Effect of the anatomical site on telomere length and pref-1 gene expression in bovine adipose tissues.

Adipose tissue growth is associated with preadipocyte proliferation and differentiation. Telomere length is a biological marker for cell proliferation. Preadipocyte factor-1 (pref-1) is specifically expressed in preadipocytes and acts as a molecular gatekeeper of adipogenesis. In the present study, we investigated the fat depot-specific differences in telomere length and pref-1 gene expression in various anatomical sites (subcutaneous, intramuscular and visceral) of fattening Wagyu cattle. Visceral adipose tissue expressed higher pref-1 mRNA than did subcutaneous and intramuscular adipose tissues. The telomere length in visceral adipose tissue tended to be longer than that of subcutaneous and intramuscular adipose tissues. The telomere length of adipose tissue was not associated with adipocyte size from three anatomical sites. No significant correlation was found between the pref-1 mRNA level and the subcutaneous adipocyte size. In contrast, the pref-1 mRNA level was negatively correlated with the intramuscular and visceral adipocyte size. These results suggest that anatomical sites of adipose tissue affect the telomere length and expression pattern of the pref-1 gene in a fat depot-specific manner.

Fat depot-specific differences in pref-1 gene expression and adipocyte cellularity between Wagyu and Holstein cattle.

Preadipocyte factor-1 (pref-1) is specifically expressed in preadipocytes and acts as a gatekeeper of adipogenesis by maintaining the preadipocyte state and preventing adipocyte differentiation. We hypothesized that the breed differences of adipogenic capacity in cattle could be explained by the expression level of pref-1. In this experiment, we studied the expression level of the pref-1 gene and adipocyte cellularity in subcutaneous and mesenteric adipose tissues of Japanese Black (Wagyu) and Holstein fattening cattle. In subcutaneous adipose tissue, there were no significant differences in the pref-1 gene expression levels and adipocyte sizes between the breeds. In contrast, the expression level of the pref-1 gene in mesenteric adipose tissue of Holsteins was significantly higher than that of Wagyu. In addition, the size of mesenteric adipocytes in Holsteins was significantly smaller than that of Wagyu. These results indicate that the breed differences of fattening cattle affect the expression pattern of the pref-1 gene and adipocyte cellularity in a fat depot-specific manner.

Prediction of methane emissions from fattening cattle using the methane-to-carbon dioxide ratio.

This study aimed to develop a prediction equation for methane (CH4 ) emissions from fattening cattle based on the CH4 /carbon dioxide (CO2 ) ratio and validate the predictive ability of the developed equation. The prediction equation was developed using the CH4 /CO2 ratio combined with oxygen consumption and respiratory quotient estimations that were theoretically calculated from the relation between gas emissions and energy metabolism. To validate the prediction equation, gas measurements in the headboxes were conducted using eight Japanese Black steers. The predictive ability of the developed equation was compared with that of two previously reported equations. As a result, the developed and reported equations had significant (P < 0.01) linear relationships between the observed and predicted CH4 emissions. Notably, only the developed equation had a significant (P < 0.01) linear relationship between the observed and predicted CH4 emissions when expressed per unit of dry matter intake. The results suggest that the developed prediction equation has a higher predictive ability than previously reported equations, particularly in evaluating the efficiency of CH4 emissions. Although further validation is required, the equation developed in this study can be a valuable tool for on-farm estimations of individual CH4 emissions from fattening cattle.

Metabolomic analysis of plasma and intramuscular adipose tissue between Wagyu and Holstein cattle.

In this experiment, we studied the effects of breed differences in intramuscular adipogenic capacity on the metabolomic profiles of plasma and intramuscular adipose tissue between Wagyu (high intramuscular adipogenic capacity) and Holstein (low intramuscular adipogenic capacity) using capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). We showed that the intramuscular fat content, intramuscular adipocyte size and the expression of adipogenic transcription factors (C/EBPß and C/EBPα) of Wagyu were significantly higher than those of Holstein. Metabolites detected at significantly higher levels in Wagyu plasma were related to the tricarboxylic acid cycle, lipid synthesis, fatty acid metabolism, diabetes, and glucose homeostasis. In contrast, metabolites detected at significantly higher levels in Holstein plasma were related to choline metabolism, the ethanolamine pathway, glutathione homeostasis, nucleic acid metabolism, and amino acid metabolism. Metabolites detected at significantly higher levels in Holstein intramuscular adipose tissue were related to nucleic acid metabolism, amino acid metabolism, amino sugar metabolism, beta oxidation, and the ethanolamine pathway. There were no metabolites significantly higher levels in Wagyu intramuscular adipose tissue. These results indicate candidate biomarkers of breed differences in intramuscular adipogenic capacity between Wagyu and Holstein.

Effects of low-crude protein diets supplemented with rumen-protected lysine and methionine on fattening performance and nitrogen excretion of Holstein steers.

The objective of this experiment was to investigate the effects of low-crude protein (CP) diets supplemented with rumen-protected lysine and methionine on growth performance, nitrogen excretion, and carcass traits in Holstein steers. Steers consumed the following diets: (1) 17.2% CP on a dry-matter basis during the early period (from 7 to 10 months of age) and 14.5% CP during the late period (from 10 to 18 months of age; CON, n = 4, initial body weight [BW] 238 kg), and (2) 14.4% CP during the early period and 11.4% CP during the late period (AA, n = 4, initial BW 243 kg). The AA diet contains rumen-protected lysine and methionine. Except for CP intake, feed intake and body weight gain were not affected by dietary CP content. Total nitrogen excretion per metabolic BW tended to be lower (p < .10) in the early period and significantly lower (p < .05) in the late period with decreasing the feed CP content. Plasma urea nitrogen concentrations were lower in AA than CON. Carcass traits and total free amino acid contents of the longissimus thoracis muscle were not affected by dietary CP content. Adding rumen-protected lysine and methionine to a low-CP diet would reduce nitrogen excretion in fattening Holstein steers without affecting productivity.

Breed differences in macrophage infiltration and senescence state in adipose tissues of Wagyu and Holsteins.

Obesity stimulates the macrophage infiltration and senescence state in adipose tissues of humans and rodents. The adipogenesis capacity of Japanese Black cattle (Wagyu) is higher than that of Holsteins. We hypothesized that breed differences between Wagyu and Holsteins may affect the level of macrophage infiltration and senescence state in adipose tissues. The macrophage infiltration, senescence marker gene expression and activity of senescence-associated ß-galactosidase (SA-ßgal) in visceral and intramuscular adipose tissue of Wagyu were higher than those of Holsteins. In contrast, there were no differences in macrophage infiltration, senescence marker gene expression and activity of SA-ßgal in subcutaneous adipose tissue between the breeds. Expression of p53 gene, the master regulator of macrophage infiltration and senescence state, in visceral and intramuscular adipose tissue of Wagyu was higher than that of Holsteins. In contrast, there was no difference in the expression of p53 gene in subcutaneous adipose tissue between the breeds. These results suggest that breed differences in macrophage infiltration and senescence state in adipose tissues of Wagyu and Holsteins are affected by p53 expression.

Fat depot-specific effects of body fat distribution and adipocyte size on intramuscular fat accumulation in Wagyu cattle.

Ectopic fats have been recognized as a new risk factor for metabolic syndrome. In obese humans, ectopic fat accumulations are affected by body fat distribution. Intramuscular adipose tissue is categorized as one of the ectopic fats. Japanese black cattle (Wagyu) are characterized by the ability to accumulate high amounts of intramuscular adipose tissue. In Japan, the marbling level is indicated by the beef marbling standard number (BMS No.), which reflects the intramuscular fat content of longissimus muscle. We hypothesized that the intramuscular fat accumulation is affected by the body fat distribution in Wagyu cattle. In this study, we showed that the BMS No. was not correlated with the subcutaneous and visceral adipocyte diameter. In contrast, the BMS No. was positively correlated with intramuscular adipocyte diameter. These results indicate that the intramuscular adipocyte diameter of Wagyu is hypertrophied with an increase in the intramuscular fat accumulation. In addition, we showed that the BMS No. was positively correlated with the subcutaneous fat percentage. In contrast, the BMS No. was negatively correlated with the visceral fat percentage. These results indicate that highly marbled Wagyu cattle have a higher percentage of subcutaneous fat and a lower percentage of visceral fat.

Influence of dietary crude protein content on fattening performance and nitrogen excretion of Holstein steers.

The objective was to investigate the influence of crude protein (CP) content in a fattening diet on feed intake, body weight gain, nitrogen excretion, and carcass traits in Holstein steers. Steers (initial body weight 241 ± 26 kg) consumed feed with the following CP content: (a) 17.7% during the early period (from 7 to 10 months of age) and 13.9% during the late period (from 11 to 18 months of age) (HIGH, n = 3), and (b) 16.2% during the early period and 12.2% during the late period (LOW, n = 4). The CP intake was lower in the LOW than the HIGH group. Urinary and total nitrogen excretion in the late period tended to be lower (p < .10) in the LOW than the HIGH group. However, growth performance and carcass traits were not affected by dietary CP content. Free histidine and total amino acid contents in the longissimus thoracis muscle tended to be higher (p < .10) in the HIGH than the LOW group, however, the CP contents were not affected by dietary CP content. The results of this experiment suggest that decreasing dietary CP to 16% (early period) or 12% (late period) of dry matter would reduce nitrogen excretion from Holstein fattening farms without affecting productivity.

Gas Chromatography-Mass Spectrometry-Based Metabolomic Analysis of Wagyu and Holstein Beef.

Japanese Black cattle (Wagyu) beef is characterized by high intramuscular fat content and has a characteristic sweet taste. However, the chemical components for characterizing the sweet taste of Wagyu beef have been unclear. In this experiment, we conducted a metabolomic analysis of the longissimus muscle (sirloin) in Wagyu and Holstein cattle to determine the key components associated with beef taste using gas chromatography-mass spectrometry (GC-MS). Holstein sirloin beef was characterized by the abundance of components such as glutamine, ribose-5-phosphate, uric acid, inosine monophosphate, 5-oxoproline, and glycine. In contrast, Wagyu sirloin beef was characterized by the abundance of sugar components (maltose and xylitol). Dietary fat is known to increase the intensity of sweet taste. These results suggest that the sweet taste of Wagyu beef is due to the synergetic effects of higher sugar components and intramuscular fat.

Fat depot-specific differences of macrophage infiltration and cellular senescence in obese bovine adipose tissues.

Obesity is associated with the chronic inflammation and senescence of adipose tissues. Macrophage is a key mediator of chronic inflammation that infiltrates obese adipose tissue and stimulates metabolic disorders. However, the fat depot-specific differences of macrophage infiltration and senescence, especially the influence on intramuscular adipose tissue, have remained unclear. We investigated the fat depot-specific differences of macrophage infiltration and senescence in obese bovine adipose tissue from three different anatomical sites (subcutaneous, intramuscular and visceral). Macrophage infiltrations and crown-like structures were observed in visceral adipose tissue, although there were few macrophages in subcutaneous and intramuscular adipose tissues. The positive reaction of senescence marker SA-ßgal activity was observed in visceral adipose tissue. In contrast, the activity of SA-ßgal in subcutaneous and intramuscular adipose tissues were low. The expression of p53 gene, the master regulator of cellular senescence, in visceral adipose tissue was higher than that of subcutaneous and intramuscular adipose tissue. At the cellular level, p53 gene expression was negatively correlated with the size of subcutaneous adipocytes. In contrast, p53 gene expressions were positively correlated with the size of intramuscular and visceral adipocytes. These results indicate that anatomical sites of obese adipose tissue affect macrophage infiltration and the senescent state in a fat depot-specific manner.

Plasma 8-isoprostane concentrations and adipogenic and adipokine gene expression patterns in subcutaneous and mesenteric adipose tissues of fattening Wagyu cattle.

We hypothesized that fattening Wagyu cattle fed conventional low-vitamin fattening diets are exposed to oxidative stress. In this experiment, we studied the plasma concentrations of 8-isoprostane and the fat depot-specific effects of the diet-induced adipogenic (C/EBPß, C/EBPδ, C/EBPα and PPARγ2) and adipokine (VEGF, FGF-2, leptin and adiponectin) gene expressions in fattening Wagyu steers. Animals were fed a high-vitamin (α-tocopherol and ß-carotene) diet (HV) or a control diet (CT) during the fattening period (from 10 to 30 months of age). The plasma concentrations of 8-isoprostane, a marker of oxidative stress, were significantly lower in the HV group than in the CT group. In mesenteric adipose tissue, the expressions of the adipogenic and adipokine genes in the HV group were significantly lower than those in the CT group. In contrast, there were no differences in the expression of the adipogenic and adipokine genes in subcutaneous adipose tissue between groups. These results suggest that higher intake of dietary α-tocopherol and ß-carotene affects the expression patterns of adipogenic and adipokine genes in a fat depot-specific manner with the reduction of plasma 8-isoprostane concentrations.

Accurate analysis of taurine, anserine, carnosine and free amino acids in a cattle muscle biopsy sample.

We have established an analysis method for some free amino acids (FAAs), as well as taurine (Tau), anserine (Ans) and carnosine (Car), in a fresh biopsy sample from cattle muscle. A series of model biopsy samples, corresponding to the mixtures of lean meat, fat and connective tissue, was prepared and showed high correlation coefficients between the compound concentration and the 3-methylhistidine (3-MeHis) content derived from hydrolysis of the biopsy sample (r = 0.74-0.95, P < 0.01). Interference from blood contamination could not be neglected, because the concentration of some FAAs in blood was comparable to that in muscle. However, it was possible to control the contamination of Tau, Ans, Car, glutamic acid, glutamine, asparatic acid and alanine to less than 5.0% when the blood contamination was controlled to less than 23%.These results suggest the necessity of measuring 3-MeHis as an index of lean meat and hemoglobin as an index of blood contamination when compounds in muscle biopsy samples are evaluated. We have carried out a series of these analyses using one biopsy sample and reveal differences in Tau, Ans, Car and some FAAs in beef muscle after different feeding regimes.