Markers For Tenderness Research Articles

Hepatic Transcriptome Analysis Reveals Genes, Polymorphisms, and Molecules Related to Lamb Tenderness.

Tenderness is a key meat quality trait that determines the public acceptance of lamb consumption, so genetic improvement toward lamb with higher tenderness is pivotal for a sustainable sheep industry. However, unravelling the genomics controlling the tenderness is the first step. Therefore, this study aimed to identify the transcriptome signatures and polymorphisms related to divergent lamb tenderness using RNA deep sequencing. Since the molecules and enzymes that control muscle growth and tenderness are metabolized and synthesized in the liver, hepatic tissues of ten sheep with divergent phenotypes: five high- and five low-lamb tenderness samples were applied for deep sequencing. Sequence analysis identified the number of reads ranged from 21.37 to 25.37 million bases with a mean value of 22.90 million bases. In total, 328 genes are detected as differentially expressed (DEGs) including 110 and 218 genes that were up- and down-regulated, respectively. Pathway analysis showed steroid hormone biosynthesis as the dominant pathway behind the lamb tenderness. Gene expression analysis identified the top high (such as TP53INP1, CYP2E1, HSD17B13, ADH1C, and LPIN1) and low (such as ANGPTL2, IGFBP7, FABP5, OLFML3, and THOC5) expressed candidate genes. Polymorphism and association analysis revealed that mutation in OLFML3, ANGPTL2, and THOC5 genes could be potential candidate markers for tenderness in sheep. The genes and pathways identified in this study cause variation in tenderness, thus could be potential genetic markers to improve meat quality in sheep. However, further validation is needed to confirm the effect of these markers in different sheep populations so that these could be used in a selection program for lamb with high tenderness.

Relationship between protein markers and the sensory/physicochemical parameters of ovine meat during refrigerated storage

Description of the subject. Commercialization requires the storage of meat for a period of time according to the distribution and supply chain, usually under refrigeration and vacuum packaging. During this stage, many biochemical reactions can modify the sensory and physicochemical properties of meat. Proteolysis is one of the most important of these reactions as proteins are the building blocks of muscle, and their degradation affects tenderness. Objectives. The objectives of this study were to use multiple factorial analyses to correlate the physicochemical, sensory, and proteolytic changes in ovine meat during refrigeration. Method. Each loin was separated and randomly assigned to day 3, 5, or 14. Sensory panel evaluated the meat for appearance, odor and texture parameters. Warner-Bratzler shear force and texture profile analysis were evaluated. Similarly, protease activity was evaluated. Samples for 2D-electrophoresis and western blotting were collected at days 3, 5, and 14 post mortem. The results were analyzed by multiple factorial analyses. Results. The relative protein levels of desmin, vinculin, and myosin were found to decrease with time. Multiple factor analysis showed that the Warner-Bratzler shear force (WBSF) was highly correlated with the relative desmin concentration, the relative vinculin concentration, and acidic protease activity. The activities of acidic and neutral proteases were positively correlated with myosin level on day 3, but were negatively correlated with manual tenderness and myosin level on days 5 and 14. Conclusions. Therefore, desmin, myosin and vinculin, which correlate with sensory and physicochemical parameters during refrigerated storage, may prove useful as markers for tenderness.

Determination of Protein Markers for Beef Tenderness in U.S. Select Beef

ObjectivesBeef tenderness is a primary quality feature that defines the consumer’s perception of the product quality. Therefore definition of chemical characteristics that contribute to variation in tenderness within a specific USDA grade can help refine our understanding of meat tenderness. The objective of this experiment was to identify novel protein markers for tenderness in Longissimus dorsi sarcoplasmic fractions utilizing proteomics and a valuable library of beef samples. Materials and MethodsSteaks (USDA Select grade) used in this trial were classified as low shear force (LSF, n = 12; 11.3 ± 1.6 kg) and high shear force (HSF, n = 12; 34.3 ± 4.7 kg), according to their slice shear force (SSF, kg) measurements at 14 d postmortem. Western blotting of whole-muscle samples was used to determine troponin-T degradation (indicator of overall proteolysis). Sarcoplasmic protein samples from d 2 postmortem were prepared for two-dimensional difference in gel electrophoresis (2D-DIGE). Samples from LSF and HSF were sorted according to SSF values and were matched to a pooled internal reference (representative of all samples). Protein samples were loaded (75 µg) on 24 cm strips, pH 6 to 9, and focused in immobilized pH gradient in the first dimension. For the second dimension the strips were run on 12.5% acrylamide gels (n = 12). After 2D-DIGE, protein spots found to be different in relative abundance between HSF and LSF samples, and with other proteins of interest, were identified using Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer (LC-MS/MS). Immunoblots were performed to confirm protein identity. ResultsThe LSF samples exhibited greater troponin-T degradation (as evidenced by abundance of a 30 kDa fragment) than the HSF samples (LSF ratio to reference was 1.8, HSF ratio to reference was < 0.01; P < 0.01). A total of 31 spots showed a significant difference (P < 0.1) in relative abundance across groups (HSF vs. LSF). Among these, 18 spots were identified as 8 different proteins (Table 1). Proteins that were more abundant in the LSF samples are metabolic enzymes involved in the glycolytic pathway (glyceraldehyde-3-phosphate dehydrogenase, fructose bisphosphate aldolase A). Among the six proteins that showed greater abundance in the HSF samples, four belong to the oxidative energy metabolism pathways (malate dehydrogenase, creatine kinase M-type, myoglobin, carbonic anhydrase 3), one is involved in redox pathways (peroxiredoxin-1) and another one in protein turnover (polyubiquitin). The greater abundance of spots corresponding to oxidative metabolism enzymes suggests that aerobic metabolism may decrease potential for postmortem proteolysis. Table 1Differentially abundant proteins in high and low slice shear force groups Protein% CoverageRatio (LSF/HSF)P-valueGlyceraldehyde-3-phosphate dehydrogenase (GAPDH)26.731.980.060Fructose-bisphosphate aldolase A62.361.79< 0.050Polyubiquitin (Fragment)21.22–1.180.100Malate dehydrogenase, mitochondrial63.31–1.18< 0.050Creatine kinase M-type70.87–1.190.080Myoglobin99.35–1.290.095Carbonic anhydrase 379.12–1.56< 0.010Perociredoxin-170.85–1.240.055ConclusionThe results demonstrate that variation in tenderness within U.S. Select Top loin steaks is due in large part to variation in proteolysis. The variation in tenderness and proteolysis in these defined groups may be partially explained by a greater abundance of metabolic proteins involved in oxidative metabolism. This project was funded in part by grants from the Beef Checkoff (Denver, CO) and the Iowa Beef Industry Council (Ames, IA). The scholarship for the first author was granted by CNPq-Brazil.

Determination of Protein Markers for Beef Tenderness in U.S. Select Beef

ObjectivesBeef tenderness is a primary quality feature that defines the consumer’s perception of the product quality. Therefore definition of chemical characteristics that contribute to variation in tenderness within a specific USDA grade can help refine our understanding of meat tenderness. The objective of this experiment was to identify novel protein markers for tenderness in Longissimus dorsi sarcoplasmic fractions utilizing proteomics and a valuable library of beef samples. Materials and MethodsSteaks (USDA Select grade) used in this trial were classified as low shear force (LSF, n = 12; 11.3 ± 1.6 kg) and high shear force (HSF, n = 12; 34.3 ± 4.7 kg), according to their slice shear force (SSF, kg) measurements at 14 d postmortem. Western blotting of whole-muscle samples was used to determine troponin-T degradation (indicator of overall proteolysis). Sarcoplasmic protein samples from d 2 postmortem were prepared for two-dimensional difference in gel electrophoresis (2D-DIGE). Samples from LSF and HSF were sorted according to SSF values and were matched to a pooled internal reference (representative of all samples). Protein samples were loaded (75 µg) on 24 cm strips, pH 6 to 9, and focused in immobilized pH gradient in the first dimension. For the second dimension the strips were run on 12.5% acrylamide gels (n = 12). After 2D-DIGE, protein spots found to be different in relative abundance between HSF and LSF samples, and with other proteins of interest, were identified using Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer (LC-MS/MS). Immunoblots were performed to confirm protein identity. ResultsThe LSF samples exhibited greater troponin-T degradation (as evidenced by abundance of a 30 kDa fragment) than the HSF samples (LSF ratio to reference was 1.8, HSF ratio to reference was < 0.01; P < 0.01). A total of 31 spots showed a significant difference (P < 0.1) in relative abundance across groups (HSF vs. LSF). Among these, 18 spots were identified as 8 different proteins (Table 1). Proteins that were more abundant in the LSF samples are metabolic enzymes involved in the glycolytic pathway (glyceraldehyde-3-phosphate dehydrogenase, fructose bisphosphate aldolase A). Among the six proteins that showed greater abundance in the HSF samples, four belong to the oxidative energy metabolism pathways (malate dehydrogenase, creatine kinase M-type, myoglobin, carbonic anhydrase 3), one is involved in redox pathways (peroxiredoxin-1) and another one in protein turnover (polyubiquitin). The greater abundance of spots corresponding to oxidative metabolism enzymes suggests that aerobic metabolism may decrease potential for postmortem proteolysis. Table 1Differentially abundant proteins in high and low slice shear force groups Protein% CoverageRatio (LSF/HSF)P-valueGlyceraldehyde-3-phosphate dehydrogenase (GAPDH)26.731.980.060Fructose-bisphosphate aldolase A62.361.79< 0.050Polyubiquitin (Fragment)21.22–1.180.100Malate dehydrogenase, mitochondrial63.31–1.18< 0.050Creatine kinase M-type70.87–1.190.080Myoglobin99.35–1.290.095Carbonic anhydrase 379.12–1.56< 0.010Perociredoxin-170.85–1.240.055ConclusionThe results demonstrate that variation in tenderness within U.S. Select Top loin steaks is due in large part to variation in proteolysis. The variation in tenderness and proteolysis in these defined groups may be partially explained by a greater abundance of metabolic proteins involved in oxidative metabolism. This project was funded in part by grants from the Beef Checkoff (Denver, CO) and the Iowa Beef Industry Council (Ames, IA). The scholarship for the first author was granted by CNPq-Brazil.

Integration of genomic information into beef cattle and sheep genetic evaluations in Australia

Genomic information has the potential to change the way beef cattle and sheep are selected and to substantially increase genetic gains. Ideally, genomic data will be used in combination with pedigree and phenotypic data to increase the accuracy of estimated breeding values (EBVs) and selection indexes. The first example of this in Australia was the integration of four markers for tenderness into beef cattle breeding values. Subsequently, the availability of high-density single nucleotide polymorphism (SNP) panels has made selection using genomic information possible, while at the same time creating significant challenges for genetic evaluation with regard to both data management and statistical modelling. Reference populations have been established in both the beef cattle and sheep industries, in which an extensive range of phenotypes have been collected and animals genotyped mainly using 50K SNP panels. From this information, genomic predictions of breeding value have been developed, albeit with varying levels of accuracy. These predictions have been incorporated into routine genetic evaluations using three approaches and trial results are now available to breeders. In the first, genomic predictions have been included in genetic evaluation models as additional traits. The challenges with this method have been the construction of consistent genetic covariance matrices, and a significant increase in computing time. The second approach has been to use a selection index procedure to blend genomic predictions with existing EBVs. This method has been shown to produce very similar results, and has the advantage of being simple to implement and fast to operate, although consistent genetic covariance matrices are still required. Third, in sheep a single-step analysis combining a genomic relationship matrix with a standard pedigree-based relationship matrix has been used to estimate breeding values for carcass and eating-quality traits. It is likely that this procedure or one similar will be incorporated into routine evaluations in the near future. While significant progress has been made in implementing methods of integrating genomic information in both beef and sheep evaluations in Australia, the major challenges for the future will be to continue to collect the phenotypes needed to derive accurate genomic predictions, and in managing much larger volumes of genomic data as the number of animals genotyped and the density of markers increase.

Functional analysis of beef tenderness

Meat tenderness represents a complex assembly of different cellular pathways. As a consequence, genomics studies have revealed many different proteins considered as tenderness markers. So it is difficult to have an overview of tenderness in terms of cellular pathways. In this work cellular pathways of tenderness were analyzed, an interactome of 330 proteins was designed, and explanations of tenderization processes were proposed.

Effects of genetic markers and implant strategy on longissimus and gluteus muscle tenderness of calf-fed steers and heifers1

Effects of genotype (GEN) and implant program (IMP) on LM and gluteus muscle (GM) tenderization were investigated using crossbred steer (n = 185) and heifer (n = 158) calves. The 3-marker GeneSTAR Tenderness panel [CAST (calpastatin), CAPN1 316 (µ-calpain), and CAPN1 4751 (µ-calpain)] was used to determine the GEN of each animal (reported as total number of favorable alleles, 0 through 6). Calves were randomly assigned to 1 of 2 IMP, conventional (CNV) or delayed. Cattle in the CNV group were implanted at the beginning of the finishing period with Revalor-IS or Revalor-IH (Intervet Inc., Millsboro, DE), and then reimplanted 59 d later with Revalor-S or Revalor-H (Intervet Inc.). Calves in the delayed group received a single terminal implant (Revalor-S or Revalor-H) administered 45 d after initiation of the finishing period. Warner-Bratzler shear force (WBSF) was measured on LM and GM steaks at 3, 7, 14, 21, and 28 d postmortem. No interactions between the main effects of sex, IMP, or GEN were detected (P > 0.05) for WBSF. An IMP × postmortem aging (age) interaction was detected (P < 0.05) for LM and GM WBSF. For both muscles, steaks from CNV cattle had WBSF values that were approximately 0.2 kg greater (P < 0.05) than the values for steaks from delayed animals, but only during the early postmortem period (3 to 7 d). A linear effect of GEN on WBSF was detected (P < 0.05) for LM and GM steaks. Within each muscle, steaks from cattle with 6 favorable alleles had WBSF values 0.33 kg less than the values for steaks from cattle with 1 favorable allele. The GEN × age interaction was not significant for LM (P = 0.14) or GM (P = 0.20), but a numerical trend was observed for the effect of GEN on WBSF to diminish as age increased. To investigate how genetic markers could be interfaced with current beef carcass quality grading, cattle were sorted into 2 gene marker groups (GMG), ≤3 vs. ≥4 favorable alleles. For both muscles, GMG was effective only at identifying tenderness differences within the Select grade. When aged ≤14 d, Select LM steaks from cattle with ≥4 alleles had smaller (P < 0.05) WBSF values than did LM steaks from animals with ≤3 alleles. Preslaughter factors (sex, IMP, and GMG) controlled in the present study each accounted for less than 7% of the explained variation in tenderness of the test population. Results from this study suggest that the 3 GeneSTAR Tenderness markers were associated with small differences (0.33 kg) in WBSF and may be useful for increasing the consistency of Select beef, but these specific markers accounted for only a minor amount of variation in beef tenderness.

Production and processing studies on calpain-system gene markers for tenderness in Brahman cattle: 1. Growth, efficiency, temperament, and carcass characteristics1

Experiments were conducted concurrently at 2 locations to quantify effects and interactions of calpain-system tenderness gene markers on growth, efficiency, temperament, and carcass traits of Brahman cattle. Cattle were selected at weaning from commercial and research herds based on their genotype for commercially available calpastatin (CAST) and calpain 3 (CAPN3) gene markers for beef tenderness. Genotypes for mu-calpain gene markers (CAPN1-4751 and CAPN1-316) were also determined and included in statistical analyses. The New South Wales (NSW) herd was composed of 82 heifers and 82 castrated male cattle with 0 or 2 favorable alleles for CAST and CAPN3. The Western Australia (WA) herd was composed of 173 castrated male cattle with 0, 1, or 2 favorable alleles for CAST and CAPN3. One-half of the cattle at each site were implanted with a hormonal growth promotant (HGP: Revalor-H) during grain finishing. Cattle were backgrounded at pasture for 6 to 8 mo and grain-fed for 117 d (NSW) or 80 d (WA) before slaughter. Individually, or in combination with each other and with CAPN1-4751 status, CAST and CAPN3 status had no significant (all P > 0.05) effects on BW, growth, feed efficiency, or temperament traits. The only significant effect of CAST or CAPN3 on carcass characteristics was a small increase in rib fat with increasing number of favorable CAST alleles (P = 0.042) in the WA herd. There were no significant interactions (all P > 0.05) between the markers, or between the markers and sex or HGP treatment apart from CAST x HGP for area of the M. longissimus lumborum (P = 0.024) in the NSW experiment. Favorable CAST or CAPN3 alleles appear unlikely to have detrimental effects on growth, efficiency, temperament, or carcass characteristics of Brahman cattle; however, some effects evident for CAPN1 status indicate the need for further production studies on effects of these markers. Overall, the findings of the present study indicate that calpain-system gene markers are suitable for use in marker-assisted selection to improve meat tenderness in Brahman cattle without negative effects on other production and carcass characteristics.

Production and processing studies on calpain-system gene markers for tenderness in Brahman cattle: 2. Objective meat quality1

Effects and interactions of calpain-system tenderness gene markers on objective meat quality traits of Brahman (Bos indicus) cattle were quantified within 2 concurrent experiments at different locations. Cattle were selected for study from commercial and research herds at weaning based on their genotype for calpastatin (CAST) and calpain 3 (CAPN3) gene markers for beef tenderness. Gene marker status for mu-calpain (CAPN1-4751 and CAPN1-316) was also determined for inclusion in statistical analyses. Eighty-two heifer and 82 castrated male cattle with 0 or 2 favorable alleles for CAST and CAPN3 were studied in New South Wales (NSW), and 143 castrated male cattle with 0, 1, or 2 favorable alleles for CAST and CAPN3 were studied in Western Australia (WA). The cattle were backgrounded for 6 to 8 mo and grain-fed for 117 d (NSW) or 80 d (WA) before slaughter. One-half the cattle in each experiment were implanted with a hormonal growth promotant during feedlotting. One side of each carcass was suspended from the Achilles tendon (AT) and the other from the pelvis (tenderstretch). The M. longissimus lumborum from both sides and the M. semitendinosus from the AT side were collected; then samples of each were aged at 1 degrees C for 1 or 7 d. Favorable alleles for one or more markers reduced shear force, with little effect on other meat quality traits. The size of effects of individual markers varied with site, muscle, method of carcass suspension, and aging period. Individual marker effects were additive as evident in cattle with 4 favorable alleles for CAST and CAPN3 markers, which had shear force reductions of 12.2 N (P < 0.001, NSW) and 9.3 N (P = 0.002, WA) in AT 7 d aged M. longissimus lumborum compared with those with no favorable alleles. There was no evidence (all P > 0.05) of interactions between the gene markers, or between the hormonal growth promotant and gene markers for any meat quality traits. This study provides further evidence that selection based on the CAST or CAPN3 gene markers improves meat tenderness in Brahman cattle, with little if any detrimental effects on other meat quality traits. The CAPN1-4751 gene marker also improved beef tenderness without affecting other objective meat quality traits in heterozygous cattle compared with homozygotes for the unfavorable allele.

Estimated gene frequencies of GeneSTAR markers and their size of effects on meat tenderness, marbling, and feed efficiency in temperate and tropical beef cattle breeds across a range of production systems1

The objectives of this study were to use genotypes for 12 commercially available GeneSTAR gene markers on 12,330 animals to estimate gene frequencies of the markers across a range of beef cattle breeds and to determine the effects of these markers on target traits using a subset of animals with both genotypic and phenotypic data (n = 9,414) for at least one trait. Tenderness markers (T1, T2, T3, T4) were assessed against shear force of 2 muscles, marbling markers (M1, M2, M3, M4) were assessed against intramuscular fat percent and marbling score, and the feed-efficiency markers (N1, N2, N3, N4) were assessed on daily feed intake and residual feed intake. Animals used were from 5 beef cattle research populations: Beef Cooperative Research Centre 1 (CRC1) temperate breeds (Angus, Hereford, Murray Grey, Shorthorn; n = 3,721), Beef CRC1 tropical breeds (Brahman, Santa Gertrudis, Belmont Red; n = 3,899), Beef CRC2 tropically adapted genotypes (Brahman and Tropical Composite; n = 4,446), and progeny test programs in Angus (n = 742) and Shorthorn (n = 347). Gene frequencies varied significantly across breeds and markers, with 86% of the markers estimated to be in Hardy-Weinberg equilibrium. Tenderness markers T1 and T2 had significant effects (P < 0.0001) on shear force, with the size and direction of effects consistent across the range of breeds for the 3 populations with phenotypes. However, sizes of marker effects differed across muscles and reduced upon tenderstretch hanging. Marker T3 was not significant (P > 0.05) in CRC1 temperate breeds but was significant (P < 0.0001) in tropically adapted breeds, with a large effect in Brahman. Marker T4 was significant for shear force in 2 CRC1 tropical breeds but with a different favorable allele. The 4 marbling markers were generally not significantly associated with intramuscular fat percent or marbling score across the 5 populations studied. Feed-efficiency markers N3 and N4 were significantly associated with residual feed intake and daily feed intake in the CRC1 temperate data set, in which a subset of the CRC1 data was used in their discovery, but were not significant in the other 4 populations. Markers N1 and N2 were generally not significant, but when they were significant, their direction of effects differed. The 12 GeneSTAR markers were studied in populations consisting of different breeds and genetic variability and showed gene frequencies and estimated effects that varied greatly across traits, suggesting large differences between the markers for their utility as selection tools in these populations.

La révolution génomique au service de la filière viande bovine

Genomics has brought with it a true biological revolution and has a great number of potential applications in all areas of life sciences. In animal sciences, it will become possible to improve the production of milk or meat according to consumer demands. Concerning food safety, molecular tools have already been commercialized for the detection of contaminants in meat. Genetic markers i.e. DNA markers, have also been marketed to detect the origins of individual bovine animals and their breed. For the genetic selection of cattle, DNA markers likely to be employed for the selection of tenderness or marbling traits of beef have been patented by Australian scientists. Several ongoing projects in France, Europe and the USA aim at studying the relevance of these markers and finding new ones. In muscle biology, tracking gene expression with DNA chips or by physically separating the expressed proteins has led to the identification of genes that play an important role in muscle development. These genes could in turn serve as new biological markers for tenderness, marbling or rearing practices. Large-scale projects both publicly and privately funded, in Europe, the South Sea Islands, and especially in North America aim at exploiting to a maximum new advances in genomics in favour of beef production.

A new single nucleotide polymorphism in CAPN1 extends the current tenderness marker test to include cattle of Bos indicus, Bos taurus, and crossbred descent1

The three objectives of this study were to 1) test for the existence of beef tenderness markers in the CAPN1 gene segregating in Brahman cattle; 2) test existing CAPN1 tenderness markers in indicus-influenced crossbred cattle; and 3) produce a revised marker system for use in cattle of all subspecies backgrounds. Previously, two SNP in the CAPN1 gene have been described that could be used to guide selection in Bos taurus cattle (designated Markers 316 and 530), but neither marker segregates at high frequency in Brahman cattle. In this study, we examined three additional SNP in CAPN1 to determine whether variation in this gene could be associated with tenderness in a large, multisire American Brahman population. One marker (termed 4751) was associated with shear force on postmortem d 7 (P < 0.01), 14 (P = 0.015), and 21 (P < 0.001) in this population, demonstrating that genetic variation important for tenderness segregates in Bos indicus cattle at or near CAPN1. Marker 4751 also was associated with shear force (P < 0.01) in the same large, multisire population of cattle of strictly Bos taurus descent that was used to develop the previously reported SNP (referred to as the Germplasm Evaluation [GPE] Cycle 7 population), indicating the possibility that one marker could have wide applicability in cattle of all subspecies backgrounds. To test this hypothesis, Marker 4751 was tested in a third large, multisire cattle population of crossbred subspecies descent (including sire breeds of Brangus, Beefmaster, Bonsmara, Romosinuano, Hereford, and Angus referred to as the GPE Cycle 8 population). The highly significant association of Marker 4751 with shear force in this population (P < 0.001) confirms the usefulness of Marker 4751 in cattle of all subspecies backgrounds, including Bos taurus, Bos indicus, and crossbred descent. This wide applicability adds substantial value over previously released Markers 316 and 530. However, Marker 316, which had previously been shown to be associated with tenderness in the GPE Cycle 7 population, also was highly associated with shear force in the GPE Cycle 8 animals (P < 0.001). Thus, Marker 316 may continue to be useful in a variety of populations with a high percentage of Bos taurus backgrounds. An optimal marker strategy for CAPN1 in many cases will be to use both Markers 316 and 4751.

Proteomics approach as a new way to predict tenderness as compared to the classical South African Beef Carcass Classification System

The current South African Beef Carcass Classification System classifies carcasses using physical attributes such as age, fatness code and the conformation of the carcass as the only indicator of tenderness, implying that optimal tender meat is obtained from carcasses from animals with no permanent teeth. Research shows that if we take modern technologies (use of beta-agonists, growth hormones and electrical stimulation amongst others) into account, this is not necessarily true. In this study m. longissimus lumborum samples were collected from Nguni breed animals, snap frozen and stored at −80 ºC for further use in search of protein markers that can be used to predict tenderness in a non-invasive manner. Proteins were extracted in 1 mL TES buffer, isoelectrically focused on strips (pH 5 - 7) and separated by 2D SDS PAGE, stained with coomassie brilliant blue G250) and imaged by a Chemi-doc Mp imager. The results showed differences in protein expression, e.g. some proteins are present at 1 hour post mortem (Day 0) but absent at 3 days post mortem (Day 3). Other proteins showed an increase in expression as ageing progresses. These proteins correlated with myofibril fragments and Warner Bratzler changes which were measured. There are several factors that may be responsible for these changes in differential expression during ageing such as proteolytic action (i.e. calpains and calpastatin system, cathepsins) or maybe apoptosis. The proteins in the profiles of Nguni breed animals which showed differential expression in response to ageing are still to be determined.Keywords: Nguni breed, protein markers for tenderness, two dimensional gel electrophoresis (2D SDS PAGE)