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An in vitro assay was developed to study protease activity during the maturation of beef postmortem. Myofibrils were purified from the semitendinosus and used as a sentinel for assessing the activity of endogenous proteases in longissimus thoracis et lumborum (LTL) and the extensor carpi radialis (ER) over time postmortem in beef carcasses. Samples were collected from each muscle at 0, 1, 2, 7, and 14 d of aging and snap frozen. Samples were powdered and added to an in vitro proteolysis assay containing buffer and purified myofibrils. Aliquots were collected at 0, 2, 120, 480, and 1440 min of incubation, and intact desmin and troponin-T were quantified. Digestions at 0 and 1 d using either muscle had little desmin degradation during the entire digestion period. In contrast, LTL muscle collected at 2, 7, and 14 d had the greatest proteolytic capacity as indicated by disappearance of intact desmin by 480 and 1440 min incubation. Though degradation ensued using powdered ER muscle, disappearance of intact proteins was limited. Degradation in vitro paralleled that observed in intact muscle. Addition of ethylene glycol tetra-acetic acid (EGTA), a cysteine protease inhibitor, and calpastatin inhibited proteolysis and suggest proteolytic activity observed in muscles and detected in our proteolysis assays is due to an active calpain protease. Collectively, our data show an active protease is minimal in bovine muscle until 48 h postmortem in the LTL muscle and suggest an in vitro assay containing purified myofibrils is a potential tool for studying temporal changes in proteolysis during the maturation and tenderization of beef across muscles., Competing Interests: Declaration of competing interest This work is supported in part by A1364 Novel Foods and Innovative Manufacturing Technology [grant no. 2020-67017-31269] from the USDA National Institute of Food and Agriculture., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Since veal production has declined in the U.S., American veal producers are currently making efforts to implement new production standards to improve product quality and animal welfare. In this study, we hypothesized that diets containing brewery grains, starch and omega-3 fatty acids could lower a blood stress indicator and improve meat quality, mostly from a nutritional value stand point. Holstein bull calves with approximately 94.67 ± 12.07 kg of body weight and two months old were randomly assigned to 1 of 3 dietary treatments. Diets were formulated with nonmedicated milk replacer, microbreweries spent grains, and a mineral mix (CONTROL); CONTROL + isolated maize starch (STARCH); and CONTROL +3% fish oil (OMEGA-3). Veal calves fed all three diets were heavier than calves of the same age from experiments reported in the existing literature. Dietary treatments did not affect carcass weights, pH, color, moisture, sensory attributes, volatile profile, and fat quality indexes. Calves fed STARCH and OMEGA-3 showed the lowest levels of blood cortisol. Veal fed CONTROL and OMEGA-3 had higher concentrations of ΣMUFA when compared with STARCH. Veal fed OMEGA-3 had the highest concentrations of EPA, DHA, and Σn-3. Veal from all treatments had very high concentrations of ΣMUFA, mostly driven by high levels of c-9 18:1 n-9 from the milk replacer. Feeding OMEGA-3 lowered blood cortisol and increased levels of EPA and DHA without harming sensory attributes. Overall, including brewery grains, starch and fish oil in liquid diets containing milk replacer can improve veal production., Competing Interests: Declaration of competing interest The authors declare no conflict of interest associated with this publication and no financial support for this work that could have influenced the outcome., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This research aimed to explore the potential influence of mitochondria on the rate of anaerobic glycolysis. We hypothesized that mitochondria could reduce the rate of anaerobic glycolysis and pH decline by metabolizing a portion of glycolytic pyruvate. We utilized an in vitro model and incorporated CPI-613 and Avidin to inhibit pyruvate dehydrogenase (PDH) and pyruvate carboxylase (PC), respectively. Four treatments were tested: 400 μM CPI-613, 1.5 U/ml Avidin, 400 μM CPI-613 + 1.5 U/ml Avidin, or control. Glycolytic metabolites and pH of the in vitro model were evaluated throughout a 1440-min incubation period. CPI-613-containing treatments, with or without Avidin, decreased pH levels and increased glycogen degradation and lactate accumulation compared to the control and Avidin treatments (P < 0.05), indicating increased glycolytic flux. In a different experiment, two treatments, 400 μM CPI-613 or control, were employed to track the fates of pyruvate using [ 13 C 6 ]glucose. CPI-613 reduced the contribution of glucose carbon to tricarboxylic acid cycle intermediates compared to control (P < 0.05). To test whether the acceleration of acidification in reactions containing CPI-613 was due to an increase in the activity of key enzymes of glycogenolysis and glycolysis, we evaluated the activities of glycogen phosphorylase, phosphofructokinase, and pyruvate kinase in the presence or absence of 400 μM CPI-613. The CPI-613 treatment did not elicit an alteration in the activity of these three enzymes. These findings indicate that inhibiting PDH increases the rate of anaerobic glycolysis and pH decline, suggesting that mitochondria are potential regulators of postmortem metabolism., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study examined the potential influence of mitochondrial calcium sequestering ability on calpain-1 autolysis and proteolysis in vitro. We first tested whether mitochondria can sequester calcium in an in vitro setting. Isolated bovine mitochondria (0, 0.5, or 2 mg/mL) were incubated in a buffer containing varying calcium levels (0, 50, or 100 μM). An inverse relationship between mitochondrial content and measured free calcium was observed (P < 0.05), confirming that mitochondria can sequester calcium within the concentration range tested. In the first in vitro experiment, intact mitochondria (0, 0.5, or 2 mg/mL) were incorporated into an in vitro model simulating postmortem muscle conditions, and calpain-1 autolysis and proteolysis were evaluated over a 168-h period. Adding intact mitochondria to the in vitro model decreased calpain-1 autolysis and proteolysis during the first 4 h of incubation (P < 0.05), likely through reducing calcium availability. However, accentuated calpain-1 autolysis and proteolysis were observed at 24 h. To further explore these effects, mitochondrial integrity was evaluated at varying pH and calcium levels. Mitochondrial integrity decreased as pH declined (P < 0.05), especially in the presence of calcium. Based on these results, we conducted a second in vitro experiment involving disrupted mitochondria. Unlike intact mitochondria, which exerted a suppressive effect on calpain-1 autolysis and proteolysis early on, disrupted mitochondria increased both parameters at most time points (P < 0.05). Overall, it appears that intact mitochondria initially cause a delay in calpain-1 autolysis and proteolysis, but as their integrity diminishes, both processes are enhanced., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Beef consumption is expected to increase worldwide, which necessitates the use of Bos indicus cattle that are well-adapted to harsher climates, like the tropics. Yet, beef from these cattle is considered inferior to that of Bos taurus breeds, primarily due to lowered tenderness values and reduced intramuscular fat content. However, the benefits of using Bos indicus genetics are numerous and undeniable. Herein, we explore how decreases in meat quality in these cattle may be offset by increases in livability. Further, we review the knowledge surrounding beef tenderness and explore the processes occurring during the early events of the transformation of muscle to meat that are different in this biological type and may be altered by stress. Growth rate, calpastatin activity and mitochondrial function will be discussed as they relate to tenderness. The opportunities of using Bos indicus cattle are of great interest to the beef industry worldwide, especially given the pressures for enhancing the overall sustainability and carbon footprint of this sector. Delivering a consistently high-quality product for consumers by exploiting Bos indicus genetics in a more sustainable manner will be proposed. Information on novel factors that influence the conversion of muscle to meat is explored to provide insights into opportunities for maximizing beef tenderization and maturation across all cattle. Exploring the use of Bos indicus cattle in modern production schemes, while addressing the mechanisms undergirding meat tenderness should provide the industry with a path forward for building greater demand through producing higher quality beef., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Reduction in muscle glycogen triggered by adverse antemortem handling events alters postmortem energy metabolism and results in a high ultimate pH and dark, firm and dry beef, often referred to as 'dark-cutting'. However, the relationship between atypical dark (AT) beef, postmortem energy metabolism and underlying tissue characteristics remains somewhat unclear. Cattle harvested in the US and Canada representing normal (pH < 5.6), AT dark (pH 5.6-5.8) and dark cutting (DC; pH > 5.8) beef were analyzed for tissue characteristics related to energy metabolism. Results show AT dark beef is more oxidative but similar to normal beef in glycolytic potential and nucleotide abundance. Mitochondria DNA content (P < 0.05, Canada; P < 0.005, US) and oxidative enzymes for DC and AT dark beef were greater (P < 0.01; Canada and US) compared to normal beef. Myoglobin tracked (P < 0.01) with color classification. These findings show both DC and AT beef are inherently more oxidative and raise the possibility that more oxidative muscle may be more prone to develop dark beef., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Studies investigating the effect of scald time on pork quality are confounded with time of dehairing. To understand better pork quality development and two-toning in hams, twenty-four carcasses were assigned to an 8- or 16-min dwell time prior to the dehairing, with or without scalding (n = 6 per trt). Semimembranosus (SM) muscles were collected following dehairing and at 24 h postmortem. Protracted time to dehair improved ultimate pH (pH u ; P < 0.005) and reduced (P < 0.05) color variation. One hundred forty-two carcasses were then subjected to protracted (control, 10-min) dwell times (15-min, or 20-min) in an industrial setting. Lightness was improved with 15-min dwell times compared to control, however 20-min dwell decreased the pH u (P < 0.001), increased lightness (P < 0.05), and percent purge (P < 0.001) in the SM. Also, lightness of the longissimus muscle (LM) increased (P < 0.001) with dwell time. These data show time to dehairing impacts pork quality development and suggest dehairing may be critical to quality development in a muscle-dependent manner., Competing Interests: Declaration of Competing Interest None., (Published by Elsevier Ltd.)

The objective of this study was to determine if ractopamine (RAC) impacts postmortem muscle metabolism and subsequent pork quality in Halothane (HAL) and Rendement Napole (RN) mutant pigs. All RAC fed pigs had increased (P < 0.04) L* values. HAL and RN mutants muscle had lower (P < 0.01) pH values but RAC feeding had no effect. RN mutants had higher and lower (P < 0.05) muscle pH and temperatures, respectfully at 15 min and RN mutant pigs had greater (P < 0.0001) glycogen initially but lactate levels similar to wild type (WT) pigs at 24 h. RAC lowered (P < 0.05) glycogen in RN mutants but not in HAL mutated or WT pig muscle. These data show RAC feeding changes postmortem energy metabolism but does not change pH and pork quality hallmark of two major pig gene mutations and supports our contention that ultimate meat quality traits and their biochemical drivers may be more complex than originally reasoned., (Published by Elsevier Ltd.)

The aim of this work was to compare the lipidome and metabolome profiling in the Longissimus thoracis muscle early and late postmortem from high and normal ultimate pH (pHu) beef. Lipid profiling discriminated between high and normal pHu beef based on fatty acid metabolism and mitochondrial beta-oxidation of long chain saturated fatty acids at 30 min postmortem, and phospholipid biosynthesis at 44 h postmortem. Metabolite profiling also discriminated between high and normal pHu beef, mainly through glutathione, purine, arginine and proline, and glycine, serine and threonine metabolisms at 30 min postmortem, and glycolysis, TCA cycle, glutathione, tyrosine, and pyruvate metabolisms at 44 h postmortem. Lipid and metabolite profiles showed reduced glycolysis and increased use of alternative energy metabolic processes that were central to differentiating high and normal pHu beef. Phospholipid biosynthesis modification suggested high pHu beef experienced greater oxidative stress., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest associated with this research., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Fresh pork color is a function of pigment, and the pH and temperature conditions in the carcass postmortem. To explore the role of scald on color development, carcasses (n = 16) were subjected to either a 4- or 8-min scald. Semimembranosus (SM) muscle samples were collected before and after scalding, and at 24 h postmortem. A 50% reduction in scald time resulted in lighter color (L*) across the muscle early postmortem (P < 0.001), yet the 8-min scald treatment was lighter (P = 0.001) at 24 h. An interaction between scald time and sampling time showed in an increase in L* values at 4-min immediately following scald (P < 0.001). Two-hundred carcasses were then subjected to a modified scald time (6.5 min, or 7.5 min) in an industrial setting. Lowering scald time failed to recapitulate results. In fact, darker meat (L* value; P = 0.0166) was noted in the SM across longer scalds. These data suggest modest changes in scald time may not be responsible for changes in pork quality development., Competing Interests: Declaration of Competing Interest There is no conflict of interest by any authors with any outside organization on the paper entitled: “Reduced scald time does not influence ultimate pork quality” for publication., (Copyright © 2022. Published by Elsevier Ltd.)

Variations in color, though a quality frustration, are common across the face of fresh and processed hams. Herein, we measured objective color across the semimembranosus (SM) muscle early postmortem and at 1440 min, then compared these differences against biochemical and metabolic characteristics responsible for pork quality development. Color (L*, a*) differed (P < 0.001) by zone and time but no interaction was evident. Lactate content and pH were highly correlated (R 2  = 0.92) at 30 min, but weakened (R 2  = 0.161412) by 1440 min. Lactate anaplerosis was not responsible for this lack of relationship. Glycolytic potential also differed across zone (P < 0.001) and time (P < 0.005). Differences in myoglobin expression and abundance, as well as mitochondrial DNA were notable (P < 0.05) across zone. These data suggest inherent differences in SM muscle are key determinants of ham color variation, while postmortem metabolism may play a lesser role in driving this quality attribute., (Copyright © 2021. Published by Elsevier Ltd.)

The impact of growth rate (GR) and finishing regime (FR) on growth and meat quality traits of Angus x Nellore crossbred steers, harvested at a constant body weight (530 ± 20 kg) or time on feed (140 days), was evaluated. Treatments were: 1) feedlot, high GR; 2) feedlot, low GR; 3) pasture, high GR and 4) pasture, low GR. Live body composition, carcass and meat quality traits were evaluated. High GR had greater impact on muscle and fat deposition in feedlot-finished, but not in pasture-finished animals. Feedlot animals had higher Longissimus muscle area, backfat thickness, meat luminosity and tenderness when compared to pasture groups. Moreover, pasture- and feedlot-finished animals with similar GR did not differ in the chromatic attributes of non-aged meat, regardless of endpoint. Thus, GR appeared to be the main factor driving beef chromatic parameters, while FR had a major impact on achromatic attributes and tenderness of meat., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of this study was to explore the use of TD-NMR relaxometry and 1 H NMR spectroscopy-based for detecting differences in meat quality attributes. There was limited association between various TD-NMR signals and any physicochemical parameters of fresh and aged meat differing in tenderness ratings. Samples were then divided into three groups based on statistical changes in metabolite concentration. Group A samples possessed near linear increases in metabolite concentration over aging time; whereas samples assigned to Groups B and C were characterized by increases in metabolites that peaked between 7 and 14 days, and up to 14 days aging, respectively. 1 H NMR spectroscopy discriminated meat quality using changes in metabolites reflective of glycolysis, the citric acid cycle, protein degradation, amino acid generation and purine metabolisms. These data suggest segregation of meat quality is possible using both NMR technologies but additional work is necessary to understand fully their utility in a commercial industry setting., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Pork quality is a product of the rate and extent of muscle pH decline paced by carbohydrate metabolism postmortem. The beta-adrenergic agonist ractopamine (RAC) alters muscle metabolism but has little impact on pork quality. The objective of this study was to determine how feeding RAC alters postmortem carbohydrate metabolism in muscle. Muscle pH was higher early postmortem in pigs fed RAC for 2 wks compared to control, while other time points and temperatures were largely unaffected. Early postmortem, muscle lactate levels were reduced (P < 0.05) after feeding RAC for 1 and 2 wks. Similarly, pigs fed RAC for 4 wks had reduced (P < 0.05) glycogen levels early postmortem compared to control pigs, but unexpectedly, L* values (lightness) increased (P < 0.05) after inclusion of RAC in the diet for 4 wk. These data show RAC feeding reduces glycogen content and changes lactate accumulation postmortem, but raise questions about the role glycolytic flux has in driving pork quality development., (Published by Elsevier Ltd.)

The purpose of this study was to test mitochondrial functionality under conditions simulating postmortem metabolism. Isolated mitochondria from porcine longissimus lumborum (LLM) and masseter (MM) muscles were incorporated into an in vitro model that mimics postmortem metabolism. pH and 13 C-enrichment of glycolytic and tricarboxylic acid (TCA) cycle intermediates were evaluated at 0, 15, 30, 120, 240, and 1440 min. Addition of mitochondria to the in vitro model lowered its pH at 240 min compared with control. Reactions containing mitochondria had lower pyruvate and lactate [M + 2] and [M + 3] isotopomers at 240 and 1440 min than controls. Furthermore, LLM lowered the enrichment of [M + 2], [M + 3], and [M + 4]α-ketoglutarate at 1440 min compared with MM and control. Succinate [M + 2] and [M + 3] were greater in MM than the control and LLM. [M + 3]fumarate was greater in control at 240 and 1440 min than LLM and MM treatments. Our data indicated that mitochondria are capable of mobilizing pyruvate generated though glycolysis under conditions simulating muscle postmortem metabolism., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Insufficient acidification results in dark, firm, and dry beef. While this defect is often indicative of a stress event antemortem, muscle tissue may change in response to feeding regime. Longissimus dorsi muscle samples from 10 grain-fed and 10 grass-fed market weight, angus-crossbred beef cattle were collected postmortem. Lower (P < .05) L* and a* values were recorded for steaks from grass-fed cattle. Higher (P < .05) ultimate pH values were noted in lean of grass-fed cattle compared to grain-fed cattle, yet differences in lactate, glycogen and glucose were not detected. Further, increased (P < .05) ultimate pH values and lower (P < .05) lactate accumulations were noted when samples from grass-fed cattle were subjected to an in vitro glycolysis system. Muscle from grass-fed beef possessed nearly two-fold more (P < .05) succinate dehydrogenase and (P < .001) myoglobin than that of grain-fed cattle. These data show lean from grass-fed beef has greater enzymes reflective of oxidative metabolism and suggest dark lean from grass-fed cattle may be a function of more oxidative metabolism rather than a stress-related event antemortem., (Copyright © 2019. Published by Elsevier Ltd.)

Muscle satellite cells (SC) are resident stem-like cells that play an integral role in skeletal muscle growth and repair. Understanding how SC maintain their identities and dynamic properties is critical to animal growth. However, the genetic and environmental factors governing SC behaviors and the underpinning mechanisms remain unknown. To explore whether genetic selection influences SC behaviors, we used 2 lines of chickens selected for over 50 generations with over a 10-fold difference in body weight at 56 d of age-the Virginia high weight selection (HWS) and low weight selection (LWS) lines. To study these 2 lines, we performed both in vivo and in vitro experiments. In vivo, we studied the abundance of SC in normal physiological settings and tested their functional roles in muscle regeneration using a muscle injury model. In vitro, we isolated SC from chicken skeletal muscle and assayed their ability to proliferate and differentiate under cultured conditions. Immunohistochemical staining of breast muscle (pectoralis major) revealed that muscle fibers from HWS chickens possessed more SC than those from LWS. Further analysis showed that the SC pool from HWS muscles contained a higher percentage of activated SC compared to that of LWS. When isolated SC from HWS and LWS muscles were cultured, HWS SC exhibited greater abilities to proliferate and differentiate than those SC from LWS. To test whether the observed in vitro differences in SC properties could be confirmed in vivo, we subjected chicken breast muscle to barium chloride to induce muscle injury and regeneration. Consistent with in vitro data, breast muscle in HWS chicken experienced a faster and more robust recovery than that of LWS, as evidenced by quicker regeneration and larger muscle fiber size. Taken together, these findings suggest divergent selection for body weight not only results in correlated responses in SC number, but also changes SC growth kinetics. Further dissection of the molecular mechanism will aid the identification of the target molecules for growth intervention in chickens.

During postmortem metabolism, muscle pH gradually declines to reach an ultimate pH near 5.6 across most meat species. Yet, broiler pectoralis major (P. major) muscle generates meat with high ultimate pH (pH ∼ 5.9). For better understanding of the underlying mechanism responsible for this phenomenon, we evaluated the involvement of breast muscle chilling on the extent of postmortem metabolism. Broiler breast muscles were either subjected to chilling treatment (control) or left at room temperature (RT) for 120 min. P. major muscle from the RT treatment had lower ultimate pH, greater glycogen degradation and lactate accumulation. While these findings suggest that carcass chilling can contribute to the premature termination of postmortem metabolism, chilling did not fully explain the high ultimate pH of P. major muscle. Our results also revealed that glucose-6-phosphate (G6P) was very low at 24 h, and therefore we hypothesized that G6P was limiting. To test this hypothesis, muscle samples from P. major and porcine longissimus lumborum (LL) muscle were homogenized into a reaction buffer that mimics postmortem glycolysis with or without 0.5 mg/mL isolated mitochondria. While samples containing porcine LL muscle reached the normal level of ultimate pH, P. major muscle samples reached a value similar to that observed in vivo even in the presence of excess G6P, indicating that G6P was not limiting. Mitochondria enhanced the glycolytic flux and pH decline in systems containing muscle from both species. More importantly, however, was that in vitro system containing chicken with mitochondria reached pH value similar to that of samples containing LL muscle without mitochondria. To investigate further, phosphofructokinase (PFK) activity was compared in broiler P. major and porcine LL muscle at different pH values. PFK activity was lower in P. major muscle at pH 7, 6.5, and 6.2 than LL muscle. In conclusion, carcass chilling can partially contribute to the high ultimate pH of broiler P. major muscle, while low PFK activity and mitochondria content limit the flux through glycolysis.

Anaerobic glycolysis dominates energy metabolism postmortem. Even so, however, recent studies suggest mitochondria can modify postmortem energy metabolism and may contribute to pH decline, possibly affecting the transformation of muscle to meat and fresh meat quality development. Because oxygen is a necessary component of mitochondrial function, oxygenation of porcine and bovine longissimus thoracis et lumborum was determined postmortem using NIR spectroscopy. The ratio of oxy- to deoxymyoglobin decreased with time postmortem in both species. Metabolic analyses of muscle samples collected over the same timeframe also revealed fluctuations in TCA intermediates. Finally, mitochondria collected from muscle of electrically stimulated carcasses differed from those of non-stimulated muscle. Collectively, these data support the thesis that muscle mitochondria function early postmortem and may play a more active part in pH decline and possibly meat quality development., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The experiment was conducted to identify the mitochondrial protein responsible for enhancing glycolytic flux. We hypothesized that mitochondrial F 1 -ATPase promotes ATP hydrolysis and thereby the flux through glycolysis. Porcine longissimus muscle mitochondria were incorporated into an in vitro system designed to recapitulate postmortem glycolysis with or without Na-azide to specifically inhibit the β-subunit of mitochondrial F 1 -ATPase that catalyzes ATP hydrolysis. Addition of mitochondria enhanced ATP hydrolysis, glycogen degradation, lactate accumulation, and pH decline in the in vitro system. However, the majority of mitochondria-mediated enhancement in glycolytic flux was abolished in the presence of Na-azide. To investigate further, myofibrillar and mitochondrial proteins were added to the in vitro system after 240min from the initiation of the reaction. Greater pH decline and lactate accumulation were observed in system containing mitochondrial protein compared to their myofibrillar counterpart. In conclusion, mitochondrial F 1 -ATPase is capable of increasing glycolytic flux through promoting greater ATP hydrolysis at lower pH., (Copyright © 2017. Published by Elsevier Ltd.)

The purpose of this study was to determine the role of mitochondria in postmortem muscle metabolism. Isolated mitochondria were incorporated into a reaction buffer that mimics postmortem glycolysis with or without mitochondrial electron transport chain inhibitors. Addition of mitochondria lowered pH values at 240 and 1440min regardless of inhibitors. Reduction in pH was accompanied by enhanced glycogen degradation and lactate accumulation. To explore the mechanism responsible for this exaggerated metabolism, mitochondrial preparations were mechanically disrupted and centrifuged. Resulting supernatants and pellets each were added to the in vitro model. Mitochondrial supernatants produced similar effects as those including intact mitochondria. To narrow further our target of investigation, mitochondrial supernatants were deproteinized with perchloric acid. The effect of mitochondrial supernatant was lost after perchloric acid treatment. These data indicate that a mitochondrial-based protein is capable of increasing glycolytic flux in an in vitro model and may partially explain acid meat development in highly oxidative AMPKγ3 R200Q mutated pigs., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Satellite cells (SCs) reside between the sarcolemma and basal lamina of muscle fibers and are the primary contributor of DNA for post-hatch muscle growth and repair. Alterations in SC content or properties by intrinsic and extrinsic factors can have detrimental effects on muscle health and function, and ultimately meat quality. We hypothesized that disrupted SC homeostasis may account in part for the increased breast myopathies observed in growing broilers. To test this hypothesis, we selected broilers with different body weights at comparable ages and studied SC characteristics in vitro and in vivo. Data shows that SC numbers in the breast muscles decrease (P < 0.001) and their inherent abilities to proliferate and differentiate diminish (P < 0.001) with age and size. Further, when breast muscle is presented with an insult, muscle of larger broilers regenerates more slowly than their smaller, age-matched counterparts arguing that SC quality changes with size and age. Together, our studies show that birds with greater muscle hypertrophy have less SCs with diminished ability to function, and suggest that aggressive selection for breast growth in broilers may exhaust SC pools when birds are grown to heavier processing weights. These findings provide new insights into a possible mechanism leading to breast myopathies in the poultry industry and provide targets for mitigating adverse fresh breast quality., (© 2017 Poultry Science Association Inc.)

Fresh hams display significant lean color variation that persists through further processing and contributes to a less desirable cured product. In an attempt to understand the underlying cause of this color disparity, we evaluated the differences in muscle characteristics and energy metabolites across semimembranosus (SM) muscles differing in color variation. The L* (lightness) and a* (redness) values were highest and lowest (P<0.001), respectfully in the most caudal aspects of the muscle while the ultimate pH was the lowest (P<0.001). Correspondingly, this region possessed highest (P<0.01) glycolytic potential (GP) and lactate dehydrogenase (LDH) levels but did not differ in the amount of myoglobin or myosin heavy chain type I isoform. These data show that differences in muscle may contribute to ham color variation but suggest other factors may mitigate or exacerbate these variances., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Fresh turkey meat color is determined by many factors that include muscle fiber type composition and heme protein concentrations. These factors either are affected by or influence biochemical events occurring postmortem. Deviations in the processing environment also can result in aberrant fresh meat quality and may ultimately change the quality characteristics of further processed products. Our objective was to describe the underlying cause and significance of the two-toning color defect in fresh turkey breast. In the first experiment, pectoralis major muscles were collected, classified as single- or two-toned, and analyzed using image processing to characterize fresh turkey color. Samples from the large and small lobes of the pectoralis major muscle were collected for pH, glycolytic intermediates, protein abundance, mRNA expression, and quality characteristics. In the second experiment, time from stun to exsanguination was tested as a promoter of fresh turkey color. Results from the first experiment showed that the turkey breast possesses two distinct lobes. The large lobe had greater (P < 0.05) glycolytic potential, lactate content, lactate dehydrogenase (LDH) abundance, and centrifugal drip loss, while pH, myoglobin mRNA expression, and soluble protein levels were lower (P < 0.05) compared to the small lobe. Results from the second experiment showed that reducing time from stun to exsanguination enhanced (P < 0.05) fresh turkey color by mitigating the differences between the two lobes. Our results also showed that birds exsanguinated first had greater (P < 0.05) muscle pH values and body temperatures. These results show inherent differences in breast muscle and processing conditions interact to establish variations in fresh turkey color., (© 2016 Poultry Science Association Inc.)

Skeletal muscle glycogen content can impact the extent of postmortem pH decline. Compared to glycolytic muscles, oxidative muscles contain lower glycogen levels antemortem which may contribute to the higher ultimate pH. In an effort to explore further the participation of glycogen in postmortem metabolism, we postulated that increasing the availability of glycogen would drive additional pH decline in oxidative muscles to equivalent pH values similar to the ultimate pH of glycolytic muscles. Glycolysis and pH declines were compared in porcine longissimus lumborum (glycolytic) and masseter (oxidative) muscles using an in vitro system in the presence of excess glycogen. The ultimate pH of the system containing longissimus lumborum reached a value similar to that observed in intact muscle. The pH decline of the system containing masseter samples stopped prematurely resulting in a higher ultimate pH which was similar to that of intact masseter muscle. To investigate further, we titrated powdered longissimus lumborum and masseter samples in the reaction buffer. As the percentage of glycolytic sample increased, the ultimate pH decreased. These data show that oxidative muscle produces meat with a high ultimate pH regardless of glycogen content and suggest that inherent muscle factors associated with glycolytic muscle control the extent of pH decline in pig muscles., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Postmortem lactate accumulation in skeletal muscle is linearly associated with the extent of pH decline. Yet, pigs harboring the AMPKγ3(R200Q) mutation produce meat with similar lactate levels to that of wild-type pigs but have a lower ultimate pH. We hypothesized that lower initial lactate levels and (or) lower buffering capacity in muscle of these pigs may help explain this discrepancy. Longissimus lumborum muscle samples were harvested at 0 and 1440 min postmortem from AMPKγ3(R200Q) and wild-type pigs. As expected, AMPKγ3(R200Q) muscle exhibited a lower ultimate pH but similar lactate levels to that of wild-type pigs at 1440 min postmortem. However, the total net lactate produced postmortem was greater in the AMPKγ3(R200Q) muscle due to lower initial lactate levels at 0 min postmortem. Buffering capacity measured over the pH range of 5.5-7.0 was also lower in AMPKγ3(R200Q) muscle. Greater net lactate accumulation postmortem (i.e., glycolytic flux) coupled with a lower buffering capacity explains the lower ultimate pH of meat from AMPKγ3(R200Q) pigs., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Our objective was to determine the influence of mitochondria on metabolites and pH decline using an in vitro model of postmortem muscle metabolism. Mitochondria were isolated from porcine longissimus lumborum and added (0, 0.5, or 2.0mg) to powdered muscle in reaction media containing either a combination of inhibitors for mitochondria complexes (I, IV, and V) or diluent (without inhibitors). In the absence of inhibitors, adding mitochondria (0.5 and 2.0mg) reduced ATP loss from 30 to 120 min, but did not alter glycogen or lactate during this time. In reactions with mitochondria, inhibitors decreased ATP levels by 30 min and increased glycogen degradation by 60 min. Regardless of mitochondria content, inhibitors enhanced lactate accumulation from 15 to 240 min, and decreased pH from 15 min to 1440 min. In the in vitro model, mitochondria influence the maintenance of ATP, and inhibition of mitochondria enzyme activity contributes to accelerated metabolism and pH decline., (Copyright © 2015. Published by Elsevier Ltd.)

Each skeletal muscle contains a fixed ratio of fast and slow myofibers that are distributed in a stereotyped pattern to achieve a specific motor function. How myofibers are specified during development and regeneration is poorly understood. Here we address this question using transgenic reporter mice that indelibly mark the myofiber lineages based on activation of fast or slow myosin. Lineage tracing indicates that during development all muscles have activated the fast myosin gene Myl1, but not the slow myosin gene Myh7, which is activated in all slow but a subset of fast myofibers. Similarly, most nascent myofibers do not activate Myh7 during fast muscle regeneration, but the ratio and pattern of fast and slow myofibers are restored at the completion of regeneration. At the single myofiber level, most mature fast myofibers are heterogeneous in nuclear composition, manifested by mosaic activation of Myh7. Strikingly, Myh7 is activated in a subpopulation of proliferating myoblasts that co-express the myogenic progenitor marker Pax7. When induced to differentiate, the Myh7-activated myoblasts differentiate more readily than the non-activated myoblasts, and have a higher tendency, but not restricted, to become slow myotubes. Together, our data reveal significant nuclear heterogeneity within a single myofiber, and challenge the conventional view that myosin genes are only expressed after myogenic differentiation. These results provide novel insights into the regulation of muscle fiber type specification., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Postmortem energy metabolism drives hydrogen accumulation in muscle and results in a fairly constant ultimate pH. Extended glycolysis results in adverse pork quality and may be possible with greater adenonucleotide availability postmortem. We hypothesized that slowing adenonucleotide removal by reducing AMP deaminase activity would extend glycolysis and lower the ultimate pH of muscle. Longissimus muscle samples were incorporated into an in vitro system that mimics postmortem glycolysis with or without pentostatin, an AMP deaminase inhibitor. Pentostatin lowered ultimate pH and increased lactate and glucose 6-phosphate with time. Based on these results and that AMPK γ3(R200Q) mutated pigs (RN⁻) produce low ultimate pH pork, we hypothesized AMP deaminase abundance and activity would be lower in RN⁻ muscle than wild-type. RN⁻ muscle contained lower AMP deaminase abundance and activity. These data show that altering adenonucleotide availability postmortem can extend postmortem pH decline and suggest that AMP deaminase activity may, in part, contribute to the low ultimate pH observed in RN⁻ pork., (Published by Elsevier Ltd.)

Fresh meat quality development is influenced by pH decline that results from muscle glycolyzing energy substrates postmortem. The exact reason why glycolysis stops in the presence of residual glycogen remains unclear. We hypothesized that a critical glycolytic enzyme loses activity near the ultimate pH of meat. Porcine longissimus muscle samples were subjected to an in vitro system that mimics postmortem anaerobic metabolism at buffered pH values (7.0, 6.5, 6.0, 5.5 or 5.0). At pH7.0, 6.5, and 6.0, glycogenolysis and glycolysis proceeded normally while pH5.5 stopped lactate formation. Additional experimentation indicated that phosphofructokinase lost activity at pH5.5 while all other glycolytic enzymes remained active. A similar inactivation of phosphofructokinase was observed when using chicken and beef muscle. Elevated temperature hastened pH decline and phosphofructokinase activity loss. Thus, pH inactivates phosphofructokinase and arrests postmortem glycolysis, which may explain the similar ultimate pH across meat of different species., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Water-holding capacity is the ability of meat to hold moisture and is subject to postmortem metabolism. The objective of this study was to characterize the loss of moisture from muscle postmortem and investigate whether these losses are useful in predicting the ultimate drip loss of fresh pork. Cotton-rayon absorptive-based devices were inserted in the longissimus dorsi muscles of pork carcasses (n = 51) postmortem and removed at various intervals for 24h. Greatest moisture absorption was observed at 105 min post exsanguination. Drip loss varied (0.6-15.3%) across carcasses. Individual absorption at 75 min correlated (r = 0.33) with final drip loss. Correlations improved using individual absorption values at 90 min (r = 0.48) and accumulated absorption values at 150 min (r = 0.41). Results show that significant moisture is lost from muscle tissue early postmortem and suggest that capture of this moisture may be useful in predicting final drip loss of fresh meat., (© 2013.)

Pigs with the AMP-activated protein kinase γ3 R200Q (AMPKγ3(R200Q)) mutation generate pork with low ultimate pH (pHu). We hypothesized that reducing muscle creatine (Cr) and phosphocreatine (PCr) may accelerate postmortem ATP consumption and prevent extended pH decline in AMPKγ3(R200Q) longissimus muscle. Wild type and AMPKγ3(R200Q) pigs were assigned to control diet or diet supplemented with the creatine analog β-guanidinopropionic acid (β-GPA, 1%) for 2 wk. β-GPA reduced muscle PCr (P = 0.006) and total Cr (P<0.0001). In general, AMPKγ3(R200Q)+β-GPA exhibited more rapid metabolism than control, AMPKγ3(R200Q), and β-GPA treatment, evidenced by more rapid loss of ATP, more rapid increase in IMP, and decreased pH during the first 90 min postmortem. Overall, pHu was similar despite elevated glycogen (AMPKγ3(R200Q)), reduced total Cr (β-GPA) or both (AMPKγ3(R200Q)+β-GPA). Thus, reducing muscle phosphagens did not affect pHu in AMPKγ3(R200Q) muscle, but it hastened ATP depletion and pH decline., (© 2013. Published by Elsevier Ltd. All rights reserved.)

Meat quality development, or the transformation of muscle to meat, involves a myriad of biochemical pathways that are largely well-studied in living muscle tissue. However, these pathways are less predictable when homeostatic ranges are violated. In addition, there is far less known about how various management or environmental stimuli impact these pathways, either by substrate load or altered cellular environment. Likewise, it is largely accepted that oxygen plays little to no role in the conversion of muscle to meat, as anaerobic metabolism predominates in the muscle tissue. Even so, the oxygen tension within the tissues does not fall precipitously at exsanguination. Therefore, transition to an anaerobic environment may impact energy metabolism postmortem. Antemortem handling, on the other hand, clearly impacts meat quality development, yet the exact mechanisms remain a mystery. In this paper, we will attempt to review those factors known to affect postmortem energy metabolism in muscle and explore those areas where additional work may be fruitful., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Rate and extent of postmortem metabolism control pork quality development. Our objective was to evaluate the role of the phosphagen system (phosphocreatine, PCr; and creatine, Cr) on metabolism and pork quality. Muscle PCr and Cr were manipulated by feeding pigs the creatine analogue, β-guanidinopropionic acid (β-GPA). In experiment 1, pigs received standard (control) diet or β-GPA supplemented (2%) diet (1 wk or 2 wk). Supplementation with β-GPA (2 wk) decreased total Cr (PCr+Cr; P=0.02) and improved pork color (decreased reflectance, P=0.003); however, β-GPA supplementation reduced growth performance (P=0.007). To separate effects of phosphagen system and growth, a second experiment was conducted with control, pair-fed, and 2 wk β-GPA (1%) supplementation; pigs were also offered a control or β-GPA supplemented flavored beverage. Neither treatment influenced pork quality. Immediately postmortem, ATP/ADP was higher in control compared to pair-fed (P<0.05); subsequently, ATP/ADP was similar among all groups. Loss of the phosphagen system may lead to adaptive changes that promote conservation of cellular ATP., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Extent of postmortem pH decline influences meat quality development. To better understand physiological determination of ultimate pH (pHu), we utilized female and castrated male pigs from a line whose selection index includes differentiated pHu. All genotypes of AMP-activated protein kinase γ3 subunit (AMPKγ3) V199I site were present. The mutant 199II genotype increased pHu, but only in castrated males. Genotype affected glycolytic potential (GP), but GP was weakly associated with pHu. A subset of animals was selected based on low (-Gly) and high (+Gly) residual glycogen content, and compared with AMPKγ3 200Q, which is associated with low pHu. Both +Gly and 200Q muscle contained glycolytic substrate at 24h; however, 200Q muscle generated low pHu and greater lactate compared to +Gly. Additionally,-Gly and +Gly groups exhibited similar pHu despite a large difference in GP. In conclusion, high GP does not appear to directly impact the extent of postmortem pH decline., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

AMP-activated protein kinase (AMPK) is activated by upstream kinases and negatively regulated by protein phosphatases. Intracellular calcium mediates protein phosphatase 2A (PP2A), which is in a heterotrimeric complex with the PR72 subunit. The PR72 subunit contains two calcium-binding sites formed by EF hands. Our previous study has shown that chronic calcium exposure decreases AMPK activity. To define the specific molecular mechanism whereby calcium can deactivate AMPK, activities of AMPK and PP2A were analyzed in C2C12 muscle cell cultures and skeletal muscle tissues from mutant pigs possessing the AMPKγ3-mutation or the ryanodine receptor (RyR1) calcium gating mutation, or both. C2C12 myotubes treated with calcium releasing agent (caffeine) for 10h decreased (P<0.05) AICAR-induced AMPK activity to control levels and this negative effect was eliminated by ryanodine receptor stabilizer, dantrolene. Interestingly, muscle from pigs with the RyR1 mutation and C2C12 cells administered with 10h caffeine showed higher (P<0.05) PP2A activity compared to controls. More importantly, the inhibitory effect of caffeine on AMPK activity was attenuated by the PP2A inhibitor, calyculin A or siRNA induced knockdown of PP2A. These data show the inhibitory effect of chronic calcium on AMPK activity is exerted through the activation of PP2A., (Published by Elsevier India Pvt Ltd.)

The development of pork quality attributes - including color, water holding capacity, and texture - are controlled largely by the rate and extent of postmortem pH decline. In turn, pH decline is thought to follow the anaerobic degradation of glycogen to "lactic acid". The "Hampshire effect" or Rendement Napole (RN(-)) pig, which has elevated muscle glycogen, propagated the widespread use of "glycolytic potential" to describe muscle's capacity for postmortem glycolysis. Since the evolution of glycolytic potential, there have been great strides in molecular and technical capabilities. Hence, it must be appreciated that the RN(-) pig possesses a mutation in the gene encoding the AMP-activated protein kinase (AMPK) γ3 subunit, which results in a non-conservative amino acid substitution (R200Q). AMPK, a major energy sensor in skeletal muscle, influences enzyme activity, gene and protein expression, fiber type, and mitochondrial biogenesis. The utility of glycolytic potential as indicator of ultimate pH should be reevaluated in the context of the metabolic differences between AMPKγ3 mutated and normal muscle. Understanding the metabolism and energetics in AMPKγ3 mutated muscle may provide insight into the mechanisms influencing the rate and extent of postmortem metabolism., (Published by Elsevier Ltd.)

Calcium is important for muscle contraction and controls many cellular processes. Although there is evidence that calcium-mediated signals regulate AMP-activated protein kinase (AMPK) activity, the molecular mechanisms by which calcium regulates AMPK are poorly understood. To compare the function of sustained vs. intermittent calcium oscillations on AMPK activity and define specific signals in this pathway, we administered mice with aminoimidazole-carboxamide-ribonucleotide (AICAR) and caffeine with or without dantrolene. AMPK activity was increased by 10 d AICAR treatment (P < 0.01). Ten day caffeine treatment decreased AICAR-induced AMPK activity to control level. This repressed AMPK activity was blocked by dantrolene. Different calcium frequencies were simulated in C2C12 myotubes by alternating media containing caffeine and dantrolene. Intermittent calcium oscillation increased AMPK activity compared to control (P < 0.05), whereas sustained calcium oscillation decreases AICAR-induced AMPK activity to control level. This result suggests a biphasic control of AMPK activity by calcium. Knockdown of CaMKII expression by short-hairpin RNA resulted in increased AMPK phosphorylation by AICAR even in the presence of caffeine. These data show different calcium oscillations elicit distinct responses in muscle cells suggesting that the negative effects of chronic calcium treatment on AMPK activity is partly mediated through the CaMKII signals., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The objective of this study was to investigate the potential of a novel optical reflectance imaging method to predict beef tenderness. Two-dimensional (2D) optical reflectance in beef muscles induced by a point incident light was acquired. A set of five parameters were extracted from each reflectance image to describe quantitatively the reflectance profiles. Two parameters, q and B, were derived by numerically fitting the equi-intensity contours of the reflectance pattern. Two spatial gradients were calculated along the directions parallel and perpendicular to muscle fibers and total scattering intensity was obtained by excluding the specular reflectance. This method was applied to analyze 2D images of optical diffuse reflectance in 336 beef samples obtained from 14 steers in which large variations in tenderness were generated by altering animal genetics, suspension method and aging time as well as utilizing muscles varying in their inherent tenderness. Tenderness was evaluated using Warner-Bratzler shear force (WBSF). The effects of animal breed, muscle, types of suspension, and aging were investigated and results indicate that the scattering intensity measured at 1-d was correlated (R(2)=0.50 at lambda=720 nm) with 10-d WBSF in M. longissimus dorsi muscles; and the q parameters measured at 1-d was correlated (R(2)=0.46 at lambda=720 nm) with 10-d WBSF in M. psoas major muscles. These results show analyzing 2D reflectance images of meat surfaces provides valuable information regarding the physical characteristics of meat that are responsible for beef tenderness., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Pale, soft, and exudative (PSE) meat has been recognized for decades and causes huge economic loss to the meat industry due to its inferior quality. Although it has been well established that fast and excessive glycolysis combined with high temperature in muscle early postmortem is the cause of PSE meat, the molecular mechanisms associated with this abnormal glycolysis remain poorly defined. Our previous studies with mice and pigs suggest that AMPK regulates muscle glycolysis postmortem. To confirm further the role of AMPK in the regulation of postmortem glycolysis, we investigated the effects of intraperitoneal injection of compound C, a specific AMPK inhibitor, on AMPK activation and glycolysis in postmortem longissimus dorsi (LD) muscle of mice. Data showed that intraperitoneal injection of compound C inhibited AMPK activation in postmortem mouse LD muscle. Simultaneously, injection of compound C inhibited glycolysis and increased muscle pH corroborating of our previous observations that postmortem glycolysis is inhibited in AMPK knockout mice. This study firmly supports that AMPK regulates glycolysis in postmortem skeletal muscle and suggests that AMPK can be a target to control postmortem glycolysis, preventing incidence of PSE meat.

Pale, soft and exudative (PSE) pork represents considerable economic losses for the industry due to its limited functionality and undesirable appearance. During the past several decades, exhaustive research covering various aspects of the food chain has established genotyping procedures, recommended handling practices, and quality indicators in order to reduce the incidence of inferior pork quality. Despite these efforts, there is still a relatively high occurrence of PSE pork. Development of pork quality attributes is largely governed by the rate and extent of postmortem pH decline. The combination of high temperature at low pH or abnormally low ultimate pH causes denaturation of sarcoplasmic and myofibrillar proteins, resulting in paler color and reduced water holding capacity. The pH decline is closely related to muscle energy availability and demand at or around slaughter. The postmortem degradation of glycogen through glycogenolysis and glycolysis provides ATP to help meet energy demand and decreases pH by generating lactate and H+. Therefore, the flux of metabolites through glycolysis, the involvement of energy signaling pathways that modulate glycolytic activity, and the inherent metabolism of different fiber types are critical factors influencing pH decline and pork quality. Further, recent work implicates adenosine monophosphate-activated protein kinase (AMPK) as a major energy sensor for the cell, and thus may be involved in the control of postmortem metabolism. The intent of this paper is to review the biochemistry controlling postmortem energy metabolism in pig muscle and explore new information generated using genetic mutations in order to define the fundamental mechanisms controlling the transformation of muscle to meat.

Several methods using bactericides, hydrostatic pressure, and post-package pasteurization technologies to control Listeria monocytogenes (LM) in ready-to-eat meats have been attempted. In addition to controlling LM contamination, any newly developed technology must have minimal effects on organoleptic properties. The objectives of this study were to: (1) determine the heat resistance of LM in two brands (A and B) of bologna differing in formulations, and, (2) evaluate the effects of post-package pasteurization on product quality. Fat content did not affect LM heat resistance in bologna at 55, 60, and 65°C; however, Brand B bologna had a numerically lower inactivation rate. Microbial heat resistance differed (P<0.05) with changes in pasteurization temperature. Time and temperature affected (P<0.05) cook-loss and L(∗) Hunter color value for both bologna brands. These data show that post-package pasteurization is effective but suggest that meat formulations may need modification to prevent development of negative quality characteristics.

The objective of this study was to determine the effects of the halothane (HAL) and Rendement Napole (RN) genes on the rate-limiting reactions of glycolysis and their relationship to pork quality development. Samples were collected from the longissimus muscle at 0, 30, 60, and 120min and 24h postmortem from homozygous HAL and RN pigs (NN/rn(+)rn(+), NN/RN(-)RN(-), nn/rn(+)rn(+), nn/RN(-)RN(-)). Muscle pH was recorded at 0, 15, and 45min, and 24h postmortem. HAL mutants, compared with HAL normal, had lower (P<0.05) ATP concentrations at 0 and 30min, lower (P<0.05) pH at 45min, and hastened glycogen degradation and lactate accumulation in the first 120min postmortem (HAL×time, P<0.0001). RN mutants had greater (P<0.0001) glycolytic potentials than RN normal, and lower (P<0.05) 24h pH compared with the normal genotype, but not the HAL mutant genotype. The HAL mutation negatively affected (P<0.05) L∗, b∗ and color scores whereas both HAL and RN mutations independently decreased (P<0.05) firmness, marbling and water holding capacity. RN mutant genotypes had higher (P<0.0001) phosphocreatine concentrations than other genotypes at 0min. Compared with HAL normal, HAL mutants had elevated (P<0.05) muscle glucose concentrations at 30, 60, and 120min, and 24h. RN mutants had higher (P<0.05) glucose levels than RN normal after 60min. Glucose-6-phosphate (G6P) concentrations decreased (P<0.05) during the first hour postmortem except in HAL/RN mutants, which had higher (P<0.01) G6P concentrations than all other genotypes at 30min. From 60min to 24h postmortem, G6P increased (P<0.05) in HAL normal genotypes. Altogether, these data show that elevated muscle glycogen content does not further aggravate rapid early postmortem metabolism.

The objectives of this study were to determine the influence of pH and MyHC isoforms on myofibrillar and actin-activated myosin subfragment 1 (S1) ATPase activity and the protective effect of actin. Red (RST) semitendinosus and white (WST) semitendinosus myofibrils were incubated at pH 7, 6, or 5.5 with 0 or 2mM ATP. RST and WST S1 isolates were incubated at pH 7, 6, or 5.5 in the presence or absence of actin. Maximum calcium-activated myofibrillar and actin-activated S1-ATPase activity were then assayed at pH 7. Incubation of myofibrils with ATP caused ATPase activity of myofibrils to decrease (p<0.05) with the pH of the incubation. RST myofibrils maintained a higher (p<0.0001) relative activity than WST myofibrils after incubation at pH 6 with ATP. Myofibrils incubated without ATP exhibited higher (p<0.001) activities than those incubated with ATP following pH 5.5 treatments. WST myofibrils had a lower (p<0.05) relative activity than RST following incubation at pH 5.5 without ATP. S1 ATPase activities decreased (p<0.05) with incubation pH in WST samples, but not in RST samples. WST S1 activity was higher (p<0.01) in samples exposed to pH 6 and 5.5 with actin bound compared to those incubated without actin. RST S1 exhibited a higher (p<0.01) relative activity than WST samples following pH 5.5 treatment with bound actin. These data show that low pH inactivates myofibrils by altering actin-activated S1 ATPase. Furthermore, these results suggest that muscles with high proportions of fast fibers are more susceptible to pH inactivation of ATPase activity and that the protective effect of actin binding to myosin is less in fast fibers.

The aim of this study was to examine whether exogenous somatotropin (ST) can alter the insulin-like growth factor (IGF) axis in the porcine epitheliochorial placenta. Crossbred gilts were injected either 6 mg of recombinant porcine ST or vehicle from days 10 to 27 after artificial insemination (term day 116). Control and ST-treated gilts were euthanized on day 28 (8 control/5 treated), day 37 (4 control/6 treated), and day 62 (4 control/6 treated) of gestation. Endometrium and placental tissue samples were collected and subjected to mRNA analyses. In control gilts, somatotropin receptor (STR) and IGF-I mRNA abundance in the endometrium decreased with gestation. Conversely, the amounts of IGF-II mRNA and of IGF binding protein (BP)-2 and -3 mRNA, which were analyzed in endometrium and placental chorion, increased with gestation. The endometrium contained less IGF-II mRNA but more IGFBP-2 and-3 mRNA than the placental chorion. In response to pST treatment, the amounts of endometrial STR and IGF-I mRNA were lower at days 28 and 37, but higher at day 62 of gestation. The content of IGF-II mRNA was higher in the endometrium of pST-treated than control gilts on day 37. The amount of IGFBP-2 mRNA was increased on day 37 in endometrium and placenta of pST-treated gilts, whereas no changes in IGFBP-3 mRNA were observed. The IGF-II/IGFBP-2 ratio was higher in the placenta in response to pST on day 28 of gestation. Results show that pST treatment of pregnant gilts during early gestation alters IGF axis in maternal and fetal placental tissues and suggest pST may exert an effect on fetal growth by altering the relative amount of IGFBPs and IGFs at the fetal-maternal interface.

The objective of this study was to determine the effects of postmortem muscle pH and temperature declines on the actomyosin ATPase activity of muscle fibers expressing different MyHC isoforms. Using a quantitative histochemical procedure to determine ATPase activity, the maximum actomyosin ATPase activity was determined on individual fibers classified by MyHC expression. Samples were collected from the red (RST) and white (WST) semitendinosus muscles at 3 min and 24 h postmortem from electrically stimulated (ES) and control (NS) pork carcasses. In samples taken at 3 min postmortem, type I fibers had the lowest ATPase activity staining and type 2X and 2B had the highest activity staining, with type 2A fibers intermediate. Postmortem time and carcass treatment did not influence the ATPase activity staining of type I muscle fibers. ATPase activity staining of 2A fibers was lower (p<0.001) in 24 h samples than in 3 min samples from ES carcasses. In 3 min and NS-24 h samples, RST type 2A fibers had lower (p<0.05) activities than type 2A fibers from the WST. In type 2X fibers, ATPase activity staining decreased (p<0.01) from 3 min to 24 h postmortem in ES carcasses. This decrease was more severe in WST 2X fibers compared to RST 2X fibers. ATPase activity staining in type 2B fibers did not decrease from 3 min to 24 h postmortem in NS carcasses. In ES carcasses, activity staining of 2B fibers decreased (p<0.0001) with time postmortem. The results of the experiment indicate that fibers expressing fast MyHC isoforms have a higher ATPase activity early postmortem than slow muscle fibers but are more prone to inactivation by a rapid pH decline.

The pH and Ca(2+) sensitivity of myofibrillar ATPase activity plays an integral role in regulating postmortem muscle ATP utilization and likely paces postmortem glycolysis. The objective of this study was to determine the influence of pH and Ca(2+) concentration on the ATPase activity of myofibrils from red semitendinosus (RST) and white semitendinosus (WST) porcine muscles. Myofibrillar ATPase was measured at 39 °C over a pH range 5-7.5 and a [Ca(2+)] range pCa 4-9 (10(-4)-10(-9)M). At maximum Ca(2+)-dependent activation (pCa 4), RST myofibrils had lower (p<0.0001) ATPase activity than WST myofibrils. This maximum activity of myofibrils from both muscle regions was not influenced from pH 7.5 to 6.5, declined between pH 6.5 and 5.75 (Hill coefficient, n(H)=2.7-3.4; pH at half maximum activity, pH(50)=5.97) and was near zero at pH 5.5. At pH 7, pCa-activity relationships showed that RST required less Ca(2+) for half-maximum activation (higher pCa(50); 6.50) than WST myofibrils (pCa(50)=6.35) but had no difference in n(H). At pH 7, both RST and WST myofibrils had maximum Ca(2+)-dependent, actin-activated ATPase activity at pCa ⩽6 and Ca(2+)-independent myosin ATPase activity at pCa ⩾6.75. pCa-activity relationships at different pH levels indicated that pCa(50) decreased with pH from pH 6.5 to 6.125 in both RST and WST myofibrils. At pH <5.75, [Ca(2+)] did not influence ATPase activity in RST or WST myofibrils. These data show that myofibrils with predominantly fast MyHC (WST) have a higher actin-activated myosin ATPase activity than myofibrils with primarily slow MyHC isoforms (RST) at Ca(2+) concentrations and pH values characteristic of postmortem muscle.

The objective of this study was to determine the influence of myofibril isolation procedures and myosin heavy chain (MyHC) isoform composition on myofibrillar ATPase activity as related to postmortem muscle metabolism. Myofibrils from the red (RST) and white (WST) portions of semitendinosus muscles were isolated using two different methods (A and B) at 3 min and 24 h postmortem in control (NS) and electrically stimulated (ES) pork carcasses. Comparison of the relative MyHC isoform profiles between the two different myofibril isolation methods and myosin extracts from the RST and WST at 3 min showed that method B myofibrils were more similar to the myosin extract than method A. Myofibrillar ATPase activity remained constant or increased (P<0.01) from 3 min to 24 h postmortem in NS carcasses and decreased (P<0.0001) in ES carcasses. From the RST, method A myofibrils had higher (P<0.0001) ATPase activity compared to method B across sampling time and carcass treatment. In the WST, method A myofibrils had lower (P<0.01) activity at 3 min, were not different at 24 h in NS carcasses, but had higher (P<0.05) activity at 24 h in ES carcasses versus method B myofibrils. Compared to method B, isolation method A biased the isoform profile of myofibril samples more towards faster MyHC (2A and 2X) in the RST and towards MyHC 2X in the WST. Results suggest that the ATPase activity and MyHC isoform profile of isolated myofibril samples are influenced by method of myofibril isolation, postmortem sampling time, and the rate of postmortem metabolism. Thus, differences in MyHC isoform profile and method of myofibril isolation must be taken into account to determine accurately the relationship between myofibrillar ATPase activity and rate of postmortem metabolism.

Color image features were computed to characterize the skeletal maturity of beef carcasses based on cartilage ossification in the thoracic vertebrae. A trained neural network was tested for predicting USDA beef maturity grades from image features of ossification. A feature curve was defined to characterize the color variations of an isolated cartilage-bone object. Both RGB and HSL color systems were used to derive image features. The maturity grades were assigned by an official USDA grader. Two sets of samples were obtained from two different meat-processing plants. The first set contained samples of only A and B maturity grades whereas the second set had all five maturity classifications (A through E). The hue value was the most useful color feature. The mean hue values of cartilage differed (P<0.05) among the maturity grades and the feature curve based on the hue value was used as neural network input for maturity prediction. The accuracy of prediction was 75% for the first set of samples and 65.9% for the second set of samples. The results data show the potential of computer vision techniques for beef maturity assessment.

Carcasses from 64 gilts were subjected to electrical stimulation (ES) at 3, 15, 25, 35, 45, and 55 min postmortem or were untreated (NS). Temperature and pH of longissimus muscles were recorded at 1, 7, 14, 20, 30, 40, 50, and 60 min, and 24 h postmortem. Muscle samples were collected at 1, 30 and 60 min, and 24 h for determining glycolytic metabolite concentrations. ES at 3, 15, and 25 min resulted in lower (P<0.05) muscle pH, but stimulation after 25 min had no effect on muscle pH. Likewise, ES prior to 25 min resulted in greater (P<0.05) muscle temperatures. Muscle lactate concentrations were greater (P<0.05) in carcasses stimulated before 45 min postmortem. Glucose 6-phosphate concentration decreased (P<0.05) during the first hr postmortem and increased (P<0.05) thereafter. ES of carcasses at 45 and 55 min resulted in higher (P<0.05) concentrations of muscle glucose 6-phosphate at 24 h compared with NS and early-stimulated carcasses. Muscle glycogen concentrations at 30 min in carcasses stimulated at 3, 15 and 25 min were lower (P<0.05) than NS carcasses. Carcasses stimulated at 3 and 15 min exhibited lower (P<0.05) concentrations of muscle glycogen at 60 min than NS carcasses. Carcasses stimulated at 3 and 15 min postmortem exhibited lower (P<0.05) color and firmness scores, while ES at 3 and 25 min postmortem resulted in lower (P<0.05) water holding capacity. ES had no significant effect on CIE L(∗), a(∗), b(∗), or 24 h muscle pH. These data show that ES of pork carcasses during the first 25 min postmortem creates PSE-like quality characteristics and suggest that ES is a potential model for studying pork quality development.