Association of a single nucleotide polymorphism in the calpastatin gene with carcass and meat quality traits of beef cattle

Calpastatin (CAST) is a naturally occurring protein that inhibits the normal tenderization of meat as it ages postmortem. A SNP was identified in the CAST gene (a G to C substitution) and genotyped on crossbred commercially fed heifers (n = 163), steers (n = 226), and bulls (n = 61) from beef feedlots, and steers (n = 178) from a University of Guelph feeding trial. The association of the CAST SNP with carcass and meat quality traits was studied. Carcass traits included fat, lean, and bone yield; grade fat; LM area; and HCW. Meat quality traits included marbling grade; i.m. fat content of LM; tenderness evaluation of LM (Warner-Bratzler shear force) at 2, 7, 14, and 21 d of postmortem aging; and tenderness evaluation of semitendinosus muscle at 7 d of postmortem aging. The mixed model used in the analyses included fixed effects of CAST genotype, sex, slaughter group, and breed composition (linear covariate); sire was a random effect. For the analysis of shear force, i.m. fat content of LM was also included in the model as a linear covariate. Shear force measures were analyzed within days of postmortem aging and by repeated measures analysis. The CAST SNP allele C was more frequent (63%) in the crossbred population than allele G. The CAST SNP was associated with shear force across days of postmortem aging (P = 0.005); genotype CC yielded beef that was more tender than GG (-0.32 kg +/- 0.13), and CG had intermediate tenderness. The corresponding average allele substitution effect (G to C substitution) was also highly significant (-0.15 +/- 0.05 kg, P = 0.002). A lower percentage of unacceptably tough steaks (shear force > 5.7 kg) at 2 and 7 d postmortem was associated with an increasing number of C alleles (P < or = 0.05). At 7 d postmortem, the percentage of unacceptably tough steaks decreased by 24 and 35%, respectively, for animals carrying 1 and 2 copies of the C allele relative to animals with no C alleles. However, genotype CC had a greater fat yield (+1.44 +/- 0.56%; P = 0.037) than genotype GG, with a corresponding allele substitution effect of 0.67 +/- 0.27% (P = 0.015). Therefore, the CAST SNP allele C was associated with increased LM tenderness across days of postmortem aging and, importantly for the beef industry, had a significant reduction in the percentage of steaks rated unacceptably tough by consumers based on an assumed threshold level.     

Predicting beef tenderness using near-infrared spectroscopy

The objective of this multiple-phase study was to determine the accuracy of an on-line near-infrared (NIR) spectral reflectance system to predict 14-d-aged cooked beef tenderness. In phase I, 292 carcasses (140 US Select, 152 US Choice) were selected (d 2) from 2 commercial beef processing facilities. After carcass selection, longissimus lumborum (LL) muscle sections (ribs 9 to 12) were individually identified, vacuum-packaged, and transported to the Oklahoma State University Meats Laboratory, where a 2.54-cm-thick steak (n = 1) was fabricated and stored in refrigerated conditions (1 degrees C +/- 1). Following a 30-min oxygenation period, a NIR spectral scan was obtained on the 12th-rib LL steak. Steaks (d 3) were individually vacuum-packaged and aged at 4 degrees C for a total of 14 d before cooking slice shear force (SSF) analysis. In phases II and III, 476 carcasses (258 US Select, 218 US Choice) were immediately NIR scanned after carcass presentation to in-plant USDA grading personnel. In a similar fashion, all LL steaks were aged (1 degrees C +/- 1) for 14 d before cooking (70 degrees C) and conducting SSF. Of the phase I and II samples, 39 (6.77%) were categorized as being tough (i.e., >/= 25 kg of SSF after the 14-d postmortem aging period). Of these 39 tough samples, 20 (3.7% error rate) were correctly placed in the 90% certification level. Another 10 tough samples were placed in the 80% certification level (2.0% error rate). The overall NIR certified tender group was 1.67 kg more tender (P < 0.05) than LL samples from the noncertified samples. When the NIR predicted samples to be tough, 10% of the samples were eliminated from the phase I and II LL populations at 90% certification. The population SSF mean improved in excess of 6.5 kg. For phase III, SSF evaluation by an independent third party indicated the NIR system was able to successfully sort tough from tender LL samples to 70% certification levels. It was concluded that NIR scanning offers an in-plant opportunity to sort carcasses into tenderness outcome groups for guaranteed-tender branded beef programs.     

Effect of postmortem storage on activity of mu- and m-calpain in five bovine muscles

An in situ system involving incubation of 60- to 80-g pieces of muscle at 4 degrees C under different conditions was used to determine the effects of time of postmortem storage, of pH, and of temperature on activities of mu- and m-calpain activity in bovine skeletal muscle. Casein zymograms were used to allow measurement of calpain activity with a minimum of sample preparation and to ensure that the calpains were not exposed to ionic strengths of 100 or greater before assay of their activities. In 4 of the 5 muscles (longissimus dorsi, lumbar; longissimus dorsi, thoracic; psoas major; semimembranosus; and triceps brachii) studied, mu-calpain activity decreased nearly to zero within 48 h postmortem. Activity of m-calpain also decreased in the in situ system used but at a much slower rate. Activities of both mu- and m-calpain decreased more slowly in the triceps brachii muscle than in the other 4 muscles during postmortem storage. Although previous studies have indicated that mu-calpain but not m-calpain is proteolytically active at pH 5.8, these studies have used calpains obtained from muscle at death. Both mu- and m-calpain are proteolytically inactive if their activities are measured at pH 5.8 and after incubating the muscle pieces for 24 h at pH 5.8. Western analysis suggested that neither the large 80-kDa subunit nor the small 28-kDa subunit of m-calpain was autolyzed during postmortem storage of the muscle pieces. As has been reported previously, the 80-kDa subunit of mu-calpain was autolyzed to 78- and then to a 76-kDa polypeptide after 7 d postmortem, but the 28-kDa small subunit was not autolyzed; hence, the autolyzed mu-calpain molecule in postmortem muscle is a 76-/28-kDa molecule and not a 76-/18-kDa molecule as previously assumed. Because both subunits were present in the postmortem calpains, loss of mu-calpain activity during postmortem storage is not due to dissociation of the 2 subunits and inactivation. Although previous studies have shown that the 76-/18-kDa mu-calpain molecule is completely active proteolytically, it is possible that the 76-/28-kDa mu-calpain molecule in postmortem muscle is proteolytically inactive and that this accounts for the loss of mu-calpain activity during postmortem storage. Because neither mu- nor m-calpain is proteolytically active at pH 5.8 after being incubated at pH 5.8 for 24 h, other proteolytic systems such as the caspases may contribute to postmortem proteolysis in addition to the calpains.     

Properties of myofibril-bound calpain activity in longissimus muscle of callipyge and normal sheep

Properties of the calpain bound to myofibrils in longissimus muscle from callipyge or noncallipyge sheep were examined after 0, 1, 3, and 10 d of postmortem storage at 4 degrees C. Western analysis has shown that most of this calpain is mu-calpain, although the sensitivity of the antibodies used in the earlier studies could not eliminate the possibility that up to 10% of the calpain was m-calpain. The calpain is bound tightly, and very little is removed by washing with the detergent Triton X-100; hence, it is not bound to phospholipids in the myofibril. Over 25% of total mu-calpain was bound to myofibrils from at-death muscle, and this increased to approximately 40% after 1 d postmortem. The amount of myofibril-bound mu-calpain increased only slightly between 1 and 10 d of postmortem storage. The percentage of autolyzed mu-calpain increases with time postmortem until after 10 d postmortem, when all myofibril-bound mu-calpain is autolyzed. The specific activity of the myofibril-bound calpain is very low and is only 6 to 13% as high as the specific activity of extractable mu-calpain from the same muscle. It is unclear whether this low specific activity is the result of unavailability of the active site of the myofibril-bound calpain to exogenous substrate. The myofibril-bound calpain degrades desmin, nebulin, titin, and troponin T in the myofibrils, and also releases undegraded alpha-actinin and undergoes additional autolysis when incubated with Ca2+; all these activities occurred slowly considering the amount of myofibril-bound calpain. Activity of the myofibril-bound calpain was partly (58 to 67%) inhibited by the calpain inhibitors, E-64 and iodoacetate; was more effectively inhibited by a broader-based protease inhibitor, leupeptin (84 to 89%); and was poorly inhibited (43 to 45%) by calpastatin. Release of undegraded alpha-actinin and autolysis are properties specific to the calpains, and it is unclear whether some of the myofibril-bound proteolytic activity originates from proteases other than the calpains or whether the active site of myofibril-bound calpain is shielded from the inhibitors. Activities and properties of the myofibril-bound calpain were identical in longissimus muscle from callipyge and normal sheep, although previous studies had indicated that the "normal" longissimus was much more tender than the callipyge longissimus. Hence, it seems unlikely that the myofibril-bound calpain has a significant role in postmortem tenderization of ovine longissimus.     

Changes in caspase activity during the postmortem conditioning period and its relationship to shear force in porcine longissimus muscle

The objective of this study was to investigate the protease family caspases in skeletal muscle and their potential contribution to postmortem proteolysis and meat tenderization. Ten Large White gilts were slaughtered, and samples of LM were taken at 0, 2, 4, 8, 16, 32, and 192 h after slaughter and immediately snap frozen in liquid nitrogen. Samples were subsequently analyzed for caspase 3/7 and caspase 9 activity, protein levels of known caspase substrates, alpha II spectrin and poly (ADP-ribose) polymerase (PARP), as well as, at 192 h, shear force. Specific degradation products of alpha II spectrin and PARP, which are known indicators of caspase activity, and apoptosis were detected on immunoblots of muscle samples taken over the postmortem period. The relationships between the changes observed in caspase activities and protein levels of PARP and spectrin across the entire postmortem conditioning period were investigated (n = 70). Protein levels of alpha II spectrin cleavage products across the conditioning period were found to correlate positively to caspase 3/7 activity (r = 0.38, P = 0.003) and caspase 9 activity (r = 0.32, P = 0.012), indicating that caspase-mediated cleavage was occurring in situ. There was a negative relationship between shear force and the 0 to 32 h ratio of caspase 3/7 (r = -0.62, P = 0.053) and caspase 9 activities (r = -0.68, P = 0.044). In addition, there was also a negative relationship between shear force and the level of the caspase-generated alpha II spectrin 120 kDa degradation product (r = -0.75, P = 0.012). The findings of this study indicate that changes in caspase activity and caspase-mediated cleavage take place in muscle during the conditioning period, and this could be associated with the development of tender meat.     

Combined effect of epinephrine and exercise on calpain/calpastatin and cathepsin B and L activity in porcine longissimus muscle.

The objective of the study was to improve the understanding of the relationship between the effect of epinephrine plus exercise and meat tenderness. The calpain, calpastatin, and cathepsin B + L activities and postmortem proteolysis in porcine longissimus muscle were studied. The muscle glycogen stores were depleted in five pigs by s.c. injection of epinephrine (.3 mg/kg) at 15 h antemortem and exercise on a treadmill (5 min, 3.8 km/h) immediately before slaughter. Antemortem injection of epinephrine and treadmill exercise resulted in higher ultimate pH (6.32 vs 5.66 in control) and decreased (P < .05) thaw loss, cooking loss, and shear force values. The muscle energy depletion treatment increased (P < .05) the muscle mu-calpain activity measured 42 min postmortem, and at 24 h mu-calpain activity was still approximately 50% greater in the high ultimate pH group. Also, as the ratio of mu-calpain to calpastatin increased (P < .01), the overall proteolytic potential of the calpain system were greater. These observations suggest that the muscle energy level may influence the activity of the calpain system in the living animal. The high ultimate pH group showed lower (P < .001) cathepsin B + L activity in the myofibrillar and the soluble fractions after 8 d of storage, suggesting that the increased ultimate pH increased the stability of the lysosomal membrane and thereby reduced the release of cathepsins from the lysosomes during storage. The SDS-PAGE showed increased (P < .001) degradation of a 39-kDa band in the epinephrine and exercise-treated samples. Degradation products at 30, 31, and 32 kDa were labeled by troponin-T antibody in western blot. An appearing 24-kDa band was identified as a troponin-I degradation product in western blot. The proteolytic degradation pattern of myofibrillar proteins during storage differed in control and treated samples, supporting the hypothesis that calpain-mediated proteolysis was affected after treatment, resulting in meat with high ultimate pH.     

Effect of two dietary concentrate levels on tenderness, calpain and calpastatin activities, and carcass merit in Waguli and Brahman steers

The objective of this study was to compare carcass characteristics of a newly introduced breed, the Waguli (Wagyu x Tuli), with the carcass characteristics of the Brahman breed. Brahman cattle are used extensively in the Southwest of the United States because of their tolerance to adverse environmental conditions. However, Brahman carcasses are discounted according to the height of their humps because of meat tenderness issues. The Waguli was developed in an attempt to obtain a breed that retained the heat tolerance of the Brahman but had meat quality attributes similar to the Wagyu. Twenty-four animals were used. Six steers from each breed were fed a 94% concentrate diet and 6 steers from each breed were fed an 86% concentrate diet. Eight steers, 2 from each group, were harvested after 128 d, after 142 d, and after 156 d on feed. Waguli steers had larger LM, greater backfat thickness, greater marbling scores, and greater quality grades than the Brahman steers (P < 0.05). The Japanese Wagyu breed is well known for its highly marbled and tender meat, and these traits are also present in the Waguli. The Waguli had significantly lower Warner-Bratzler shear force values than the Brahman steers after 7 and 10 d of postmortem aging (P < 0.05); this difference decreased after 14 d postmortem (P = 0.2), when tenderness of the slower aging Brahman had increased to acceptable levels. Toughness of the Brahman has been associated with high levels of calpastatin in Brahman muscle, and the Waguli LM had significantly less calpastatin activity (P = 0.02) at 0 h postmortem than the Brahman LM. At 0-h postmortem, the total LM calpain activity did not differ between the Brahman and Waguli (P = 0.57). Neither diet nor days on feed had any significant effect on the 0-h postmortem calpain or at 0-h postmortem calpastatin activity, nor an effect on Warner-Bratzler shear-force values. In conclusion, LM muscle from the Waguli steers had a high degree of marbling, lower shear force values, and low calpastatin activity, all of which are related to more tender meat.     

Tenderness classification of beef: II. Design and analysis of a system to measure beef longissimus shear force under commercial processing conditions.

The objectives of this study were to evaluate the efficacy of a system for classifying beef for tenderness based on a rapid, simple method of measuring cooked longissimus shear force. Longissimus steaks (2.54 cm thick) were trimmed free of s.c. fat and bone and rapidly cooked using a belt grill. A 1-cm-thick, 5-cm-long slice was removed from the cooked longissimus parallel with the muscle fibers for measurement of shear force. Slices were sheared with a flat, blunt-end blade using an electronic testing machine. The entire process was completed in less than 10 min. Therefore, in commercial application, this process could be completed during the 10- to 15-min period that carcasses are normally held to allow the ribeye to bloom for quality grading. In Exp. 1, the repeatability of slice shear force (SSF), as determined by evaluation of duplicate samples from 204 A-maturity carcasses, was .89. In Exp. 2, A-maturity carcasses (n = 483) were classified into three groups based on SSF (< 23, 23 to 40, and > 40 kg) at 3 d postmortem that differed (P < .001) in mean trained sensory panel tenderness ratings (7.3 +/- .04, 6.4 +/- .06, and 4.4 +/- .20) and the percentages (100, 91, and 28%) of samples rated "Slightly Tender" or higher at 14 d postmortem. Therefore, this tenderness classification system could be used to accurately segregate beef carcasses into expected tenderness groups. Further research is needed to test the feasibility and accuracy of this system under a variety of commercial processing conditions.     

Altered calpain levels in longissimus muscle from normal pigs and heterozygotes with the ryanodine receptor mutation

The calpain proteolytic system was examined in the longissimus muscle (LD) of heterozygote pigs carrying a single copy of a mutation in the skeletal muscle ryanodine receptor gene (RyR1) that is associated with porcine stress syndrome and reduced meat quality. Conventional British White-type pigs (n = 30) were selected from a commercial line on the basis of slaughter weight, backfat depth, and pH at 45 min postmortem > 6.0; based on DNA analysis, 11 were heterozygous RyR1 mutants (Nn), and 19 were normal genotype (NN). The LD samples were taken from carcasses at 2, 4, and 24 h postmortem for calpain analysis with enzyme assay and immunoblotting, using specific antisera raised against recombinant polypeptides derived from calpain large subunits and calpastatin. Shear force (SF) was measured after conditioning for 8 d at 2 degrees C and did not differ between Nn and NN groups. The extractable activity of mu-calpain decreased over 24 h postmortem (P < .001), with no significant difference in activity between NN and Nn animals at any time. The activity of m-calpain also decreased with time (P < .001), but it was lower at all times in Nn than in normal genotypes (P < .001). After Western blotting, the immunoreactivity of mu- and m-calpain large subunit bands declined over 24 h postmortem (P < .001); values for mu-calpain were higher (P < .05) and for m-calpain were lower (P < .001) in heterozygotes than in normal animals at each sampling time. The calpastatin antibody detected a major band of 135 kDa that declined with time postmortem but did not differ between Nn and NN genotypes at any sampling time. These data indicate that the levels of extractable mu- and m-calpain, but not calpastatin, may be different in pigs that carry the RyR1 mutation.     

Using the near-infrared system to sort various beef middle and end muscle cuts into tenderness categories

The objectives of this study were to determine the effectiveness of a visible-near-infrared (VIS-NIR) system to predict the ultimate tenderness rating of various beef muscles and conclude if a relationship exists between predicted LM shear force and tenderness of other subprimal cuts. Carcasses (n = 768) were scanned with the VIS-NIR system in 2 commercial beef-processing facilities. Carcasses were categorized based on their predicted 14-d LM slice shear force value. After carcass scanning, 100 carcasses were randomly selected based on their tenderness classification, and subprimals (ribeye rolls, clods, knuckles, top sirloins, inside rounds, and eye of rounds) were removed, vacuum-packaged, and transported to the Oklahoma State University Food and Agricultural Products Research Center, where 2.54-cm steaks (n = 6) were fabricated and stored in refrigerated conditions (1 degrees C +/- 1) and aged for 14 d. The center steak from right-side subprimals was designated for slice shear force (LM) or Warner-Bratzler shear force (all other subprimals) analysis. The remaining steaks were categorized based on predicted tenderness taken at 2 d postmortem with the VIS-NIR spectrophotometer and used in a consumer taste study. The test population of carcasses (n = 100) scanned in-plant predicted 27 carcasses as tender, 45 carcasses as intermediate, and 28 carcasses as tough. The VIS-NIR system correctly classified 26 of the 28 (92.9% accuracy) tough carcasses. Overall consumer satisfaction was greatest (P < 0.05) for steaks classified as tender and was intermediate compared with the steaks classified as tough. It was concluded that in-plant VIS-NIR scanning can properly identify and sort carcasses into tenderness groups, which may lead to the development of certified not-tough programs.     

The effect of postmortem time of injection and freezing on the effectiveness of calcium chloride for improving beef tenderness.

Three experiments were conducted to determine the effect of freezing and time postmortem on the effectiveness of injecting CaCl2 to tenderize beef. In Exp. 1, longissimus muscle treatments included 1) control 0 h, 2) CaCl2-injected 0 h, 3) control 24 h, and 4) CaCl2-injected 24 h. Injection consisted of .3 M CaCl2 at 10% by weight. Injecting CaCl2 at 24 h postmortem reduced (P < .05) shear force requirements compared with the 24 h control but did not (P < .05) tenderize meat as much as injecting at 0 h. In Exp. 2, longissimus muscle treatments included the following: 1) aged 2 d; 2) aged 7 d; 3) frozen d 1, thawed, aged 6 d; 4) CaCl2-injected d 1, aged 6 d; 5) frozen d 1, thawed, CaCl2-injected, aged 6 d; and 6) CaCl2-injected d 1, frozen, thawed, aged 6 d. Injection alone at d 1 or freezing, then thawing and injecting resulted in the lowest (P < .05) shear force requirements. In Exp. 3, longissimus muscle treatments included the following: 1) aged 1 d; 2) aged 7 d; 3) CaCl2-injected 0 h, aged 7 d; 4) CaCl2-injected d 1, aged 6 d; 5) frozen d 1, thawed, aged 6 d; and 6) frozen, thawed, CaCl2-injected, aged 6 d. Both d-1 injection alone and freezing, thawing, then injecting resulted in meat with shear force requirements similar to those of 0-h injected meat. The effect of treatments on cooking loss was inconsistent. Treatments that reduced shear force also reduced (P < .05) calpain and calpastatin activity proportionately.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and characterization of mu-calpain, m-calpain, and calpastatin from postmortem muscle. I. Initial steps

Evidence has indicated that mu-calpain, m-calpain, and calpastatin have important roles in the proteolytic degradation that results in postmortem tenderization. Simple assays of these 3 proteins at different times postmortem, however, has shown that calpastatin and mu-calpain both rapidly lose their activity during postmortem storage, so that proteolytic activity of mu-calpain is nearly zero after 3 d postmortem, even when assayed at pH 7.5 and 25 degrees C, and ability of calpastatin to inhibit the calpains is 30% or less of its ability when assayed at death. m-Calpain, however, retains much of its proteolytic activity during postmortem storage, but the Ca(2+) requirement of m-calpain is much higher than that reported to exist in postmortem muscle. Consequently, it is unclear how the calpain system functions in postmortem muscle. To clarify this issue, we have initiated attempts to purify the 2 calpains and calpastatin from bovine semitendinosus muscle after 11-13 d postmortem. The known properties of the calpains and calpastatin in postmortem muscle have important effects on approaches that can be used to purify them. A hexyl-TSK hydrophobic interaction column is a critical first step in separating calpastatin from the 2 calpains in postmortem muscle. Dot-blot assays were used to detect proteolytically inactive mu-calpain. After 2 column chromatographic steps, 5 fractions can be identified: 1) calpastatin I that does not bind to an anion-exchange matrix, that does not completely inhibit the calpains, and that consists of small polypeptides <60 kDa; 2) calpastatin II that binds weakly to an anion-exchange matrix and that contains polypeptides <60 kDa; all these polypeptides are smaller than the native 115- to 125-kDa skeletal muscle calpastatin; 3) proteolytically active mu-calpain even though very little mu-calpain activity can be detected in zymogram assays of muscle extracts from 11- to 13-d postmortem muscle; this mu-calpain has an autolyzed 76-kDa large subunit but the small subunit consists of 24-, 26- and a small amount of unautolyzed 28-kDa polypeptides; 4) proteolytically active m-calpain that is not autolyzed; and 5) proteolytically inactive mu-calpain whose large subunit is autolyzed to a 76-kDa polypeptide and whose small subunit contains polypeptides similar to the proteolytically active mu-calpain. Hence, loss of calpastatin activity in postmortem muscle is due to its degradation, but the cause of the loss of mu-calpain activity remains unknown.     

Dose-response relationships between porcine somatotropin, muscle composition, muscle fiber characteristics and pork quality

The dose-dependent effects of porcine somatotropin (pST) on cellular aspects of skeletal muscle growth, muscle composition and measures of pork quality were investigated in growing barrows and gilts. Eighty crossbred pigs weighing 46 kg were assigned randomly to receive daily subcutaneous injections of 0, 30, 60, 120 or 200 micrograms pST/kg BW until they weighed 100 kg. Semitendinosus muscle weights were increased with pST dose (linear, P less than .001) by 21%. Percentage of type I and type II muscle fibers was not changed with pST, but cross-sectional area of type I and type II fibers was increased in parallel with muscle weight. Percentage of moisture increased (P less than .01) and percentage of lipid decreased (P less than .01) as pST dose increased. The pH of the longissimus 24 h postmortem increased (P less than .01) .1 to .2 units with increasing pST dose, but subjective evaluation for color, firmness and wateriness of the longissimus indicated no discernible treatment effect. Gardner color difference meter "Rd" and "A" values decreased (P less than .01) with a pST dose of 60 micrograms/kg or more, signifying a slightly darker and less red color, respectively, of the longissimus muscle. Weight loss of loin chops 2.54 cm thick cooked to 71 degrees C (20.3% to 23.7%) and shear force of cores 1.27 cm in diameter (2.89 to 3.76 kg) were not related to pST treatment or dose.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationships among glycolytic potential,dark cutting (dark,firm, and dry) beef,and cooked beef palatability

One hundred beef carcasses were selected at three packing plants and were used to determine the relationship between glycolytic potential (GP) and dark, firm, and dry (DFD) beef and to determine the effects of DFD status and GP on cooked beef palatability. Eight individual muscles were excised from one hindquarter of each carcass at d 7 postmortem: longissimus lumborum, psoas major, gluteus medius, tensor fasciae latae, rectus femoris, semimembranosus, biceps femoris, and semitendinosus. Ultimate pH, colorimeter readings, and Warner-Bratzler shear force were determined for all eight muscles at d 7 postmortem. A nine-member trained sensory panel evaluated cooked longissimus lumborum, gluteus medius, and semimembranosus steaks. Traits determined solely for the longissimus lumborum were GP (2 x [glycogen + glucose + glucose-6-phosphate] + lactate) and ether-extractable fat. A curvilinear relationship existed between GP and ultimate pH within the longissimus muscle. There appeared to be a GP threshold at approximately 100 micromol/g, below which lower GP was associated with higher ultimate pH and above which GP had no effect on ultimate pH. The greatest pH and muscle color differences between normal and DFD carcasses were observed in the longissimus lumborum, gluteus medius, semimembranosus, and semitendinosus muscles. Cooked longissimus from DFD carcasses had higher shear force values (46% greater) and more shear force variation (2.3 times greater variation) than those from normal carcasses. Dark cutting carcasses also had higher shear force values for gluteus medius (33% greater) and semimembranosus (36% greater) than normal carcasses. Sensory panel tenderness of longissimus, gluteus medius, and semimembranosus was lower for DFD carcasses than for normal carcasses. Longissimus and gluteus medius flavor desirability scores were lower for DFD than for normal carcasses. Steaks from DFD carcasses had more off-flavor comments than steaks from normal carcasses, specifically more "peanutty," "sour," and "bitter" flavors. The DFD effect of higher shear force values was approximately five times greater (+3.11 kg vs +0.63 kg) for carcasses with "slight" marbling scores than for carcasses with "small" marbling scores. In general, higher GP was associated with increased tenderness, even among normal carcasses. In conclusion, low GP was associated with DFD beef and resulted in substantially less-palatable cooked steaks.     

Changes in protein abundance between tender and tough meat from bovine longissimus thoracis muscle assessed by isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis analysis

The aim of this study was to find potential biomarkers for meat tenderness in bovine Longissimus thoracis muscle and to compare results from isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis (2-DE) analysis. The experiment included 4 tender and 4 tough samples, based on shear force measurements at 7 d postmortem, from young Norwegian red (NRF) bulls, taken at 1 h postmortem. A number of the proteins which have previously been related to tenderness were found to change in abundance between tender and tough samples, both in iTRAQ (P < 0.1) and 2-DE analysis (P < 0.05). Furthermore, 3 proteins that have not previously been related to tenderness were found to change significantly in abundance between tender and tough meat samples in the present study. These include proteins related to control of flux through the tricarboxylate cycle [2-oxoglutarate dehydrogenase complex component E2 (OGDC-E2)], apoptosis (galectin-1) and regulatory role in the release of Ca(2+) from intracellular stores (annexin A6). Even though the overlap in significantly changing proteins was relatively low between iTRAQ and 2-DE analysis, certain proteins predicted to have the same function were found in both analyses and showed similar changes between the groups, such as structural proteins and proteins related to apoptosis and energy metabolism.     

Technical note: comparison of myofibril fragmentation index from fresh and frozen pork and lamb longissimus

The myofibril fragmentation index (MFI) is strongly associated with indices of meat tenderness, such as Warner-Bratzler shear force and sensory tenderness. The MFI is normally determined on fresh muscle. It is not known whether this index can be determined on frozen muscle. The objective of this experiment was, therefore, to determine whether there is a difference between MFI values of fresh and frozen lamb and pork longissimus. To compare the effect of freezing on MFI, longissimus samples were obtained from eight lamb carcasses at 1, 3, and 15 d postmortem and longissimus samples were obtained from 12 pork carcasses at 3 d postmortem. For each sample, MFI was conducted on both fresh muscle and snap-frozen muscle (frozen in liquid nitrogen and stored 23 to 26 d at -70 degrees C). The R2 between MFI of fresh and frozen muscle was 0.94 and 0.92 for lamb and pork longissimus, respectively. The differences between fresh and frozen MFI were not significant for either species (P > 0.05). These results indicate that it is not necessary to determine MFI on fresh muscle.     

Calcium-activated tenderization of strip loin, top sirloin, and top round steaks in diverse genotypes of cattle

Steers of known percentage Brahman (B) and Angus (A) breeding (100% A, n = 6; F1 B x A, n = 6; and 100% B, n = 6) were used to determine the effect of calcium chloride injection on the calpain proteinase system and meat tenderness. The steers were slaughtered in six replications (at either 9 or 14 mm of backfat, determined ultrasonically), with each breed type represented. Calpains and calpastatin activities were measured on fresh, prerigor longissimus muscle samples. Carcass data were collected after a 24-h chill, and the short loin (IMPS #180), top sirloin (IMPS #184), and top round (IMPS #168) were removed from both sides of each carcass. The cuts from the right side were then injected at 5% (wt/wt) with CaCl2 solution (2.2%). Longissimus muscle calpain and calpastatin activities were also measured at 48 h postmortem from the injected and control sides of each carcass. Warner-Bratzler shear force was measured on steaks from the three subprimals aged 1, 2, 5, 15, or 31 d. Marbling scores and USDA quality grades were higher (P<.05) in A than in F1 B x A and B carcasses. Calpastatin activity was higher (P<.05) in muscle from B than in muscle from A and F1 B x A steers, and postmortem storage (O vs 48 h) and CaCl2 injection reduced (P<.05) the activity of the calpains and calpastatin. Strip loin and top sirloin steaks from A and F1 B x A steers were more tender (P<.05) than steaks from B steers; however, top round steak tenderness did not differ (P>.05) across breed type. Calcium injection improved strip loin and top sirloin steak tenderness, but it did not affect top round steak tenderness. Collectively, these data show that CaC12 injection can be used to improve meat tenderness, with similar responses shown in cattle containing 0, 50, and 100% B inheritance. However, even with CaCl2 injection, B steaks are less tender than their A and F1 B x A counterparts.