Metabolites play essential roles in biological systems, but detailed identities and significance of the seminal plasma metabolome related to bull fertility are still unknown. The objectives of this study were to determine the comprehensive metabolome of seminal plasma from Holstein bulls and to ascertain the potential of metabolites as biomarkers of bull fertility. The seminal plasma metabolome from 16 Holstein bulls with two fertility rates were determined by gas chromatography-mass spectrometry (GC-MS). Multivariate and univariate analyses of the data were performed, and the pathways associated with the seminal plasma metabolome were identified using bioinformatics approaches. Sixty-three metabolites were identified in the seminal plasma of all bulls. Fructose was the most abundant metabolite in the seminal fluid, followed for citric acid, lactic acid, urea and phosphoric acid. Androstenedione, 4-ketoglucose, D-xylofuranose, 2-oxoglutaric acid and erythronic acid represented the least predominant metabolites. Partial-Least Squares Discriminant Analysis (PLSDA) revealed a distinct separation between high and low fertility bulls. The metabolites with the greatest Variable Importance in Projection score (VIP > 2) were 2-oxoglutaric acid and fructose. Heat-map analysis, based on VIP score, and univariate analysis indicated that 2-oxoglutaric acid was less (P = 0.02); whereas fructose was greater (P = 0.02) in high fertility than in low fertility bulls. The current study is the first to describe the metabolome of bull seminal plasma using GC-MS and presented metabolites such as 2-oxoglutaric acid and fructose as potential biomarkers of bull fertility.
Available data for cholesterol content of beef, pork, poultry, and processed meat products were reported. Although the cholesterol concentration in meat and poultry can be influenced by various factors, effects of animal species, muscle fiber type, and muscle fat content are focused on in this review. Oxidative red muscles tend to have greater total lipid and cholesterol contents, although differences in the same types of muscles or cuts have been reported. Moreover, contradictory results among various studies suggest that unless there are pronounced changes in muscle structure and composition, cholesterol content is unlikely to be affected. Second, multiple issues in cholesterol analysis, including sample preparation, detection, and quantification, were evaluated. Cholesterol content of meat and poultry has been determined mostly by colorimetry and chromatography, although the latter has become predominant because of technological advances and method performance. Direct saponification has been the preferred method for hydrolyzing samples because of cost‐ and time‐effectiveness. The extraction solvent varies, but toluene seems to provide sufficient recovery in a single extraction, although the possible formation of an emulsion associated with using toluene requires experience in postsaponification manipulation. The most commonly used internal standard is 5α‐cholestane, although its behavior is not identical to that of cholesterol. Cholesterol can be analyzed routinely by gas chromatography (GC)‐flame ionization detector without derivatization; however, other methods, especially high‐performance liquid chromatography (HPLC) coupled with different detectors, can also be used. For research purposes, HPLC‐ultraviolet/Visible/photodiode array detector with nondestructiveness is preferred, especially when cholesterol must be separated from other coexisting compounds such as tocopherols. More advanced methods, such as GC/HPLC‐isotope dilution/mass spectrometry, are primarily used for quality control purposes.
The objective of this study was to compare the fatty acid (FA) composition of intramuscular fat from the LM of 3 divergent breeds of cattle: Angus (AN, n = 9), Brahman (BR, n = 7), and Romosinuano (RM, n = 11). Cattle were blocked by breed and finished 129 d before slaughter in one year and 157 d in the next year. Longissimus muscle samples were collected from each carcass between the 10th and 13th ribs, trimmed of external fat, frozen in liquid nitrogen, homogenized, and used for fat extraction, using a modified Folch procedure. Extracted fat was analyzed for FA by using a GLC system with an HP-88 capillary column. Fatty acid composition was expressed using both a normalized percentage (%) and gravimetric calculation (mg/g of fresh muscle tissue) in relation to degree of saturation, which was determined using a saturation index (ratio of total SFA to total unsaturated FA). Crude fat determination revealed that LM from AN purebred cattle had the greatest amount of intramuscular fat (7.08%; P = 0.001). Although intramuscular fat of LM from RM contained a reduced percentage of total SFA (P = 0.002) compared with AN, it had the greatest percentage of total PUFA (P < 0.001 and P = 0.020). The percentages of total MUFA were similar among the 3 breeds (P = 0.675). The gravimetric calculation, a measure of actual FA concentration, showed significantly greater concentrations of SFA (26.67 mg/g), MUFA (26.50 mg/g), and PUFA (2.37 mg/g) in LM from AN cattle, as compared with LM from BR and RM cattle (P < 0.001). Interestingly, BR purebreds had the least PUFA concentration (1.49 mg/g; P
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