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.

Post-mortem muscle proteome of crossbred bulls and steers: Relationships with carcass and meat quality.

This study investigated the skeletal muscle proteome of crossbred bulls and steers with the aim of explaining the differences in carcass and meat quality traits. Therefore, 640 post-weaning Angus-Nellore calves were fed a high-energy diet for a period of 180 days. In the feedlot trial, comparisons of steers (n = 320) and bulls (n = 320) showed lower (P < 0.01) average daily gain (1.38 vs. 1.60 ± 0.05 kg/d), final body weight (547.4 vs. 585.1 ± 9.3 kg), which resulted in lower hot carcass weight (298.4 vs. 333.7 ± 7.7 kg) and ribeye area (68.6 vs. 81.0 ± 2.56 cm2). Steers had higher (P < 0.01) carcass fatness, meat color parameters (L*, a*, b*, chroma (C*), hue (h°)) and lower ultimate pH. Moreover, lower (P < 0.01) Warner-Bratzler shear force (WBSF) were observed in steers compared to bulls (WBSF = 3.68 vs. 4.97 ± 0.08 kg; and 3.19 vs. 4.08 ± 0.08 kg). The proteomic approach using two-dimensional electrophoresis, mass spectrometry and bioinformatics procedures revealed several differentially expressed proteins between steers and bulls (P < 0.05). Interconnected pathways and substantial changes were revealed in biological processes, molecular functions, and cellular components between the post-mortem muscle proteomes of the compared animals. Steers had increased (P < 0.05) abundance of proteins related to energy metabolism (CKM, ALDOA, and GAPDH), and bulls had greater abundance of proteins associated with catabolic (glycolysis) processes (PGM1); oxidative stress (HSP60, HSPA8 and GSTP1); and muscle structure and contraction (TNNI2 and TNNT3). The better carcass (fatness and marbling degree) and meat quality traits (tenderness and color parameters) of steers were associated with higher abundance of key proteins of energy metabolism and lower abundance of enzymes related to catabolic processes, oxidative stress, and proteins of muscle contraction SIGNIFICANCE: Sexual condition of cattle is known to be an important factor affecting animal performances and growth as well as the carcass and meat quality traits. The investigation of skeletal muscle proteome help a better understanding of the origin of the differences in quality traits between bulls and steers. The inferior meat quality of bulls was found to be due to the greater expression of proteins associated with primary and catabolic processes, oxidative stress, and muscle contraction. Steers had greater expression of proteins, from which several are known biomarkers of beef quality (mainly tenderness).

Proteomics Unveils Post-Mortem Changes in Beef Muscle Proteins and Provides Insight into Variations in Meat Quality Traits of Crossbred Young Steers and Heifers Raised in Feedlot.

Proteomics has been widely used to study muscle biology and meat quality traits from different species including beef. Beef proteomics studies allow a better understanding of the biological processes related to meat quality trait determination. This study aimed to decipher by means of two-dimensional electrophoresis (2D-PAGE), mass spectrometry and bioinformatics the changes in post-mortem muscle with a focus on proteins differentially expressed in the Longissimus thoracis (LT) muscle of immunocastrated young heifers and steers. Carcass traits, chemical composition, pH, instrumental color (L*, a*, b*), cooking loss and Warner-Bratzler shear force (WBSF) of meat from F1 Montana-Nellore cattle were also evaluated. Backfat thickness (BFT) and intramuscular fat content (IMF) were 46.8% and 63.6% higher in heifers (p < 0.05), respectively, while evaporation losses (EL) were 10.22% lower compared to steers. No differences (p > 0.05) were observed for tenderness evaluated by WBSF (3, 10, and 17 days post-mortem), pH, and color traits (L*, a* and b*) between the experimental groups. The study revealed several proteins to be differentially expressed proteins in heifers compared steers (p < 0.05). In heifers, proteins involved in nutrient transport (TF, ALB, and MB), energy metabolism (ALDOA, GAPDH, and PKM), and oxidative stress and response to stress (HSPA8 and CA3) were associated with a greater BFT and IMF deposition. The higher expression of these proteins indicated greater oxidative capacity and lower glycolytic activity in the LT muscle of heifers. In steers, there was greater abundance of protein expression related to muscle contraction and proteins of structure (ACTA1, TPM2 and TNNT3), energy metabolism (ENO1, ENO3, PYGM, PGM1 and TPI1) and ATP metabolism (ATP5F1B, PEBP1 and AK1), indicating greater glycogenolysis in LT muscle, suggesting a shift in the glycolytic/oxidative fibers of steers.

Impact of Cattle Feeding Strategy on the Beef Metabolome.

The present study explored changes in the meat metabolome of animals subjected to different finishing systems and growth rates. Thirty-six Angus × Nellore crossbred steers were used in a completely randomized design with four treatments: (1) feedlot system with high average daily gain (ADG; FH); (2) feedlot system with low ADG (FL); (3) pasture system with high ADG (PH); and (4) pasture system with low ADG (PL). After harvest and chilling, Longissimus thoracis (LT) muscle samples were taken for metabolite profile analysis using nuclear magnetic resonance. Spectrum was analyzed using chenomx software, and multi- and mega-variate data analyses were performed. The PLS-DA showed clear separation between FH and PL groups and overlap among treatments with different finishing systems but similar for matching ADG (FL and PH) treatments. Using a VIP cut-off of around 1.0, ATP and fumarate were shown to be greater in meat from PL cattle, while succinate, leucine, AMP, glutamate, carnosine, inosine, methionine, G1P, and choline were greater in meat from FH. Comparing FL and PH treatments, glutamine, carnosine, urea, NAD+, malonate, lactate, isoleucine, and alanine were greater in the meat of PH cattle, while G6P and betaine were elevated in that of FL cattle. Relevant pathways were also identified by differences in growth rate (FH versus PL) and finishing system were also noted. Growth rate caused a clear difference in meat metabolism that was highlighted by energy metabolism and associated pathways, while the feeding system tended to alter protein and lipid metabolism.

Meat Quality and Muscle Tissue Proteome of Crossbred Bulls Finished under Feedlot Using Wet Distiller Grains By-Product.

Wet distiller grains (WDG) are a corn by-product rich in protein and fiber that can be used in feedlot diets. This study evaluated F1 Angus-Nellore bulls fed on a control diet vs. WDG (n = 25/treatment). After a period of 129 days on these feeds, the animals were slaughtered and Longissimusthoracis samples were collected for both a meat quality evaluation and gel-based proteomic analyses. A greater ribeye area (99.47 cm²) and higher carcass weight (333.6 kg) (p < 0.05) were observed in the WDG-finished cattle compared to the control (80.7 cm²; 306.3 kg). Furthermore, there were differences (p < 0.05) in the intramuscular fat between the WDG and control animals (IMF = 2.77 vs. 4.19%), which led to a significant decrease (p < 0.05) in saturated fatty acids (FA). However, no differences (p > 0.10) were observed in terms of tenderness, evaluated using Warner-Bratzler shear force (WBSF). The proteomic and bioinformatic analyses revealed substantial changes in the biological processes, molecular functions, and cellular components of the WDG-finished cattle compared to the control. Proteins related to a myriad of interconnected pathways, such as contractile and structural pathways, energy metabolism, oxidative stress and cell redox homeostasis, and transport and signaling. In this experiment, the use of WDG supplementation influenced the protein expression of several proteins, some of which are known biomarkers of beef quality (tenderness and color), as well as the protein-protein interactions that can act as the origins of increases in muscle growth and reductions in IMF deposition. However, despite the effects on the proteome, the tenderness, evaluated by WBSF, and fatty acid profile were not compromised by WDG supplementation.

Inclusion of Healthy Oils for Improving the Nutritional Characteristics of Dry-Fermented Deer Sausage.

The influence of partial replacement of animal fat by healthy oils on composition, physicochemical, volatile, and sensory properties of dry-fermented deer sausage was evaluated. Four different batches were manufactured: the control was formulated with animal fat (18.2%), while in the reformulated batches the 50% of animal fat was substituted by olive, canola, and soy oil emulsions immobilized in Prosella gel. The reformulation resulted in a decrease of moisture and fat contents and an increase of protein and ash amount. Moreover, reformulated sausages were harder, darker, and had higher pH values. This fact is related to the lower moisture content in these samples. As expected, the fatty acid composition was changed by the reformulation. The use of soy and canola oils increased polyunsaturated fatty acids and omega-3 content and decreased n-6/n-3 ratio and saturated fatty acids. Thus, the use of these two oils presented the best nutritional benefits. The changes observed in the fatty acids reflected the fatty acid composition of the oils employed in the emulsions. Regarding volatile compounds (VOC), the replacement of animal fat by healthy emulsion gels increased the content of both total VOC and most of individual VOC. However, the lipid-derived VOC did not show this trend. Generally speaking, the control samples presented similar or higher VOC derived from lipid oxidation processes, which could be related to the natural antioxidant compounds present in the vegetable oils. Finally, all reformulated sausages presented higher consumer acceptability than control samples. In fact, the sausage reformulated with soy oil emulsion gel was the most preferred. Thus, as a general conclusion, the reformulation of deer sausages with soy emulsion gel improves both composition and sensory quality of the final product, which could be an excellent strategy to the elaboration of healthy fermented sausages.