Association of a single nucleotide polymorphism in ribosomal protein L27a gene with marbling in Japanese Black beef cattle

Marbling, defined by the amount and distribution of intramuscular fat, is an economically important trait of beef cattle in Japan. The c2‐11#2 expressed sequence tag (EST) has been previously shown to possess expression difference in musculus longissimus muscle between low‐marbled and high‐marbled steer groups, and to be located within genomic region of a quantitative trait locus for marbling. Thus, the ribosomal protein L27a (RPL27A) gene containing the c2‐11#2 EST sequence was considered as a positional candidate for the gene responsible for marbling. In the present study, a single nucleotide polymorphism (SNP) in the promoter region of the RPL27A, referred to as g.3109537C>T, was detected between the 2 steer groups. The SNP was associated with the predicted breeding value for beef marbling standard number by the analyses using Japanese Black beef cattle population. The effect of genotypes of the SNP on the predicted breeding value for subcutaneous fat thickness was not statistically significant. These findings suggest that the RPL27A SNP may be useful for effective marker‐assisted selection to increase the levels of marbling in Japanese Black beef cattle.     

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.     

Micronutrients affecting adipogenesis in beef cattle

Beef marbling is an important trait of meat quality and beef marbling influences the tenderness and flavor of beef, which contributes directly to the value of beef especially in the Japanese market. The lipid content of beef depends on the strain, sex, diet and fattening period of the animal. Japanese Black cattle (Wagyu) are well‐known for their ability to produce marbling beef and this is a popular strain in Japan. The development of beef marbling was closely associated with an increase in the number of adipocytes, that is, adipocyte differentiation in the skeletal muscle. This review article describes our experiment and related reports on micronutrients, especially vitamins and minerals, affecting adipogenesis in beef cattle. We pursue the possibility that manipulating the level of dietary micronutrients may become a new technique to promote beef marbling.     

Novel SNP in 5' flanking region of EDG1 associated with marbling in Japanese Black beef cattle

Marbling, defined by the amount and distribution of intramuscular fat, is an economically important trait of beef cattle in Japan. The endothelial differentiation , sphingolipid G-protein-coupled receptor , 1 ( EDG1 ) gene has been considered as a positional functional candidate for the gene responsible for marbling. We have recently reported that 2 single nucleotide polymorphisms (SNPs), c.-312A > G in the 5' untranslated region (UTR) and c.*446G > A in the 3' UTR in EDG1 were associated with marbling in Japanese Black beef cattle, but this was not functional and a causal mutation for marbling. In the present study, we detected 2 novel SNPs, referred to as g.1475435G > A and g.1471620G > T , in the 5' flanking region of the EDG1 between low-marbled and high-marbled steer groups, which were previously shown to have EDG1 expression differences in musculus longissimus muscle. The g.1475435G > A SNP seemed not to segregate in Japanese Black beef cattle. The g.1471620G > T SNP was associated with the predicted breeding value for beef marbling standard number by the analyses using Japanese Black beef cattle population. Based on these findings, we hypothesized that the g.1471620G > T SNP might have an impact on EDG1 expression and also marbling.     

利用荷斯坦阉牛生产大理石纹牛肉的研究

本文主要从荷斯坦牛肌内脂肪沉积相关基因的研究入手,主要阐述荷斯坦牛大理石纹的影响因素,去势对荷斯坦牛肉品质(大理石纹和牛肉嫩度)影响及营养(营养浓度和维生素A成分)调控对荷斯坦阉牛肌内脂肪沉积的影响展开综述,以期为利用荷斯坦阉牛生产大理石纹牛肉提供参考。     

Comprehensive evaluation of growth performance and meat characteristics of a fattening system combining grazing with feeding rice whole‐crop silage in Japanese Black steers

We investigated the effect of a system for fattening steers combining grazing with feeding rice whole‐crop silage (rWCS) on growth performance, meat characteristics, and the expression of genes involved in skeletal muscle growth. Steers were randomly assigned to grazing or concentrate‐fed groups (CT). The grazing group (GZ) was fed rWCS after grazing until 16 months of age. The final body weight was the same in the two groups, but the dressed weight was lower in the GZ than in the CT. The beef color standard was higher in the GZ than in the CT. Although beef marbling did not differ between the two groups, there was less intramuscular fat and subcutaneous fat in the GZ than in the CT. The α‐tocopherol and β‐carotene contents in the muscle were higher in the GZ than in the CT. The GZ showed a lower daily gain (DG) during the grazing period, which may have resulted from decelerating skeletal muscle growth caused by the increased expression of genes encoding myostatin and atrogin‐1. However, the DG and feed efficiency of the GZ increased after grazing. The two groups exhibited a similar level of beef productivity.     

Validation of endogenous reference genes for expression profiling of RAW264.7 cells infected with Mycobacterium avium subsp. paratuberculosis by quantitative PCR

Reference genes are frequently used to normalize between different biological samples the levels of mRNA measured using quantitative PCR (qPCR). The expression level of many commonly used reference genes has been shown to vary between tissues or cells, or following exposure to various treatments including infection with microbes. The selection of an appropriate reference gene for an individual experiment is therefore a crucial step in the process of accurately determining changes in gene expression. For this purpose, we analyzed the expression of nine commonly used reference genes in a murine macrophage cell line, RAW264.7, for their potential use in the analysis of differential gene expression by quantitative polymerase chain reaction (qPCR) following experimental infection with Mycobacterium avium subsp. paratuberculosis. Only one of nine putative reference genes tested, casc3a, was found to be suitable, and combinations of two or more reference genes were disadvantageous. Based on data from the study, we recommend an approach for selection of reference genes, conducting assays with technical replicates in duplicate rather than triplicate, determining decision-limit quality control criteria for technical replicates and assessing the significance of gene expression fold differences using DeltaDeltaC(t) based on knowledge of the variation in the reference gene.     

Estimation of beef marbling in the Longissimus muscle with computer image analysis of ultrasonic pictures of the Iliocostalis muscle area

This study investigated an objective method for estimating beef marbling using ultrasonic images of the Iliocostalis muscle and the Lomgissimus muscle area sections. Thirty‐one Japanese Black cattle steers were used in this study. The end of the left side shoulder blade bone was scanned using an ultrasonic device. Ultrasonic images were captured of the Longissimus muscle area and that around the Iliocostalis muscle area. Twenty items were measured in the two images using computer image analysis software. The level of beef marbling was measured according to the Beef Marbling Standard (BMS) for carcass grading, and the percentage of ether‐extractable fat content in the Longissimus muscle (EE). The difference in the gray level between the Iliocostalis muscle and intermuscular fat (X10) was used to estimate the BMS and the EE, which were highly correlated (r² = 67.72% and 61.30%). An equation was developed using four parameters from the two ultrasonic images, which could estimate the BMS (r² = 85.88%). This equation could also estimate the EE (r² = 68.98%). The equations used to estimate beef marbling were based on one to four parameters that included X10. Thus, ultrasonic images of the Iliocostalis muscle area section are important for estimating beef marbling.     

Relationships among glycolytic potential,dark cutting (dark,firm, and dry) beef,and cooked beef palatability

One hundred beef carcasses were selected at three packing plants and were used to determine the relationship between glycolytic potential (GP) and dark, firm, and dry (DFD) beef and to determine the effects of DFD status and GP on cooked beef palatability. Eight individual muscles were excised from one hindquarter of each carcass at d 7 postmortem: longissimus lumborum, psoas major, gluteus medius, tensor fasciae latae, rectus femoris, semimembranosus, biceps femoris, and semitendinosus. Ultimate pH, colorimeter readings, and Warner-Bratzler shear force were determined for all eight muscles at d 7 postmortem. A nine-member trained sensory panel evaluated cooked longissimus lumborum, gluteus medius, and semimembranosus steaks. Traits determined solely for the longissimus lumborum were GP (2 x [glycogen + glucose + glucose-6-phosphate] + lactate) and ether-extractable fat. A curvilinear relationship existed between GP and ultimate pH within the longissimus muscle. There appeared to be a GP threshold at approximately 100 micromol/g, below which lower GP was associated with higher ultimate pH and above which GP had no effect on ultimate pH. The greatest pH and muscle color differences between normal and DFD carcasses were observed in the longissimus lumborum, gluteus medius, semimembranosus, and semitendinosus muscles. Cooked longissimus from DFD carcasses had higher shear force values (46% greater) and more shear force variation (2.3 times greater variation) than those from normal carcasses. Dark cutting carcasses also had higher shear force values for gluteus medius (33% greater) and semimembranosus (36% greater) than normal carcasses. Sensory panel tenderness of longissimus, gluteus medius, and semimembranosus was lower for DFD carcasses than for normal carcasses. Longissimus and gluteus medius flavor desirability scores were lower for DFD than for normal carcasses. Steaks from DFD carcasses had more off-flavor comments than steaks from normal carcasses, specifically more "peanutty," "sour," and "bitter" flavors. The DFD effect of higher shear force values was approximately five times greater (+3.11 kg vs +0.63 kg) for carcasses with "slight" marbling scores than for carcasses with "small" marbling scores. In general, higher GP was associated with increased tenderness, even among normal carcasses. In conclusion, low GP was associated with DFD beef and resulted in substantially less-palatable cooked steaks.     

Exploring novel candidate genes from the Mouse Genome Informatics database: Potential implications for avian migration research

To search for genes associated with migratory phenotypes in songbirds, we selected candidate genes through annotations from the Mouse Genome Informatics database and assembled an extensive candidate‐gene library. Then, we implemented a next‐generation sequencing approach to obtain DNA sequences from the Painted Bunting genome. We focused on those sequences that were conserved across avian species and that aligned with candidate genes in our mouse library. We genotyped short sequence repeats from the following candidate genes: ADRA1d, ANKRD17, CISH and MYH7. We studied the possible correlations between allelic variations occurring in these novel candidate migration genes and avian migratory phenotypes available from the published literature. We found that allele variation at MYH7 correlated with a calculated index of speed of migration (km/day) across 11 species of songbirds. We highlight the potential of the Mouse Genome Informatics database in providing new candidate genes that might play a crucial role in regulating migration in birds and possibly in other taxa. Our research effort shows the benefits and limitations of working with extensive genomic datasets and offers a snapshot of the challenges related to cross‐species validation in behavioral and molecular ecology studies.     

维生素A对肉牛脂肪沉积及其调控的研究进展

维生素A是肉牛维持正常生理功能、生化代谢及生长发育所必需的一类脂溶性维生素，机体自身不能合成，必须由饲粮供给。饲粮中维生素A不仅影响肉牛视觉和骨骼发育，也对肉牛脂肪沉积和肌肉大理石花纹形成等发挥着重要调控作用。在生产中，肉牛处在不同生理阶段对维生素A的需要量也不同，胎牛和犊牛阶段补充维生素A可增强肌内脂肪细胞发育和脂肪细胞增生，促进肌内脂肪沉积；育肥期限饲维生素A可提高肉牛的肌内脂肪沉积和大理石花纹等级。大理石花纹与牛肉的嫩度和风味密切相关，是衡量牛肉品质的重要指标。维生素A在肉牛体内通过其活性代谢产物视黄醇、视黄醛和视黄酸促进脂肪的形成，并在成脂定型、成脂分化和脂质蓄积的每个阶段都发挥重要作用。脂肪沉积过程受转录因子过氧化物酶体增殖物激活受体（peroxisome proliferator-activated receptors，PPARs）、CCAAT增强子结合蛋白（CCAAT-enhancer binding proteins，C/EBPs）和Janus激酶-信号转导及转录激活因子（JAK-STAT）信号转导通路等调控。表观遗传修饰的DNA甲基化和去甲基化也可通过调控脂肪形成过程中相关基因的表达参与脂肪细胞的分化和脂肪组织的生长发育，从而影响肉牛的脂肪沉积。作者主要介绍了脂肪组织不同阶段的形成过程和维生素A的生物学功能；重点阐述了肉牛在不同生理阶段补充和限饲维生素A，通过转录因子的表达、表观遗传修饰等途径来影响成脂相关信号通路调控脂肪沉积的机理，以期为促进维生素A改善肉牛品质和高档牛肉生产提供参考。     

Influence of rice whole‐crop silage diet on growth performance,carcass and meat characteristics and muscle‐related gene expression in Japanese Black steers

The present study investigated the influence of a diet largely comprising rice whole‐crop silage (rWCS) on growth performance, carcass and meat characteristics, and expression of genes involved in muscle growth of Japanese Black steers. Steers were randomly separated into rWCS‐fed (rWCS ad libitum and restricted feeding of concentrate) and concentrate‐fed groups. Total digestible nutrient intake and daily gain (DG) decreased in rWCS‐fed steers in comparison with concentrate‐fed steers, whereas dressed carcass weight and final body weight did not significantly differ between the groups. Decreases in drip loss in the muscle of rWCS‐fed steers may be caused by α‐tocopherol and β‐carotene in muscle. Feeding large amounts of rWCS to steers may maintain quantitative productivity of beef steers equally to a concentrate‐based diet, and improve the qualitative productivity. Results of gene expression suggest that activation of skeletal muscle growth in rWCS‐fed steers may occur at the late fattening period owing to a decrease in myostatin and increase in myosin heavy chain gene expression. Preadipocyte factor‐1 and myostatin genes may be strongly involved in the control of lipid accumulation. This rearing system would allow beef production to switch to rWCS‐based diets from concentrate‐based diets.     

On-line prediction of yield grade,longissimus muscle area,preliminary yield grade,adjusted preliminary yield grade,and marbling score using the MARC beef carcass image analysis system

The present experiment was conducted to evaluate the ability of the U.S. Meat Animal Research Center's beef carcass image analysis system to predict calculated yield grade, longissimus muscle area, preliminary yield grade, adjusted preliminary yield grade, and marbling score under commercial beef processing conditions. In two commercial beef-processing facilities, image analysis was conducted on 800 carcasses on the beef-grading chain immediately after the conventional USDA beef quality and yield grades were applied. Carcasses were blocked by plant and observed calculated yield grade. The carcasses were then separated, with 400 carcasses assigned to a calibration data set that was used to develop regression equations, and the remaining 400 carcasses assigned to a prediction data set used to validate the regression equations. Prediction equations, which included image analysis variables and hot carcass weight, accounted for 90, 88, 90, 88, and 76% of the variation in calculated yield grade, longissimus muscle area, preliminary yield grade, adjusted preliminary yield grade, and marbling score, respectively, in the prediction data set. In comparison, the official USDA yield grade as applied by online graders accounted for 73% of the variation in calculated yield grade. The technology described herein could be used by the beef industry to more accurately determine beef yield grades; however, this system does not provide an accurate enough prediction of marbling score to be used without USDA grader interaction for USDA quality grading.     

Flavor characterization of top-blade, top-sirloin, and tenderloin steaks as affected by pH, maturity, and marbling

Little information is available in the literature on the interrelationships and interactions among pH, aging time, marbling, and maturity on the flavor profile of some beef muscles commonly used for steaks. To investigate these effects on beef flavor, the infraspinatus (top-blade steak) from the chuck clod, the gluteus medius (top-sirloin steak) from the sirloin, and the psoas major (tenderloin steak) from the loin were obtained from A- (n = 80) and B-maturity (n = 60) carcasses with either Slight (n = 68) or Small (n = 72) marbling, and with either normal (< or = 5.7; n = 80) or high (> or = 6.0; n = 60) pH. Muscles were selected from two commercial processing plants at six different sampling times to evaluate factors that affect the flavor profile of cooked beef steaks. Muscles were vacuum-aged for 7, 14, 21, or 35 d, and a highly trained, flavor-profile sensory panel evaluated charbroiled steaks from these muscles. Numerous statistical interactions (P < 0.05) were detected for flavor attributes of the different muscles. In general, muscles from high pH (dark cutting) carcasses had less typical beef flavor identity and less brown-roasted flavor than those from carcasses with normal pH. Aging longer than 21 d generally decreased beef flavor identity. Top-blade steaks generally had less intense beef flavor identity and more intense bloody/serumy flavor than did top-sirloin and tenderloin steaks. Tenderloin and top-sirloin steaks of normal pH generally had the most brown-roasted flavor, especially when aged 21 d or less. Small degree of marbling generally resulted in a more rancid flavor compared with Slight marbling, but marbling had no other appreciable effects on the flavor profile. Aging steaks for 35 d increased (P < 0.05) the metallic flavor compared with aging for only 7 or 14 d. Top-sirloin steaks had a more intense (P < 0.05) sour flavor than did top-blade steaks, and steaks from carcasses with a high pH were more rancid (P < 0.05) than steaks from carcasses with normal pH. Vacuum-aging top-blade, top-sirloin, and tenderloin steaks to 21 or 35 d postmortem generally increased metallic and rancid flavors and increased sour flavor in top-sirloin steaks that were high in pH.     

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.