D. E. Gerrard

Mechanisms controlling pork quality development: The biochemistry controlling postmortem energy metabolism

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.

Evaluation of pork color by using computer vision.

The objective of this study was to determine the potential of computer vision technology for evaluating fresh pork loin color. Software was developed to segment pork loin images into background, muscle and fat. Color image features were then extracted from segmented images. Features used in this study included mean and standard deviation of red, green, and blue bands of the segmented muscle area. Sensory scores were obtained for the color characteristics of the lean meat from a trained panel using a 5-point color scale. The scores were based on visual perception and ranged from 1 to 5. Both statistical and neural network models were employed to predict the color scores by using the image features as inputs. The statistical model used partial least squares technique to derive latent variables. The latent variables were subsequently used in a multiple linear regression. The neural network used a back-propagation learning algorithm. Correlation coefficients between predicted and original sensory scores were 0.75 and 0.52 for neural network and statistical models, respectively. Prediction error was the difference between average sensory score and the predicted color score. An error of 0.6 or lower was considered negligible from a practical viewpoint. For 93.2% of the 44 pork loin samples, prediction error was lower than 0.6 in neural network modeling. In addition, 84.1% of the samples gave an error lower than 0.6 in the statistical predictions. Results of this study showed that an image processing system in conjunction with a neural network is an effective tool for evaluating fresh pork color.

Pleiotropic effects in Hereford, Limousin, and Piedmontese F2 crossbred calves of genes controlling muscularity including the Piedmontese myostatin allele.

Objectives were to determine 1) effects on traits measured from birth to slaughter in F2 cross calves from sire breeds that differ in potential for lean tissue growth but have similar mature BW and 2) the gene action of the mutant Piedmontese myostatin allele. Hereford (normal muscling, H), Limousin (moderate increase in muscling, L), and Piedmontese (muscular hypertrophy, P) sires (20 to 25 per breed) were bred at random to crossbred cows to produce F1 calves that were inter se-mated within sire breed to produce F2 calves that were grown out, finished, and slaughtered. Piedmontese-cross calves were genotyped for the G-A transition mutation at the myostatin locus characteristic of P (msP). Genotypes were classified on the basis of having zero (P0), one (P1), or two (P2) copies of msP (H, n = 227; L, n = 207; P0, n = 40; P1, n = 107; and P2, n = 37). Limousin-cross F2 calves had heavier birth (but dystocia was not affected) and weaning weights, gained faster, had more muscle, less fat, larger pelvic area, and more efficient feed conversion than Hereford-cross F2 calves. Normal-muscled Piedmontese-cross F2 calves (P0) were similar to Hereford-cross F2 calves except that they required less assistance at birth in heifer dams, had less fat, gained slower, were less efficient, and had larger pelvic area. Addition of msP alleles (P1 and P2) consistently increased muscle through hyperplasia, decreased fat, and increased adjusted efficiency, but many of those changes were not linear. Residual variances for breed were heterogeneous for most traits related to muscularity. This heterogeneity was caused by increased variances for L and P and(or) lower variances for H. Accounting for the msP alleles decreased the variance for P in most traits, but heterogeneity remained for most traits among the five genotypes because L remained high, H was low, and(or) P2 was low. We conclude that differences in muscularity affect most traits, and when differences in muscularity include the msP allele, there is an incremental, but not equal, change in most traits with the addition of each copy of the msP allele. Advantages of L could be captured through normal crossbreeding and selection schemes but with some caution because of potential problems from increased variability. Advantages of P could be best captured through more complex breeding and selection programs that would lessen potential negative impacts and through marketing systems that do not penalize for very low fat.

Mitogen-activated protein kinase signaling is necessary for the maintenance of skeletal muscle mass

The signal transduction cascades that maintain muscle mass remain to be fully defined. Herein, we report that inhibition of extracellular signal-regulated kinase 1/2 (ERK1/2) signaling in vitro decreases myotube size and protein content after 3-day treatment with a MEK inhibitor. Neither p38 nor JNK inhibitors had any effect on myotube size or morphology. ERK1/2 inhibition also upregulated gene transcription of atrogin-1 and muscle-specific RING finger protein 1 and downregulated the phosphorylation of Akt and its downstream kinases. Forced expression of enhanced green fluorescent protein-tagged MAPK phosphatase 1 (MKP-1) in soleus and gastrocnemius muscles decreased both fiber size and reporter activity. This atrophic effect of MKP-1 was time dependent. Analysis of the reporter activity in vivo revealed that the activities of nuclear factor-kappaB and 26S proteasome were differentially activated in slow and fast muscles, suggesting muscle type-specific mechanisms may be utilized. Together, these findings suggest that MAPK signaling is necessary for the maintenance of skeletal muscle mass because inhibition of these signaling cascades elicits muscle atrophy in vitro and in vivo.

Myosin heavy chain isoforms account for variation in pork quality

The objective of this study was to determine the impact of myosin heavy chain (MyHC) isoforms (I, IIB, IIA and IIX) on pork quality traits of halothane (HAL)-negative (NN) and halothane-carrier (Nn) pigs. Gilts (n=32) were assigned to a 2×2 factorial of genetic population (GP) and slaughter weight (WT; 120 and 135 kg). Classical meat quality characteristics were collected and MyHC content was determined on muscle samples. Regression equations for pork quality and carcass composition traits were determined. Only I/IIB accounted for variation in drip loss of NN gilts (R(2)=0.18), while GP related to drip loss in Nn gilts (R(2)=0.70). Type I MyHC content explained variation in ultimate (24 h) muscle pH of NN gilts (R(2)=0.09), while I/IIB, I/IIX and IIB/IIX were significant for Nn gilts (R(2)=0.99). I/IIA, I/IIX, IIB/IIX and GP accounted for variation in Hunter Color a (redness) values of NN gilts (R(2)=0.69), while IIB, IIA, IIB/IIA and GP were significant for Nn gilts (R(2)=0.97). Overall, fiber type composition accounts for a larger proportion of variation in the quality traits of Nn compared to NN gilts.

Modulation of skeletal muscle fiber type by mitogen-activated protein kinase signaling

Skeletal muscle is composed of diverse fiber types, yet the underlying molecular mechanisms responsible for this diversification remain unclear. Herein, we report that the extracellular signal‐regulated kinase (ERK) 1/2 pathway, but not p38 or c‐Jun NH2‐terminal kinase (JNK), is preferentially activated in fast‐twitch muscles. Pharmacological blocking of ERK1/2 pathway increased slow‐twitch fiber type‐specific reporter activity and repressed those associated with the fast‐twitch fiber phenotype in vitro. Overexpression of a constitutively active ERK2 had an opposite effect. Inhibition of ERK signaling in cultured myotubes increased slow‐twitch fiber‐specific protein accumulation while repressing those characteristic of fast‐twitch fibers. Overexpression of MAP kinase phosphatase‐1 (MKP1) in mouse and rat muscle fibers containing almost exclusively type IIb or IIx fast myosin heavy chain (MyHC) isoforms induced de novo synthesis of the slower, more oxidative type IIa and I MyHCs in a time‐dependent manner. Conversion to the slower phenotype was confirmed by up‐regulation of slow reporter gene activity and down‐regulation of fast reporter activities in response to forced MKP1 expression in vivo. In addition, activation of ERK2 signaling induced up‐regulation of fast‐twitch fiber program in soleus. These data suggest that the MAPK signaling, most likely the ERK1/2 pathway, is necessary to preserve the fast‐twitch fiber phenotype with a concomitant repression of slow‐twitch fiber program.—Shi, H., Scheffler, J. M., Pleitner, J. M., Zeng, C., Park, S., Hannon, K. M., Grant, A. L., Gerrard, D. E. Modulation of skeletal muscle fiber type by mitogen‐activated protein kinase signaling. FASEB J. 22, 2990–3000 (2008)

Chronic elevated calcium blocks AMPK-induced GLUT-4 expression in skeletal muscle

Muscle contraction stimulates glucose transport independent of insulin. Glucose uptake into muscle cells is positively related to skeletal muscle-specific glucose transporter (GLUT-4) expression. Therefore, our objective was to determine the effects of the contraction-mediated signals, calcium and AMP-activated protein kinase (AMPK), on glucose uptake and GLUT-4 expression under acute and chronic conditions. To accomplish this, we used pharmacological agents, cell culture, and pigs possessing genetic mutations for increased cytosolic calcium and constitutively active AMPK. In C2C12 myotubes, caffeine, a sarcoplasmic reticulum calcium-releasing agent, had a biphasic effect on GLUT-4 expression and glucose uptake. Low-concentration (1.25 to 2 mM) or short-term (4 h) caffeine treatment together with the AMPK activator, 5-aminoimidazole-4-carboxamide-1-beta-D-ribonucleoside (AICAR), had an additive effect on GLUT-4 expression. However, high-concentration (2.5 to 5 mM) or long-term (4 to 30 h) caffeine treatment decreased AMPK-induced GLUT-4 expression without affecting cell viability. The negative effect of caffeine on AICAR-induced GLUT-4 expression was reduced by dantrolene, which desensitizes the ryanodine receptor. Consistent with cell culture data, increases in GLUT-4 mRNA and protein expression induced by AMPK were blunted in pigs possessing genetic mutations for both increased cytosolic calcium and constitutively active AMPK. Altogether, these data suggest that chronic exposure to elevated cytosolic calcium concentration blocks AMPK-induced GLUT-4 expression in skeletal muscle.

Early postmortem electrical stimulation simulates PSE pork development

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.

Effects of electrical stimulation on early postmortem muscle pH and temperature declines in pigs from different genetic lines and halothane genotypes

The objective of this study was to determine if electrical stimulation (ES) early postmortem is an effective method to generate PSE-like meat. One hundred and thirty-eight gilts (85-125 kg) from heavy muscled (HM), normal muscled (NM), and light muscled (LM) porcine genetic lines were subjected to one of two treatments: ES (26 pulses of 500 V, 60 Hz) at 3 min postmortem or non-stimulated (NS). Pigs from HM line were further characterized as halothane (HAL) carriers (Nn) or non-carriers (NN). ES carcasses had lower (P<0.0001) pH values and higher (P<0.0001) temperature than NS carcasses during the first 56 min postmortem. ES carcasses had lower (P<0.0001) a*-values, and color and firmness scores, as well as higher (P<0.0001) drip loss and L*-values. No significant interactions were found between treatment and genetic line or HAL gene status with regard to pH, temperature, or quality characteristics. Temperature and pH declines within the first hour postmortem were not affected by genetic line, but slight (P<0.01) quality differences were observed. Nn and NN did not differ in pH or temperature within the first hour postmortem, but Nn carcasses had lower (P<0.01) color and firmness scores, and higher (P<0.05) drip loss. These results show that ES early postmortem is an effective method for simulating PSE development in pigs of different muscling and HAL gene status, and suggest that pH and temperature decline alone cannot explain all aspects of pork quality.

Effects of dietary fat and crude protein on feedlot performance, carcass characteristics, and meat quality in finishing steers fed differing levels of dried distillers grains with solubles.

The objective of this study was to evaluate the influence of dietary protein and fat from distillers dried grains with solubles (DDGS) on feedlot performance, carcass characteristics, and meat quality in finishing steers. Angus-cross steers (n = 105; 443 +/- 20 kg of BW) were blocked by BW and randomly assigned to 1 of 5 dietary treatments: 1) corn-based diet with DDGS included at 25% of DM (CON), 2) CON with DDGS included at twice the amount of CON (50% of DM; 50DDGS), 3) CON with added corn protein to equal the CP in the 50DDGS diet (CON+CP), 4) CON with added vegetable oil to equal the fat in the 50DDGS diet (CON+VO), and 5) CON with protein and fat added to equal the CP and fat in the 50DDGS diet (CON+CPVO). Steers were fed to a common 12th-rib fat depth endpoint (1.3 +/- 0.2 cm; 68 to 125 d on trial). Loins and rounds were collected from 44 carcasses for Warner-Bratzler shear force (WBSF), ether extract, and case-life analyses. Data were analyzed using the MIXED procedure of SAS. Contrasts between 1) CON vs. elevated CP diets (50DDGS, CON+CP, and CON+CPVO; EP), 2) CON vs. elevated fat diets (50DDGS, CON+VO, and CON+CPVO; EF) and 3) CON vs. diets with elevated CP and fat (50DDGS and CON+CPVO; EPF) were analyzed. There were no differences in days on feed or DMI among treatments. Steers fed CON had greater ADG (P <or= 0.03) than EP, EF, and EPF diets. Steers fed CON also had greater G:F (P <or= 0.04) than EP and EPF steers. Final BW was greater for CON than EP and EPF diets (P <or= 0.03). Likewise, CON steers had heavier HCW than EPF steers (P = 0.04). Dressing percent, 12th-rib fat depth, LM area, KPH, and yield grade were not affected by treatment (P >or= 0.06). Steers fed the CON diet had greater marbling scores (P <or= 0.03) and quality grades (P <or= 0.02) compared with those fed EP, EF, and EPF diets. There were no differences in WBSF, ether extract, or lipid oxidation due to treatment (P >or= 0.44). However, CON steers had greater (P = 0.02) L* values than EF-fed steers and greater b* values than EP, EF, and EPF steers (P <or= 0.02) during retail display of ground product. Data from this study illustrate that live animal performance, marbling and quality grade, and color stability of ground product during retail display are negatively affected when DDGS are increased from 25 to 50% of the diet DM. This response appears to be due to elevated dietary fat, elevated CP, and a combination of elevated fat and protein within in the diet.