Identification by isoelectric focusing of creatine kinase-MM isoforms in the plasma and striated muscle of pigs

Total creatine kinase (CK) and CK-MM isoforms were determined in plasma and longissimus dorsi muscle extracts from normal pigs. Based on their total CK activity, the pigs were divided into two groups. Pigs of group 1 (n=16) had a mean plasma total CK of 298±16 U/L and the distribution of the CK-MM isoforms was 65.7±2.5% CK-MM₃, 18.9±1.6% CK-MM₂ and 15.3±1.5% CK-MM. In group 2 (n=18; 826±75 U/L total CK) four isoforms were observed: 3.1±0.9% CK-MM, 67.9±3.0% CK-MM₃, 21.5±2.3% CK-MM₂ and 7.5±1.3% CK-MM₁. The differences between the two groups of pigs were significant (p<0.001) for CK-MM₁ and the presence of CK-MM. Four CK-MM isoforms were also detected in longissimus dorsi muscle homogenates: 45.6±8.1% CK-MM, 32.6±11.7% CK-MM₃, 16.6±2.3% CK-MM₂ and 5.1±2.8% CK-MM₁. The release of CK-MM isoforms from muscle into plasma seems to be unrelated to the concentration of these isoforms in striated muscle.     

Effects of I.M. injections on serum creatine phosphokinase (CPK) values in dogs

Intramuscular injections of compounds of low irritancy (bicillin, valium, saline) were found to cause significant elevations in serum CPK in dogs. This route of treatment must therefore be avoided in clinical conditions where CPK values have diagnostic or prognostic importance.     

Value of measurements of the serum activities of creatine phosphokinase,aspartate aminotransferase and lactate dehydrogenase for predicting whether recumbent dairy cows will recover

Over a period of seven years, 262 recumbent dairy cows were investigated, and serum samples were analysed for creatine phosphokinase (CPK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities. The results from cows for which all three measurements were available, were analysed statistically by the non-parametric receiver operating characteristic (ROC) method, to evaluate the value of the serum enzyme activities for predicting a failure to recover. The sensitivity and specificity of the activity levels were calculated over all cut-off points, and ROC curves were created by plotting sensitivity as a function of 1--specificity at each cut-off point for samples grouped by the interval after the cows became recumbent. The predictive values of the tests were compared by calculating the areas under the curves, and the positive and negative predictive values of the tests were calculated and plotted against the prevalence of a failure to recover. The results from each of the enzymes were significantly better than from a random test with no discriminatory ability. The optimal cut-off points maximising the sensitivity and specificity of the tests were 2330, 2225 and 171 U/litre for CPK, LDH and AST, respectively. The predictive value of AST was significantly better than that of CPK or LDH, and measures made on the second and third day of recumbency were significantly better with optimal cut-off points of 128 and 189 U/litre, respectively.     

Effects of exposure to a 50 Hz electric field on plasma levels of lactate, glucose, free Fatty acids, triglycerides and creatine phosphokinase activity in hind-limb ischemic rats

We previously reported that extremely low frequency electric fields (ELF-EFs) affect energy metabolism in stressed conditions. To further confirm this, the effect of exposure to ELF-EFs on the experimental ischemic rat was examined. The test was based on a comparison of rats treated with EF alone, ischemic surgery alone, the combination of EF with ischemic surgery, or no treatment (double sham). The EF condition used in this study was an alternating current of 50 Hz EF at 17 500 V/m intensity for 15 min per day. The exposure to EF in ischemic rats significantly decreased plasma levels of free fatty acids and triglycerides, compared to those of the no treatment or EF alone group. The plasma lactate levels of two ischemic groups peaked on experimental day-4 and gradually decreased until the end of the study. The changes in the lactate levels induced by ischemia did not show any difference between rats treated with ischemia alone or a combination of ischemia with an EF. Any changes in plasma levels of glucose and creatine phosphokinase activity were not influenced by EF treatment. These results indicate that the EF effect on glycolysis parameters, plasma lactate or glucose levels, does not appear in a highly stressed condition and that EF effects varied dependent on the condition of organism but ELF-EF used in this study have impact on lipid metabolism parameter in a hind-limb ischemic rat. However, further studies are needed to elucidate the association of ELF-EF with the lipid metabolism system.     

Changes in heart rate, mean arterial pressure, blood biochemistry, plasma glucose, plasma lactate and some plasma enzymes during sufentanil/halothane anaesthesia in horses

Nine horses were each anaesthetised for 40 min using SufentaniVhalothane. No surgery was performed. After premedication (detomidine 5 pgkg bwt iv) induction of anaesthesia was achieved by a combination of guaiphenesinlthiopentone. Anaesthesia was maintained by inhalation of halothane (0.8%) in oxygen. Six horses (Group 1) received 1 pgkg bwt sufentanil followed by a second injection (1 pg/kg bwt) after 20 min. Three horses (Group 2) received 2 pg/kg bwt sufentanil also followed by a second injection (2 pg/kg bwt) after 20 min. Each sufentanil injection produced a slight decrease in mean arterial blood pressure with a gradual return to the initial pressure. Bradycardia was also observed. Sufentanil injection induced apnoea needing artificial ventilation. Arterial blood was sampled for analysis during the anaesthetic procedure. At the end of anaesthesia, 1 h and 24 h after rising, venous blood was sampled to determine concentrations of lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and creatine phosphokinase (CPK). Values obtained were compared with values in blood taken before premedication. Plasma glucose and lactate concentrations just before sufentanil administration, at the end of anaesthesia and 1 h after rising were compared to control values. Plasma glucose concentration increased significantly during anaesthesia but returned to normal values 1 h after rising. All other parameters stayed within physiological ranges. In both groups spontaneous respiration returned 20–25 min after the second sufentanil injection. Recovery was uneventful.     

Utilization of the halothane test in the detection of the malignant hypertension syndrome [MHS] in pigs]

By the halothane test we determined the development of the syndrome of malignant hyperthermia in pigs of the Belgian Landrace breed with body weight of 20 kg. We found that from 27 animals that were subjected to the test 12 positively reacted to Narcotan - Spofa anaesthesis. Pigs with the syndrome of malignant hyperthermia had considerably higher rectal temperature after anaesthesis, lower PH of venose blood, higher concentration of lactic acid and glucosis in the blood plasma. Halothane-negative animals did not produce and conspicuous changes of these indices. Most differences were statisticaly highly significant.     

Effects of experimenters and different blood sampling procedures on blood metabolite values in growing pigs.

The aims of the present study were firstly to verify if sera obtained by jugular venipuncture and those obtained from an anterior vena cava cannula were comparable and secondly to observe the effect of different experimenters on the biochemical profile of pigs. In the first experiment, 16 Yorkshire pigs with a mean weight of 48.9 +/- 2.3 kg were venipunctured using either a vacuum tube system or a syringe. The experiment was conducted on two separate days, seven days apart, in a crossover design experiment. At the time of obtaining blood via venipuncture, blood was also withdrawn from the jugular cannula. The tip of the latter was in the anterior vena cava. No effect of the sampling system was observed. However, a significant decrease in serum concentrations of Na, P and CO2 and a significant increase in anion gap, total protein, albumin, globulin, total bilirubin, aspartate amniotransferase, L-gamma glutamyl-transferase and creatine kinase were observed from blood samples obtained by venipuncture. In the second experiment, ten pigs from the first experiment were sampled by an inexperienced manipulator. Variations of the same magnitude as observed in the first experiment were observed in sera obtained by venipuncture and anterior vena cava but 18 of the 22 biochemical variables of sera obtained from the anterior vena cava were significantly different than those from the first experiment. The results show that blood samples obtained by jugular venipuncture in pigs can differ from samples obtained from an anterior vena cava cannula, specially in enzyme concentrations, and the dexterity of the experimenter may have a major effect on blood values.     

The relationship between respiratory exchange ratio, plasma lactate and muscle lactate concentrations in exercising horses using a valved gas collection system.

A valved gas collection system for horses was validated, then used to examine the relationship between the respiratory exchange ratio (RER), and plasma and muscle lactate in exercising horses. Four healthy Standardbred horses were trained to breathe through the apparatus while exercising on a treadmill. Comparisons of arterial blood gas tensions were made at 3 work levels for each horse, without (control), and with the gas collection system present. At the highest work level, the arterial oxygen tension (PaO2) was significantly lower (P < 0.05), and the arterial carbon dioxide tension (PaCO2) was significantly higher (P < 0.05), than control levels when the apparatus was present; however arterial oxygen content remained unchanged. The horses completed a standardized incremental treadmill test on 4 occasions to determine the repeatability of measurements of oxygen consumption (VO2), carbon dioxide production (VCO2), inspired minute ventilation (VI), respiratory exchange ratio (RER), ventilatory equivalent for oxygen (VI/VO2), tidal volume (VT), and ventilatory frequency (VF). All gas exchange and respiratory measurements showed good reproducibility with the mean coefficient of variation of the 4 horses ranging from 3.8 to 12%. We examined the relationship between 3 indices of energy metabolism in horses performing treadmill exercise: respiratory exchange ratio (RER), central venous plasma and muscle lactate concentrations. A relationship between RER and plasma lactate concentration was established. To compare muscle and plasma lactate concentrations, the horses completed a discontinuous exercise test without the gas collection apparatus present. Significant relationships (P < 0.05), between plasma lactate concentration and RER, and between plasma and muscle lactate concentration, were described for each horse. The valved gas collection system produced a measurable but tolerable degree of interference to respiration, and provided reproducible measurements of gas exchange and ventilatory measurements. It was concluded that measurements of both gas exchange and blood lactate may be used to indicate increased glycolytic activity within exercising skeletal muscle.     

Stability of vitamin E in blood and plasma from cattle, sheep, and pigs.

The stability of alpha-tocopherol concentrations in sheep, cattle, and pig blood and plasma stored at different temperatures was examined. For all species, the vitamin was stable for at least 6 days in plasma stored at -20 C, 4 C, and 25 C and in blood stored at 4 C and 25 C. For sheep and cattle, the vitamin was stable for at least 6 days in plasma stored at 37 C, but it was unstable in blood from all species stored at 37 C and in pig plasma stored at 37 C.     

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.     

The effect of halothane anesthesia on the function of the adrenal cortex and some metabolites in the blood plasma of pigs not susceptible to malignant hyperthermia]

The effects were investigated of a 25-minute inhalation of halothane with oxygen on three to four months old pigs of the Large White breed. Symptoms of malignant hyperthermia did not occur. The actual total anesthesia, which causes slight hypoproteinemia, hypoglycemia and hypocholesterolemia without significant changes in the content of non-esterified fatty acids (NEFA) and urea, induced only a slight increase of circulating 11-hydroxycorticosteroids (11-OHCS). The combination of anesthesia with castration of gilts or barrows significantly increased the concentration of 11-OHCS but did not reach the level recorded after the application of ACTH. The higher levels of 11-OHCS were accompanied by higher concentrations of NEFA and glucose. The treatment of the animals lasting half an hour prior to inhalation of halothane at maximum doses or one hour in the control unanesthetized pigs produced an effect, mainly on the 11-OHCS concentration and on the activity of creatine kinase in the plasma. The results indicate that the adrenocortical response to the effect of halothane is not stronger than the response to simple handling connected with excitement and muscular activity of the animals. Therefore there is no reason of considering halothane anesthesia as a factor causing great stress and pigs which in its course do not respond with malignant hyperthermia as animals insensitive to stress. The aptness of denotation of clinical manifestations of genetically defective muscles in pigs is discussed.     

Effect of creatine supplementation on muscle metabolic response to a maximal treadmill exercise test in Standardbred horses

The aim of the present study was to investigate the effect of creatine (Cr) supplementation on muscle metabolic response in connection with a maximal treadmill exercise test, known to cause a marked anaerobic metabolic response and adenine nucleotide degradation. First, 6 Standardbred trotters performed a standardised maximal exercise test until fatigue (baseline test). The test used was an inclined incremental treadmill test in which the speed was increased by 1 m/s, starting at 7 m/s, every 60 s until the horse could no longer keep pace with the treadmill. After this baseline test, the horses were separated into 2 equal groups. One half received a dose of 25 g creatine monohydrate twice daily, and the other group were given the same dose of lactose (placebo). The supplementation period was 6.5 days, after which the maximal treadmill exercise test was performed again. A washout period of 14 days was allowed before treatments were switched between groups and a new supplementation period started. After this second supplementation period a new maximal exercise test was performed. After supplementation with creatine or placebo, horses were stopped after performing the same number of speed steps and duration of exercise as they had in the baseline test. Blood samples for analysis of plasma lactate, creatine (Cr), creatinine, hypoxanthine, xanthine and uric acid concentrations were collected at rest, during each speed step and during recovery. The total blood volume (TBV) was also determined. Muscle biopsies for analysis of muscle metabolites (adenosine triphosphate [ATP], adenosine diphosphate [ADP], adenosine monophosphate [AMP], inosine monophosphate [IMP], creatine phosphate [CP], lactate [La] and glycogen) were taken at rest, immediately post exercise and after 15 min recovery. The results showed no significant increase in plasma Cr or muscle total creatine concentration (TCr) after supplementation with Cr. At the end of exercise ATP and CP concentrations had decreased and IMP and lactate concentrations increased in muscle in all groups. Plasma lactate concentration increased during exercise and recovery and plasma uric acid concentration increased during recovery in all groups. No influence could be found in TBV after supplementation with creatine. These results show that creatine supplementation in the dosage used in this study had no influence on muscle metabolic response or TBV.     

Effect of ration and exercise on plasma creatine kinase activity and lactate concentration in Thoroughbred horses with recurrent exertional rhabdomyolysis

OBJECTIVE: To determine the effects of 3 rations (low grain, fat, high grain) on plasma creatine kinase (CK) activity and lactate concentration in Thoroughbred horses with recurrent exertional rhabdomyolysis (RER). ANIMALS: 5 Thoroughbreds with RER and 3 healthy Thoroughbreds (control horses). PROCEDURES: Rations were formulated to meet (low-grain and fat rations) or exceed (high-grain ration) daily energy requirements. Each ration was fed to horses in a crossover design for 3 weeks. Horses were exercised on a treadmill Monday through Friday; maximum speed on Monday and Friday was 11 m/s (6% slope), on Tuesday and Thursday was 9 m/s, and on Wednesday was 4.5 m/s. Plasma CK activity and lactate concentration were determined before and after exercise. RESULTS: Horses with RER fed the high-grain ration had significantly greater CK activity and change in CK activity 4 hours after exercise, compared with those fed the low-grain ration. Horses with RER exercised at the trot or canter had significantly greater increases in CK activity, compared with those exercised at the gallop. Plasma lactate concentrations after exercise were similar in control and affected horses. Lactate concentration and CK activity were not correlated in horses with RER. CONCLUSIONS AND CLINICAL RELEVANCE: Rations high in grain and formulated to exceed daily energy requirements may increase episodes of rhabdomyolysis in thoroughbred horses susceptible to RER.     

Stress resistant and stress susceptible landrace pigs: comparison of blood variables after exposure to halothane or exercise on a treadmill

Stress susceptible pigs develop pale, soft, exudative pork as a consequence of exposure to natural stress or exposure to halothane. Hyperstimulation of glycolysis is a major component of the stress reaction. Whether natural and artificial stress have other similarities was investigated by exposing pigs to halothane or to physical exercise, and measuring their blood composition. There were many similarities between the two groups but the significant differences suggest that although both halothane and exercise induce glycolysis, the mechanisms involved are different.