Application of proteomic technologies to assess the quality of raw pork and pork products: an overview from farm-to-fork

The quality assurance of pork meat and products includes the study of factors prior to slaughter such as handling practices, diet and castration, and others during the post-mortem period such as aging, storage, and cooking. The development over the last two decades of high-throughput techniques such as proteomics offer great opportunities to examine the molecular mechanisms and study a priori the proteins in the living pigs and main post-mortem changes and post-translational modifications during the conversion of the muscle into the meat. When the most traditional crossbreeding and rearing strategies to improve pork quality were assessed, the main findings indicate that metabolic pathways early post-mortem were affected. Among the factors, it is well documented that pre-slaughter stress provokes substantial changes in the pork proteome that led to defective meat, and consequently, novel protein biomarkers should be identified and validated. Additionally, modifications in pork proteins had a strong effect on the sensory attributes due to the impact of processing, either physical or chemical. Maillard compounds and protein oxidation should be monitored in order to control proteolysis and volatile compounds. Beyond this, the search of bioactive peptides is becoming a paramount goal of the food and nutraceutical industry. In this regard, peptidomics is a major tool to identify and quantify these peptides with beneficial effects for human health.CYTED | Ref. 119RT0568Axencia Galega de Innovación | Ref. IN607A2019/0

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

peer-reviewedThis 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 2 muscles whatever the evaluation method of tenderness, but some relationships with tenderness for others (MYH1, TNNT3, HSPB6) are inversed. Muscle specificities were revealed in this meta-proteomic study. 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 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

Evaluation of the Antioxidant and Antimicrobial Activities of Porcine Liver Protein Hydrolysates Obtained Using Alcalase, Bromelain, and Papain

peer-reviewedIn order to make the by-products generated from the porcine industry more valuable, pig livers were used in this trial to obtain protein hydrolysates. Three proteases (alcalase, bromelain, and papain) were utilized for enzymatic hydrolysis with two different durations, 4 and 8 hours. Ultrafiltration process was used for the recovery of the extracts, employing three different membrane pore sizes (30, 10, and 5 kDa). The porcine livers contained considerable amounts of protein (19.0%), considering they are almost composed of water (74.1%). The antioxidant activity of the obtained hydrolysates was investigated using four antioxidant methods (2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2-2′-Azino-di-[3-ethylbenzthiazoline sulfonate] (ABTS) radical scavenging activity, ferric reducing antioxidant power assay (FRAP), and oxygen radical absorbance capacity assay (ORAC)). Antibacterial properties were also measured against Gram-negative and Gram-positive bacteria. Results indicated that the three studied factors (type of enzyme, membrane pore size, and time) significantly affected the parameters evaluated. Hydrolysates obtained at 8 hours with alcalase had the best antioxidant properties. The 30 kDa alcalase extracts exhibited the highest DPPH (562 µg Trolox/g), FRAP (82.9 µmol Fe2+/100 g), and ORAC (53.2 mg Trolox/g) activities, while for ABTS the 10 kDa alcalase showed the higher values (1068 mg ascorbic acid/100 g). Concerning the antibacterial activity, 30 kDa hydrolysates obtained with bromelain for 4 hours exhibited the highest antimicrobial capacity, providing an inhibition of 91.7%

Application of Proteomic Technologies to Assess the Quality of Raw Pork and Pork Products: An Overview from Farm-To-Fork

The quality assurance of pork meat and products includes the study of factors prior to slaughter such as handling practices, diet and castration, and others during the post-mortem period such as aging, storage, and cooking. The development over the last two decades of high-throughput techniques such as proteomics offer great opportunities to examine the molecular mechanisms and study a priori the proteins in the living pigs and main post-mortem changes and post-translational modifications during the conversion of the muscle into the meat. When the most traditional crossbreeding and rearing strategies to improve pork quality were assessed, the main findings indicate that metabolic pathways early post-mortem were affected. Among the factors, it is well documented that pre-slaughter stress provokes substantial changes in the pork proteome that led to defective meat, and consequently, novel protein biomarkers should be identified and validated. Additionally, modifications in pork proteins had a strong effect on the sensory attributes due to the impact of processing, either physical or chemical. Maillard compounds and protein oxidation should be monitored in order to control proteolysis and volatile compounds. Beyond this, the search of bioactive peptides is becoming a paramount goal of the food and nutraceutical industry. In this regard, peptidomics is a major tool to identify and quantify these peptides with beneficial effects for human healthJosé M. Lorenzo and Daniel Franco are members of the HealthyMeat network, funded by CYTED (Programa Iberoamericano de Ciencia y Tecnología para el Desarrollo) (ref. 119RT0568). Our thanks go to GAIN (Axencia Galega de Innovación, Xunta de Galicia, Spain) for supporting this research (grant number IN607A2019/01). Mohammed Gagaoua is grateful to the funding received from Marie Skłodowska-Curie, grant agreement no. 713654S

Influence of Three Probiotics Strains, Lactobacillus rhamnosus GG, Bifidobacterium animalis subsp. Lactis BB-12 and Saccharomyces boulardii CNCM I-745 on the Biochemical and Haematological Profiles and Body Weight of Healthy Rabbits

Currently, probiotics are used as growth promoters on a large scale to improve the productivity of several animals’ species within the aim of reducing the presence of antibiotic residues in animal products consumed by humans. Several reports evidenced the positive effect of probiotic supplementation on the growth performances and health of rabbits, mainly through the balance of the intestinal microbiota of the host animal. Therefore, certain probiotics, including Lactobacilli, Bifidobacteria, Saccharomyces, can improve the biochemical and haematological profiles, especially in production animals. In this context, this study was performed on rabbits for the economic importance they play as a source of meat proteins in developing countries and their use as experimental models in research and biomedicine. This study then aimed to evaluate the effect of three strains of probiotics: Lactobacillus rhamnosus GG Bifidobacterium animalis subsp. Lactis BB-12 and Saccharomyces boulardii CNCM I-745, on the biochemical and haematological parameters and their influence on the rabbit’s weight of the ITELV2006 strain. The findings evidenced that the probiotic strain affected the biochemical and haematological parameters. Further, the strains showed a positive effect on the weight gain of the rabbits.This study aimed to investigate the effects of three strains of probiotics, these being Lactobacillus rhamnosus GG, Bifidobacterium animalis subsp. Lactis BB-12 and Saccharomyces boulardii CNCM I-745, on the body weight, animal performances and blood parameters of rabbits (male and female) of the ITELV2006 strain. The supplementation of the feed of the rabbits with the three probiotic strains allowed observing positive effects on most of the biochemical and haematological parameters investigated during a period of 60 days (30 days of supplementation and 30 days without treatment). Further, there was a significant improvement in the body weight of the rabbits at the end of the experiment. The effect of the three probiotics investigated in this trial was found to be related to the sex of the rabbits and to the intake period (duration). Ultimately, these findings raise the possibility of using probiotics to investigate in an in-depth and specific manner based on fixed factors such as the strain, the gender and age of the animals, the main underlying mechanisms and effects, which would allow achieving optimal and adapted health benefits and sustainable production. In the context of animal production, it is worth investigating in a targeted study the effect of the three strains on muscle growth and development and finding evidence of the possible consequences on meat quality traits of the rabbits supplemented with probiotics

A Comprehensive Review on Lipid Oxidation in Meat and Meat Products

peer-reviewedMeat and meat products are a fundamental part of the human diet. The protein and vitamin content, as well as essential fatty acids, gives them an appropriate composition to complete the nutritional requirements. However, meat constituents are susceptible to degradation processes. Among them, the most important, after microbial deterioration, are oxidative processes, which affect lipids, pigments, proteins and vitamins. During these reactions a sensory degradation of the product occurs, causing consumer rejection. In addition, there is a nutritional loss that leads to the formation of toxic substances, so the control of oxidative processes is of vital importance for the meat industry. Nonetheless, despite lipid oxidation being widely investigated for decades, the complex reactions involved in the process, as well as the different pathways and factors that influenced them, make that lipid oxidation mechanisms have not yet been completely understood. Thus, this article reviews the fundamental mechanisms of lipid oxidation, the most important oxidative reactions, the main factors that influence lipid oxidation, and the routine methods to measure compounds derived from lipid oxidation in meat

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

peer-reviewedThis 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: β-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

Insights on meat quality from combining traditional studies and proteomics

peer-reviewedFollowing a century of major discoveries on the mechanisms determining meat colour and tenderness using traditional scientific methods, further research into complex and interactive factors contributing to variations in meat quality is increasingly being based on data-driven “omics” approaches such as proteomics. Using two recent meta-analyses of proteomics studies on beef colour and tenderness, this review examines how knowledge of the mechanisms and factors underlying variations in these meat qualities can be both confirmed and extended by data-driven approaches. While proteomics seems to overlook some sources of variations in beef toughness, it highlights the role of post-mortem energy metabolism in setting the conditions for development of meat colour and tenderness, and also points to the complex interplay of energy metabolism, calcium regulation and mitochondrial metabolism. In using proteomics as a future tool for explaining variations in meat quality, the need for confirmation by further hypothesis-driven experimental studies of post-hoc explanations of why certain proteins are biomarkers of beef quality in data-driven studies is emphasised

A proteomic approach to identify biomarkers of foal meat quality: A focus on tenderness, color and intramuscular fat traits

Foal meat is considered a healthy alternative to other meat sources and more environmentally sustainable. However, its quality is highly variable and there is lack of knowledge about the molecular mechanisms underlying its determination. Genotype and diet play a relevant role as the main factors that can allow a control of the final quality and the use of high-throughput analytical methods such as proteomics is a way to achieve this lofty goal. This research aimed to study-two breeds (Burguete and Jaca Navarra) supplemented with two different finishing diets: conventional concentrate and straw (C) vs silage and organic feed (S). The proteomic approach built a library of 294 proteins that were subjected to several statistical and bioinformatic analyses. Burguete breed finished with concentrate produced higher meat quality in terms of tenderness, intramuscular fat and color lightness mainly due to the high abundance of energy metabolic proteins. Tenderness was correlated to myofibrillar proteins (ACTA1, MYBPH, MYL1 and TNNC1) and energy metabolic proteins (ALDOA, CKM, TPI1 and PGMA2). Regarding color, the main pathways were energy metabolism, involving several glycolytic enzymes (ALDOA, PKM, PFKM and CKM). Oxidative stress and response to stress proteins (HSPA1A, SOD2 and PRDX2) were further involved in color variation. Moreover, we revealed that several proteins were related to the intramuscular fat accordingly to the breed. This study proposed several candidate protein biomarkers for foal meat quality that are worthy to evaluate in the futureThis research was funded by Interreg V SUDOE, through OPEN2PRESERVE project, Grant No. SOE2/P5/E0804S

Application of pomegranate by-products in muscle foods: oxidative indices, colour stability, shelf life and health benefits

In recent years, considerable importance is given to the use of agrifood wastes as they contain several groups of substances that are useful for development of functional foods. As muscle foods are prone to lipid and protein oxidation and perishable in nature, the industry is in constant search of synthetic free additives that help in retarding the oxidation process, leading to the development of healthier and shelf stable products. The by-products or residues of pomegranate fruit (seeds, pomace, and peel) are reported to contain bioactive compounds, including phenolic and polyphenolic compounds, dietary fibre, complex polysaccharides, minerals, vitamins, etc. Such compounds extracted from the by-products of pomegranate can be used as functional ingredients or food additives to harness the antioxidant, antimicrobial potential, or as substitutes for fat, and protein in various muscle food products. Besides, these natural additives are reported to improve the quality, safety, and extend the shelf life of different types of food products, including meat and fish. Although studies on application of pomegranate by-products on various foods are available, their effect on the physicochemical, oxidative changes, microbial, colour stabilizing, sensory acceptability, and shelf life of muscle foods are not comprehensively discussed previously. In this review, we vividly discuss these issues, and highlight the benefits of pomegranate by-products and their phenolic composition on human health.Axencia Galega de Innovación | Ref. IN607A2019 / 01CYTED | Ref. 119RT056