Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
CREATINE E. R. MILLER, ]3. V. IN THE DIET OF THE B A B Y P I G 1, 2 H. D. STOWE, D. E. U L L R E Y , J. A. HOEFER AND R. W. LUECKE a Michigan State University, East Lansing BALTZER, recent years interest has increased in the I Ndietary administration of creatine or creafine precursors to experimental animals to study their effects upon growth and creafine content of muscle and upon bodyweight gain. Fisher et al. (1956a,b) have shown that the muscle creatine level of 1-day old chicks, on a 25% casein purified diet, increases linearly over a 4-week period. They have shown that with such diets arginine is the most limiting amino acid and that by supplying both arginine and glycine to the 25% casein purified diet, growth equal to that of chicks on practical diets is obtainable. Monson et al. (1955), Wietlake et al. (1954) and Edwards et al. (1958) obtained similar results. Poutsiaka (1956) and Baron (1958) increased muscle creatine and growth rate of rats when their 12% casein purified diet was supplemented with glycocyamine and methionine. Baker et al. (1961) have recently reported failure to increase growth or muscle creatine by parenteral administration of creatine or creatine precursors to the suckling pig. The present study was conducted to determine if the addition of creatine or creatine precursors to a purified diet containing 30% casein for the baby pig will improve his rate of growth and feed economy and increase the size and creatine content of muscle. Experimental Two trials were conducted utilizing 24 baby pigs. Pigs were taken from the sow at 3 or 4 days of age and housed individually in wire-bottomed steel cages. The diet was fed as a homogenized synthetic milk similar to that utilized by Miller et al. (1954). After a 4-day adjustment period the pigs were allotted to groups and started on trial. The control lot received the complete synthetic milk 1 Published with the approval of the Director of the _Michigan Agricultural Experiment Station as Journal Article No. 2922. 2 A prelinlinary report has been presented in J. Animal Sci. 17:1161, 1958 (abstract). a Departments of Animal Husbandry, Biochemistry and Veterinary Pathology. 458 diet, the creatine-fed lot received the same diet supplemented with 0.2% creatine monohydrate, and a third lot received the control diet supplemented with 0.28% L-arginine hydrochloride plus 0.1% glycine. The length of the experimental feeding period was 32 and 42 days in trials 1 and 2, respectively. Pig weights were taken every 4 days. Twenty-four hour urine samples were collected weekly from all pigs. Creatine and creatinine determinations were made on the urine samples using the procedure of Clark and Thompson (1949). Two pigs from each regime were sacrificed at the conclusion of the trials to make gross pathological inspection and to obtain certain organ and muscle weights. The creatine content of the right superficial digital flexor and gastrocnemius muscles was determined by the method of Poutsiaka (1956). All data were analyzed using the multiple range test of Duncan (1955). Results and D i s c u s s i o n In both trials the pigs were about 1 week old when experimental feeding was commenced. The rate of food consumption was very satisfactory throughout the feeding periods. A comparison of growth and the efficiency of food utilization of pigs on the three dietary regimes is presented in table 1. In the first trial the creatine-fed pigs gained 14% faster on 8% less feed than the control group but these differences were not statistically significant ( P > . 0 5 ) . The performance of pigs receiving added arginine and glycine was similar to that of the control pigs. Since it has been shown that the younger chick (Fisher et al., 1956a,b) and pig (Baker et al., 1961) have lower muscle creatine levels than other animals, it w a s considered advisable in the present study to utilize smaller' pigs in the second trial believing that the physiologically younger animals would receive a greater benefit from dietary creatine or creatine precursors. Such was not the case. As C R E A T I N E I N D I E T OF P I G 459 TABLE 1. G R O W T H AND FOOD U T I L I Z A T I O N OF BABY PIGS R E C E I V I N G S Y N T H E T I C M I L K D I E T S S U P P L E M E N T E D W I T H C R E A T I N E OR C R E A T I N E PRECURSORS Trial 1 Trial 2 Arginine + Control Creatine a glycine b Control Creatine" glycine b 4 32 5.0 -+0.5 0.50-+0.06 4 32 5.0 -+0.2 0.57-+0.02 4 32 5.0 -+0.2 0.51-+0.02 4 42 3.5 -+0.1 0.43__+0.01 4 42 3.5 • 0.43-+0.01 4 42 3.5 ___0.1 0.43-+0.02 0.59-+0.05 1.20-+0.06 0.62-+0.05 1.10-+-0.01 0.57-+0.02 1.12-+0.05 0.57__+0.01 1.31-+-0.03 0.57-+0.01 1.31-+0.01 0.57-+0.01 1.31-+0.06 Item No. of pigs Days on experiment Av. initialwt.,lb. Av. daily gain, lb. Av. daily solids consumed, lb. Solids/gain Arginine + a 0.2 % creatine monohydrate added to control diet. b 0.28% L-arginine hydrochloride and 0.1% glycine added to diet. a group each of the lots ate and gained identically over a 42-day experimental feeding period (table 1, trial 2). Twenty-four hour urine collections were made weekly on each pig in both trials. D a t a from both trials were combined and are summarized in table 2. Daily urine volume as well as urinary creatine and creatinine excretion were highly variable. Urinary excretion of creatine was significantly greater by those pigs receiving creatine in the diet. Urinary creatinine excretion by these pigs was also somewhat greater. There was no increase in urinary excretion of creatine or creatinine T A B L E 2. DALLY U R I N E V O L U M E AND U R I NARY E X C R E T I O N OF C R E A T I N E AND C R E A T I N I N E Regimen Item No. of pigs Control 8 24 hour urine volume (ml.) 1st week 229-+33 2nd " 396-+26 3rd " 599• 4th " 579-+38 5th " 828-+232 Creatine 8 184-+31 361+37 520-+34 657-+82 845-+146 Arginine + glycine 8 221-+36 381-+20 439+47 401+__87 947-+236 24 hour creafine excretion (mg.) lstweek 19.2-+ 4.1 21.5-+ 5.8 2nd " 19.7-+ 2.3 49.3-+__ 9.9** 3rd " 36.6-+ 1.6 64.3+13.9"* 4th " 37.5----- 3.1 70.5-+20.8* 5th " 61.2-+12.7 129.2-+17.7"* 18.5-+1.7 20.9-++-1.5 29.3-+-2.4 29.6• 54.2-+7.0 24 hour creatinine excretion (rag.) 1st week 60-+9 53-+9 2nd " 97--+-_7 100-+8 3rd " 147-+12 160-+25 4th " 148-+13 212+__16" 5th " 244-+34 397+__88 48-+9 98-+9 152_+16 113_____26 299-+-58 * Significantly greater than other regimen mean values (P~ .05). *~ P<.OI. by pigs receiving additional arginine and glycine in the diet indicating little or no creatine biosynthesis from the added precursors. Examination of the pigs which were sacrificed at the end of the two trials did not reveal any abnormal findings. Relative organ weights were similar for pigs from each regimen. At no time during the course of the trials was there any indication of toxicity, reduced appetite or impaired growth from the inclusion of creatine in the diet. Muscle creatine data presented in table 3 indicate a slight increase in concentration and total muscle creatine in those pigs receiving dietary creatine, however, neither the increase in size nor creatine content of muscle was statistically significant. Furthermore, dietary creatine precursors (arginine and glycine) were likewise ineffective in increasing muscle creatine. Results obtained in this study indicate that creatine supplemented to a 30% casein diet is largely excreted by the baby pig and does not increase muscle creatine content or growth. Failure of dietary creatine to increase muscle creatine can be well explained by the recent work of Walker (1960, 1961), Fitch et al. (1960) and Coleman (1961) showing a marked repression of mouse kidney transamidinase or chick and duck liver transamidinase activity when either creatine or guanidoacetate is included in the diet. As a consequence, creatine biosynthesis is depressed when an exogenous source of creatine is available. Van Pilsum (1961) has expressed the belief that creatine competes with glycine for attachment on the site of transamidinase synthesis. Walker (1960, 1961) and Fitch et al. (1960) have explained the reduction of transamidinase activity as resulting from a feedback mechanism since there is no MILLER, ET AL. 460 TABLE 3. MUSCLE SIZE AND CREATINE CONTENT Regimen creatine or, creatinine. The supplemental creafine precursors, arginine and glycine, apparently were not utilized in creatine biosynthesis. Argini~e + Item No. of pigs Control Creatine 2 2 glycine 2 Bodyweight (kg.) 10.1 -+- 1.9 10.7 • 10.1 + 0.8 Muscle weight (gin.) ~ 48.2-+- 5.0 56.1 __+1.3 46.3 ~ 3.8 Muscle creatine b Total muscle creatine (rag.) 3.02+ 0.24 146.7 +26.8 3.11__+0.10 2.90~ 0.12 174.7 ___9.2 135.0 ~16.3 a Right superficial digital flexor and gastrocnemius muscles, fresh tissue weight. b Milligrams per gram of fresh tissue. enzyme activity reduction when creatinine replaces creatine in the diet. The addition of the amino acid precursors of guanidoacetate, arginine and glycine, to a diet already adequate in the amino acids as in the present study does not appear to influence the rate of creafine biosynthesis. Baker et al. (1961) have obtained similar results when creatine precursors were administered intraperitoneally to pigs nursing their dams. The two studies indicate that in the baby pig creatine biosynthesis is largely unaffected by supplemental creatine precursors whether supplied parenterally or in the diet. Summary Twenty-four baby pigs receiving a synthetic milk diet containing 30% casein were utilized to determine if the inclusion of 0.2% creatine monohydrate or an equimolar amount of the creatine precursors, arginine and glycine, would improve growth rate, muscle size and creafine content. Neither of these regimens was effective in improving any of these criteria. Dietary creatine which was absorbed appeared to be largely excreted as urinary Literature Cited Baker, J. P., D. E. Becker, A. H. Jensen and S. W. Terrill. 1961. Muscle creatine in the neonatal pig. J. Animal Sci. 20:276. Baron, H. 1958. Some effects of DL-methionine and glycocyamine on growth and nitrogen retention in rats. J. Nutr. 64:229. Clark, L., Jr. and H. Thompson. 1949. Determination of creatine and ereatinine in urine. Anal. Chem. 21:1218. Coleman, D. L. 1961. Effects of dietary creatine and glycine on transamidinase activity in dystrophic mice. Arch. Biochem. Biophys. 94:183. Duncan, D. B. 1955. Multiple range and multiple F tests. Biometrics 11 : 1. Edwards, H. M., Jr., R. J. Young and M. B. Gillis. 1958. Studies on arginine deficiency in chicks. J. Nutr. 64:271. Fisher, H., R. C. Salander and M. W. Taylor. 1956a. Growth and creatine biosynthesis in the chick as affected by the amino acid deficiencies of casein. J. Nutr. 58:459. Fisher, H., R. C. Salander and M. W. Taylor. 1956b. The influence of creatine biosynthesis on the arginine requirement of the chick. J. Nutr. 59:491. Fitch, C. D., C. Hsu and J. S. Dinning. 1960. Some factors affecting kidney transamidinase activity in rats. J. Biol. Chem. 235:2362. Miller, E. R., R. L. Johnston, J. A. Hoefer and R. W. Luecke. 1954. The riboflavin requirement of the baby pig. J. Nutr. 52:405. Monson, W. J., A. E. Harper, D. A. Benton, M. Winje and C. A. Elvehjem. 1955. Effect of arginine and glycine on the growth of chicks receiving complete purified diets. Poul. Sci. 34:186. Poutsiaka, J. W. 1956. Relationship between growth and muscle creatine levels in young rats fed supplementary folacin, B~ and methylating agents with and without glycocyamine. Am. J. Physiol. 185 : 564. Van Pilsum, J. F. 1961. Dietary control of kidney transamidinase. Fed. Proc. 20:225. Walker, J. B. 1960. Metabolic control of creatine biosynthesis. I. Effect of dietary creatine. J. Biol. Chem. 235:2357. Walker, J. B. 1961. Metabolic control of creatine biosynthesis. II. Restoration of transamidinase activity following creatine repression. J. Biol. Chem. 236:493. Wietlake, A. W., A. G. Hogan, B. L. O'dell and H. L. Kempster. 1954. Amino acid deficiencies of casein as a source of protein for the chick. J. Nutr. 52:311.