The effect of physical training on skeletal muscle enzyme composition in pigs.

Running exercise in pigs results in an elevation of lactic acid in blood. This elevation in blood lactate does not occur in physically conditioned pigs. Activities of succinic dehydrogenase, fructose-l,6-diphosphate aldolase, lactate dehydrogenase and creatine Phosphokinase as well as the myoglobin content were determined in m. gastrocnemius from 6 ergometer-trained and 4 untrained pigs. The succinic dehydrogenase and myoglobin contents were significantly higher (P<0.01) in trained animals, whereas no changes were noted in the aldolase and creatine Phosphokinase contents. The lactate dehydrogenase showed somewhat reduced levels in the trained pigs. This was accompanied by an increased H/M subunit ratio. The results provide evidence for an increase in the maximal aerobic metabolism in trained pigs and that trained pigs to a higher extent can rely on an aerobic energy metabolism during running exercise.     

Effects of exercise on skeletal muscle and serum enzyme activities in pigs

In order to determine skeletal muscle and serum enzyme activities following exercise, six adult Landrace pigs were submitted to 10 min running on a treadmill (0.5 m/s, on a 12% gradient) and compared to six controls. Blood samples were obtained just before the exercise, immediately after, and 24 h, 48 h and 144 h after exercise. Muscle biopsies were taken from the longissimus dorsi and biceps femoris 24 h after exercise. Total lactic dehydrogenase (LDH) and LDH isoenzymes in muscle and serum were unchanged. In muscle homogenates, there was no difference in total creatine phosphokinase (CK) activity between the two groups. Total CK activity in the biceps femoris muscle represented only 59% of that observed in the longissimus dorsi muscle. A mean CK-MB value of 2.5% was found in the control group for both muscles but after exercise it was 9.5% (p<0.05) for the biceps femoris and 12.2% (p<0.01) of the longissimus dorsi muscle. In serum, the total CK (p<0.05), CK-MM (p<0.05) and CK-BB (p<0.05) increased immediately after the exercise, followed by a progressive decrease.     

Dose-response relationships between porcine somatotropin, muscle composition, muscle fiber characteristics and pork quality

The dose-dependent effects of porcine somatotropin (pST) on cellular aspects of skeletal muscle growth, muscle composition and measures of pork quality were investigated in growing barrows and gilts. Eighty crossbred pigs weighing 46 kg were assigned randomly to receive daily subcutaneous injections of 0, 30, 60, 120 or 200 micrograms pST/kg BW until they weighed 100 kg. Semitendinosus muscle weights were increased with pST dose (linear, P less than .001) by 21%. Percentage of type I and type II muscle fibers was not changed with pST, but cross-sectional area of type I and type II fibers was increased in parallel with muscle weight. Percentage of moisture increased (P less than .01) and percentage of lipid decreased (P less than .01) as pST dose increased. The pH of the longissimus 24 h postmortem increased (P less than .01) .1 to .2 units with increasing pST dose, but subjective evaluation for color, firmness and wateriness of the longissimus indicated no discernible treatment effect. Gardner color difference meter "Rd" and "A" values decreased (P less than .01) with a pST dose of 60 micrograms/kg or more, signifying a slightly darker and less red color, respectively, of the longissimus muscle. Weight loss of loin chops 2.54 cm thick cooked to 71 degrees C (20.3% to 23.7%) and shear force of cores 1.27 cm in diameter (2.89 to 3.76 kg) were not related to pST treatment or dose.(ABSTRACT TRUNCATED AT 250 WORDS)     

Simultaneous histochemical determination of three fiber types in single sections of ovine, bovine and porcine skeletal muscle

Classification of muscle fiber types involves tedious comparisons of serial muscle tissue sections. Procedures that would incorporate pertinent histochemical properties into one simplified assay for the successful differentiation and determination of fiber types were evaluated using muscle samples from three different species (ovine, bovine and porcine). For ovine and porcine muscle tissue, the best staining procedure involved initial preincubation of the tissue section at 4.35 (for ovine) and 4.30 (for porcine) followed by incubating for succinate dehydrogenase (SDH) activity, then staining for myofibrillar adenosine triphosphatase (ATPase) activity after an acid preincubation and, finally, counterstaining with hematoxylin. For bovine muscle tissue, the procedure involves first incubating for SDH activity, after which the section is incubated at pH 4.15 and then stained for myofibrillar ATPase activity after an acid preincubation, with a concluding counterstaining using hematoxylin. These procedures permit successful differentiation and identification of three fiber types (beta R, alpha R and alpha W) from a single section of muscle tissue with approximately a 65% saving in time and materials. The results of histochemically typing muscle fibers in a single tissue section stained with these techniques were similar to those obtained with the traditional methods comparing serial sections.     

Effects of a submaximal treadmill training programme on histochemical properties, enzyme activities and glycogen utilisation of skeletal muscle in the horse

The effects of training on skeletal muscle composition were studied in four Standardbred geldings given a seven week submaximal treadmill training programme. Before the start of training, muscle biopsies were collected from the left middle gluteal muscle for the determination of muscle fibre types, oxidative capacity and capillary numbers using histochemical techniques. The concentrations of citrate synthase, 3-hydroxyacyl-CoA dehydrogenase (HAD), lactate dehydrogenase and total muscle glycogen were measured using fluorometric methods. Muscle biopsy samples were repeated after one, three, five and seven weeks of training and the same measurements performed. No significant changes were found in muscle fibre types or capillary numbers as a result of training, although there was an increase in the oxidative capacity of the Type IIB fibres after seven weeks training. After seven weeks of training there were also increases in the concentrations of citrate synthase and HAD and a decrease in lactate dehydrogenase. Before commencement of training the horses underwent a standardised submaximal exercise test and muscle glycogen concentrations were measured before and immediately after the exercise. This procedure was repeated after one, three, five and seven weeks of training. A progressive decrease in the rate of glycogen utilisation occurred throughout the period of training, which by seven weeks was 36 per cent lower than that before training.     

Myostatin allelic status interacts with level of nutrition to affect growth, composition, and myofiber characteristics of lambs

The objective of this experiment was to determine if growth, carcass composition, and myofiber characteristics of lambs were affected by heterozygosity for a myostatin mutation (g+6723G>A) when offered differing allowances of feed administered with or without ractopamine. Heterozygote [MSTN A/G (n = 40)] and homozygote wildtype [MSTN G/G (n = 39)] castrate male lambs were individually fed ad libitum (HI; 1.8 × estimated ME(m)) or a restricted allowance (LO; 1.1 × estimated ME(m)) of a diet (191 g of CP/kg of DM and 12 MJ of ME/kg of DM), supplemented with (0.4 mg/kg of BW) or without the β-adrenergic agonist ractopamine (RAC or NO RAC) for 47 d. The lambs were scanned by computed tomography at the beginning and completion of the feeding experiment to calculate composition of lean, fat, and bone in the carcass component of the body. The MSTN A/G HI intake lambs had significantly greater total daily carcass growth (P = 0.045) and loin eye depth (P = 0.022) and tended to have a greater daily growth of lean (P = 0.09) in the carcass, compared with MSTN G/G HI intake lambs. Conversely, MSTN A/G LO intake lambs tended to have less daily lean deposition (P = 0.09), significantly less total daily carcass growth (P = 0.045), and had a greater percentage of type IIX myofibers (P < 0.01) and total myofiber area (relative area) of type IIX myofibers (P = 0.013). The inclusion of RAC increased final BW (P = 0.03) and ADG (P = 0.02), percentage of type IIC (P < 0.001) and IIA (P = 0.012) myofibers, cross-sectional area of types I (P = 0.04) and IIAX (P = 0.04) fibers, and the relative area of type IIC (P = 0.003) and IIA (P = 0.01) myofibers in the LM. The experiment demonstrated that including RAC in the diet of lambs increased final BW and ADG, but not HCW, and increased proportion of type IIC and IIA myofibers and cross-sectional area of type I and IIAX myofibers. Our data suggest that RAC and the heterozygous myostatin mutation act together to increase growth of muscle on a high plane of nutrition. The experiment also demonstrated that poor nutritional background of lambs heterozygous for the myostatin mutation may negatively influence their growth rates and myofiber characteristics.     

Effect of exogenous porcine somatotropin on the functional and textural characteristics of the porcine semimembranosus muscle

Sixteen barrows and gilts (sex effects were balanced within treatment, but not included in the model for analysis) were used in a 2 (porcine somatotropin; 2 mg.animal-1.d-1 vs control) X 2 (10% dietary fat vs control) X 4 (time postmortem; 0, 3, 6, and 24 h) factorial treatment array to evaluate the effects of porcine somatotropin (pST) administration and level of dietary fat intake on the functional and textural characteristics of the semimembranosus muscle during the first 24 h postmortem. Porcine somatotropin administration resulted in a decrease (P less than .05) in muscle tenderness, an increase (P less than .05) in chilled carcass weight, and an increase (P less than .08) in longissimus muscle area. The pH values were lower over time with elevated levels of dietary fat, but pST resulted in no alterations in muscle pH. The R-values (the ratio of inosine to adenine nucleotides) were unaffected by pST or by level of dietary fat, but the combination of the two resulted in an increase in moisture binding capabilities. It could be concluded from the present study that pST and increased level of dietary fat result in an alteration of certain functional (tenderness) and textural (water binding ability and cooking losses) characteristics of the porcine semimembranosus muscle.     

Dietary manipulation of pro- and macroglycogen in porcine skeletal muscle

The aim of the study was to investigate how feeding-induced changes in muscle glycogen stores affect the ratio of between the glycogen pools, pro- and macroglycogen. Pro- and macroglycogen content were determined in longissumus muscle from slaughter pigs subjected to a feeding strategy known to reduce total glycogen stores. Furthermore, early postslaughter glycolysis of the two glycogen forms was determined. The feeding strategy involved a diet (GLYRED diet) with a low digestible carbohydrate (5%)/high fat (18%) content, which was fed to the pigs the last 3 wk before harvest. A control group was fed a standard pig diet (49% digestible carbohydrate/5% fat). Total glycogen was reduced by 48 micromol/g dry weight (d.w.) in GLYRED pigs during the 3-wk feeding period. This was mainly due to a reduction in macroglycogen of 42 micromol/g d.w. During postmortem glycolysis the proglycogen appeared to be degraded in favor of macroglycogen. Moreover, total glycogen was degraded to a larger extent in muscle from the control pigs compared with muscle from GLYRED pigs. This difference was due to a significantly greater degradation of proglycogen in the control pigs. In conclusion, the results support earlier studies suggesting that proglycogen and macroglycogen are different glycogen pools that have different functions. Furthermore, the results show that the muscle glycogen pools can be manipulated through diet and that proglycogen is degraded in favor of macroglycogen under the anaerobic conditions postmortem.     

Histochemical properties of muscle fibres types and enzyme activities in skeletal muscles of Standardbred trotters of different ages

Fibre characteristics and enzyme activities were determined for the gluteus, semitendinosus, vastus lateralis and triceps brachii muscles of 55 Standardbred trotters of different ages. Four fibre types (I, IIA, IIB, IIC) were demonstrated by histochemical staining of myofibrillar adenosine triphosphatase after preincubation at different pH values. Type II fibres predominated in all the muscles and the type IIA/IIB ratio was higher in horses over 5 years than in younger horses, except in the vastus in which the IIA/IIB ratio did not change with age. The vastus had the highest proportion of type IIA fibres and the semitendinosus the highest proportion of type IIB fibres. Histochemical demonstration of NADH dehydrogenase disclosed that almost 100 per cent of the type IIA and many of the type I and IIB fibres were medium-stained; the remaining type I fibres were darkly stained and the type IIB fibres lightly stained. In older horses more fibres were stained for NADH dehydrogenase. The activity of triosephosphate dehydrogenase decreased that that of 3-hydroxy-acyl-coA dehydrogenase and citrate synthase increased in all the muscles except the vastus with increasing age. The greatest increase in oxidative capacity occurred in the gluteus and triceps. Training, rather than age, was regarded as the factor inducing these changes. The results emphasise that histochemical data are only semiquantitative, and there are apparent discrepancies in the intensities of histochemical staining and the biochemical evaluation of various enzymes.     

Effects of age and castration on activities of calpains and calpastatin in sheep skeletal muscle.

The objectives of this experiment were to assess effects of animal age and castration on activities of calpain I, calpain II, and calpastatin in sheep skeletal muscle. Ten newborn male lambs (2.9 kg), six weaned wethers (23.2 kg), six weaned rams (22.2 kg), six market wethers (55.4 kg), and six market rams (60.2 kg) were slaughtered and samples of biceps femoris were taken for assay of calpain I (micromolar calcium-dependent proteinase), calpain II (millimolar calcium-dependent proteinase), and calpastatin. Preweaning weight gain was similar for rams and wethers; however, postweaning ram growth exceeded (P less than .05) that of wethers. Ram biceps femoris weights at market were greater than those of wethers (P less than .05). Irrespective of age or gender, activity of calpain II was two- to threefold greater than that of calpain I. Muscle calpastatin activity was severa fold higher than calpain I and II activities. Activities of calpains and calpastatin declined (P less than .05) between birth and weaning. A portion of these losses were due to a dilution effect caused by accumulation of muscle proteins. Neonatal attenuation of calpain activities may underlie age-related attenuation of fractional rates of muscle protein degradation. Although ram muscle growth exceeded that of wethers, no differences (P greater than .05) in activities of muscle calpains or calpastatin were detected between these two groups at weaning or at market weight. Hence, castration did not influence lamb muscle growth by altering muscle calpain or calpastatin activities.     

Effect of maternal diet on skeletal muscle composition and metabolism and on bone dimensions and composition of the fetal pig

The purpose of the present study was to determine the effect of addition of fat to sow diets on selected muscle and bone traits of the fetus. Crossbred dams were assigned to receive one of four dietary treatments at 80 d of gestation. Percentage added poultry fat in the diet was 0, 2.3, 12.7 or 29.6%. Isocaloric-isonitrogenous diets (7,000 kcal ME) were fed once daily. At 110 d of gestation, sows were anesthetized with fluothane and nitrous oxide. Fetuses were removed by Caesarean section. Fetal body weight, biceps femoris muscles and femur wet weights and blood (concentration of protein, free fatty acids and triacylglycerol), muscle (percentage dry matter, concentration of DNA, RNA, protein and lipid) and bone (percentage dry matter, lipid, ash and length) traits were not influenced by maternal dietary fat content. The rates of oxidation of palmitate and of lipid to CO2 and the rates of incorporation of palmitate into triacylglycerol and into phospholipid by fetal pig muscle also were not influenced by maternal dietary fat content. It was suggested that fetal skeletal muscle can mobilize glycogen and, to a limited extent, lipid stores.     

Effect of age, muscle type, and insulin-like growth factor-II genotype on muscle proteolytic and lipolytic enzyme activities in boars

Recently, a paternally expressed quantitative trait nucleotide (QTN) in the regulatory sequence of the IGF-II gene with effects on muscle growth and fat deposition was discovered in the pig. This QTN is also known as the IGF-II intron3 G3072A mutation. The aim of the current study was to determine the effects of age, muscle type, and IGF-II genotype (Apat, mutant allele vs. Gpat, wild-type allele) on muscle proteolytic and lipolytic enzyme activities. At approximately 4, 8, 16, and 26 wk of age, boars (n = 6 to 15 per genotype x age group) were slaughtered and mu- and m-calpain (CALP), calpastatin (CAST), cathepsins (CATH) B+L and H, acid lipase, and phospholipase activities were measured in Longissimus thoracis et lumborum, Semimembranosus, and Triceps brachii muscle samples taken soon after slaughter. Activities of CATH B+L and H, mu- and m-CALP, and acid lipase were not affected by the IGF-II genotype. Activity of CAST was greater (P < 0.005) and m-CALP:CAST was less (P < 0.05) in Apat animals. Because CAST activity and m-CALP:CAST are known to be related to protein degradation, satellite cell fusion, or both, it is likely that differences in proteolytic enzyme activities are involved in the greater percentage of muscle mass in Apat animals. Age and muscle type influenced proteolytic and lipolytic enzyme activities (P < 0.05), except for mu- and m-CALP (no effect of muscle) and acid lipase (no effect of age). The same pattern in mu-CALP, CAST, and m-CALP:CAST with age was found during growth for the 3 muscles, although clear differences (P < 0.05) between muscles existed. In general, and in agreement with previous reports, greater enzyme activities were found in the more oxidative Triceps brachii muscle compared with the other 2 muscles. A remarkable increase (P < 0.05) from 16 to 26 wk of age in mu-CALP, CAST, mu-CALP:CAST, and CATH H and a large decrease (P < 0.05) in acid phospholipase and m-CALP:CAST was found. For m-CALP and CATH B+L, a gradual decrease (P < 0.05) was found with age. Although age effects on enzyme activities could only partly be interpreted biologically in relation to the muscle growth rate, this study showed that proteolytic and lipolytic enzyme activities change during growth.     

Composition of intermediate filament subunit proteins in embryonic, neonatal and postnatal porcine skeletal muscle

The intermediate (10-nm) filament subunit proteins (desmin and vimentin) in samples obtained from embryonic, neonatal, and postnatal porcine skeletal muscle were examined by two-dimensional electrophoresis (isoelectric focusing/sodium dodecylsulfate polyacrylamide gel electrophoresis). The skeletal muscle samples were taken from pig embryos at 45, 73 and 102 d of gestation; from neonatal pigs and from postnatal pigs at 1, 6 and 30 mo of age. Three fractions (namely, whole homogenized muscle, purified myofibrils and myofibrillar-protein-extracted residues) were prepared from each skeletal muscle sample for analysis. Vimentin was the major (approximately 75% vimentin: 25% desmin) 10-nm filament protein present in skeletal muscle samples obtained from the 45-d-old pig embryos. The relative proportion of vimentin decreased progressively during embryogenesis. At birth, the vimentin comprised approximately 15%, and desmin, 85%, of the 10-nm filament protein. The proportional amount of vimentin continued to decline postnatally, with the 10-nm filament protein of samples from the 30-mo-old animals consisting of less than approximately 5% vimentin and over 95% desmin. These results show a developmental stage-dependent pattern in the expression of vimentin and desmin intermediate filament subunit proteins in mammalian skeletal muscle. In the adult mammal, desmin is the significant 10-nm filament protein present.     

Effect of clenbuterol on growth,carcase and skeletal muscle characteristics in broiler chickens

1. Male and female broiler chickens (144 in total) were given diets supplemented with clenbuterol (CB) at 0 (control) and at 1 mg/kg between 28 and 49 d of age to study the effect of CB on growth, carcase and skeletal muscle.

2. CB improved growth in males by increasing daily weight gain and final live weight and by lowering food conversion ratio. In females it changed the carcase composition by reducing abdominal fat pad and by increasing the proportion of protein. Consequently, carcase protein gain was increased in both sexes (11% and 16%, respectively).

3. Skeletal muscle weights were enhanced by between 6% and 22%. Muscle fibre diameters were increased in extensor hallucis longus (EHL) but not in gastrocnemius (GAS) muscle. This increase was more pronounced in females. EHL total muscle fibre number remained unchanged. The proportion of fast‐twitch glycolytic fibres was increased at the expense of fast‐twitch oxidative fibres in males only. Nuclear/cytoplasm and DNA/protein ratios tended to be decreased by CB.

4. From the elevated EHL muscle RNA/DNA, unchanged protein/RNA and translation activity it is suggested that CB stimulated protein synthesis at the pretranslational level. Reduced protein degradation is deduced from decreased neutral calcium‐dependent proteolytic activity.

5. It is concluded that broiler chickens respond to long‐term CB treatment as has been shown in various mammals. However, the sex‐specific response in growth, carcase composition and skeletal muscle cellularity is more clearly apparent in broiler chickens.     

Effect of exogenous porcine somatotropin administration on nitrogen and energy metabolism in three genotypes of pigs

Effects of exogenous administration of porcine recombinant somatotropin (rpST) on protein gain and metabolic rate were measured in three genotypes (castrated males) of pigs (Pietrain, Duroc and a crossbreed between Dutch Yorkshire and Dutch Landrace). Six pigs of each genotype were assigned at approximately 60 kg to receive pST doses of either 0 (C) or 14 mg (T) administered i.m. twice weekly over 10 wk. Pigs were housed in individual metabolism cages at a room temperature of 20 to 22 degrees C and received feed at 2.6 times maintenance. Protein gain (N x 6.25) was measured over the final 6 wk of the 10-wk test period. For 2 wk in the test period (wk 2 and wk 5), six pigs of each treatment x genotype group were placed in a large respiration chamber and energy balances (in protein and fat) and metabolic rate were measured. Rate of weight gain measured over the final 6 wk of the experimental period increased by 105 g/d (13%) with rpST administration (P less than .01). Daily protein gain over 6 wk was increased by 48 g/animal with application of rpST (P less than .001). There was a genotype x treatment interaction (P less than .01) for protein gain. Daily protein gain in Durocs with pST treatment was increased (39%) more than in crossbreds (31%). Daily fat gain was decreased by 42 g/animal (P less than .001) by T. Daily heat production with rpST was increased by 12 kcal/kg.75, which is comparable to a 12% increase in the maintenance energy requirement.     

Effect of the repartitioning agent cimaterol on growth, carcass and skeletal muscle characteristics in lambs

Ten crossbred (Suffolk X Rambouillet) whether lambs were randomly assigned to receive 0 or 10 ppm cimaterol (CIM) in a completely mixed high-concentrate diet for 8 wk. Total weight gain and feed efficiency were improved 29% (P less than .05) and 14%, respectively, in the CIM-fed group. CIM also improved (P less than .01) dressing percent by 4.9 percentage points and improved yield grade by one grade. CIM increased longissimus muscle (LD) area 38% (P less than .01) and the yield of four lean cuts 28% (P less than .01). No difference was found in the proportion of type I (slow-contracting, oxidative) and type II (fast-contracting, mixed glycolytic/oxidative) fibers in LD and semitendinosus (ST) muscles between control and CIM groups, indicating no change in fiber type. The cross-sectional area of type II fibers in LD and ST muscles of the CIM group was 2,081 and 1,951 micron 2 as compared with 1,391 and 1,296 micron2 of the control group, respectively. The increase was approximately 50% (P less than .01). No difference was found in cross-sectional area of type I fibers, indicating that the increase of muscle mass was due to hypertrophy of type II fibers only. DNA concentration (micrograms/g wet muscle or microgram/g protein) of CIM muscle was much lower (P less than .01) than that of control muscle, suggesting that the protein accretion in muscle was accomplished without additional incorporation of nuclei from satellite cells.