In-depth exploration of the high and normal pH beef proteome: First insights emphasizing the dynamic protein changes in Longissimus thoracis muscle from pasture-finished Nellore bulls over different postmortem times.

This study aimed to evaluate for the first time the temporal dynamic changes in early postmortem proteome of normal and high ultimate pH (pHu) beef samples from the same cattle using a shotgun proteomics approach. Ten selected carcasses classified as normal (pHu < 5.8; n = 5) or high (pHu ≥ 6.2; n = 5) pHu beef from pasture-finished Nellore (Bos taurus indicus) bulls were sampled from Longissimus thoracis muscle at 30 min, 9 h and 44 h postmortem for proteome comparison. The temporal proteomics profiling quantified 863 proteins, from which 251 were differentially abundant (DAPs) between high and normal pHu at 30 min (n = 33), 9 h (n = 181) and 44 h (n = 37). Among the myriad interconnected pathways regulating pH decline during postmortem metabolism, this study revealed the pivotal role of energy metabolism, cellular response to stress, oxidoreductase activity and muscle system process pathways throughout the early postmortem. Twenty-three proteins overlap among postmortem times and may be suggested as candidate biomarkers to the dark-cutting condition development. The study further evidenced for the first time the central role of ribosomal proteins and histones in the first minutes after animal bleeding. Moreover, this study revealed the disparity in the mechanisms underpinning the development of dark-cutting beef condition among postmortem times, emphasizing multiple dynamic changes in the muscle proteome. Therefore, this study revealed important insights regarding the temporal dynamic changes that occur in early postmortem of high and normal muscle pHu beef, proposing specific pathways to determine the biological mechanisms behind dark-cutting determination.

Feeding strategies impact animal growth and beef color and tenderness.

The impact of growth rate (GR) and finishing regime (FR) on growth and meat quality traits of Angus x Nellore crossbred steers, harvested at a constant body weight (530 ± 20 kg) or time on feed (140 days), was evaluated. Treatments were: 1) feedlot, high GR; 2) feedlot, low GR; 3) pasture, high GR and 4) pasture, low GR. Live body composition, carcass and meat quality traits were evaluated. High GR had greater impact on muscle and fat deposition in feedlot-finished, but not in pasture-finished animals. Feedlot animals had higher Longissimus muscle area, backfat thickness, meat luminosity and tenderness when compared to pasture groups. Moreover, pasture- and feedlot-finished animals with similar GR did not differ in the chromatic attributes of non-aged meat, regardless of endpoint. Thus, GR appeared to be the main factor driving beef chromatic parameters, while FR had a major impact on achromatic attributes and tenderness of meat.

Proteome basis for the biological variations in color and tenderness of longissimus thoracis muscle from beef cattle differing in growth rate and feeding regime.

The proteome basis for the biological variations in color and tenderness of longissimus thoracis muscle from ½ Angus (Bos taurus taurus) × ½ Nellore (Bos taurus indicus) crossbred steers was evaluated in a completely randomized experimental design consisting of four treatments (n = 9 per treatment): 1) feedlot finished, high growth rate (FH); 2) feedlot finished, low growth rate (FL); 3) pasture finished, high growth rate (PH); and 4) pasture finished, low growth rate (PL). The following comparisons were made to evaluate the effects of finishing systems and growth rates on muscle proteome: 1) FH × PL; 2) FL × PH; 3) FH × FL; and 4) PH × PL. Sixteen protein spots were differentially abundant among these comparisons (P ≤ 0.05), which were distinguished in two major clusters, energy metabolism- and muscle structure-related proteins that impacted glycolysis, carbon metabolism, amino acid biosynthesis and muscle contraction pathways (FDR ≤ 0.05). For FH × PL comparison, triosephosphate isomerase (TPI), phosphoglucomutase-1 (PGM1) and phosphoglycerate kinase 1 (PGK1) were overabundant in FH beef whereas troponin T (TNNT3), α-actin (ACTA1) and myosin regulatory light chain 2 (MYLPF) were overabundant in PL beef. For the FL × PH comparison, PGM1, phosphoglycerate mutase 2 (PGAM2) and annexin 2 (ANXA2) were overabundant in PH beef. For the FH × FL comparison, AMP deaminase (AMPD1) and serum albumin (ALB) were overabundant in FH beef whereas glycogen phosphorylase (PYGM) was overabundant in FL beef. For the PH × PL comparison, myoglobin (MB) was overabundant in PH beef whereas PYGM and MYLPF were overabundant in PL beef. In non-aged beef, L* was positively correlated with PGM1 (r = 0.54) while tenderness was negatively correlated with PGAM2 (r = -0.74) and ANXA2 (r = -0.60). In 7-d aged beef, color attributes (L*, a* and b*) were positively correlated with PGM1 (r = 0.67, 0.64 and 0.64, respectively) while tenderness was negatively correlated with TNNT3 (r = -0.57), PGK1 (r = -0.52) and MYLPF (r = -0.66). Therefore, finishing systems and growth rate affected the muscle proteome profile, which was related to beef color and tenderness. Additionally, these results suggest potential biomarkers for beef color (PGM1 and PGAM2) and tenderness (ANXA2, MYLPF, PGK1 and TNNT3).

Exploratory lipidome and metabolome profiling contributes to understanding differences in high and normal ultimate pH beef.

The aim of this work was to compare the lipidome and metabolome profiling in the Longissimus thoracis muscle early and late postmortem from high and normal ultimate pH (pHu) beef. Lipid profiling discriminated between high and normal pHu beef based on fatty acid metabolism and mitochondrial beta-oxidation of long chain saturated fatty acids at 30 min postmortem, and phospholipid biosynthesis at 44 h postmortem. Metabolite profiling also discriminated between high and normal pHu beef, mainly through glutathione, purine, arginine and proline, and glycine, serine and threonine metabolisms at 30 min postmortem, and glycolysis, TCA cycle, glutathione, tyrosine, and pyruvate metabolisms at 44 h postmortem. Lipid and metabolite profiles showed reduced glycolysis and increased use of alternative energy metabolic processes that were central to differentiating high and normal pHu beef. Phospholipid biosynthesis modification suggested high pHu beef experienced greater oxidative stress.

Broiler genetics influences proteome profiles of normal and woody breast muscle.

Wooden or woody breast (WB) is a myopathy of the pectoralis major in fast-growing broilers that influences the quality of breast meat and causes an economic loss in the poultry industry. The objective of this study was to evaluate growth and proteome differences between 5 genetic strains of broilers that yield WB and normal breast (NB) meat. Eight-week-old broilers were evaluated for the WB myopathy and divided into NB and WB groups. Differential expression of proteins was analyzed using 2-dimensional gel electrophoresis and LC-MS/MS to elucidate the mechanism behind the breast myopathy because of the genetic backgrounds of the birds. The percentages of birds with WB were 61.3, 68.8, 46.9, 45.2, and 87.5% for strains 1-5, respectively, indicating variability in WB myopathy among broiler strains. Birds from strains 1, 3, and 5 in the WB group were heavier than those in the NB group (P < 0.05). Woody breast meat from all strains were heavier than NB meat (P < 0.05). Within WB, strain 5 had a greater breast yield than strains 1, 3, and 4 (P < 0.0001). Woody breast from strains 2, 3, 4, and 5 had a greater breast yield than NB (P < 0.05). Six proteins were more abundant in NB of strain 5 than those of strains 2, 3, and 4, and these proteins were related to muscle growth, regeneration, contraction, apoptosis, and oxidative stress. Within WB, 14 proteins were differentially expressed between strain 5 and other strains, suggesting high protein synthesis, weak structural integrity, intense contraction, and oxidative stress in strain 5 birds. The differences between WB from strain 3 and strains 1, 2, and 4 were mainly glycolytic. In conclusion, protein profiles of broiler breast differed because of both broiler genetics and the presence of WB myopathy.

Early Postmortem Proteome Changes in Normal and Woody Broiler Breast Muscles.

Early postmortem changes in the whole muscle proteome from normal broiler (NB) and woody broiler (WB) breasts at 0 min, 15 min, 4 h, and 24 h after slaughter were analyzed using two-dimensional gel electrophoresis (2DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Elongation factor 2, EH domain-containing protein 2, phosphoglycerate mutase 1 (PGAM1), and T-complex protein 1 subunit gamma were differentially abundant in both NB and WB muscles during the early postmortem storage. Twenty additional proteins were differentially abundant among four postmortem time points in either NB or WB muscles. In the postmortem WB, changes in protein degradation were observed, including the degradation of desmin fragments, ovotransferrin chain A, and troponin I chain I. Additionally, a few glycolytic proteins in the WB might have undergone post-translational modification, including enolase, phosphoglucomutase-1, PGAM1, and pyruvate kinase. These changes in protein biomarkers highlight the impact of WB myopathy on postmortem proteome changes and increase our understanding of the relationship between WB conditions, postmortem biochemistry, and meat quality.

Metabolite profile and consumer sensory acceptability of meat from lean Nellore and Angus × Nellore crossbreed cattle fed soybean oil.

Thirty each Nellore (NEL) and crossbred Angus × Nellore (AxN) were used to evaluate the effect of feeding soybean oil (SBO) and breed on meat sensory acceptability and its relation to muscle metabolite profiles. Cattle were fed for 133 d on two different diets: 1) basal feedlot diet (CON) and 2) CON diet with 3.5% added SBO. No interactions between diet and genetic group were detected for any traits measured. Meat from animals fed SBO diet had lower overall liking, flavor, tenderness and juiciness scores compared to meat from animals fed CON diet. The four most important compounds differing between animals fed CON and SBO diets were betaine, glycerol, fumarate, and carnosine, suggesting that metabolic pathways such as glycerolipid metabolism; glycine, serine and threonine metabolism; glutamine and glutamate metabolism; valine, leucine and isoleucine biosynthesis; and alanine, aspartate and glutamate metabolism were affected by diets. Nellore beef had a higher overall liking and meat flavor scores than AxN beef. The four most important compounds differing between breeds were glycine, glucose, alanine, and carnosine, which may indicate that metabolic pathways such as glutathione metabolism; primary bile acid biosynthesis; alanine, aspartate and glutamate metabolism; and valine, leucine and isoleucine biosynthesis were affected by genetic groups. Meat carnosine, inosine monophosphate, glutamate, betaine, glycerol and creatinine levels were correlated with sensory acceptability scores. Meat metabolite profiles and sensory acceptability were differentially impacted by diet and breed.