Biological tenderness differences between longissimus muscles (LM) from Bos indicus and Bos taurus breeds were evaluated. Steers and heifers of Hereford x Angus (H x A, n = 10), 3/8 Sahiwal x H, A or H x A (3/8 SAH, n = 6) and 5/8 Sahiwal x H, A or H x A (5/8 SAH, n = 11) crosses were utilized. Muscle temperature and pH were monitored every 3 h for the first 12 h and at 24 h. Samples were obtained within 1 h and at 24 h postmortem from the LM for determination of calcium-dependent protease (CDP) -I and -II and CDP inhibitor (INH) activities. At 1 and 14 d postmortem, LM samples were removed for determining cathepsin B and B + L activity, soluble and total collagen, sarcomere length, muscle-fiber histochemistry, shear force and sensory-panel traits. Data were analyzed using least squares procedures with fixed effects of breed cross, sex and their interaction. No significant breed cross effects were observed for carcass traits or rates of pH and temperature decline. Steaks from H x A had lower (P less than .05) shear-force values and higher (P less than .05) sensory scores for tenderness at 1 and 14 d postmortem than steaks from 3/8 and 5/8 SAH. Correspondingly, 5/8 SAH had lower (P less than .05) myofibril fragmentation indices than H x A at 1, 3, 7 and 14 d postmortem. Breed cross effects were not significant for sarcomere length, fiber types, soluble and total collagen, cathepsin B and B + L specific activity, CDP-I and -II activity and INH activity within 1 h postmortem. However, INH total activity/100 g of muscle was greater (P less than .01) at 24 h postmortem for 5/8 SAH (208.8 +/- 14.8) and 3/8 SAH (195.6 +/- 19.3) than for H x A (136.3 +/- 14.9). For H x A, SDS-PAGE revealed that by d 1 desmin had been subjected to proteolysis, and by d 14 desmin could not be detected, but a 30,000-dalton component was clearly evident. However, in 5/8 SAH, desmin remained visible at d 14 without a 30,000-dalton component appearing. This reduced protein hydrolysis may account for less tender meat in SAH; INH apparently influences this process.

Postmortem proteolysis in skeletal muscle and factors affecting this process were examined in pork, lamb and beef longissimus muscles (LM) to determine the cause of differences in meat tenderness among these species. Fat thickness differed among species in the following order: pork greater than beef greater than lamb. The following patterns were observed for rate of temperature and pH decline: lamb greater than pork greater than beef and pork greater than beef greater than lamb, respectively. At 1 d postmortem, pork was the most tender, followed by beef and lamb, respectively. Between 1 and 14 d of postmortem storage, lamb LM was the most improved in tenderness, followed by beef and pork, respectively. Species did not differ (P greater than .05) in LM collagen solubility. Pork LM from fed pigs had the highest (P less than .05) level of cathepsins B + L and cystatin(s) activities, whereas no differences (P greater than .05) were observed among the species for cathepsin B activity. The lowest (P less than .01) Ca2(+)-dependent protease (CDP)-II and CDP inhibitor activities were observed in pork LM. Beef LM had the highest CDP inhibitor activity (P less than .05) but was intermediate in CDP-II activity. No differences were observed among species for CDP-I activity. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of myofibrils isolated at 0, 1 and 14 d postmortem indicated that by d 1, desmin hydrolysis was most extensive in pork muscle, followed by lamb and beef.(ABSTRACT TRUNCATED AT 250 WORDS)

Spring-born purebred Brahman bull calves (n = 467) with known pedigrees, sired by 68 bulls in 17 private herds in Louisiana, were purchased at weaning from 1996 through 2000 to study variation in growth, carcass, and tenderness traits. After purchase, calves were processed for stocker grazing on ryegrass, fed in a south Texas feedlot, and processed in a commercial facility. Carcass data were recorded 24 h postmortem. Muscle samples and primal ribs were taken to measure calpastatin activity and shear force. An animal model was used to estimate heritability, genetic correlations, and sire EPD. Relatively high heritability estimates were found for BW at slaughter (0.59 +/- 0.16), HCW (0.57 +/- 0.15), LM area (0.50 +/- 0.16), yield grade (0.46 +/- 0.17), calpastatin enzyme activity (0.45 +/- 0.17), and carcass quality grade (0.42 +/- 0.16); moderate heritability estimates were found for hump height (0.38 +/- 0.16), marbling score (0.37 +/- 0.16), backfat thickness (0.36 +/- 0.17), feedlot ADG (0.33 +/- 0.14), 7-d shear force (0.29 +/- 0.14), and 14-d shear force (0.20 +/- 0.11); relatively low heritability estimates were found for skeletal maturity (0.10 +/- 0.10), lean maturity (0.00 +/- 0.07), and percent KPH (0.00 +/- 0.07). Most genetic correlations were between -0.50 and +0.50. Other genetic correlations were 0.74 +/- 0.27 between calpastatin activity and 7-d shear force, 0.72 +/- 0.25 between calpastatin activity and 14-d shear force, (0.90 +/- 0.30 between yield grade and 7-d shear force, and -0.82 +/- 0.27 between backfat thickness and 7-d shear force. Heritability estimates and genetic correlations for most traits were similar to estimates reported in the literature. Sire EPD ranges for carcass traits approached those reported for sires in other breeds. The magnitude of heritability estimates suggests that improvement in carcass yield, carcass quality, and consumer acceptance traits can be made within the Brahman population.

Our objective was to determine the predictive value of various biochemical and histological traits for tenderness of the longissimus muscle. Data collected from 27 crossbred cattle included longissimus pH, temperature, sarcomere length, total and percentage of soluble collagen, muscle-fiber type and area, cathepsin B and B + L activities, calcium-dependent protease (CDP)-I, -II and inhibitor activities, myofibril fragmentation indices (MFI), Warner-Bratzler shear (WBS) force, sensory-panel tenderness (SPT) ratings and carcass traits. Stepwise regression analyses were performed among breeds or pooled within breeds with WBS and SPT as dependent variables. When MFI were included in the analysis, MFI at d 7 explained 50% of the variation in WBS and SPT at d 14. An additional 19% of SPT was accounted for by the addition of CDP inhibitor d 1 activity and percentage-area of alpha R fibers to the model. However, because variation in MFI was not significant within breed subclasses and MFI could be classified more as a dependent variable, it was removed from the model. This resulted in CDP inhibitor d 1 activity explaining 44% of the variation in WBS and SPT at d 14. Also, percentage-area of beta R fibers, 6 h pH and cathepsin B + L d 14 activity appeared in the model. In addition, CDP inhibitor activity was the only variable to be significant within breed groups. These data suggest that d 7 MFI could be used as a single predictor of d 14 longissimus muscle tenderness; however, CDP inhibitor d 1 activity (a biological event) also may be useful in predicting tenderness.

Equations were developed to predict beef longissimus dorsi (LD) tenderness after postmortem refrigerated aging. Warner-Bratzler shear force (WBSF) and myofibril fragmentation indices (MFI) were determined'at 1, 3, 7 and-14 days postmortem on LD of Angus-Hereford (AH. n = 8) and 5/8 Brahman crossbred (n = 8) heifer carcasses. Correlation coefficients between WBSF and MFI w&e -0.91, -0.74, -0.63, and -0.40 at 1, 3, 7 and 14 days postmortem, respectively. None of the traits measured correlated significantly with 1Cday WBSF (P > 0.051. A three-variable mediction eauation that included 24-hr calcium-dependent protease (CDP) inhibitor activity, 0-hr CDP-I activity and 24-hr cystatin activity accounted for 63% of the variation in 14-day WBSF.

The objectives of this study were to examine the amount of postmortem proteolysis in three different lamb muscles at different ages and to determine whether a relationship exists between the extent of myofibrillar degradation and certain endogenous proteinase activities. Wether lambs were slaughtered at 8 (n = 6) and 26 (n = 6) wk of age. Samples were taken from the longissimus (LM), gluteus medius (GM), and supraspinatus (SS) muscles for determining myofibrillar fragmentation index (MFI) at 1 and 7 d postmortem, cathepsins B and B + L, cystatin(s), mu- and m-calpain and calpastatin 24-h activities, and muscle fiber type and area. Muscle samples were removed for SDS-PAGE analysis at 0, 1, and 7 d postmortem. The SS muscle consisted of more (P less than .05) oxidative fibers, whereas no age effects (P greater than .10) were observed for fiber type. The younger lambs had higher (P less than .01) cathepsins B and B + L, cystatin, and calpastatin 24-h activities but less (P less than .01) m-calpain activity than did the 26-wk-old lambs. Within each age, the SS muscle had the highest (P less than .05) mu-calpain, m-calpain, and calpastatin specific activities. The MFI values and SDS-PAGE results indicate that less proteolysis occurred in the SS muscle. Samples from 26-wk-old lambs tended to have greater MFI values at 1 and 7 d postmortem, especially for the LM and GM muscles. From these results, it seems that less postmortem proteolysis occurs in younger and more oxidative muscles and this may be attributed to the greater calpastatin 24-h activity.

A study was conducted to examine the effects that physiological levels of m-calpain (calpain requiring millimolar concentrations of Ca2+) extract and a lysosomal extract have on myofibrillar proteins in vitro, and the effects that zinc has on inhibiting proteolysis by these extracts. During a 22-h incubation period, the lysosomal extract degraded myosin heavy chain, alpha-actinin, desmin, troponin-I, and myosin light chains 1 and 2. The effectiveness of the lysosomal extract to degrade myofibrillar proteins was significantly affected by the presence or absence of EDTA. Zinc, which is a potent inhibitor of cysteine proteinases, prevented most, but not all, of the lysosomal extract-induced myofibrillar protein degradation. Incubation of myofibrils with m-calpain resulted in the hydrolysis of troponin-T, desmin, and a 58-kDa molecular weight protein, possibly vimentin, and 5 mM ZnCl2 completely blocked these changes. Results from this study indicate that the degradation by the lysosomal extract is far more extensive than the degradation that occurs with normal postmortem storage and that possibly a non-cysteine protease is present that is capable of hydrolyzing some myofibrillar proteins under this in vitro condition, because Zn2+ did not block all proteolysis. However, similar changes were induced by m-calpain incubation and postmortem storage.

Because freezing samples decreases calpastatin activity and the application of exogenous calcium activates the calpain proteolytic system, thereby improving tenderness, the objective of this study was to determine whether freezing would enhance the effects of CaCl2 marination on the tenderness of beef steaks. Longissimus steaks were obtained from 10 beef steers 6 d postmortem. One-half of the steaks were frozen at -30 degrees C for 6 wk. The remaining steaks were treated fresh; one-half were subjected to a 150 mM CaCl2 marinade for 48 h. Frozen steaks were thawed and subjected to the same treatment. Treatments consisted of 1) fresh control, 2) fresh marinated, 3) frozen control, and 4) frozen marinated. Samples were taken before and after treatment (6 and 8 d) for calpastatin activity determination and d 8 for SDS-PAGE. Warner-Bratzler shear force values were measured 8 d postmortem. Data were analyzed using a paired comparison t-test procedure. Results showed that freezing and marination significantly decreased calpastatin activity. A .35-kg improvement (P = .07) in Warner-Bratzler shear force was observed with freezing, whereas a .78-kg improvement (P less than .01) in tenderness was observed with marination. However, prior freezing enhanced the effects of marination. Therefore, the decrease in calpastatin activity seemed to allow greater proteolysis by the calpains with the application of Ca2+. The SDS-PAGE of myofibril preparations indicated that more small polypeptide fragments (28 to 32 kDa) appeared and a 95-kDa fragment was more intense in the marinated samples than in control samples, indicating that proteolysis was enhanced.(ABSTRACT TRUNCATED AT 250 WORDS)