Comparison of postmortem proteolysis between Pekin and Muscovy duck breast muscles.

BACKGROUND: Duck muscle is a popular source of red meat in Asia. However, information regarding the postmortem proteolysis of skeletal muscle between duck species is very limited. Therefore, the purpose of this study was to compare the postmortem calpain and desmin degradations between Pekin (PD) and Muscovy (MD) duck breast muscles stored at 5°C. RESULTS: The pH and µ/m-calpain activity were not different (P > 0.05) between PD and MD postmortem muscles. However, µ-calpain activity and desmin content decreased more rapidly (P < 0.05) in PD than in MD samples. CONCLUSION: Therefore, our results suggest that the postmortem proteolysis is more rapid and extensive in breast muscle from PD compared to MD.

Underlying formation mechanisms of ultrasound-assisted brined porcine meat: The role of physicochemical modification, myofiber fragmentation and histological organization.

Ultrasound treatment has been a good hurdle technique for meat curing processing, where both physical and chemical consequences can be involved towards final quality of obtained products. However, the specific correlation between ultrasound parameters and muscle fiber fragmentation and myofibrillar microstructural changes during curing deserve further evaluation. In present study, we comparatively studied the effect of ultrasound-assisted brining (UAB) and static brining (SB) on the muscle proteolysis events and microstructural/morphological variation of porcine meat as well as the physicochemical indices and histological characteristics. The results showed that UAB (20 kHz, 315 W for 1 h) could markedly enhance the muscle proteolysis with higher free-/peptide-bound alpha-amino-nitrogen (α-NH2-N) content (P < 0.05) than SB treatment and greatly improved the fragmentation of muscle fiber tissues of cured meat. Meanwhile, UAB processing favored more opening structures of myofibrillar proteins with more hydrophobic groups being exposed. The quantitative histological analysis revealed that, compared with SB treatment, UAB could significantly increase the gap between muscle fibers and the swelling of the perimysium (P < 0.01), proving an efficient curing process with better textural and water holding properties.

Sarcoplasmic and myofibril-bound calpains during storage of pork longissimus muscle: New insights on protein degradation.

The subcellular distribution of calpain-1 and -2 and the proteolytical activity of myofibril-bound calpains in pork were investigated during 12 days cold storage. The content of sarcoplasmic calpain-1 decreased during storage while myofibril-bound calpain-1 content first increased (P < 0.05) to 17% of that of 12 h-sarcoplasmic calpain-1 on day 6 followed by a gradual decrease with subsequent storage, suggesting that calpain-1 gradually translocated from sarcoplasm to myofibrils during the initial 6 days of postmortem storage. Intact desmin decreased (P < 0.05) after incubation of myofibrils with 0.05 mM Ca2+, and this was more pronounced with 5 mM Ca2+ (P < 0.05). Ca2+ titration curves of day 6 myofibrils showed two distinct proteolytic activities becoming activated in the range 0.03 to 0.06 mM and 0.4 to 0.8 mM Ca2+, respectively. The results suggest that both calpain-1 and calpain-2 binds to myofibrils during storage and subsequently degrade structural proteins including desmin.

Post-Harvest Strategies to Improve Tenderness of Underutilized Mature Beef: A Review.

Beef muscles from mature cows and bulls, especially those originating from the extremities of the carcass, are considered as underutilized due to unsatisfactory palatability. However, beef from culled animals comprises a substantial proportion of the total slaughter in the US and globally. Modern consumers typically favor cuts suitable for fast, dry-heat cookery, thereby creating challenges for the industry to market inherently tough muscles. In general, cull cow beef would be categorized as having a lower extent of postmortem proteolysis compared to youthful carcasses, coupled with a high amount of background toughness. The extent of cross-linking and resulting insolubility of intramuscular connective tissues typically serves as the limiting factor for tenderness development of mature beef. Thus, numerous post-harvest strategies have been developed to improve the quality and palatability attributes, often aimed at overcoming deficiencies in tenderness through enhancing the degradation of myofibrillar and stromal proteins or physically disrupting the tissue structure. The aim of this review is to highlight existing and recent innovations in the field that have been demonstrated as effective to enhance the tenderness and palatability traits of mature beef during the chilling and postmortem aging processes, as well as the use of physical interventions and enhancement.

Tumbling and subsequent aging improves tenderness of beef longissimus lumborum and semitendinosus steaks by disrupting myofibrillar structure and enhancing proteolysis.

Tenderness is an important sensory attribute to the overall eating experience of beef. Identifying novel methods to ensure consistent tenderness, especially in inherently tough cuts, is critical for the industry. This study investigated if tumbling without brine inclusion could be an effective method to improve the quality and palatability attributes of beef longissimus lumborum (LL) and semitendinosus (ST) steaks. Furthermore, interactions with postmortem aging were evaluated to determine how tumbling might affect protein degradation and muscle ultrastructure. At 5 d postmortem, pairs of LL and ST muscles from beef carcasses (n = 16) were bisected, vacuum packaged, and tumbled for 0, 40, 80, or 120 min. Sections were divided and subsequently aged an additional 0 or 10 d at 2 °C. Tumbling for any duration improved instrumental tenderness of LL (P < 0.001) but not ST (P > 0.05) steaks, regardless of aging time. Tumbling exacerbated moisture loss in both muscles shown by greater purge and cooking losses (P < 0.05). Myofibrillar fragmentation was induced through tumbling in both muscles (P < 0.001), which was supported by transmission electron microscopy images. Tumbling for 120 min followed by 10 d of aging resulted in less abundant intact troponin-T in both LL and ST (P < 0.05), as well as less intact desmin in ST (P < 0.05); however, calpain-1 autolysis was not affected by tumbling (P > 0.05). No effects of tumbling, aging, nor the interaction were found for the content and solubility of collagen (P > 0.05). Consumer panelists (n = 120/muscle) rated LL steaks tumbled for any duration higher for tenderness and overall liking compared to control steaks (P < 0.05). For ST, significant interactions were found for consumer liking of tenderness and juiciness. In general, tumbling without subsequent aging resulted in poorer juiciness than non-tumbled (P < 0.05), while at 10 d no differences in juiciness were found between treatments (P > 0.05). For ST steaks that were aged 10 d, 120 min of tumbling resulted in greater tenderness liking than non-tumbled steaks (P < 0.05). These results suggest that tumbling would result in myofibrillar fragmentation and may benefit the degradation of myofibrillar proteins; however, there would be negligible impacts on collagen. Accordingly, tumbling without brine inclusion alone may be sufficient to improve tenderness and overall liking of LL steaks, while combined tumbling with subsequent postmortem aging would be necessary to improve tenderness liking of ST.

Tenderness is a primary driver to the overall eating experience of beef. Postmortem aging is applied to improve beef tenderness through the activity of endogenous proteolytic enzymes; however, certain inherently tough cuts may not reach acceptable levels of tenderness through aging alone. This study investigated if tumbling in the absence of a brine solution, coupled with subsequent postmortem aging, would be an effective strategy to improve the quality and palatability attributes of fresh beef loin (longissimus lumborum; LL) and eye of round (semitendinosus; ST) steaks. Regardless of aging, tumbling for 40, 80, or 120 min improved instrumental tenderness of LL but not ST steaks. Consumer panelists rated tumbled LL steaks to be more tender with greater overall liking compared to those that were not tumbled. Tumbling alone was insufficient to affect consumer liking of tenderness of ST steaks, although tumbling combined with subsequent aging improved tenderness liking. The results supported that tumbling would physically disrupt and fragment the myofibrillar structure, which may aid in the degradation of myofibrillar proteins during postmortem aging.

Biochemical and Physicochemical Changes in Spaghetti Meat During Refrigerated Storage of Chicken Breast.

This study investigated postmortem muscle protein degradation and myowater properties in broiler breasts afflicted with the Spaghetti Meat (SM) myopathy during 7 days of storage. Severe SM and unaffected (NORM) breast fillets were analyzed at days 0, 3, and 7 postmortem for TD-NMR myowater traits, myofibrillar protein profiles, calpain activity, free calcium, and desmin and troponin-T degradation patterns. Only at day 0, muscle histology, fiber size and sarcomere length were assessed on multiple fillet portions. In SM breasts, the intramyofibrillar water population exhibited longer relaxation times (p = 0.0172) and a lower proportion (p = 0.0118) compared to NORM. SM had a greater proportion of extramyofibrillar water (p = 0.0080) possessing a longer relaxation time (p = 0.0001). Overall, the SM myopathy had only a minor impact on the myofibrillar proteins profiles and did not affect either free calcium concentration, calpain activity, or the degradation of desmin and TnT, while storage time strongly affected all the traits measured. At microscopic level, muscle tissue from SM fillets exhibited the typical indicators of myodegeneration mostly in the superficial-cranial portion of the breast, while fiber size and sarcomere length were similar between the two muscle conditions irrespectively from the portion considered. The lack of overall significant interaction effects between muscle condition and storage period suggested that SM and NORM breast meat experience similar proteolytic and physical changes during the postmortem period.

Calpain activation and proteolysis in postmortem goose muscles.

Meat tenderness is considered as the most important criterion for meat quality by consumers and can be improved by the actions of endogenous proteases, mainly calpains, during postmortem storage at 0-5°C. The purpose of this study, therefore, was to examine the postmortem calpain activation and proteolysis in breast (BM) and leg and thigh (LM) muscles of White Roman goose. BM and LM were taken from goose carcasses (n = 15) at 0 (10-15 min postmortem), 1, 3, and 7 days of storage at 5°C. The decrease in postmortem pH, calpain-1 and -11 activities, and contents of the calpain-1 80 kDa subunit and desmin was more rapid (p < .05) in BM than in LM. Our results show that postmortem proteolysis was more extensive in BM than in LM of White Roman goose, not only because the difference in fiber type composition between two muscles, but because the rate and extent of calpain activation were greater in BM as well. These results may provide useful information to optimize meat processing for different muscles in goose industry.

Postmortem role of calpain in Chinese and Wuzong goose muscles.

The purpose of this study was to compare postmortem proteolysis and tenderization between Chinese and Wuzong goose breast muscles. Four months old Chinese (CG, n = 15) and Wuzong (WZ, n = 15) goose carcasses were vacuum-packaged 10 to 15 min postmortem and stored at 5°C. Breast (Pectoralis major) samples from each carcass were sampled at 0 (∼10 min postmortem), 1, 3, and 7 D of storage. Our results showed that the decrease in pH and calpain-1 activity was not different in CG and WG samples. However, the decrease in calpain-11 activity, desmin content, and shear force were more rapid (P < 0.05) in WZ than in CG samples. Our results indicate that postmortem proteolysis and tenderization of goose breast muscle were more extensive in WZ than in CG goose muscle.

Effect of age on calpain changes in postmortem goose muscle.

Meat tenderization can be affected by a variety of factors including animal age. However, results obtained from goose, chicken, turkey, and ostrich studies have shown that the effects of age on meat tenderization were insignificant, which may be due to a very limited age difference in birds used in those studies. Therefore, the purpose of this study was to investigate the effects of animal age on postmortem proteolysis and tenderization of breast muscle from developing and mature White Roman geese. Goose carcasses from mature (50 mo old, n = 10) and young (3 mo old, n = 10) geese were vacuum-packaged and stored at 5°C within 10 min postmortem. Breast (pectoralis major) samples were taken at 0 (∼10 min postmortem), 1, 3, and 7 D of storage. Our results showed that the decrease in pH, calpain-1 and -11 activities, desmin content and shear force, as well as the increase in myofibrillar fragmentation index were more rapid (P < 0.05) in young than in mature goose breast. These results suggest that postmortem proteolysis and tenderization of goose muscle are extensively affected by age.

Structural Equation Modeling and Whole-Genome Scans Uncover Chromosome Regions and Enriched Pathways for Carcass and Meat Quality in Beef.

Structural equation models involving latent variables are useful tools for formulating hypothesized models defined by theoretical variables and causal links between these variables. The objectives of this study were: (1) to identify latent variables underlying carcass and meat quality traits and (2) to perform whole-genome scans for these latent variables in order to identify genomic regions and individual genes with both direct and indirect effects. A total of 726 steers from an Angus-Brahman multibreed population with records for 22 phenotypes were used. A total of 480 animals were genotyped with the GGP Bovine F-250. The single-step genomic best linear unbiased prediction method was used to estimate the amount of genetic variance explained for each latent variable by chromosome regions of 20 adjacent SNP-windows across the genome. Three types of genetic effects were considered: (1) direct effects on a single latent phenotype; (2) direct effects on two latent phenotypes simultaneously; and (3) indirect effects. The final structural model included carcass quality as an independent latent variable and meat quality as a dependent latent variable. Carcass quality was defined by quality grade, fat over the ribeye and marbling, while the meat quality was described by juiciness, tenderness and connective tissue, all of them measured through a taste panel. From 571 associated genomic regions (643 genes), each one explaining at least 0.05% of the additive variance, 159 regions (179 genes) were associated with carcass quality, 106 regions (114 genes) were associated with both carcass and meat quality, 242 regions (266 genes) were associated with meat quality, and 64 regions (84 genes) were associated with carcass quality, having an indirect effect on meat quality. Three biological mechanisms emerged from these findings: postmortem proteolysis of structural proteins and cellular compartmentalization, cellular proliferation and differentiation of adipocytes, and fat deposition.

A new insight into meat toughness of callipyge lamb loins - The relevance of anti-apoptotic systems to decreased proteolysis.

The objective of this study was to determine associations of small heat shock proteins (sHSPs) in tenderness development of loins from callipyge and normal genotype lambs. Loins (M. longissimus lumborum) from sixteen lambs across four genotypes were collected throughout 9 days of postmortem aging. The loins from callipyge lambs had more intact desmin and troponin T throughout aging periods, as well as less µ-calpain autolysis and more calpastatin compared to loins from other genotypes (P < 0.05). Delayed onset of apoptosis was found in the callipyge loins indicated by less cytochrome c and more inactive procaspase-3 compared to normal lamb loins (P < 0.05). Less degraded HSP27 was also consistently found in the callipyge loins compared with loins from normal lambs (P < 0.001). The results found up-regulation of anti-apoptotic activities coincided with toughness in callipyge loins, which suggest apoptosis is likely involved in postmortem proteolysis and subsequent meat tenderization.

Postmortem role of calpain-11 in ostrich skeletal muscle.

The postmortem calpain-11 role in ostrich muscle was investigated. Pairs of ostrich muscle (Iliotibialis cranialis) were excised from 32 ostrich carcasses in 3-h postmortem and randomly assigned into four treatments. The muscle was cut into 2.5-cm thick meat cores. The cores were incubated in 30 mM CaCl2, 30 mM EDTA, 90 mM NaCl, or control. The cores from the left-side carcasses were sampled after 0, 1, 2, and 3 days of incubation at 5 °C, while the right-side meat cores were taken at 1-day and 3-day incubation for shear force measurements. The results showed that the decrease in unautolyzed and total activities of calpain-11, desmin content and shear force was more rapid in CaCl2-incubated samples than in control, NaCl- and EDTA-incubated samples. Thus, present results suggest that in the absence of calpain-1, calpain-11 with an extensive activation by adding exogenous Ca2+ could enhance the postmortem proteolysis and tenderization of ostrich muscle.

A comparison of intact and degraded desmin in cooked and uncooked pork longissimus thoracis and their relationship to pork quality.

The objective was to compare desmin abundance (intact, degraded, and the ratio of intact desmin to degraded desmin) in samples from cooked and uncooked pork and establish its relationship to pork quality traits. Pork chops (2.54-cm thick) from twenty-four pork loins were randomly assigned to treatment (cooked or uncooked). Intact and degraded desmin in cooked and uncooked chops aged 1d were weakly correlated (r<|0.35|) with Warner-Bratzler shear force at 1 and 14d postmortem aging. Intact and degraded desmin in cooked and uncooked chops aged 14d were moderately correlated (|0.35|

Postmortem calpain changes in ostrich skeletal muscle.

The objective of this study was to study the postmortem calpain change in ostrich muscle. Iliotibialis cranialis and Obturatorius medialis muscles were removed from the both sides of carcasses (n=8). The muscles from the left side were sampled after 0, 1, 2, 3, and 7days of storage at 5°C, while the right-side muscles were taken at 1-, 3-, and 7-day postmortem for shear force measurements. The results showed that the calpain-1 activity was not detected in ostrich muscle during the entire 7-day postmortem storage period, while the calpain-11 was. The unautolyzed calpain-11 activity decreased and the autolyzed calpain-11 activity increased with time postmortem. Desmin content and shear force did not change during postmortem storage although a minor degradation of desmin was observed. Therefore, our results suggest that limited postmortem proteolysis (as suggested by the limited degradation of desmin) and tenderization might be due to the lack of calpain-1 and/or insufficient calpain-11 activity present in ostrich muscle.

Postmortem proteolysis in three muscles from growing and mature beef cattle.

The objective of this study was to determine calpain system activity and postmortem protein degradation in three muscles from growing (n=6, 7.3 ± 0.5 months) and mature (n=6, 106.7 ± 43.1 months) beef cattle. The ratio of µ-calpain:total calpastatin activity tended to be lower in mature animals (P=0.08), suggesting reduced potential for proteolysis. Additionally, muscles from the mature group had greater calpastatin activity compared to calves at 6 days postmortem and had less µ-calpain autolysis and troponin-T and titin degradation during the aging period (P<0.01). Between the longissimus, semimembranosus, and triceps brachii muscles, the triceps brachii had the least postmortem proteolysis, with greater calpastatin activity and less troponin-T and titin degradation compared to other muscles (P<0.01). These data suggest that calpastatin activity in muscle from older animals is more persistent postmortem. This difference may contribute to the decreased protein degradation and increased toughness of beef from mature cattle, even after aging.