Biomarkers of meat tenderness: Present knowledge and perspectives in regards to our current understanding of the mechanisms involved

Biomarkers of the meat quality are of prime importance for meat industry, which has to satisfy consumers' expectations and, for them, meat tenderness is and will remain the primary and most important quality attribute. The tenderization of meat starts immediately after animal death with the onset of apoptosis followed by a cooperative action of endogenous proteolytic systems. Before consideration of the biomarkers identified so far, we present here some new features on the apoptotic process. Among them, the most important is the recent discovery of a complex family of serpins capable to inhibit, in a pseudo-irreversible manner, caspases, the major enzymes responsible of cell dismantling during apoptosis. The biomarkers so far identified have been then sorted and grouped according to their common biological functions. All of them refer to a series of biological pathways including glycolytic and oxidative energy production, cell detoxification, protease inhibition and production of Heat Shock Proteins. Some unusual biomarkers are also presented: annexins, galectins and peroxiredoxins. On this basis, a detailed analysis of these metabolic pathways allowed us to identify some domains of interest for future investigations. It was thus emphasized that mitochondria, an important organelle in the production of energy from carbohydrates, lipids and proteins are a central element in the initiation and development of apoptosis. It was therefore stressed forward that, in fact, very little is known about the postmortem fate of these organelles and their multiple associated activities. Other topics discussed here would provide avenues for the future in the context of identifying reliable predictors of the ultimate meat tenderness.     

Control of fresh meat quality through manipulation of muscle fiber characteristics

Variations of fresh meat quality exist because the quality traits are affected by various intrinsic and extrinsic factors. Because the meat quality is basically dependent on muscle fiber characteristics, numerous studies have reported the relationship between quality traits and fiber characteristics. Despite intensive research, the relationship is yet to be fully established, however, the present knowledge suggests several potential ways to manipulate muscle fiber characteristics to improve meat quality. The present paper reviews the definition of fresh meat quality, meat quality traits and variations of meat quality. Also, this review presents recent knowledge underlying the relationship between fresh meat quality traits and muscle fiber characteristics. Finally, the present work proposes several potential factors including breed, genotype, sex, hormone, growth performance, diet, muscle location, exercise and ambient temperature that can be used to manipulate muscle fiber characteristics and subsequently meat quality in animals.     

Proteins in longissimus muscle of Korean native cattle and their relationship to meat quality

Proteomic profiling by two-dimensional gel electrophoresis and mass spectrometry of longissimus dorsi muscle tissue from Korean native cattle identified seven proteins that are differentially expressed in animals producing low and high quality grade beef. The expression level of alpha actin is increased in high quality grade beef and the expression levels of T-complex protein 1 (TCP-1), heat shock protein beta-1 (HSP27), and inositol 1,4,5-triphosphate receptor type1 (IP3R1), a new protein to be associated with meat quality, are increased in low quality grade beef. In particular, the quantitation of HSP27 and IP3R1 by both silver staining and immunoblotting correlated well with intramuscular fat content, meat tenderness, and free calcium levels. The data suggest that HSP27 and IP3R1 are potential meat quality biomarkers and their identification provides new insight into the molecular mechanisms and pathways associated with overall beef quality.     

Influence of muscle type on physicochemical and sensory properties of foal meat

The effect of muscle type on physico-chemical properties and sensory characteristics of foal meat was investigated. Six muscles: longissimus dorsi (LD), semimembranosus (SM), semitendinosus (ST), biceps femoris (BF), triceps brachii (TB) and psoas major & minor (PM) from twelve foals slaughtered at 15 months from an extensive production system in freedom regimen were extracted for this study. Regarding chemical composition, intramuscular fat content showed significant differences among muscle where PM presented higher values (0.7%) than the other ones. The heme-iron content also presented significant differences ranging between 1.5 and 2.4 mg/100 g fresh meat. ST was the lightest, while TB and PM presented a more intense redness. The muscles that had a greater capacity to hold water were ST and BF, which presented lower drip loss (2 and 2.2%) and cooking loss (17.3 and 17.2%), respectively. Textural traits established the following scale in response to the tenderness: LD > PM > ST > TB > BF > SM. Finally, sensory analyses revealed that color and texture traits (hardness, juiciness and fibrousness) were significantly influenced by muscle type.     

Skeletal muscle fiber type and myofibrillar proteins in relation to meat quality

Although numerous studies have reported the relationships among muscle fiber characteristics, lean meat content and meat quality, controversial perspectives still remain. Conventional histochemical classifications may be involved in a high level of error, subjectivity and it could not clearly explain variety of myofibrillar protein isoforms. Therefore, more information is needed on how different factors, such as species, breeds, gender, nutrient conditions, physiological state of animals, and environment factors, affect ultimate meat quality in order to evaluate these uncertainness. Unfortunately, there is little information that completely covers with relationship among the muscle fiber types, myofibrillar proteins and enzymatic proteolysis. In addition to the perspective of postmortem metabolism, protein quality control in skeletal muscle and proteolytic degradation of muscle proteins during postmortem period could help to clarify this relationship. Therefore, the present review will focus on muscle fiber types, typing methods, muscle proteins and meat quality, and will summarize aspects of enzymatic view of proteasome.     

Impact of carcass scalding and chilling on muscle proteins and meat quality of broiler breast fillets

The objective of this study was to determine the effects of broiler carcass scalding and chilling methods on meat quality and muscle proteins. During processing, carcasses were hard (60 °C, 1.5 min) or soft (52.8 °C, 3 min) scalded, and either immersion chilled (IC: 0.5 °C, 40 min) or air chilled (AC: 0.5 °C, 120 min). Breast fillets were deboned at 4 h postmortem and used for measuring meat quality and muscle protein characteristics. Scalding by chilling treatment interaction effects on meat quality were not observed. Air chilled carcasses had greater pH₂₄, and reduced drip loss and shear force compared to IC carcasses. Cook yield, color (L*a*b*), and moisture content were not different between chilling treatments. Scalding treatments did not influence quality traits. Sarcoplasmic protein solubility was not influenced by chilling treatment, but was greater in hard versus soft scalded carcasses. Myofibrillar protein solubility was greater in fillets from soft scalded IC carcasses. Alterations in the electrophoretic profiles of the myofibrillar and sarcoplasmic proteins due to treatments indicated minor changes in protein degradation and solubility. Data suggest that while only chilling method influenced meat quality, both scalding and chilling methods influenced protein solubility and degradation in breast fillets deboned 4 h postmortem.     

Water distribution and mobility in meat during the conversion of muscle to meat and ageing and the impacts on fresh meat quality attributes--a review

This paper reviews current knowledge on the distribution and mobility of water in muscle (myowater) ante- and post mortem and factors affecting these in relation to fresh meat quality parameters; water-holding capacity (WHC), tenderness and juiciness. NMR transverse relaxometry (T(2)) using bench-top Low-Field Nuclear Magnetic Resonance (LF-NMR) has characterised myowater distribution and mobility as well as structural features in meat which directly affect WHC. The current literature demonstrates that WHC is correlated to the water located outside the myofibrillar network (extra-myofibrillar). This review identifies the critical stages which affect the translocation of water into the extra-myofibrillar space and thus the potential for decreased WHC during proteolysis (the conversion of muscle to meat). This review discusses how the intrinsic properties of the water held within the meat could contribute to juiciness and tenderness. Tenderness has been shown to correlate to T(2), however breed and species differences made it difficult to draw firm conclusions. Further understanding of the inherent water properties of fresh meat and the factors affecting water distribution and mobility using NMR technologies will increase the understanding of WHC and tenderisation of fresh meat.     

Prediction of the time evolution of pH in meat

In this paper we compare experimental data on pH decline in carcasses and predictions using a model, based on earlier work of Vetharaniam and coworkers. This model is extended in order to cope with the varying temperatures in the slaughterhouse. We have measured initial glycogen, and the pH and temperature at multiple times in the production chain. We have obtained good comparison between model predictions and our measurements. Furthermore, the correlation between initial glycogen content and ultimate pH as predicted by the model, follows closely experimental data reported earlier in literature. Being able to predict pH decline and ultimate pH, one can obtain reliable indications of final meat quality.     

Mitochondrial respiratory activity in porcine longissimus muscle fibers of different pig genetics in relation to their meat quality

The pig genetics of Duroc, Pietrain (MHS homozygote negative, PiNN), Pietrain (MHS homozygote positive, PiPP) and a F2-Duroc-Pietrain cross-breed were analyzed. The animals had comparable (P > 0.05) carcass weights, but the PiPP pigs had higher carcass yield and lean meat values (P < 0.05). Considering the meat quality characteristics, the PiPP showed a faster pH drop and higher electrical conductivity, drip loss, shear force as well as lightness and redness values (P < 0.05). The PiPP animals had less slow-twitch-oxidative (STO) and more fast-twitch-glycolytic (FTG) muscle fibers, whereas the results of the Duroc animals were converse (P < 0.05). The STO and FTG fibers of the PiPP animals were larger than those of the other genetics (P < 0.05). The analysis of the mitochondrial respiratory activity (MRA) using permeabilized longissimus muscle fibers resulted in no differences between the pig genetics before and immediately after slaughter. During chilling the MRA decreased in all pigs but to a higher extent in the PiPP pigs (P < 0.05).     

Effects of dietary daidzein supplementation of pregnant sows on carcass and meat quality and skeletal muscle cellularity of the progeny

The effects of dietary daidzein during late gestation on maternal performance, neonatal body composition, carcass and meat quality at market weight, and skeletal muscle cellularity were studied. Multiparous sows received a soybean-free diet (n=8, control) or the same diet supplemented with 1mg daidzein/kg body weight (n=7) daily from d 85 of gestation to parturition. Litter size, litter weight, and birth weight remained unaffected by daidzein feeding. In newborn piglets from litters >15, the proportions of muscle tissue and skin tended to be decreased (P=0.09) or increased (P=0.03), respectively, after gestational daidzein feeding. The body fat percentage was higher in response to maternal daidzein (P=0.04). Postnatal growth and carcass composition at 180d of age were not affected, however, maternal daidzein supplementation led to increases in longissimus muscle pH(45) (P=0.02) and pH(end) (P=0.11) in pigs from large litters. The proportion of fast-twitch glycolytic fibres in semitendinosus muscle was increased (P=0.06) by maternal daidzein feeding. The results suggest that supplemental daidzein in the maternal diet during late gestation marginally affects meat quality and skeletal muscle cellularity of the progeny.     

Muscle and meat quality characteristics of Holstein and Salers cull cows

Muscle characteristics and sensory rating of meat were determined in M. longissimus thoracis (LT), M. semimembranosus (SM), M. semitendinosus (ST) and M. triceps brachii (TB) from seven Holstein (HO, dairy breed) and six Salers (SA, beef breed) cull cows slaughtered at 6–7 years of age at the same fat score. Significant differences (P < 0.001) among muscle types were observed: ST was the more glycolytic and TB the more oxidative; total collagen: ST > SM = TB > LT; initial and overall tenderness: LT > TB = SM > ST, juiciness: TB > LT = SM > ST. Flavour differed only between breeds: HO > SA (P < 0.01). Three tenderness classes (high, intermediate, low) were determined from scores for sensory overall tenderness for all 52 meats: the lower total and insoluble collagen contents, the more oxidative metabolism, the more tender was the meat. Muscle type, and not breed explained most of the variability of meat quality from dairy and beef cull cows slaughtered at the same age and fat score.     

Biochemistry of postmortem muscle — Lessons on mechanisms of meat tenderization

It is certain that meat tenderness is a highly valued consumer trait and thus definition of the multiple processes that influence meat tenderness will provide clues toward improving meat quality and value. The natural process by which meat becomes tender is complex. Tenderness development is dependent on the architecture and the integrity of the skeletal muscle cell and on events that modify those proteins and their interaction. Specifically protein degradation and protein oxidation have been identified as processes that modify proteins as well as the tenderness of meat. The intracellular environment is a major factor that controls these events. Ultimately, the interplay between these events determines the rate and extent of tenderization. Given the intricacy of the structure of the muscle cell, coupled with the complexity of the regulation of protein modification and the ever-changing intracellular environment it is not surprising that this area of research is a very dynamic field. Just as the overall integrity and function of muscle cells does not depend on a single protein, but rather on the coordinated interaction of several proteins, the structural weakening of muscle cells during postmortem aging also must not depend on the degradation of a single myofibrillar or other cytoskeletal protein. The proteins mentioned in this review are located in different regions of the muscle cell, and most have been implicated in some manner as being important in maintaining the structure and function of the muscle cell. Oxidation of myosin heavy chain, a predominant protein in the myofibril, is known to promote aggregation and toughening of meat. Degradation of proteins such as desmin, filamin, dystrophin, and talin (all located at the periphery of the Z-line) may disrupt the lateral register and integrity of the myofibril themselves as well as the attachments of the peripheral layer of myofibril to the sarcolemma. Degradation of the proteins within the myofibril that are associated with the thick and thin filaments may allow lateral movement or breaks to occur within the sarcomeres of postmortem aged samples. Titin, nebulin, and troponin-T, by their ability to directly interact with, or modulate the interaction between, major proteins of the thick and thin filaments and (or) the Z-line, play key roles in muscle cell integrity. Disruption of these proteins, especially titin and nebulin, could initiate further physicochemical and structural changes that result in myofibril fragmentation and loss of muscle cell integrity, and ultimately in tenderization of the muscle. In order to make real progress in this area, the scientific community must have a global appreciation of how both the structural proteins and the key proteases are influenced by the vast changes that occur during the conversion of muscle to meat.     

Small heat shock proteins and their role in meat tenderness: A review

The eating quality of meat is a result of complex interactions between the biological traits and biochemical processes during the conversion of muscle to meat. It was hypothesised that muscles inevitably engage towards apoptotic cell death due to the termination of oxygen and nutrient supply to the muscle following exsanguination. Thus, factors that regulate the process of apoptotic cell death of muscle cells are believed to ultimately influence meat quality. Proteomic studies have associated the regulation of small heat shock proteins (sHSPs) with various meat quality attributes including tenderness, colour, juiciness and flavour. Due to the anti-apoptotic and chaperone functions of sHSPs, they are proposed to be involved with the eating quality of meat. In this review, we discuss the possible chaperone and anti-apoptotic role of sHSPs during the conversion of muscle to meat and consider the repercussions of this on the development of meat tenderness.     

Consequences of divergent selection for residual feed intake in pigs on muscle energy metabolism and meat quality

Selection to decrease Residual Feed Intake (RFI) is a relevant way to improve feed efficiency in growing pigs. However, RFI criterion is correlated with body composition and muscle characteristics. Present study evaluated adaptive responses to divergent selection on RFI on muscle metabolism and homeostasis through AMP-activated protein kinase pathway. Consequences on technological and sensory meat quality were also analyzed in two lines of Large White pigs after six generations of divergent selection on RFI.     

A review of meat protein hydrolysates and hypertension

We tested the hypothesis that meat is a source of peptides that are effective at preventing and reducing chronic lifestyle-related diseases (CLSRDs) such as hypertension. This analysis reflects the importance of empowering hypertensive people in quality versus quantity of life issues, and in offering nutritional treatment options rather than medical alternatives. For both hypertensive and normotensive individuals, chemically based medications may have harmful side effects. Functional food rich in antioxidant vitamins, and proteins or biologically active peptides, can lower blood pressure in persons with essential hypertension, possibly by preventing an underlying cause of the condition. Deficiency in consumption of crucial nutrients such as proteins from meat origins, along with abnormalities in carbohydrate and fat metabolism, may underlie the etiology of the clinical course of hypertension. Food derived from meat rich in nutrients may provide physiologically functional peptides, as well as improve digestion, and the metabolism of carbohydrate and fats, thus lowering blood pressure, and normalizing associated biochemical and histopathological changes.     

Relationships between biochemical characteristics and meat quality of Longissimus thoracis and Semimembranosus muscles in five porcine lines

Five porcine genetic lines which represent a high proportion of European pig production were fully characterized for meat quality parameters and muscle biochemical characteristics (Longissimus thoracis, LT, and Semimembranosus, SM). The line characterisation was based on 100 animals each representing Large White, Landrace, Duroc, Piétrain (Halothane negative) and Meishan (a Meishan/Large White crossbred line) prevalent genetic backgrounds. Different meat quality parameters (pH 45min, pH ultimate, electrical conductivity, and colour measurements), as well as muscle water holding capacity, muscle metabolic and contractile traits, fibre type, size and frequencies were measured and their relationships studied. The main differences in the LT were found between the Meishan and Piétrain genetic lines, in relation to the muscle fibre size (larger in Piétrain). The Duroc line was characterized by the muscle oxidative traits and the Landrace by the high percentage of fast glycolytic fibres. In SM, Duroc and Piétrain were distinguished from Landrace and Meishan according to the metabolic and contractile characteristics of this muscle. Large White tended to lie between the other breeds for many of the traits. The measured muscle characteristics were related to differences in drip loss and marbling values and could thereby influence the eating quality of pork. Overall the results show differences between the genetic lines for a number of muscle traits which could have impact on consumer appeal and eating quality. The present findings should serve to emphasise the importance of including eating quality as a trait in breed selection.     

Relationship between muscle antioxidant status, forms of iron, polyunsaturated fatty acids and functionality (retail colour) of meat in lambs

The relationship between muscle vitamin E, forms of iron, polyunsaturated fatty acids (PUFA) and the redness of meat (retail display) at days 3 to 4 post slaughter from lambs offered 2 different diets was examined. Meat redness was positively related to vitamin E and heme iron and negatively related to total n-3, total n-6 and total PUFA content. However, after adjusting for the effects of vitamin E and heme iron content, there was no indication of any residual relationship between redness at days 3-4 of retail display and total n-3, total n-6 or total PUFA. This indicates that the relationship between PUFA and redness in meat is mediated through the effects of heme iron and vitamin E in the muscle. It appears that the level of highly oxidisable PUFAs in muscle tissues do not play a major role in maintenance of redness at days 3-4 of retail display, but the level of vitamin E and heme iron content are important.     

Effects of different lairage times after long distance transportation on carcass and meat quality characteristics of Hungarian Simmental bulls

The objective of this study was to determine the effects of three lairage times (24 h, 48 h and 72 h) on the meat quality of tame trained to lead Hungarian Simmental bulls subjected to long commercial transportation of approximately 1800 km. A total of 30 bulls, with an average age of 24 months, were used. During the lairage, bulls received 0.5 kg concentrate feed per animal per day and ad libitum access to the hay and water. As the lairage duration increased, the pH_ult decreased (P < 0.05). Bulls lairaged for 24 h had lower L*, b* and H* values than those lairaged for 48 h and 72 h (P < 0.05). The effect of lairage time on WHC, cooking loss and Warner–Bratzler shear force values was not significant. The b* value was considered the best predictor of muscle pH_ult. In conclusion, 72 h quiet lairage time is recommended after transportation in order to prevent the adverse effects of transportation on meat quality.     

Explaining the variation in the shear force of lamb meat using sarcomere length, the rate of rigor onset and pH

The temperature when the pH=6.0 (temp@pH6) impacts on the tenderness and eating quality of sheep meat. Due to the expense, sarcomere length is not routinely measured as a variable to explain variation in shear force, but whether measures such as temp@pH6 are as useful a parameter needs to be established. Measures of rigor onset in 261 carcases, including the temp@pH6, were evaluated in this study for their ability to explain some of the variation in shear force. The results show that for 1 day aged product combinations of the temp@pH6, the pH at 18 °C and the pH at 24 h provided a larger reduction (almost double) in total shear force variation than sarcomere length alone, with pH at 24 h being the single best measure. For 5 day aged product, pH at 18 °C was the single best measure. Inclusion of sarcomere length did represent some improvement, but the marginal increase would not be cost effective.     

The influence of fiber size distribution of type IIB on carcass traits and meat quality in pigs

The effects of detailed characteristics such as the size and proportion of type IIB fibers in longissimus thoracis muscle on carcass traits and pork quality were investigated. A total of 96 pigs were classified into four groups by the proportion of different IIB fiber sizes. Group NS (high proportion of both small- and normal-sized IIB fibers) had a higher total number (136.4) and density (231.31) of type IIB fibers, backfat thickness (37.20 mm) and intramuscular fat content (4.77%) than the other groups (P < 0.05), whereas Group NS had the lowest values of cross-sectional area (3413.85 μm²) and diameter (60.15 μm) of type IIB fiber among the groups (P < 0.05). Pig muscles with higher percentage of large IIB fibers exhibit tougher, lighter and more exudative meat than pig muscles with a higher proportion of small- or normal-sized IIB fibers. Therefore, an increase in the proportion of large IIB fibers causes poor quality of pork.