Effects of supplemental vitamin D3 on feed intake, carcass characteristics, tenderness, and muscle properties of beef steers.

Research was conducted to determine the effects of supplemental dietary vitamin D3 on DMI, carcass traits, Warner Bratzler shear (WBS) force, calpastatin activity, plasma minerals, pH (0, 3, 12, and 24 h after slaughter), water-holding capacity (WHC), and sensory characteristics of three muscles. Pre-slaughter vitamin D3 treatments included no supplemental vitamin D3, 6 x 106 IU (MIU) of vitamin D3 for 4 d, or 6 MIU of vitamin D3 for 6 d. Cattle were slaughtered and carcasses were chilled for 48 h before removal of steaks from the longissimus, gluteus medius, and biceps femoris muscles. Steaks were aged at 2 degrees C for 7, 14, or 21 d before cooking to a final internal temperature of 70 degrees C for WBS and sensory panel analysis. Dry matter intake was lower for steers supplemented with vitamin D3 for 4 or 6 d. Live and carcass weights were lower (P < 0.05) in steers supplemented with vitamin D3. Supplementing 6 MIU/6 d of vitamin D3 decreased (P < 0.05) WBS values of gluteus steaks (pooled over aging times). Longissimus steaks from steers supplemented with vitamin D3 for 6 d had lower (P < 0.05) WBS force values than these steaks from control steers or steers fed vitamin D3 for 4 d at 7 d postmortem. Biceps femoris steaks from steers receiving vitamin D3 for 4 d had higher WBS values than steaks from control steers at 14 and 21 d postmortem. Feeding vitamin D3 at 6 MIU for 6 d decreased (P < 0.05) the percentage of steaks that had WBS values > or = 3.86 kg for all steaks. Feeding vitamin D3 had no effect on palatability traits evaluated by trained panelists. Blood Ca concentrations were greater (P < 0.05) when vitamin D3 was fed and with increased vitamin D3 feeding time. Feeding vitamin D3 for 6 d (vs 4 d) delayed pH decline for all muscle types after 0, 3, and 12 h postmortem. Water-holding capacity was increased (P > 0.02) after 0 h, 24 h, and 21 d postmortem when vitamin D3 was fed and was greater at 0 and 24 h if vitamin D3 was fed for 6 d rather than 4 d. These data suggest that supplementing 6 MIU of vitamin D3 will decrease DMI and improve beef tenderness through increased blood plasma Ca concentrations and WHC.     

Immunoblot analysis of calpastatin degradation: evidence for cleavage by calpain in postmortem muscle.

A negative correlation exists between calpastatin activity and meat tenderness. Therefore, it is important to determine the mechanism of calpastatin inactivation in postmortem skeletal muscle. Western immunoblot analysis was performed to determine the protease(s) responsible for degradation of muscle calpastatin during postmortem storage. To accomplish this, purified calpastatin was digested with different proteases in vitro, and their pattern of calpastatin degradation was compared with that of calpastatin degradation in postmortem muscle. Polyclonal antibodies raised in mice against recombinant bovine skeletal muscle calpastatin were used to monitor calpastatin degradation. Lamb longissimus was stored at 4 degrees C and sampled at 0, 6, 12, 24, 72, 168, and 336 h postmortem. Postmortem storage produced a discrete pattern of calpastatin degradation products that included immunoreactive bands at approximately 100, 80, 65, 54, 32, and 29 kDa. Undegraded calpastatin (130 kDa) was barely detectable after 72 h of postmortem storage at 4 degrees C, and no immunoreactive calpastatin was observed by 336 h postmortem. For in vitro proteolysis, lamb longissimus calpastatin (0 h postmortem) was purified using Affi-Gel Blue chromatography. Calpastatin was digested with m-calpain, mu-calpain, cathepsin B, proteasome, trypsin, or chymotrypsin. Each of these enzymes degraded calpastatin. Immunoreactive fragments resulting from digestion of calpastatin with m- and mu-calpain were similar to each other and closely resembled those observed during postmortem aging of lamb longissimus at 4 degrees C. Digestion of calpastatin with mu-calpain reduced calpastatin activity. Degradation of calpastatin by other proteases resulted in unique patterns of immunoreactive fragments, distinct from that observed in longissimus. Thus, m- and(or) mu-calpain seem to be responsible for calpastatin degradation during postmortem storage of meat.     

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.     

Longissimus muscle quality, palatability and connective tissue histological characteristics of bulls and steers fed different energy levels and slaughtered at four ages

One hundred forty-four Angus male calves were assigned to one of five slaughter-age groups: 9, 12, 15, 18 and 24 mo of age. After weaning, one-half of the calves within slaughter age were castrated. One-half of the 12-, 15-, 18- and 24-mo age groups of each sex were further randomly allotted to a high-energy (HE) or low-energy (LE) finishing diet. Eight bulls and eight steers assigned to the 9-mo slaughter group were fed the HE diet only. Samples from the longissimus muscle (LM) were taken at the 12th rib for histological evaluations of fat deposits. Thin sections from the LM at the 11th rib were stained for lipid, collagen and elastin. Steaks from the LM were cooked and evaluated by a sensory panel and sheared by a Warner-Bratzler shear (WBS) device. The 9-10-11th rib sections were separated into muscle, fat and bone. Collagen, elastin and collagen solubility were determined chemically. Bulls had heavier carcasses, less fat and lower sensory scores for tenderness and juiciness than steers. Steers fed the HE diet had lower WBS values than bulls on either diet. Cooking losses were higher for bulls at 12, 18 and 24 mo than for steers. Histological evaluation showed that collagen content was higher for bulls than for steers and higher for cattle fed the LE diet than those fed the HE diet. Among cattle on the HE diet, the 9-mo cattle had more collagen than all older groups. Neither sex nor diet had an effect on elastin content shown by histological evaluation, but 12-mo cattle had the most elastin and the 15- and 18-mo cattle had the least elastin. Collagen content determined by chemical analysis was not affected by sex, diet or slaughter age. Correlation coefficients among histologically and chemically evaluated collagen and elastin and meat traits were too low to be of practical importance. The time-intensive histological techniques used in our study provided little additional information over that obtained from visually evaluated or chemically measured meat traits.     

Influence of zeranol implants and electrical stimulation on the palatability traits of five muscles in Angus and Limousin bulls and steers

A split-plot design was used to study effects of male status (bull vs steer), zeranol implants, breed types (Angus vs Limousin) and electrical stimulation on palatability traits of five major muscles when all animals were fed to a similar fat-thickness endpoint. Warner-Bratzler shear (WBS) determinations and taste-panel evaluations were performed on the longissimus, adductor, semimembranosus, semitendinosus and biceps femoris muscles removed 24 h postmortem. Breed was the most consistent main effect difference, with Angus cattle having lower WBS values than Limousin for the longissimus and semitendinosus muscles. Angus cattle also were favored in many sensory traits. Few significant differences were noted between bulls and steers in taste-panel traits. Differences due to electrical stimulation were most evident in the longissimus muscle, with higher panel-evaluation scores, an enhancement of marbling and lean color and a lower incidence of heat ring. The implant tended to have negative effects on palatability in this trial. An overall ranking of the five muscles based on WBS from most tender to least tender was: adductor, semimembranosus, longissimus, semitendinosus and biceps femoris. Correlations of palatability traits of one muscle to other muscles within an experimental unit were found to be low.     

Effect of the callipyge gene on muscle growth, calpastatin activity, and tenderness of three muscles across the growth curve

Changes in muscle growth, calpastatin activity, and tenderness of three muscles were assessed in 20 callipyge and 20 normal wether lambs slaughtered at live weights (LW) of 7, 20, 36, 52, and 69 kg. At 24 h postmortem, the longissimus (LM), semimembranosus (SM), and supraspinatus (SS) muscles were removed and weighed and samples were obtained for calpastatin activity (CA; 24 h) and Warner-Bratzler shear force (WBS; aged 6 d). For muscle weights and calpastatin activity, the weight group x muscle x phenotype interaction was significant (P < 0.05). Muscle weights were similar (P > 0.05) between phenotypes for all three muscles at 7 kg LW. At 20 kg LW, the LM and SM muscles from the callipyge lambs were heavier (P < 0.05) than those from normal lambs; however, the SS did not differ (P > 0.05) between phenotypes at 7, 20, or 52 kg. From 20 to 69 kg LW, the LM and SM weights were 42 and 49% heavier (P < 0.05) for callipyge than for normal lambs. Calpastatin activity of the callipyge LM was greater (P < 0.05) than that of normal LM at 36, 52, and 69 kg. In the callipyge LM, CA was similar (P > 0.05) at 20, 36, and 52 kg LW and did not differ (P > 0.05) from 7-kg or 69-kg values. Calpastatin activity declined (P < 0.05) across the growth curve for the SM and SS, but values were higher (P < 0.05) in the SM in callipyge than in normal lambs. Shear force values of the LM were lower (P < 0.05) for normal lambs at 36, 52, and 69 kg LW than for callipyge lambs. In the SM and SS, WBS values decreased (P < 0.05) across the growth curve, but values were higher (P < 0.05) for callipyge lambs in the SM only. These data indicate that the selective muscular hypertrophy of the callipyge phenotype develops during the postnatal growth period between 7 and 20 kg LW (19 and 100 d of age). Longissimus and semimembranosus muscles in the callipyge lambs were over 40% heavier from 20 to 69 kg LW; however, they also had higher levels of calpastatin activity and Warner-Bratzler shear force during this time period, indicating the need for postmortem tenderization treatments to improve palatability.     

Differences in cathepsin B + L and calcium-dependent protease activities among breed type and their relationship to beef tenderness

Activities of acidic proteases (cathepsin B + L) and neutral, calcium-dependent proteases (CDP) were quantified to determine whether differences in proteolytic activity could explain differences in meat tenderness among breed types. Steers (n = 32) of known percentage Angus (A) and Brahman (B) breeding were used to establish differences in meat tenderness (A; 3/4A-1/4B; 1/2A-1/2B; 1/4A-3/4B). Samples were removed from the longissimus muscle within 1 h postmortem and within 2 h were frozen for subsequent determination of cathepsin B + L, CDP-I, CDP-II and CDP-inhibitor activities. Warner-Bratzler shear (WBS) was assessed after 1, 5 and 10 d of postmortem aging. Taste panel evaluations, conducted on steaks that were subjected to 5 d of aging, detected no differences. At d 1, WBS did not differ among breed types; however, by d 10 of aging, steaks from Angus steers were more tender (P less than .05) than steaks from 1/2B and 3/4B steers. The Angus and 1/4B steaks had significantly more (P less than .05) cathepsin B + L activity than the 3/4B. The CDP had no relationship with WBS; however, CDP-inhibitor was positively related to d-1 WBS (r = .41, P less than .05). Cathepsin B + L activity was negatively related to WBS at d 10 (r = -.44, P less than .05). These data suggest that differences in meat tenderness among breed types may be explained partially by differences in proteolytic enzyme activity.     

Proportion of types I and III collagen in longissimus collagen from bulls and steers

The proportion of types I and III intramuscular collagen in longissimus muscles of Simmental bulls (n = 8) and steers (n = 8) 17 mo of age was studied. Longissimus samples taken 7 d after slaughter were evaluated for total collagen, types I and III collagen, heat-soluble collagen, sensory panel traits and Warner-Bratzler shear force. Intramuscular collagen (IMC) was isolated and digested with cyanogen bromide, and peptides were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Percentage of type III IMC was calculated from the total of types I and III collagen as determined from the peak area of densitometric scans of the cyanogen bromide peptides alpha 1(I)CB8 and alpha 1(III)CB8. Longissimus muscles from steers had lower (P less than .05) Warner-Bratzler shear values, less (P less than .05) sensory panel-detectable connective tissue and more (P less than .05) tender panel ratings for muscle fiber tenderness and overall tenderness. Muscles from steers had more (P less than .05) heat-soluble collagen than those from bulls, but no differences (P greater than .05) were found for total collagen and percentage of type III collagen. Some intramuscular-collagen characteristics may have contributed to the less tender muscle of bulls. However, the proportion of types I and III collagen did not account entirely for the tenderness difference between steer and bull muscles. Because there were differences in collagen solubility in muscles from steers and bulls, other collagen characteristics such as crosslinking or fiber size may have been more important than collagen type.     

The calpain system in three muscles of normal and callipyge sheep

Activities of mu- and m-calpain and of calpastatin were measured at four different times during postmortem storage (0, 1, 3, and 10 d) in three muscles from either callipyge or noncallipyge (normal) sheep. The weights of two muscles, the biceps femoris and the longissimus, are greater in the callipyge phenotype, whereas the weight of the infraspinatus is not affected. The activity of m-calpain was greater (P < 0.05) in the biceps femoris and longissimus from callipyge than in those from normal sheep, but it was the same in the infraspinatus in the two phenotypes. The extractable activity of m-calpain did not change (biceps femoris and infraspinatus) or decreased slightly (longissimus) during postmortem storage. Extractable activity of mu-calpain decreased to zero or nearly zero after 10 d postmortem in all muscles from both groups of sheep. The rate of decrease in mu-calpain activity was the same in muscles from the callipyge and normal sheep. At all time points during postmortem storage, calpastatin activity was greater (P < 0.05) in the biceps femoris and longissimus from the callipyge than from the normal sheep, but it was the same in the infraspinatus from callipyge and normal sheep. Calpastatin activity decreased (P < 0.05) in all three muscles from both phenotypes during postmortem storage; the rate of this decrease in the callipyge biceps femoris and longissimus and in the infraspinatus from both the callipyge and normal sheep was slow, especially after the first 24 h postmortem, whereas calpastatin activity in the biceps femoris and longissimus from the normal sheep decreased rapidly. During postmortem storage, the 125-kDa calpastatin polypeptide was degraded, but the 80-kDa subunit of mu-calpain was cleaved only to 76- and 78-kDa polypeptides even though extractable mu-calpain activity declined nearly to zero. Approximately 50 to 60% of total mu-calpain became associated with the nonextractable pellet after 1 d postmortem. The myofibril fragmentation index for the biceps femoris and longissimus from normal sheep increased significantly during postmortem storage. The fragmentation index for the infraspinatus from the callipyge and normal sheep increased to an intermediate extent, whereas the index for the biceps femoris and longissimus from the callipyge did not change during 10-d postmortem storage. The results suggest that postmortem tenderization is related to the rate of calpastatin degradation in postmortem muscle and that calpastatin inhibition of the calpains in postmortem muscle is modulated in some as yet unknown manner.     

Evaluation of the Tendertec beef grading instrument to predict the tenderness of steaks from beef carcasses

Four experiments were conducted, using carcasses from cattle identified for anticipated variability in tenderness (Exp. 1, 2, and 3) and carcasses selected for variability in physiological maturity and marbling score (Exp. 4), to evaluate the ability of the Tendertec Mark III Beef Grading Probe (Tendertec) to predict tenderness of steaks from beef carcasses. In Exp. 1, 2, and 3, longissimus steaks were aged for different periods of time, cooked to a medium degree of doneness (70 degrees C), and evaluated for Warner-Bratzler shear force (WBS) and trained sensory panel ratings. In Exp. 4, longissimus steaks were aged 14 d and cooked to 60, 65, 70, 75, or 80 degrees C for WBS tests and to 65 or 75 degrees C for sensory panel evaluations. Tendertec output variables were not correlated with 1) 24-h calpastatin activity, steak WBS (following 1, 4, 7, 14, 21, or 35 d of aging), or d-14 sensory panel tenderness ratings in Exp. 1 (n = 467 carcasses) or 2) 14-d WBS in Exp. 2 (n = 202 carcasses). However, in Exp. 3 (n = 29 carcasses), Tendertec output variables were correlated (P < 0.05) with tenderness of steaks aged 1, 21, 28, or 35 d, and we were able to separate carcasses into groups yielding tough, acceptable, and tender steaks. In Exp. 4 (n = 70), Tendertec output variables were correlated (P < 0.05) with steak WBS at 60 degrees C and with steak ratings for muscle fiber tenderness, connective tissue amount, and overall tenderness at 65 degrees C, but these relationships weakened (P > 0.05) as degree of doneness increased. Consequently, Tendertec output variables only were effective for stratifying carcasses according to tenderness when steaks from those carcasses in Exp. 4 were cooked to a rare or medium-rare degree of doneness. Although Tendertec was able to sort carcasses of older, mature cattle based on tenderness of steaks at some cooked end points, it failed to detect tenderness differences in steaks derived from youthful carcasses consistently, and was thus of limited value as an instrument for use in improving the quality, consistency, and uniformity of the U.S. fed-beef supply.     

Effect of vitamin D3 supplementation level on the postmortem tenderization of beef from steers

The objective of this experiment was to determine the effect of different doses of vitamin D3 (VITD) on beef feedlot performance, plasma and muscle Ca2+, tissue residues, and improvement of Warner-Bratzler shear force (WBS) and panel tenderness. A total of 167 steers were fed one of six levels of VITD. The VITD treatments (28 steers/treatment) were 0, 0.5 x 10(6), 1 x 10(6), 2.5 x 10(6), 5 x 10(6), and 7.5 x 10(6) IU/steer daily of VITD fed nine consecutive days before slaughter. Feedlot performance and plasma Ca2+ were measured during the last 21 days on feed. Warner-Bratzler shear force was measured on strip loin and top round steaks at 7, 10, 14, and 21 d postmortem. The VITD treatments of 5 and 7.5 x 10(6) IU/steer daily decreased (P < 0.05) ADG, and VITD supplementation of 2.5, 5, and 7.5 x 10(6) IU/steer daily decreased average dry matter feed intake (P < 0.05) at the end of the feeding trial. Plasma Ca2+ increased linearly with VITD treatment (P < 0.01). Calpastatin and calpain activity were not influenced by treatment (P > 0.05), but muscle Ca2+ was increased (P < 0.05) by VITD treatments of 1, 2.5, 5, and 7.5 10(6) IU/steer daily. Feeding VITD did not influence (P > 0.05) carcass quality or yield traits. Supplementing VITD at levels of 1, 2.5, 5, and 7.5 10(6) IU/steer daily increased (P < 0.05) VITD concentrations in strip loin and liver samples. Cooking liver decreased VITD concentrations 10 to 28%. Vitamin D3 treatments of 0.5 and 7.5 x 10(6) IU/d reduced strip loin steak WBS at d 7 (P < 0.05), but VITD treatments did not decrease strip loin steak WBS at any other time postmortem. The VITD treatments of 0.5, 1, and 5 x 10(6) IU/steer daily decreased top round steak WBS at 7 d, and all VITD treatments decreased 10-d top round steak WBS (P < 0.05). Supplementing steers with 0.5 x 10(6) IU/steer daily of VITD also decreased (P < 0.05) top round steak WBS at 21 d postmortem compared with controls. Sensory tenderness at 7 d postmortem was increased (P < 0.05) by all VITD treatments in top round steaks, yet strip loin tenderness scores were not affected (P > 0.05) by VITD treatment. Treatment with VITD quadratically decreased (P < 0.05) round WBS. Thus, VITD treatment will effectively improve tenderness when cattle tend to be tough and have no impact on cattle that produce tender beef. Feeding steers 0.5 x 10(6) IU of VITD daily for 9 d improved tenderness in two muscles without negatively affecting feedlot performance or tissue residues.     

Vitamin D3 supplementation of beef steers increases longissimus tenderness.

The objectives of these experiments were to determine 1) the effectiveness of supplemental vitamin D3 (VITD) on altering plasma and muscle calcium levels, 2) whether VITD supplementation improves Warner-Bratzler shear force (WBS) values of steaks from feedlot beef steers, and 3) the tenderness response curve of longissimus steaks from steers supplemented with VITD. In Exp. 1, 20 crossbred steers were assigned randomly to one of four treatment diets consisting of either 0, 2.5, 5.0, or 7.5 x 106 IU of VITD per day for 10 d. Blood samples were obtained daily during this supplementation period and 5 d thereafter (d 11 to 15). Between d 6 and 13, a linear increase (P < .01) in ionized plasma calcium concentrations was observed in steers supplemented with VITD. Compared to unsupplemented steers, serum calcium concentrations of the steers receiving 7.5 x 106 IU of VITD per day were increased 8 to 48%. In Exp. 2, longissimus samples from crossbred steers (n = 118) that were supplemented with either 0 or 5 x 106 IU of VITD per day for 7 d were obtained and aged for 7, 14, or 21 d. Following the initial 7-d postmortem aging period, VITD supplementation lowered (P < .01) WBS (.58 kg) and increased sensory tenderness rating (.6 units) compared to cuts originating from unsupplemented steers. In Exp. 3, 44 steers were supplemented with either 0 or 7.5 x 106 IU of VITD per day for 10 d immediately prior to slaughter. Results indicated that plasma and longissimus calcium concentration were higher (P < .05) for steers that received supplemental VITD. Compared with unsupplemented cuts, VITD supplementation improved WBS of cuts aged for either 7 or 14 d (P = .02 and P = .07, respectively). Sensory panelists rated samples from VITD supplemented steers as more tender than their unsupplemented counterparts. Activation of calpain proteases could be responsible for the observed tenderization due to the supplementation of VITD.     

Early postmortem biochemical factors influence tenderness and water-holding capacity of three porcine muscles

The objective of this study was to determine whether differences in pork tenderness and water-holding capacity could be explained by factors influencing calpain activity and proteolysis. Halothane-negative (HAL-1843 normal) Duroc pigs (n = 16) were slaughtered, and temperature and pH of the longissimus dorsi (LD), semimembranosus (SM), and psoas major (PM) were measured at 30 and 45 min and 1, 6, 12, and 24 h postmortem. Calpastatin activity; mu-calpain activity; and autolysis and proteolysis of titin, nebulin, desmin, and troponin-T were determined on muscle samples from the LD, SM, and PM at early times postmortem. Myofibrils from each muscle were purified to assess myofibril-bound (mu-calpain. Percentage drip loss was determined, and Warner-Bratzler shear (WBS) force was analyzed. Myosin heavy-chain (MHC) isoforms were examined using SDS-PAGE. The pH of PM was lower (P < 0.01) than the pH of LD and SM at 30 and 45 min and 1 h postmortem. The PM had a higher (P < 0.01) percentage of the MHC type IIa/IIx isoforms than the LD. The-LD had the greatest proportion of (P < 0.01) MHC IIb isoforms of any of the muscles. The PM had the lowest (P < 0.01) percentage of MHC IIb isoforms and a greater (P < 0.05) percentage of type I MHC isoforms than the LD and SM. The PM had less (P < 0.01) drip loss after 96 h of storage than the SM and LD. The PM had more desmin degradation (P < 0.01) than the LD and SM at 45 min and 6 h postmortem. Degradation of titin occurred earlier in the PM than the LD and SM. At 45 min postmortem, the PM consistently had some autolysis of mu-calpain, whereas the LD and SM did not. At 6 h postmortem, some autolysis of mu-calpain (80-kDa subunit) was observed in all three muscles. The rapid pH decline and increased rate of autolysis in the PM paralleled an earlier appearance of myofibril-bound mu-calpain. The SM had higher calpastatin activity (P < 0.05) at 45 min, 6 h, and 24 h and had higher WBS values at 48 h (P < 0.01) and 120 h (P < 0.05) postmortem than the LD. At 48 and 120 h postmortem, more degradation of desmin, titin, and nebulin were observed in the LD than in the SM. These results show that mu-calpain activity, mu-calpain autolysis, and protein degradation are associated with differences in pork tenderness and water-holding capacity observed in different muscles.     

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.     

Electrical stimulation effects on tenderness of five muscles from Hampshire x Rambouillet crossbred lambs with the callipyge phenotype

The objective of this study was to determine effects of electrical stimulation (ES) on muscle quality and sensory traits of 12 Hampshire x Rambouillet callipyge lambs. One side of each carcass was randomly assigned to an ES treatment of 550 V and 60 Hz of electricity for 2 s on and 2 s off 15 times. The other side was a nonstimulated control (NES). Heated calpastatin, sarcomere length, myofibrillar fragmentation index (MFI), Warner-Bratzler shear (WBS), and trained sensory panel values were measured on the semitendinosus (ST), semimembranosus (SM), longissimus (ML), supraspinatus (SP), and triceps brachii (TB) muscles. Electrically stimulating the carcass sides induced a more rapid (P = .001) pH decline in the longissimus muscle, and ES sides had a brighter (P = .001) red color of loineye than nonstimulated sides. At d 14 of storage (2 degrees C), the TB had the highest (P < .05) MFI value, indicating more protein degradation, and the ST and ML muscles had the lowest MFI (P = .008). Regardless of ES treatment, SM and ML had the highest (P < .05) WBS values. The ST muscle had higher (P < .05) WBS values than the SP but did not differ (P > .05) from the TB muscle. Electrical stimulation had no effect on WBS or any trained sensory panel values (P > .05). The percentage of loin chops rated slightly tender or better was improved 30 to 34% by electrical stimulation (P < .05). The ML muscle was scored lower (P < .05) in sustained juiciness compared with the SM, SP, and TB but did not differ (P > .05) from the ST muscle. The SM and ML muscles were rated lower (P < .05) in initial and sustained tenderness scores than other muscles. Tenderness scores were higher (P < .05) for the TB than for the SP but did not differ (P > .05) from the ST muscle. Electrically stimulating callipyge carcasses improves the tenderness of loin chops by increasing the percentage of chops rated from slightly tough to slightly tender.     

Caspase 3 is not likely involved in the postmortem tenderization of beef muscle

Postmortem proteolysis is an important determinant of beef tenderness. Caspase 3 is a protease that functions in apoptosis and has been shown to degrade myofibrillar proteins. Our objective was to evaluate whether caspase 3 activity is related to beef tenderness and muscle growth, and whether caspase 3 is activated in postmortem beef muscle. In experiment 1, longissimus thoracis (LT) and sternomandibularis muscle samples were obtained at 0, 0.25, 1, 3, 24, 72, and 240 h postmortem from 5 steers. In experiment 2, a group of 40 beef cattle was slaughtered at the University of Wyoming Meat Lab with 10 steers of different tenderness and growth characteristics chosen for the analysis of caspase 3 activity in the LT. In experiment 3, 10 steers with different tenderness but matched growth characteristics were chosen for analyses. In experiment 1, no significant activation (P = 0.70) of caspase 3 activity was detected; only a decreased activity at 72 (P = 0.05) and 240 h (P = 0.02) postmortem was observed. Western blot analysis of both muscle samples showed only the pro-caspase 3 form and failed to detect the activated enzyme. In experiment 2, caspase 3 activity in the LT immediately postmortem was greater (P = 0.05) for the cattle with increased Warner-Bratzler shear force values. No difference in caspase 3 activity was detected for experiment 3. Our results demonstrate that caspase 3 activity is not activated, with its activity decreasing with time postmortem, and caspase 3 activity is not associated with Warner-Bratzler shear force at slaughter. Therefore, caspase 3 is not anticipated to be involved in postmortem tenderization of beef.     

Technical note: Sampling methodology for relating sarcomere length,collagen concentration,and the extent of postmortem proteolysis to beef and pork longissimus tenderness

The objective of this study was to determine the effect of sampling methodology on the relationship between longissimus tenderness and measures of biochemical meat traits. Sampling methodology included measurements of sarcomere length, collagen concentration, and postmortem desmin proteolysis on raw samples and measurements of these same traits on the same cooked meat used for shear force measurement. Twenty crossbred steers and 20 crossbred barrows were used for these studies. The beef longissimus thoracis were vacuum-packaged, stored at 2 degrees C until 14 d postmortem, then frozen and stored at -30 degrees C. The pork longissimus thoracis et lumborum were vacuum-packaged, stored at 2 degrees C until 7 d postmortem, then frozen and stored at -30 degrees C. Trained sensory panel tenderness rating ranged from 3.1 to 7.6 for beef and 4.1 to 7.4 for pork. The coefficient of variation was lower for sarcomere length than for all other traits. Simple correlation coefficients between measurements on raw and cooked samples were 0.58 (beef) and 0.11 (pork) for sarcomere length, 0.66 (beef) and 0.59 (pork) for collagen, and 0.74 (beef) and 0.76 (pork) for desmin degradation. Simple correlation coefficients between biochemical traits and measures of tenderness (Warner-Bratzler shear force and trained sensory tenderness rating) were higher or not different for cooked compared to raw samples. Correlation coefficients between biochemical traits and tenderness rating were 0.38 (raw) and 0.22 (cooked) for sarcomere length, -0.12 (raw) and -0.45 (cooked) for collagen, and 0.48 (raw) and 0.80 (cooked) for desmin degradation in beef longissimus and 0.14 (raw) and 0.15 (cooked) for sarcomere length, -0.38 (raw) and -0.33 (cooked) for collagen, and 0.53 (raw) and 0.67 (cooked) for desmin degradation in pork longissimus. The coefficients of determination for explaining variation in tenderness rating using sarcomere length, collagen concentration, and desmin degradation for raw and cooked samples were 0.43 and 0.73 (beef) and 0.48 and 0.57 (pork), respectively. This study indicates that measurements of biochemical traits on the same cooked meat as used for shear force determination account for more of the variation in measures of tenderness than biochemical measurements made on a separate raw sample.     

Influence of dental carcass maturity classification on carcass traits and tenderness of longissimus steaks from commercially fed cattle

Two hundred beef carcasses were randomly selected by dental classification (zero, two, four, six, or eight permanent incisors) from a population of 11,136 carcasses harvested by a large commercial beef processor. Warner-Bratzler shear force and trained sensory panel evaluations of longissimus thoracis steaks as well as cooking and carcass traits were evaluated for differences among dental classes. No differences in Warner-Bratzler shear force (P = 0.60), sensory panel evaluations (P = 0.64) for tenderness, or percentage of total cooking loss (P = 0.73) were found among the five dental classes. Longissimus muscle color, USDA marbling score, hot carcass weight, adjusted fat thickness, longissimus muscle area, and USDA yield grade did not differ among the five dental classes. A significant dental classification x sex interaction indicated that heifers advanced in skeletal and overall maturity at a much faster rate than steers. An increase of intramuscular fat was associated (P < 0.05) with decreased shear force (r = -0.31), whereas darkening of the lean (r = 0.16), advancing lean maturity (r = 0.21), and increased evaporative cooking loss (r = 0.39) were associated (P < 0.05) with increased shear force values. Warner-Bratzler shear force measurements were not related to sensory panel overall tenderness scores. Carcass traits accounted for a relatively small proportion of the variation in tenderness of longissimus steaks, and dental classification was not related to tenderness.     

The use of vitamin D3 to improve beef tenderness

An experiment was designed to test the hypothesis that short-term oral administration of dietary vitamin D3 to beef cattle before slaughter would increase beef tenderness through greater calcium-activated calpain activity in postmortem aged skeletal muscle. Thirty continental crossbred steers were allotted randomly to three treatment groups housed in one pen. One group served as a control; two other groups were administered boluses with either 5 x 10(6) or 7.5 x 10(6) IU of vitamin D3 daily for 9 d. Cattle were slaughtered 1 d later. The longissimus lumborum was excised from each carcass 72 h postmortem and steaks removed at 3, 7, 14, and 21 d postmortem. The semimembranosus muscle (top round) was excised from each carcass 72 h postmortem and steaks removed at 7, 14, and 21 d postmortem. Blood plasma calcium concentration of cattle treated with 5 or 7.5 x 10(6) IU of vitamin D3 was higher (P < .05) than that of controls. Strip loin and top loin steaks from cattle fed supplemental doses of vitamin D3 had lower (P < .05) Warner-Bratzler (W-B) shear values at 14 d postmortem but were not significantly different from controls at 3, 7, or 21 d (strip loins) or 7 or 21 d (top rounds). No significant difference in strip loin steak tenderness was observed by sensory panel at 14 d postmortem (P < .17) between steaks from control and vitamin D3-treated steers. At 14 d postmortem, strip loin and top round steaks from cattle fed 5 x 10(6) IU of vitamin D3, but not from those given 7.5 x 10(6) IU, showed more proteolysis (P < .05) than did steaks from control cattle, based on Western blotting analysis. Therefore, the use of supplemental dietary vitamin D3 given daily for 9 d before slaughter did improve tenderness (lower W-B shear values) of 14-d postmortem aged beef. Increased proteolysis seems to be the mechanism of tenderization.