINE-1 revealed predominant LINE-1 presence in European breeds (0.917∼0.991), absence in the Bos taurus indicus populations (0.000∼0.017), and intermediate presence in Northeast Asian cattle (0.417∼0.522). From genetic features of LINE families, LINE-1 of GHR could be attributed to the same origin in both European and Asian cattle, and Asian LINE-1 may not be derived from recent introgression. This result suggested that LINE-1 in bovine GHR gene could have arisen in an ancestral population of Bos taurus taurus .