Relationships between the abundance of 29 proteins and several meat or carcass quality traits in two bovine muscles revealed by a combination of univariate and multivariate analyses.

We aimed to evaluate the relationships between meat or carcass properties and the abundance of 29 proteins quantified in two muscles, Longissimus thoracis and Rectus abdominis, of Rouge des Prés cows. The relative abundance of the proteins was evaluated using a high throughput immunological method: the Reverse Phase Protein array. A combination of univariate and multivariate analyses has shown that small HSPs (CRYAB, HSPB6), fast glycolytic metabolic and structural proteins (MYH1, ENO3, ENO1, TPI1) when assayed both in RA and LT, were related to meat tenderness, marbling, ultimate pH, as well as carcass fat-to-lean ratio or conformation score. In addition to some small HSP, ALDH1A1 and TRIM72 contributed to the molecular signature of muscular and carcass adiposity. MYH1 and HSPA1A were among the top proteins related to carcass traits. We thus shortened the list to 10 putative biomarkers to be considered in future tools to manage both meat and carcass properties. SIGNIFICANCE: In three aspects this manuscript is notable. First, this is the first proteomics study that aims to evaluate putative biomarkers of both meat and carcass qualities that are of economic importance for the beef industry. Second, the relationship between the abundance of proteins and the carcass or meat traits were evaluated by a combination of univariate and multivariate analyses on 48 cows that are representative of the biological variability of the traits. Third, we provide a short list of ten proteins to be tested in a larger population to feed the pipeline of biomarker discovery.

Effects of Rearing Management Applied throughout the Charolais Young Bulls' Life on Carcass and Meat Quality.

The aim of this work was to study, for the first time, the effects of the rearing management (from birth to slaughter) applied throughout the life of young bulls on carcass and meat quality. Five rearing managements were defined statistically, from a combination of 30 rearing factors, using a hierarchical clustering on principal components. This study considered the individual data of 179 Charolais young bulls from commercial farms. The carcass traits were more sensitive to rearing management than the meat traits. Rearing management had an effect mainly on fat and overall meat grain for the carcass, and on color and tenderness for the longissimus meat. However, it was possible to produce carcass and/or meat with similar properties from different rearing managements. Among the five rearing managements defined in this study, two were identified as allowing the best trade-off to produce simultaneously high carcass and meat quality. The first management was characterized by absence of growth period and a short fattening duration, with a wrapped haylage or corn silage-based diet. The second management was characterized by short pre-weaning and growth periods, and a long fattening period.

Does the Rearing Management following by Charolais Cull Cows Influence the Qualities of Carcass and Beef Meat?

This study characterized, for the first time, the rearing managements (from birth to slaughter) applied throughout the cull cows' life and observed the effect of these managements on the carcass and meat properties. From the individual data of 371 Charolais cull cows, three rearing managements were defined and characterized with 60 rearing factors. The results showed that the rearing managements had low effects on the carcass and meat properties. For the carcass traits, only the carcass weight, and fat and longissimus (LM) colors at the level of the sixth rib were impacted. Concerning the meat, only the red color intensity, the fat aroma, the flavor intensity and persistence were affected. According to our results, this study confirmed that it is possible to produce carcass or meat with similar properties; consequently, it is difficult to favor a rearing management. However, to manage jointly both carcass and meat qualities, trade-offs are needed.

Does Finishing at Pasture Influence the Colour of Muscle from Suckler Bulls and Can Colour Be Used to Authenticate Their Pre-Slaughter Diet?

The primary objective of this study was to compare the colour of muscle from bulls finished at pasture or indoors on a high concentrate diet. The ancillary objectives were to identify possible explanations for any differences in the colour observed and the potential of muscle colour to discriminate between bull beef from different production systems. Growth, longissimus muscle colour, fibre type composition and metabolic profile were measured in late-maturing breed sired suckler bulls slaughtered at 19 months of age after 199 days at pasture (G0), 100 days indoors after 98 days at pasture (G0AL) and indoors for 199 days (AL). When compared to bulls finished indoors and offered a high concentrate ration, the carcass weight of G0 bulls was lower, their carcasses were leaner, and their longissimus muscle was similar in lightness but less red and had a lower glycolytic metabolism. The temperature at which the longissimus muscle reached pH 6.0 was lower (19.7 °C) for G0 than for G0AL (29.9 °C) and AL (31.6 °C), which did not differ. Co-variate adjustment for this variable removed the differences in redness. Adjusting the chill settings appears to be a practical strategy for abattoirs to minimise early post-mortem differences in muscle colour between lighter grass-fed and heavier concentrate-fed carcasses. The preliminary results demonstrate the potential of both L*, a*, b* values and the visible reflectance spectra of muscle to discriminate between grass- finished and concentrate-finished bull beef, but further refinement and validation of the models is required.

Characterization of Four Rearing Managements and Their Influence on Carcass and Meat Qualities in Charolais Heifers.

The study aim was to identify the effects of the rearing management applied throughout the heifers' life on the carcass (e.g., conformation, marbling, fat) and meat (color, texture, and sensory profiles) properties. From the individual data of 171 heifers from 25 commercial farms, a typology of four rearing managements was established from 50 rearing factors. The managements had an effect on the conformation, the color (fat and muscle), and the rhomboideus grain meat, for the carcass, and the lightness, the atypical flavor, and the overall acceptability for the longissimus (LM) meat. The carcass traits compared to the meat were more sensitive to a change of rearing management. Our results confirmed that it was possible to target the same carcass or meat quality from different managements. Moreover, according to the aims of the targeted carcass and LM meat quality, management 3 could be an interesting trade-off to jointly manage the quality of both products. For example, the carcasses that were produced had a high conformation, smooth meat grain and the LM meat was more liked. This management was intermediate compared to the other rearing managements and had a long fattening period with a diet mainly based on conserved grass and a high concentrate quantity.

Myostatin gene inactivation increases post-mortem calpain-dependent muscle proteolysis in mice.

Myostatin deficiency leads to extensive skeletal muscle hypertrophy, but its consequence on post-mortem muscle proteolysis is unknown. Here, we compared muscle myofibrillar protein degradation, and autophagy, ubiquitin-proteasome and Ca2+-dependent proteolysis relative to the energetic and redox status in wild-type (WT) and myostatin knock-out mice (KO) during early post-mortem storage. KO muscles showed higher degradation of myofibrillar proteins in the first 24 h after death, associated with preserved antioxidant status, compared with WT muscles. Analysis of key autophagy and ubiquitin-proteasome system markers indicated that these two pathways were not upregulated in post-mortem muscle (both genotypes), but basal autophagic flux and ATP content were lower in KO muscles. Proteasome and caspase activities were not different between WT and KO mice. Conversely, calpain activity was higher in KO muscles, concomitantly with higher troponin T and desmin degradation. Altogether, these results suggest that calpains but not the autophagy, proteasome and caspase systems, explain the difference in post-mortem muscle protein proteolysis between both genotypes.

Dark-cutting beef: A brief review and an integromics meta-analysis at the proteome level to decipher the underlying pathways.

Comprehensive characterization of the post-mortem muscle proteome defines a fundamental goal in meat proteomics. During the last decade, proteomics tools have been applied in the field of foodomics to help decipher factors underpinning meat quality variations and to enlighten us, through data-driven methods, on the underlying mechanisms leading to meat quality defects such as dark-cutting meat known also as dark, firm and dry (DFD) meat. In cattle, several proteomics studies have focused on the extent to which changes in the post-mortem muscle proteome relate to dark-cutting beef development. The present data-mining study firstly reviews proteomics studies which investigated dark-cutting beef, and secondly, gathers the protein biomarkers that differ between dark-cutting versus beef with normal-pH in a unique repertoire. A list of 130 proteins from eight eligible studies was curated and mined through bioinformatics for Gene Ontology annotations, molecular pathways enrichments, secretome analysis and biological pathways comparisons to normal beef color from a previous meta-analysis. The major biological pathways underpinning dark-cutting beef at the proteome level have been described and deeply discussed in this integromics study.

Combining label-free and label-based accurate quantifications with SWATH-MS: Comparison with SRM and PRM for the evaluation of bovine muscle type effects.

Mass spectrometry has proven to be a valuable tool for the accurate quantification of proteins. In this study, the performances of three targeted approaches, namely selected reaction monitoring (SRM), parallel reaction monitoring (PRM) and sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS), to accurately quantify ten potential biomarkers of beef meat tenderness or marbling in a cohort of 64 muscle samples were evaluated. So as to get the most benefit out of the complete MS2 maps that are acquired in SWATH-MS, an original label-free quantification method to estimate protein amounts using an I-spline regression model was developed. Overall, SWATH-MS outperformed SRM in terms of sensitivity and dynamic range, while PRM still performed the best, and all three strategies showed similar quantification accuracies and precisions for the absolute quantification of targets of interest. This targeted picture was extended by 585 additional proteins for which amounts were estimated using the label-free approach on SWATH-MS; thus, offering a more global profiling of muscle proteomes and further insights into muscle type effect on candidate biomarkers of beef meat qualities as well as muscle metabolism.

The Blonde d'Aquitaine T3811>G3811 mutation in the myostatin gene: association with growth, carcass, and muscle phenotypes in veal calves.

The mutation T3811 → G3811 (TG3811) discovered in the myostatin gene of the Blonde d'Aquitaine breed is suspected of contributing to the outstanding muscularity of this breed. An experiment was designed to estimate the effect of this mutation in an F2 and back-cross Blonde d'Aquitaine × Holstein population. By genotyping all known mutations in the myostatin gene, it was ensured that the TG3811 mutation was indeed the only known mutation segregating in this population. Fifty-six calves (43 F2, 13 back-cross) were intensively fattened and slaughtered at 24.0 ± 1.4 wk of age. The effects of the mutation were estimated by comparing the calves with the [T/T] (n = 18), [T/G] (n = 30), and [G/G] (n = 8) genotypes. Highly significant substitution effects (P < 0.001), above + 1.2 phenotypic SD, were shown on carcass yield and muscularity scores. Birth weight (P < 0.001) was positively affected by the mutation (+0.8 SD) but not growth rate (P = 0.97), while carcass length (P = 0.03), and fatness (P ≤ 0.03) were negatively affected (-0.5 to -0.7 SD). The characteristics of the Triceps brachii muscle were affected by the mutation (P < 0.001), with lower ICDH activity (oxidative) and a higher proportion of myosin type 2X muscle fibers (fast twitch). The effects of the TG3811 mutation were similar to those of other known myostatin mutations, although the Blonde d'Aquitaine animals, which are predominantly [G/G] homozygous, do not exhibit extreme double muscling.

Understanding the Determination of Meat Quality Using Biochemical Characteristics of the Muscle: Stress at Slaughter and Other Missing Keys.

Despite increasingly detailed knowledge of the biochemical processes involved in the determination of meat quality traits, robust models, using biochemical characteristics of the muscle to predict future meat quality, lack. The neglecting of various aspects of the model paradigm may explain this. First, preslaughter stress has a major impact on meat quality and varies according to slaughter context and individuals. Yet, it is rarely taken into account in meat quality models. Second, phenotypic similarity does not imply similarity in the underlying biological causes, and several models may be needed to explain a given phenotype. Finally, the implications of the complexity of biological systems are discussed: a homeostatic equilibrium can be reached in countless ways, involving thousands of interacting processes and molecules at different levels of the organism, changing over time and differing between animals. Consequently, even a robust model may explain a significant part, but not all of the variability between individuals.

Molecular signatures of beef tenderness: Underlying mechanisms based on integromics of protein biomarkers from multi-platform proteomics studies.

Over the last two decades, proteomics have been employed to decipher the underlying factors contributing to variation in the quality of muscle foods, including beef tenderness. One such approach is the application of high-throughput protein analytical platforms in the identification of meat quality biomarkers. To broaden our understanding about the biological mechanisms underpinning meat tenderization across a large number of studies, an integromics study was performed to review the current status of protein biomarker discovery targeting beef tenderness. This meta-analysis is the first to gather and propose a comprehensive list of 124 putative protein biomarkers derived from 28 independent proteomics-based experiments, from which 33 robust candidates were identified worthy of evaluation using targeted or untargeted data-independent acquisition proteomic methods. We further provide an overview of the interconnectedness of the main biological pathways impacting tenderness determination after multistep analyses including Gene Ontology annotations, pathway and process enrichment and literature mining, and specifically discuss the major proteins and pathways most often reported in proteomics research.

Protein Array-Based Approach to Evaluate Biomarkers of Beef Tenderness and Marbling in Cows: Understanding of the Underlying Mechanisms and Prediction.

This study evaluated the potential of a panel of 20 protein biomarkers, quantified by Reverse Phase Protein Array (RPPA), to explain and predict two important meat quality traits, these being beef tenderness assessed by Warner-Bratzler shear force (WBSF) and the intramuscular fat (IMF) content (also termed marbling), in a large database of 188 Protected Designation of Origin (PDO) Maine-Anjou cows. Thus, the main objective was to move forward in the progression of biomarker-discovery for beef qualities by evaluating, at the same time for the two quality traits, a list of candidate proteins so far identified by proteomics and belonging to five interconnected biological pathways: (i) energy metabolic enzymes, (ii) heat shock proteins (HSPs), (iii) oxidative stress, (iv) structural proteins and (v) cell death and protein binding. Therefore, three statistical approaches were applied, these being Pearson correlations, unsupervised learning for the clustering of WBSF and IMF into quality classes, and Partial Least Squares regressions (PLS-R) to relate the phenotypes with the 20 biomarkers. Irrespective of the statistical method and quality trait, seven biomarkers were related with both WBSF and IMF, including three small HSPs (CRYAB, HSP20 and HSP27), two metabolic enzymes from the oxidative pathway (MDH1: Malate dehydrogenase and ALDH1A1: Retinal dehydrogenase 1), the structural protein MYH1 (Myosin heavy chain-IIx) and the multifunctional protein FHL1 (four and a half LIM domains 1). Further, three more proteins were retained for tenderness whatever the statistical method, among which two were structural proteins (MYL1: Myosin light chain 1/3 and TNNT1: Troponin T, slow skeletal muscle) and one was glycolytic enzyme (ENO3: ß-enolase 3). For IMF, two proteins were, in this trial, specific for marbling whatever the statistical method: TRIM72 (Tripartite motif protein 72, negative) and PRDX6 (Peroxiredoxin 6, positive). From the 20 proteins, this trial allowed us to qualify 10 and 9 proteins respectively as strongly related with beef tenderness and marbling in PDO Maine-Anjou cows.

Prediction of the Secretome and the Surfaceome: A Strategy to Decipher the Crosstalk between Adipose Tissue and Muscle during Fetal Growth.

Crosstalk between adipose and muscular tissues is hypothesized to regulate the number of muscular and adipose cells during fetal growth, with post-natal consequences on lean and fat masses. Such crosstalk largely remains, however, to be described. We hypothesized that a characterization of the proteomes of adipose and muscular tissues from bovine fetuses may enhance the understanding of the crosstalk between these tissues through the prediction of their secretomes and surfaceomes. Proteomic experiments have identified 751 and 514 proteins in fetal adipose tissue and muscle. These are mainly involved in the regulation of cell proliferation or differentiation, but also in pathways such as apoptosis, Wnt signalling, or cytokine-mediated signalling. Of the identified proteins, 51 adipokines, 11 myokines, and 37 adipomyokines were predicted, together with 26 adipose and 13 muscular cell surface proteins. Analysis of protein-protein interactions suggested 13 links between secreted and cell surface proteins that may contribute to the adipose-muscular crosstalk. Of these, an interaction between the adipokine plasminogen and the muscular cell surface alpha-enolase may regulate the fetal myogenesis. The in silico secretome and surfaceome analyzed herein exemplify a powerful strategy to enhance the elucidation of the crosstalk between cell types or tissues.

Muscle Fiber Properties in Cattle and Their Relationships with Meat Qualities: An Overview.

The control of meat quality traits constitutes an important target for any farm animal production, including cattle. Therefore, better understanding of the biochemical properties that drive muscle development and final outcomes constitutes one of the main challenging topics of animal production and meat science. Accordingly, this review has focused on skeletal muscle fibers in cattle and their relationships with beef qualities. It aimed to describe the chemical and structural properties of muscle fibers as well as a comprehensive review of their contractile and metabolic characteristics during the life of the animal. The existing methods for the classification of muscle fibers were reviewed, compared, and discussed. Then, the different stages of myogenesis in cattle were defined. The main factors regulating fetal and postnatal growth and the plasticity of muscle fibers were evidenced, especially the role of myostatin growth factor and the impact of nutritional factors. This review highlights that the knowledge about muscle fibers is paramount for a better understanding of how to control the muscle properties throughout the life of the animal for better management of the final eating qualities of beef. Accordingly, the associations between bovine muscle fibers and different meat eating qualities such as tenderness, pH decline, and color traits were further presented.

Current Advances in Meat Nutritional, Sensory and Physical Quality Improvement.

Meat is an important source of proteins, vitamins, minerals and fat, and these nutrients are important for their beneficial effects on human health. In recent years, meat quality has become a more relevant topic for consumers with regard to health and sensory characteristics, and for beef industry stakeholders because it affects their profitability. Therefore, the control of meat quality, including technological, sensory and nutritional quality traits, constitutes an important target for any farm animal production. What those qualities are and how we best evaluate them at the different levels of the continuum from the farm to fork are critical to understanding meat production and consumption patterns. However, despite the efforts of the industrials to control the eating and nutritional quality traits of meat and meat products, there remains a high level of variability, which is one reason for consumer dissatisfaction. This Special Issue focuses on the study of continuum aspects from farm to fork, which would have an impact on the control of the nutritional, sensory and technological aspects of carcass, muscle, meat and meat-product qualities. It groups fourteen original studies and one comprehensive review within five main topics that are (i) production systems and rearing practices, (ii) prediction of meat qualities, (iii) statistical approaches for meat quality prediction/management, (iv) muscle biochemistry and proteomics techniques and (v) consumer acceptability, development and characterisation of meat products.

Quantification of biomarkers for beef meat qualities using a combination of Parallel Reaction Monitoring- and antibody-based proteomics.

We and others have identified biomarker candidates of tenderness or marbling, two major attributes of bovine meat-eating qualities for consumers' satisfaction. In this study, Reverse Phase Protein Arrays (RPPA) and targeted mass spectrometry assays using Parallel Reaction Monitoring (PRM) were developed to test whether 10 proteins pass the sequential qualification and verification steps of the challenging biomarker discovery pipeline. At least MYH1, TPI1, ALDH1A1 and CRYAB were qualified by RPPA or PRM as being differentially abundant according to marbling values of longissimus thoracis and semimembranosus muscles. Significant mathematical relationships between the individual abundance of each of the four proteins and marbling values were verified by linear or logistic regressions. Four proteins, TNNT1, MDH1, PRDX6 and ENO3 were qualified and verified for tenderness, and the abundance of MDH1 explained 49% of the tenderness variability. The present PRM and RPPA results pave the way for development of useful meat industrial multiplex-proteins assays.

Label free shotgun proteomics for the identification of protein biomarkers for beef tenderness in muscle and plasma of heifers.

Meat quality prediction is a priority for the beef industry. Label free shotgun proteomics was performed on Longissimus muscle and plasma from 20 crossbred Charolais x Aubrac beef heifers, classified as subgroups of 5 extreme tender and 5 extreme tough meat according to sensory evaluation, Warner Bratzler shear force, and a synthetic tenderness index. This technique identified 268 proteins in muscle and 136 in plasma. Among them, 71 muscle proteins and 21 plasma proteins discriminated tender and tough groups. These proteins were analyzed to select the most correlated and explicative ones which were used in a linear regression on the 20 heifers. The results validated in heifers 33 muscle proteins previously identified as related with tenderness, and revealed 38 new candidates. Twelve are localized in shear force or tenderness score QTL. Among them ACTN2, ADSSL1, GOT1, HPX, OGDH, OGN, TNNC1 and VCL are proposed as robust candidates with 3 other proteins known to be related with tenderness (MYBPH, CAPZB, MYH1). Examination of the plasma proteome showed 8 putative biomarkers (MYH7, CFH, ENO3, PLA2G2D5, FHL1, GAPDH, MASP2 and SERPINF2). Three of them (MYH7, ENO3 and FHL1) were identified as discriminative of tenderness both in Longissimus muscle and in plasma. SIGNIFICANCE: The label free proteomic approach used in this study allowed to complete the atlas of biomarkers for tenderness of the Longissimus muscle. This innovative proteomic approach applied on plasma samples allowed to identify circulating candidate biomarkers for beef tenderness. This low-invasive approach constitutes an interesting alternative to evaluate early the "beef meat potential" of living animals in farm or of the carcass in slaughterhouses.

Contribution of connective tissue components, muscle fibres and marbling to beef tenderness variability in longissimus thoracis, rectus abdominis, semimembranosus and semitendinosus muscles.

BACKGROUND: The present study aimed to identify relationships between components of intramuscular connective tissue, proportions of the different fiber types, intramuscular fat and sensory tenderness of beef cooked at 55 °C. Accordingly, four muscles differing in their metabolic and contractile properties, as well as in their collagen content and butcher value, were obtained from dairy and beef cattle of several ages and sexes and were then used to create variability. RESULTS: Correlation analyses and/or stepwise regressions were applied on Z-scores to identify the existing and robust associations. Tenderness scores were further categorized into tender, medium and tough classes using unsupervised learning methods. The findings revealed a muscle-dependant role with respect to tenderness of total and insoluble collagen, cross-links, and type IIB + X and IIA muscle fibers. The longissimus thoracis and semitendinosus muscles that, in the present study, were found to be extreme in their tenderness potential were also very different from each other and from the rectus abdominis (RA) and semimembranosus (SM). RA and SM muscles were very similar regarding their relationship for muscle components and tenderness. A relationship between marbling and tenderness was only present when the results were analysed irrespective of all factors of variation of the experimental model relating to muscle and animal type. CONCLUSION: The statistical approaches applied in the present study using Z-scores allowed identification of the robust associations between muscle components and sensory beef tenderness and also identified discriminatory variables of beef tenderness classes. © 2020 Society of Chemical Industry.

Meta-proteomics for the discovery of protein biomarkers of beef tenderness: An overview of integrated studies.

This meta-proteomics review focused on proteins identified as candidate biomarkers of beef tenderness by comparing extreme groups of tenderness using two-dimensional electrophoresis (2-DE) associated with mass spectrometry (MS). We reviewed in this integromics study the results of 12 experiments that identified protein biomarkers from two muscles, Longissimus thoracis (LT) and Semitendinosus (ST), of different types of cattle: young bulls, steers or cows from beef breeds (Charolais, Limousin, Blond d'Aquitaine), hardy breed (Salers) or mixed breed (PDO Maine-Anjou). Comparative proteomics of groups differing in their tenderness evaluated by instrumental Warner-Bratzler shear force (WBSF) or by sensory analysis using trained panelists, revealed 61 proteins differentially abundant (P < 0.05) between tender and tough groups. A higher number of discriminative proteins was observed for LT (50 proteins) compared to ST muscle (28 proteins). The Gene Ontology annotations showed that the proteins of structure and contraction, protection against oxidative stress and apoptosis, energy metabolism, 70 family HSPs and proteasome subunits are more involved in LT tenderness than in ST. Amongst the list of candidate biomarkers of tenderness some proteins such as HSPB1 are common between the two muscles whatever the evaluation method of tenderness, but some relationships with tenderness for other proteins (MYH1, TNNT3, HSPB6) were inversed. Muscle specificities were revealed in this meta-proteomics. For example, Parvalbumin (PVALB) appeared as a robust biomarker in ST muscle whatever the evaluation method of tenderness. HSPA1B seems to be a robust candidate for LT tenderness (with WBSF) regardless the animal type. Some gender specificities were further identified including similarities between cows and steers (MSRA and HSPA9) in contrast to bulls. The comparison of the 12 proteomic studies revealed strong dissimilarities to identify generic biomarkers of beef tenderness. This integrative analysis allowed better understanding of the biological processes involved in beef tenderness in two muscles and their variations according to the main factors underlying this quality. It allowed also proposing for the first time a comprehensive list of candidate biomarkers to be evaluated deeply to validate their relationships with tenderness on a large number of cattle and breeds.