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Amino Acid Requirements of the Chick
62
G. H. ARSCOTT, J. A. HARPER, J. R. SCHUBERT AND P. H. WESWIG
of unidentified growth factors. Poultry Sci. 36: 1137. Menge, H., R. J. Lillie and C. A. Denton, 1957a. Unidentified growth factor in egg yolk. Fed. Proc. 16: 392.
Menge, H., R. J. Lillie and C. A. Denton, 1957b A chick-growth factor in egg yolk. J. Nut. 63: 499-508. Romanoff, A. L., and A. J. Romanoff, 1949..The Avian Egg. John Wiley and Sons, Inc., N. Y.
Amino Acid Requirements of the Chick I. EFFECT OF THYROXINE AND KIND OF PROTEIN ON THE ARGININE, METHIONINE AND GLYCINE REQUIREMENTS 1 H. B. FLUCKIGER 2 AND J. O. ANDERSON
(Received for publication May 13, 1958)
N
UMEROUS reports have appeared1 which indicate that arginine is essential in the diet of the chick. Almquistt (1947) estimated the chick's arginine requirement to be 1.2% in a 20% proteinl diet. A later report by Almquist and Merritt (1950) indicated the requirement to be; 6% of the dietary protein in diets withl from 15 to 30% protein. Several investigators (Young et al., 1953; Wietlake ett al., 1954; Griminger et al., 1955; Snyderr et al., 1956; Fisher et al., 1956; Krautmannl et al., 1957; and Hogan et al., 1957) havet reported that the chick's requirement forr arginine is higher than this when fed ai diet in which casein provides most of the5 protein. Snyder et al. (1956) and Krautmann et al. (1957) obtained no growthI response by the addition of arginine to) practical type corn-soybean diets although they calculated the arginine level1 of these diets to be below 1.2%. They suggest that the natural feed ingredients contain a factor which spares the arginine inx these rations. Wietlake et al. (1954), Fisher et al. (1956) and Edwards et al. 1 Approved as Utah Agricultural Experiment Station Journal Paper No. 62. 2 Present address: Mead Johnson and Company, Evansville 21, Indiana.
(1958) have reported that creatine and certain related compounds reduce the arginine requirement of the chick on casein type diets, Experiments conducted at this station showed that thiouracil increased and iodinated casein decreased the growth rate of chicks fed diets in which casein, glycine and methionine provided the protein. Following this observation, many more experiments were conducted to determine how much thiouracil and iodinated casein affect the arginine, methionine and glycine requirements of the chick, Other experiments were conducted in which graded levels of thiouracil, iodinated casein or thyroxine were fed. This paper reports the results of these experiments. PROCEDURE
All of the diets fed in these experiments contained the following, in grams per kilogram of diet: cottonseed oil, 30; mineral mix, 50; choline chloride, 2; Klotogen F (2.5 gms. menadione sodium bisulfite/lb.), 0.5; vitamins A and D supplement (10,000 I.U. vitamin A and 1,500 I. C. U. vitamin D 3 /gm.), 1; alpha-tocopherol supplement (20 gms./lb.), 1; and B-vitamin mixture,
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Poultry Department, Utah State University, Logan, Utah
63
AMINO A C I D R E Q U I R E M E N T S
TABLE 1.—Composition of the basic diets Diet number Ingredients* 1 233
— — — — —
3 10
— — —
85.5 668.5
2
3
— —
— —
147.4 60.7
147.0
—
63.9 25.4 3.0 5.0 7.2
25.5 2.2 4.3 4.4
— —
85.5 670.0
—
— •
0.5 85.5 662.5
4
5
265
156 93
— — — — —
3 10
— — —
85.5 636.5
— — — — — — — — —
85.5 665.5
6
— — — —
244
— — 5
— — —
85.5 665.5
7 233
— — — — — 3
— — 5
—
85.5 673.5
* The purified casein, wheat gluten and zein used in these experiments were purchased from Nutritional Biochemicals Corporation, Cleveland, Ohio; the blood fibrin from Armour and Company, Chicago, Illinois; and the gelatin from Swift and Company, Chicago, Illinois. t Contains the vitamins, minerals, and cottonseed oil; see text for composition. t Equal parts of sucrose and dextrin.
1. The vitamin mixture was on a carbohydrate carrier and supplied the following in milligrams to each kilogram of diet: thiamine HCl, 6; riboflavin, 10; pyridoxine HCl, 6; d-calcium pantothenate, 20; biotin, 0.3; folic acid, 2; niacin, 100; and vitamin Bi2, 0.01. The mineral mixture contained the following in grams Ca 3 (P0 4 ) 2 , 35.13; KC1, 5.55; NaCl, 5.58 MgSC-4, 2.77; MnS0 4 , 0.44; FeS0 4 , 0.44 KI, 0.022; ZnCl2, 0.022; CuS0 4 , 0.022 H3BOs, 0.011; Na 2 Mo0 4 -2H 2 0, 0.011 CoS04, 0.002. The composition of the basic diets fed in these experiments is given in Table 1. In each experiment involving the addition of one or more levels of arginine to the diets, the total amount of nitrogen was kept constant. This was done by removing 19 grams of casein from diets 1 and 4 or 19 grams of the complete protein and amino acid mixture from diets 2 and 3 for each 10 grams of arginine HCl added. Thiouracil, iodinated casein, glycine and methionine replaced an equal amount of carbohydrate when they were added to an experimental ration. All proteins and amino
acids were used as purchased from commercial sources. The arginine content of each protein was determined chemically. The protein was first hydrolyzed with 6N HCl; most of the acid was removed under reduced pressure. The samples were diluted to a standard volume and their arginine content determined by the method of Rosenberg et al. (1956). This method is not specific for arginine; it can be used to estimate other guanido compounds including creatine. The arginine contents of these proteins, as used, and as determined by this method, were casein 3.7, wheat gluten 3.4, blood fibrin 6.8, Drackett protein 6.8, and zein 1.5%. The protein contents (NX6.25) of these substances were 86, 85, 88.5, 83.2, and 83% respectively. These values were used in calculating the arginine content of the rations fed in these experiments. All chicks were reared in electrically heated battery brooders. Feed and water were offered free-choice. Four male and four female chicks from a New Hampshire X Silver Cornish cross were used in each
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Casein, purified Gelatin Wheat gluten Blood fibrin Drackett assay protein C-l Zein DL-methionine Glycine Lysine HCl (95% L) L-arginine HCl DL-tryptophan Constant ingredients! Carbohydrate:):
64
H. B. FLUCKIGER AND J. O. ANDERSON
RESULTS AND DISCUSSION
Graded Levels of Thiouracil and Io-
dinated Casein. Several experiments were conducted to study the effects of graded levels of thiouracil, iodinated casein or thyroxine on chicks fed diet 1 and diet 1 with 1% arginine HCI. The arginine level of these diets is calculated to be 0.86 and 1.62% respectively. Casein, glycine, and methionine provided the protein in diet 1. Figure 1 presents data from three fourweek experiments in which graded levels of thiouracil and iodinated casein were added to these diets. Since iodinated casein has thyroxine activity and thiouracil is known to reduce the rate thyroxine is secreted by the thyroid, it was decided to present the results in the manner shown in Figure 1. Note the labeling of the horizontal axis. It is recognized that the effect of thiouracil may not be a result of reduced thyroxine secretion. Each dot in Figure 1 gives the average daily weight gain of one group of eight chicks fed diet 1 with the indicated addition of thiouracil or iodinated casein. The solid line was X X
10 * X
^
X
^ , ' '
x
8
-x-
-
—
**
— ^ X ^ X
X X
X
x
•
X ^ X~
^
x
X ^
* 6
/'
Die* 1
/ //
•
4
x
*
Die* 1 +
HCI
"~~^~"--^
< o ii
1 * Arginine
•\^ . 2
o uj
5 1 —
0.64
i
i
0.16 PERCENT
i
i
i
0.04
1
0.01
THIOURACIL ADDITION
,. i
0
1
0.005
1
.—i
0.02
PERCENT IODINATED
1
1—
0.08 CASEIN
TO DIETS
FIG. 1. Effect of additions of graded levels of thiouracil or iodinated casein on the growth rate o: chicks fed diet 1 and diet 1 with 1% arginine HCI.
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group, unless otherwise indicated. In the experiments involving the addition of graded levels of thiouracil, iodinated casein, thyroxine or potassium iodide to a diet, the chicks were placed on the experimental diets on the day they were removed from the incubator. When the chicks were to be used in an experiment to study their requirement for an amino acid, they were started on a practical type diet for five to seven days. If the chicks were to receive an experimental diet containing thiouracil, thiouracil was added to the practical starting ration at a level of 0.08%. Also, 0.022% iodinated casein was added to the starting ration of chicks which would later be fed an experimental ration containing iodinated casein. The experimental diets in these experiments were fed from 15 to 19 days.
AMINO ACID REQUIREMENTS
The upper broken line and series of crosses in Figure 1 show the average daily gains made by chicks fed diet 1 with 1% added arginine HC1 and the indicated addition of thiouracil or iodinated casein. The addition of arginine to diet 1 produced more than a four fold increase in average daily weight gain. This gain was not increased by the addition of either thiouracil or iodinated casein; in fact, the daily gain was decreased by addition of the higher levels of these substances.
One experiment was also conducted in which L-thyroxine was added to diet 1 and diet 1 with 1% arginine HC1. The results were similar to those obtained by the addition of iodinated casein. The addition of 1.5 milligram of thyroxine per kilogram of diet 1 reduced the weight gained by 50%. Up to 0.5 milligram of thyroxine per kilogram of diet did not affect growth when 1% arginine HC1 was added; higher levels reduced growth. The results given in Figure 1 were obtained with the New Hampshire X Silver Cornish chicks. Another similar experiment was conducted using a commercial broiler chick obtained from a local hatchery. These chicks gained weight faster than the New Hampshire X Silver Cornish chicks when fed either diet 1 or a practical broiler ration. They showed less growth response to the addition of thiouracil to diet 1 and required less thiouracil to obtain maximum response. Addition of iodinated casein or thyroxine reduced growth more than with the crossbred chicks. When combinations of 0.04 to 0.32% thiouracil and either 0.03% iodinated casein or 0.5 mg. of thyroxine per kg. were added to diet 1, the rate of growth was between that obtained by the addition of either substance alone. Three experiments were conducted in which graded levels of thiouracil or iodinated casein were added to diet 2. Wheat gluten, blood fibrin, zein and amino acids provided the protein in this diet. The calculated arginine content of diet 2 was 0.95%; this level was increased to 1.71% when 1% arginine HC1 was added. The average daily weight gains are plotted in Figure 2. The daily gain made by chicks fed diet 2 was four times that made by chicks fed diet 1. The chicks fed diet 2 did not show the arginine deficiency symptoms found in chicks fed diet 1. When the higher levels of iodinated casein
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drawn through these points to indicate the general trend of the weight gains. The average daily weight gain was increased by the addition' of thiouracil to diet 1. The maximum response was obtained with 0.16% thiouracil. At this level, the rate of gain was twice that on diet 1. When the thiouracil level was increased to 0.96%, the rate of gain was less than that obtained with 0.16%. The difference was very marked during the last two weeks of the experiment. Even though thiouracil increased the rate of gain, this rate is still much less than that obtained by the addition of 1% arginine HC1 to the diet. The chicks fed diet 1 appeared to be weak and had an abnormal gait. Many of their feathers have an abnormal curvature and a ragged appearance; the barbs of the main wing feathers tend to remain close to the shaft. These symptoms have been described by Hegsted el al. (1941) and several of the groups already listed in the introduction. They have been termed arginine deficiency symptoms. When thiouracil is added to diet 1, the rate of feathering is decreased, but the arginine deficiency symptoms were less severe or they were not noted at all. The addition of iodinated casein to diet 1 reduced growth rate. Mortality was increased and the weakness and abnormal feather condition were more severe.
65
66
H. B. FLUCKIGER AND J. O. ANDERSON
IO Id
a
y s i
o
6
< O
li
4
•
Diet 2
x
Diet 2 + I * Arginint
HCI
2r
o 0.64
0.16 PERCENT
0.04
0.01
0
THIOURACIL
0.005 PERCENT
ADDITION
TO
0.02
0.08
IODINATED CASEIN
DIETS
FIG. 2. Effect of additions of graded levels of thiouracil or iodinated casein on the growth rate of chicks fed diet 2 and diet 2 with 1% arginine HCI.
were added to diet 2, these symptoms were noted and were corrected by the addition of arginine to the diet. When iodinated casein was added to diet 2, the addition of 1% arginine HCI significantly increased growth. On the other hand, when arginine was added to diets containing thiouracil, the groups attained slightly lower average weights, but the difference was not significant. Diets 1 and 2, with and without added arginine, produced about the same average daily gain when 0.64% thiouracil was added. The maximum growth on diet 2 with 1% arginine HCI was obtained with low levels of iodinated casein in the diet. High levels of iodinated casein and all levels of thiouracil decreased growth. Graded levels of thiouracil and L-thyroxine were fed in diet 3. The protein in this diet is similar to that in diet 2, except
Drackett protein replaces the blood fibrin. Diet 3 contains 0.97% arginine. The average daily gain made by chicks fed diet 3 was about 8.25 grams. This was slightly, but not significantly, increased by the addition of 0.1% arginine HCI to the diet. Levels of 0.2 and 0.3% produced no further increase. The effects of additions of thiouracil and thyroxine to diet 3 were similar to those noted with thiouracil and iodinated casein in diet 2. The addition of 1% arginine HCI decreased the average daily weight gains to four weeks of age with all levels of thyroxine and levels of thiouracil up to 0.32%. It was expected that the addition of the arginine to diet 3 with a high level of thyroxine would increase growth. This was not true of gains during the entire four week period. The highest level of thyroxine fed in these experiments was 5 milligrams per kilogram of diet. The weight
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I z < o
67
AMINO ACID REQUIREMENTS
1.00
1.26
1.59
2.00
2.51
3.16
PERCENT ARGININE IN RATION
FIG. 3. Effect of thiouracil on the arginine requirement of the chick. level was reduced enough to keep the nitrogen content constant. Figure 3 shows how the average daily gains were affected by the addition of 0.08% thiouracil to the diets containing several levels of arginine. Each dot or cross on Figure 3 represents the average daily weight gain of one group of eight chicks. The addition of 0.08% thiouracil to the diets containing the lower levels of arginine always increased weight gain; its addition to diets containing higher arginine levels decreased weight gain. The growth rate showed a definite increase as the arginine level of the diet 4 was increased up to 1.48% arginine. Smaller increases were noted beyond this point. The maximum growth rate was not obtained until the arginine content of the diet was over 2%. When 0.08% thiouracil was added to diet 4, the maximum gain was obtained with 1.38% arginine. This rate was not as high as the maximum obtained without the addition of thiouracil. Figure 4 shows the effect of the addition of 0.022% iodinated casein on the arginine requirement. The plotted weight gains for chicks fed rations without io-
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gained during the last two weeks of the experiment was increased when 1% arginine HC1 was added to this diet. No arginine deficiency symptoms were noted among chicks fed diet 3, but the addition of 2 to 5 milligrams of thyroxine to each kilogram of diet caused typical arginine deficiency symptoms to appear in the chicks. These were not noted when 1% arginine HC1 was added. Apparently, the addition of thyroxine does increase the arginine requirement as with diets 1 and 2, but it is not as evident as in the first two diets. It is possible that the addition of thyroxine to diet 3 produced a deficiency of some other nutrient besides arginine. This nutrient would then have to be added before a response to arginine would be obtained. Iodide. It seemed possible that the trace amounts of potassium iodide added to diet 1 may be iodinating the casein in the diet, and thus increasing the arginine requirement of the chicks. Two experiments were conducted to determine what effect omitting the potassium iodide or increasing it to five times the normal level would have on growth of chicks fed diet 1. When the potassium iodide was omitted, growth rate was not increased; addition of five times the normal level did not decrease growth rate. Thus, no evidence was obtained which would indicate that the casein was iodinated. Effect on Arginine Requirement. The experiments already reported indicated that thiouracil decreased and iodinated casein increased the arginine requirements of the chick. Three experiments were conducted to determine how much the arginine requirement was changed by these substances. Diet 4 was used in these experiments. The total protein in the diet 4 (NX6.25) is about 24%; its arginine content is calculated to be 0.98%. In order to increase the arginine content of the diet, arginine HC1 was added and the casein
68
H. B. FLTJCKIGER AND J. 0. ANDERSON
. o 2
Di.t 4 Wilh 0 . 0 2 2 % lodinat*d Casvin
1.26 1.59 2.00 2.51 3.16 PERCENT ARGININE IN RATION
FIG. 4. Effect of iodinated casein on the arginine requirement of the chick.
dinated casein are the same as found in Figure 3. The addition of iodinated casein reduced average gains on all the lower levels of arginine; whereas, on the higher levels of arginine, growth was slightly greater when iodinated casein was added. The weight gains showed a definite increase as the arginine level was increased up to 1.66% when 0.022% iodinated casein was added to diet 4. Again smaller increases in gain were noted when the arginine content was increased beyond this point. Over 2% arginine was required to produce maximum gains. Effect on the Glycine Requirement. Three experiments were conducted which were designed to determine how much iodinated casein and thiouracil affect the. glycine requirement of the chick. Diet 7 was used in these experiments. The results of the three experiments are presented- in Figure 5. These results have been analyzed statistically. Chicks fed diets with added glycine attained slightly higher average weights, but the increase is not significant. Addition of iodinated casein significantly reduced growth. The reduction tended to be slightly less on the two high-
D i . l 7 With 0.06 «
.
D i . l 7 Wilh 0 . 0 2 2 % lodinot.d CoMin. 0.65
0.42
1.00
Thiouracil
2.41
1.55
PERCENT GLYCINE IN DIET
FIG. 5. Effect of thiouracil and iodinated casein on the glycine requirement of the chick.
est levels of glycine. This is the only observed indication that the addition of iodinated casein affects the glycine requirement of the chick. Effect on the Methionine Requirement. Both casein-gelatin and Drackett protein type diets have been used in studies on the methionine requirement. Figure 6 presents the results of three experiments con< o ac
0
UJ
Q-IO
/*
o X u
8
-
cr Z 6 '>•
<
a.
o
~
< O r-
I
2
'
Y .
Diet 5
»
Die* 5
With 0 . 0 6 % Thiouracil
o
Die! 5
With 0 . 0 2 2 % Iodinated
u S
Casein. —1
0.54
1
1
0.64
0.78
i ._
0.94
PERCENT METHIONINE IN DIET
FIG. 6. Effect of thiouracil and iodinated casein on the methionine requirement of chicks fed a casein type diet (diet 5).
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1.00
oi.i 7
«
AMINO ACID REQUIREMENTS
69
I
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ducted with diet 5. Casein and gelatin provide the protein in this diet. The diet was > < o calculated to contain 0.53% methionine a: 10 -«-x and 0.06% cystine, using values given by ua Block and Boiling (1951). Each point plotted in Figure 6 is an average of at yi s least two values obtained in three sepa- o Jtrate experiments. Addition of 0.022% io-
I
70
H. B. FLUCKIGER AND J. 0 . ANDERSON
more methionine and cystine than the values reported by the manufacturer which were used in our calculations. Even if the higher values are correct, the total sulfur amino acid requirement of the chick is higher on the casein-gelatin diet (diet 5) than it is on the Drackett protein diet (diet 6). SUMMARY
Purified type diets containing 23 to 28% casein, 1% glycine and 0.3% DLmethionine to provide the protein, were found to be very deficient in arginine for the chick. When thiouracil was added to this type of diet, growth rate was increased, arginine deficiency symptoms were less severe, and the amount of arginine required for maximum growth was decreased. Addition of iodinated casein or thyroxine produced effects just opposite to those produced by the addition of thiouracil. Two other diets were fed in which the protein was provided by a mixture of wheat gluten, zein, amino acids and either blood fibrin or Drackett protein. These diets contained only slightly more arginine than the casein diet, but good growth was obtained without the addition of arginine. Chicks fed these diets did not show arginine deficiency symptoms unless iodinated casein or thyroxine was added. Thus, the arginine requirement of the chick was influenced by the kind of protein in the diet. The chick's methionine and glycine requirements were not affected by the addition of thiouracil or iodinated casein to their diet. ACKNOWLEDGMENTS
The authors are indebted to Merck and Company, Inc., Rahway, New Jersey, for some of the crystalline vitamins and the L-lysine HC1; Nopco Chemical Company,
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rate. Opposite effects also were noted on the arginine requirement of the chick. When both thiouracil and thyroxine were added to diet 1, the growth rate was less than that obtained by the addition of thiouracil, but more than that obtained by the addition of thyroxine. This suggests that thiouracil produces its effect on arginine requirement of the chick by reducing the rate thyroxine is secreted by the thyroid of the chick. The presence of a substance with thyroxine activity in casein might explain the high arginine requirement of chicks fed a casein type diet. These experiments offer very little evidence for the presence of such a substance in casein. Chicks gained weight four times as fast on diet 2 as they did on diet 1. When 1% arginine HC1 was added to both diets, the rate of gain was approximately equal. This would indicate that diet 1 is very deficient in arginine. The arginine content of diet 2 is only 0.09% greater than diet 1. When 0.09% arginine is added to diet 1, the rate of gain is still far less than that obtained with diet 2. These results are similar to those obtained by Snyder et al. (1956) and Krautmann et al. (1957). Their reports indicated that the chick's arginine requirement is higher on a purified casein type diet than it is on a cornsoybean diet. Apparently, some differences in the protein supplements used in diets 1 and 2 affect the arginine requirement of the chick. The methionine requirement of the chick appears to be much higher on diet 5 than it is on diet 6. Cystine can be utilized by the chick to correct the deficiency in diet 5, but it could not in diet 6. This helps to explain this difference, but the total sulfur amino acid requirement still appears to be higher on diet 5 than on diet 6. According to the data of Nelson et al. (1957) Drackett protein contains
AMINO ACID REQUIREMENTS
Harrison, New Jersey, for the vitamins A and D supplement; Lederle Laboratories Division, American Cyanamid Company, for the folic acid; and the Dow Chemical Company, Midland, Michigan, for some of the DL-methionine and glycine used in these experiments. REFERENCES
E. B. Hart, 1941. The role of arginine and glycine in chick nutrition. J. Biol. Chem. 140: 191-200. Hogan, A. G., R. W. Craghead, J. E. Savage, J. J. Cole and B. L. O'Dell, 1957. Casein as a source of protein for the chick. J. Nutrition, 62: 97-106. Krautmann, B. A., S. M. Hauge, E. T. Mertz and C. W. Carrick, 1957. The arginine level for chicks as influenced by ingredients. Poultry Sci. 36: 935-939. Nelson, T. S., L. C. Norris, F. W. Hill, M. L. Scott and G. F. Heuser, 1957. Further studies on the requirement of chicks for methionine and cystine. Proc. Cornell Nutrition Conf. 126-131. Rosenberg, H., A. H. Ennor and J. F. Morrison, 1956. The estimation of arginine. Biochem. J. 63: 153-159. Snyder, J. M., W. D. Morrison and H. M. Scott, 1956. The arginine requirement of chicks fed purified and corn-soya diets. Poultry Sci. 35: • 852-855. Vohra, P., and F. H. Kratzer, 1958. The effect of thyroprotein, thiouracil and cortisone on the feather pigmentation of turkey poults. Poultry Sci. 37: 151-153. 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. Nutrition, 52: 311-323. Young, R. J., M. B. Gillis and L. C. Norris, 1953. An unidentified factor in peanut meal required by the chick. J. Nutrition, 50: 291-297.
Variation in the Effects of Fat Supplements on Broiler Pigmentation, Growth and Feed Conversion D . S. C A R V E R
General Mills, Inc., Larro Research Farm, P. O. Box 263, Indianola, Iowa (Received for publication May 13, 1958)
T
HE range in price between prime tallow and lower grades of fat such as yellow grease or No. 2 tallow is normally f jf to If! a pound. This difference in price is very important to the feed manufacturer, because it means a substantial savings in ingredient cost can be realized if fats such as yellow grease and No. 2 tallow
can be substituted for prime tallow without reducing the feeding value of feeds. Yacowitz (1953) found that soybean oil, cottonseed oil and lard at levels of 2\ percent and 5 percent produced similar responses in growth and feed efficiency. March and Biely (1954) reported the results of feeding 5 percent to 10 percent
Downloaded from http://ps.oxfordjournals.org/ by guest on April 23, 2015
I
Almquist, H. J., 1947. Evaluation of amino acid requirements by observations on the chick. J. Nutrition, 34: 543-563. Almquist, H. J., and J. B. Merritt, 1950. Protein and arginine levels in the chick's diet. Proc. Soc. Exp. Biol, and Med. 73: 136. Block, R. J., and D. Boiling, 1951. The Amino Acid Composition of Proteins and Foods. Charles C Thomas, Publisher, Springfield, 111. 2nd Ed. 486492. Edwards, H. M., Jr., R. J. Young and M. B. Gillis, 1958. Studies on arginine deficiency in chicks. J. Nutrition, 64: 271-279. Fisher, H., R. C. Salander and M. W. Taylor, 1956. The influence of creatine biosynthesis on the arginine requirement of the chick. J. Nutrition, 59: 491-500. Griminger, P., H. Fisher and H. M. Scott, 1955. On the arginine requirement of the growing chick. Poultry Sci. 34: 1247-1249. Hegsted, D. M., G. M. Briggs, C. A. Elvehjem and
71
G. H. ARSCOTT, J. A. HARPER, J. R. SCHUBERT AND P. H. WESWIG
of unidentified growth factors. Poultry Sci. 36: 1137. Menge, H., R. J. Lillie and C. A. Denton, 1957a. Unidentified growth factor in egg yolk. Fed. Proc. 16: 392.
Menge, H., R. J. Lillie and C. A. Denton, 1957b A chick-growth factor in egg yolk. J. Nut. 63: 499-508. Romanoff, A. L., and A. J. Romanoff, 1949..The Avian Egg. John Wiley and Sons, Inc., N. Y.
Amino Acid Requirements of the Chick I. EFFECT OF THYROXINE AND KIND OF PROTEIN ON THE ARGININE, METHIONINE AND GLYCINE REQUIREMENTS 1 H. B. FLUCKIGER 2 AND J. O. ANDERSON
(Received for publication May 13, 1958)
N
UMEROUS reports have appeared1 which indicate that arginine is essential in the diet of the chick. Almquistt (1947) estimated the chick's arginine requirement to be 1.2% in a 20% proteinl diet. A later report by Almquist and Merritt (1950) indicated the requirement to be; 6% of the dietary protein in diets withl from 15 to 30% protein. Several investigators (Young et al., 1953; Wietlake ett al., 1954; Griminger et al., 1955; Snyderr et al., 1956; Fisher et al., 1956; Krautmannl et al., 1957; and Hogan et al., 1957) havet reported that the chick's requirement forr arginine is higher than this when fed ai diet in which casein provides most of the5 protein. Snyder et al. (1956) and Krautmann et al. (1957) obtained no growthI response by the addition of arginine to) practical type corn-soybean diets although they calculated the arginine level1 of these diets to be below 1.2%. They suggest that the natural feed ingredients contain a factor which spares the arginine inx these rations. Wietlake et al. (1954), Fisher et al. (1956) and Edwards et al. 1 Approved as Utah Agricultural Experiment Station Journal Paper No. 62. 2 Present address: Mead Johnson and Company, Evansville 21, Indiana.
(1958) have reported that creatine and certain related compounds reduce the arginine requirement of the chick on casein type diets, Experiments conducted at this station showed that thiouracil increased and iodinated casein decreased the growth rate of chicks fed diets in which casein, glycine and methionine provided the protein. Following this observation, many more experiments were conducted to determine how much thiouracil and iodinated casein affect the arginine, methionine and glycine requirements of the chick, Other experiments were conducted in which graded levels of thiouracil, iodinated casein or thyroxine were fed. This paper reports the results of these experiments. PROCEDURE
All of the diets fed in these experiments contained the following, in grams per kilogram of diet: cottonseed oil, 30; mineral mix, 50; choline chloride, 2; Klotogen F (2.5 gms. menadione sodium bisulfite/lb.), 0.5; vitamins A and D supplement (10,000 I.U. vitamin A and 1,500 I. C. U. vitamin D 3 /gm.), 1; alpha-tocopherol supplement (20 gms./lb.), 1; and B-vitamin mixture,
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Poultry Department, Utah State University, Logan, Utah
63
AMINO A C I D R E Q U I R E M E N T S
TABLE 1.—Composition of the basic diets Diet number Ingredients* 1 233
— — — — —
3 10
— — —
85.5 668.5
2
3
— —
— —
147.4 60.7
147.0
—
63.9 25.4 3.0 5.0 7.2
25.5 2.2 4.3 4.4
— —
85.5 670.0
—
— •
0.5 85.5 662.5
4
5
265
156 93
— — — — —
3 10
— — —
85.5 636.5
— — — — — — — — —
85.5 665.5
6
— — — —
244
— — 5
— — —
85.5 665.5
7 233
— — — — — 3
— — 5
—
85.5 673.5
* The purified casein, wheat gluten and zein used in these experiments were purchased from Nutritional Biochemicals Corporation, Cleveland, Ohio; the blood fibrin from Armour and Company, Chicago, Illinois; and the gelatin from Swift and Company, Chicago, Illinois. t Contains the vitamins, minerals, and cottonseed oil; see text for composition. t Equal parts of sucrose and dextrin.
1. The vitamin mixture was on a carbohydrate carrier and supplied the following in milligrams to each kilogram of diet: thiamine HCl, 6; riboflavin, 10; pyridoxine HCl, 6; d-calcium pantothenate, 20; biotin, 0.3; folic acid, 2; niacin, 100; and vitamin Bi2, 0.01. The mineral mixture contained the following in grams Ca 3 (P0 4 ) 2 , 35.13; KC1, 5.55; NaCl, 5.58 MgSC-4, 2.77; MnS0 4 , 0.44; FeS0 4 , 0.44 KI, 0.022; ZnCl2, 0.022; CuS0 4 , 0.022 H3BOs, 0.011; Na 2 Mo0 4 -2H 2 0, 0.011 CoS04, 0.002. The composition of the basic diets fed in these experiments is given in Table 1. In each experiment involving the addition of one or more levels of arginine to the diets, the total amount of nitrogen was kept constant. This was done by removing 19 grams of casein from diets 1 and 4 or 19 grams of the complete protein and amino acid mixture from diets 2 and 3 for each 10 grams of arginine HCl added. Thiouracil, iodinated casein, glycine and methionine replaced an equal amount of carbohydrate when they were added to an experimental ration. All proteins and amino
acids were used as purchased from commercial sources. The arginine content of each protein was determined chemically. The protein was first hydrolyzed with 6N HCl; most of the acid was removed under reduced pressure. The samples were diluted to a standard volume and their arginine content determined by the method of Rosenberg et al. (1956). This method is not specific for arginine; it can be used to estimate other guanido compounds including creatine. The arginine contents of these proteins, as used, and as determined by this method, were casein 3.7, wheat gluten 3.4, blood fibrin 6.8, Drackett protein 6.8, and zein 1.5%. The protein contents (NX6.25) of these substances were 86, 85, 88.5, 83.2, and 83% respectively. These values were used in calculating the arginine content of the rations fed in these experiments. All chicks were reared in electrically heated battery brooders. Feed and water were offered free-choice. Four male and four female chicks from a New Hampshire X Silver Cornish cross were used in each
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Casein, purified Gelatin Wheat gluten Blood fibrin Drackett assay protein C-l Zein DL-methionine Glycine Lysine HCl (95% L) L-arginine HCl DL-tryptophan Constant ingredients! Carbohydrate:):
64
H. B. FLUCKIGER AND J. O. ANDERSON
RESULTS AND DISCUSSION
Graded Levels of Thiouracil and Io-
dinated Casein. Several experiments were conducted to study the effects of graded levels of thiouracil, iodinated casein or thyroxine on chicks fed diet 1 and diet 1 with 1% arginine HCI. The arginine level of these diets is calculated to be 0.86 and 1.62% respectively. Casein, glycine, and methionine provided the protein in diet 1. Figure 1 presents data from three fourweek experiments in which graded levels of thiouracil and iodinated casein were added to these diets. Since iodinated casein has thyroxine activity and thiouracil is known to reduce the rate thyroxine is secreted by the thyroid, it was decided to present the results in the manner shown in Figure 1. Note the labeling of the horizontal axis. It is recognized that the effect of thiouracil may not be a result of reduced thyroxine secretion. Each dot in Figure 1 gives the average daily weight gain of one group of eight chicks fed diet 1 with the indicated addition of thiouracil or iodinated casein. The solid line was X X
10 * X
^
X
^ , ' '
x
8
-x-
-
—
**
— ^ X ^ X
X X
X
x
•
X ^ X~
^
x
X ^
* 6
/'
Die* 1
/ //
•
4
x
*
Die* 1 +
HCI
"~~^~"--^
< o ii
1 * Arginine
•\^ . 2
o uj
5 1 —
0.64
i
i
0.16 PERCENT
i
i
i
0.04
1
0.01
THIOURACIL ADDITION
,. i
0
1
0.005
1
.—i
0.02
PERCENT IODINATED
1
1—
0.08 CASEIN
TO DIETS
FIG. 1. Effect of additions of graded levels of thiouracil or iodinated casein on the growth rate o: chicks fed diet 1 and diet 1 with 1% arginine HCI.
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group, unless otherwise indicated. In the experiments involving the addition of graded levels of thiouracil, iodinated casein, thyroxine or potassium iodide to a diet, the chicks were placed on the experimental diets on the day they were removed from the incubator. When the chicks were to be used in an experiment to study their requirement for an amino acid, they were started on a practical type diet for five to seven days. If the chicks were to receive an experimental diet containing thiouracil, thiouracil was added to the practical starting ration at a level of 0.08%. Also, 0.022% iodinated casein was added to the starting ration of chicks which would later be fed an experimental ration containing iodinated casein. The experimental diets in these experiments were fed from 15 to 19 days.
AMINO ACID REQUIREMENTS
The upper broken line and series of crosses in Figure 1 show the average daily gains made by chicks fed diet 1 with 1% added arginine HC1 and the indicated addition of thiouracil or iodinated casein. The addition of arginine to diet 1 produced more than a four fold increase in average daily weight gain. This gain was not increased by the addition of either thiouracil or iodinated casein; in fact, the daily gain was decreased by addition of the higher levels of these substances.
One experiment was also conducted in which L-thyroxine was added to diet 1 and diet 1 with 1% arginine HC1. The results were similar to those obtained by the addition of iodinated casein. The addition of 1.5 milligram of thyroxine per kilogram of diet 1 reduced the weight gained by 50%. Up to 0.5 milligram of thyroxine per kilogram of diet did not affect growth when 1% arginine HC1 was added; higher levels reduced growth. The results given in Figure 1 were obtained with the New Hampshire X Silver Cornish chicks. Another similar experiment was conducted using a commercial broiler chick obtained from a local hatchery. These chicks gained weight faster than the New Hampshire X Silver Cornish chicks when fed either diet 1 or a practical broiler ration. They showed less growth response to the addition of thiouracil to diet 1 and required less thiouracil to obtain maximum response. Addition of iodinated casein or thyroxine reduced growth more than with the crossbred chicks. When combinations of 0.04 to 0.32% thiouracil and either 0.03% iodinated casein or 0.5 mg. of thyroxine per kg. were added to diet 1, the rate of growth was between that obtained by the addition of either substance alone. Three experiments were conducted in which graded levels of thiouracil or iodinated casein were added to diet 2. Wheat gluten, blood fibrin, zein and amino acids provided the protein in this diet. The calculated arginine content of diet 2 was 0.95%; this level was increased to 1.71% when 1% arginine HC1 was added. The average daily weight gains are plotted in Figure 2. The daily gain made by chicks fed diet 2 was four times that made by chicks fed diet 1. The chicks fed diet 2 did not show the arginine deficiency symptoms found in chicks fed diet 1. When the higher levels of iodinated casein
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drawn through these points to indicate the general trend of the weight gains. The average daily weight gain was increased by the addition' of thiouracil to diet 1. The maximum response was obtained with 0.16% thiouracil. At this level, the rate of gain was twice that on diet 1. When the thiouracil level was increased to 0.96%, the rate of gain was less than that obtained with 0.16%. The difference was very marked during the last two weeks of the experiment. Even though thiouracil increased the rate of gain, this rate is still much less than that obtained by the addition of 1% arginine HC1 to the diet. The chicks fed diet 1 appeared to be weak and had an abnormal gait. Many of their feathers have an abnormal curvature and a ragged appearance; the barbs of the main wing feathers tend to remain close to the shaft. These symptoms have been described by Hegsted el al. (1941) and several of the groups already listed in the introduction. They have been termed arginine deficiency symptoms. When thiouracil is added to diet 1, the rate of feathering is decreased, but the arginine deficiency symptoms were less severe or they were not noted at all. The addition of iodinated casein to diet 1 reduced growth rate. Mortality was increased and the weakness and abnormal feather condition were more severe.
65
66
H. B. FLUCKIGER AND J. O. ANDERSON
IO Id
a
y s i
o
6
< O
li
4
•
Diet 2
x
Diet 2 + I * Arginint
HCI
2r
o 0.64
0.16 PERCENT
0.04
0.01
0
THIOURACIL
0.005 PERCENT
ADDITION
TO
0.02
0.08
IODINATED CASEIN
DIETS
FIG. 2. Effect of additions of graded levels of thiouracil or iodinated casein on the growth rate of chicks fed diet 2 and diet 2 with 1% arginine HCI.
were added to diet 2, these symptoms were noted and were corrected by the addition of arginine to the diet. When iodinated casein was added to diet 2, the addition of 1% arginine HCI significantly increased growth. On the other hand, when arginine was added to diets containing thiouracil, the groups attained slightly lower average weights, but the difference was not significant. Diets 1 and 2, with and without added arginine, produced about the same average daily gain when 0.64% thiouracil was added. The maximum growth on diet 2 with 1% arginine HCI was obtained with low levels of iodinated casein in the diet. High levels of iodinated casein and all levels of thiouracil decreased growth. Graded levels of thiouracil and L-thyroxine were fed in diet 3. The protein in this diet is similar to that in diet 2, except
Drackett protein replaces the blood fibrin. Diet 3 contains 0.97% arginine. The average daily gain made by chicks fed diet 3 was about 8.25 grams. This was slightly, but not significantly, increased by the addition of 0.1% arginine HCI to the diet. Levels of 0.2 and 0.3% produced no further increase. The effects of additions of thiouracil and thyroxine to diet 3 were similar to those noted with thiouracil and iodinated casein in diet 2. The addition of 1% arginine HCI decreased the average daily weight gains to four weeks of age with all levels of thyroxine and levels of thiouracil up to 0.32%. It was expected that the addition of the arginine to diet 3 with a high level of thyroxine would increase growth. This was not true of gains during the entire four week period. The highest level of thyroxine fed in these experiments was 5 milligrams per kilogram of diet. The weight
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I z < o
67
AMINO ACID REQUIREMENTS
1.00
1.26
1.59
2.00
2.51
3.16
PERCENT ARGININE IN RATION
FIG. 3. Effect of thiouracil on the arginine requirement of the chick. level was reduced enough to keep the nitrogen content constant. Figure 3 shows how the average daily gains were affected by the addition of 0.08% thiouracil to the diets containing several levels of arginine. Each dot or cross on Figure 3 represents the average daily weight gain of one group of eight chicks. The addition of 0.08% thiouracil to the diets containing the lower levels of arginine always increased weight gain; its addition to diets containing higher arginine levels decreased weight gain. The growth rate showed a definite increase as the arginine level of the diet 4 was increased up to 1.48% arginine. Smaller increases were noted beyond this point. The maximum growth rate was not obtained until the arginine content of the diet was over 2%. When 0.08% thiouracil was added to diet 4, the maximum gain was obtained with 1.38% arginine. This rate was not as high as the maximum obtained without the addition of thiouracil. Figure 4 shows the effect of the addition of 0.022% iodinated casein on the arginine requirement. The plotted weight gains for chicks fed rations without io-
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gained during the last two weeks of the experiment was increased when 1% arginine HC1 was added to this diet. No arginine deficiency symptoms were noted among chicks fed diet 3, but the addition of 2 to 5 milligrams of thyroxine to each kilogram of diet caused typical arginine deficiency symptoms to appear in the chicks. These were not noted when 1% arginine HC1 was added. Apparently, the addition of thyroxine does increase the arginine requirement as with diets 1 and 2, but it is not as evident as in the first two diets. It is possible that the addition of thyroxine to diet 3 produced a deficiency of some other nutrient besides arginine. This nutrient would then have to be added before a response to arginine would be obtained. Iodide. It seemed possible that the trace amounts of potassium iodide added to diet 1 may be iodinating the casein in the diet, and thus increasing the arginine requirement of the chicks. Two experiments were conducted to determine what effect omitting the potassium iodide or increasing it to five times the normal level would have on growth of chicks fed diet 1. When the potassium iodide was omitted, growth rate was not increased; addition of five times the normal level did not decrease growth rate. Thus, no evidence was obtained which would indicate that the casein was iodinated. Effect on Arginine Requirement. The experiments already reported indicated that thiouracil decreased and iodinated casein increased the arginine requirements of the chick. Three experiments were conducted to determine how much the arginine requirement was changed by these substances. Diet 4 was used in these experiments. The total protein in the diet 4 (NX6.25) is about 24%; its arginine content is calculated to be 0.98%. In order to increase the arginine content of the diet, arginine HC1 was added and the casein
68
H. B. FLTJCKIGER AND J. 0. ANDERSON
. o 2
Di.t 4 Wilh 0 . 0 2 2 % lodinat*d Casvin
1.26 1.59 2.00 2.51 3.16 PERCENT ARGININE IN RATION
FIG. 4. Effect of iodinated casein on the arginine requirement of the chick.
dinated casein are the same as found in Figure 3. The addition of iodinated casein reduced average gains on all the lower levels of arginine; whereas, on the higher levels of arginine, growth was slightly greater when iodinated casein was added. The weight gains showed a definite increase as the arginine level was increased up to 1.66% when 0.022% iodinated casein was added to diet 4. Again smaller increases in gain were noted when the arginine content was increased beyond this point. Over 2% arginine was required to produce maximum gains. Effect on the Glycine Requirement. Three experiments were conducted which were designed to determine how much iodinated casein and thiouracil affect the. glycine requirement of the chick. Diet 7 was used in these experiments. The results of the three experiments are presented- in Figure 5. These results have been analyzed statistically. Chicks fed diets with added glycine attained slightly higher average weights, but the increase is not significant. Addition of iodinated casein significantly reduced growth. The reduction tended to be slightly less on the two high-
D i . l 7 With 0.06 «
.
D i . l 7 Wilh 0 . 0 2 2 % lodinot.d CoMin. 0.65
0.42
1.00
Thiouracil
2.41
1.55
PERCENT GLYCINE IN DIET
FIG. 5. Effect of thiouracil and iodinated casein on the glycine requirement of the chick.
est levels of glycine. This is the only observed indication that the addition of iodinated casein affects the glycine requirement of the chick. Effect on the Methionine Requirement. Both casein-gelatin and Drackett protein type diets have been used in studies on the methionine requirement. Figure 6 presents the results of three experiments con< o ac
0
UJ
Q-IO
/*
o X u
8
-
cr Z 6 '>•
<
a.
o
~
< O r-
I
2
'
Y .
Diet 5
»
Die* 5
With 0 . 0 6 % Thiouracil
o
Die! 5
With 0 . 0 2 2 % Iodinated
u S
Casein. —1
0.54
1
1
0.64
0.78
i ._
0.94
PERCENT METHIONINE IN DIET
FIG. 6. Effect of thiouracil and iodinated casein on the methionine requirement of chicks fed a casein type diet (diet 5).
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1.00
oi.i 7
«
AMINO ACID REQUIREMENTS
69
I
Downloaded from http://ps.oxfordjournals.org/ by guest on April 23, 2015
ducted with diet 5. Casein and gelatin provide the protein in this diet. The diet was > < o calculated to contain 0.53% methionine a: 10 -«-x and 0.06% cystine, using values given by ua Block and Boiling (1951). Each point plotted in Figure 6 is an average of at yi s least two values obtained in three sepa- o Jtrate experiments. Addition of 0.022% io-
I
70
H. B. FLUCKIGER AND J. 0 . ANDERSON
more methionine and cystine than the values reported by the manufacturer which were used in our calculations. Even if the higher values are correct, the total sulfur amino acid requirement of the chick is higher on the casein-gelatin diet (diet 5) than it is on the Drackett protein diet (diet 6). SUMMARY
Purified type diets containing 23 to 28% casein, 1% glycine and 0.3% DLmethionine to provide the protein, were found to be very deficient in arginine for the chick. When thiouracil was added to this type of diet, growth rate was increased, arginine deficiency symptoms were less severe, and the amount of arginine required for maximum growth was decreased. Addition of iodinated casein or thyroxine produced effects just opposite to those produced by the addition of thiouracil. Two other diets were fed in which the protein was provided by a mixture of wheat gluten, zein, amino acids and either blood fibrin or Drackett protein. These diets contained only slightly more arginine than the casein diet, but good growth was obtained without the addition of arginine. Chicks fed these diets did not show arginine deficiency symptoms unless iodinated casein or thyroxine was added. Thus, the arginine requirement of the chick was influenced by the kind of protein in the diet. The chick's methionine and glycine requirements were not affected by the addition of thiouracil or iodinated casein to their diet. ACKNOWLEDGMENTS
The authors are indebted to Merck and Company, Inc., Rahway, New Jersey, for some of the crystalline vitamins and the L-lysine HC1; Nopco Chemical Company,
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rate. Opposite effects also were noted on the arginine requirement of the chick. When both thiouracil and thyroxine were added to diet 1, the growth rate was less than that obtained by the addition of thiouracil, but more than that obtained by the addition of thyroxine. This suggests that thiouracil produces its effect on arginine requirement of the chick by reducing the rate thyroxine is secreted by the thyroid of the chick. The presence of a substance with thyroxine activity in casein might explain the high arginine requirement of chicks fed a casein type diet. These experiments offer very little evidence for the presence of such a substance in casein. Chicks gained weight four times as fast on diet 2 as they did on diet 1. When 1% arginine HC1 was added to both diets, the rate of gain was approximately equal. This would indicate that diet 1 is very deficient in arginine. The arginine content of diet 2 is only 0.09% greater than diet 1. When 0.09% arginine is added to diet 1, the rate of gain is still far less than that obtained with diet 2. These results are similar to those obtained by Snyder et al. (1956) and Krautmann et al. (1957). Their reports indicated that the chick's arginine requirement is higher on a purified casein type diet than it is on a cornsoybean diet. Apparently, some differences in the protein supplements used in diets 1 and 2 affect the arginine requirement of the chick. The methionine requirement of the chick appears to be much higher on diet 5 than it is on diet 6. Cystine can be utilized by the chick to correct the deficiency in diet 5, but it could not in diet 6. This helps to explain this difference, but the total sulfur amino acid requirement still appears to be higher on diet 5 than on diet 6. According to the data of Nelson et al. (1957) Drackett protein contains
AMINO ACID REQUIREMENTS
Harrison, New Jersey, for the vitamins A and D supplement; Lederle Laboratories Division, American Cyanamid Company, for the folic acid; and the Dow Chemical Company, Midland, Michigan, for some of the DL-methionine and glycine used in these experiments. REFERENCES
E. B. Hart, 1941. The role of arginine and glycine in chick nutrition. J. Biol. Chem. 140: 191-200. Hogan, A. G., R. W. Craghead, J. E. Savage, J. J. Cole and B. L. O'Dell, 1957. Casein as a source of protein for the chick. J. Nutrition, 62: 97-106. Krautmann, B. A., S. M. Hauge, E. T. Mertz and C. W. Carrick, 1957. The arginine level for chicks as influenced by ingredients. Poultry Sci. 36: 935-939. Nelson, T. S., L. C. Norris, F. W. Hill, M. L. Scott and G. F. Heuser, 1957. Further studies on the requirement of chicks for methionine and cystine. Proc. Cornell Nutrition Conf. 126-131. Rosenberg, H., A. H. Ennor and J. F. Morrison, 1956. The estimation of arginine. Biochem. J. 63: 153-159. Snyder, J. M., W. D. Morrison and H. M. Scott, 1956. The arginine requirement of chicks fed purified and corn-soya diets. Poultry Sci. 35: • 852-855. Vohra, P., and F. H. Kratzer, 1958. The effect of thyroprotein, thiouracil and cortisone on the feather pigmentation of turkey poults. Poultry Sci. 37: 151-153. 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. Nutrition, 52: 311-323. Young, R. J., M. B. Gillis and L. C. Norris, 1953. An unidentified factor in peanut meal required by the chick. J. Nutrition, 50: 291-297.
Variation in the Effects of Fat Supplements on Broiler Pigmentation, Growth and Feed Conversion D . S. C A R V E R
General Mills, Inc., Larro Research Farm, P. O. Box 263, Indianola, Iowa (Received for publication May 13, 1958)
T
HE range in price between prime tallow and lower grades of fat such as yellow grease or No. 2 tallow is normally f jf to If! a pound. This difference in price is very important to the feed manufacturer, because it means a substantial savings in ingredient cost can be realized if fats such as yellow grease and No. 2 tallow
can be substituted for prime tallow without reducing the feeding value of feeds. Yacowitz (1953) found that soybean oil, cottonseed oil and lard at levels of 2\ percent and 5 percent produced similar responses in growth and feed efficiency. March and Biely (1954) reported the results of feeding 5 percent to 10 percent
Downloaded from http://ps.oxfordjournals.org/ by guest on April 23, 2015
I
Almquist, H. J., 1947. Evaluation of amino acid requirements by observations on the chick. J. Nutrition, 34: 543-563. Almquist, H. J., and J. B. Merritt, 1950. Protein and arginine levels in the chick's diet. Proc. Soc. Exp. Biol, and Med. 73: 136. Block, R. J., and D. Boiling, 1951. The Amino Acid Composition of Proteins and Foods. Charles C Thomas, Publisher, Springfield, 111. 2nd Ed. 486492. Edwards, H. M., Jr., R. J. Young and M. B. Gillis, 1958. Studies on arginine deficiency in chicks. J. Nutrition, 64: 271-279. Fisher, H., R. C. Salander and M. W. Taylor, 1956. The influence of creatine biosynthesis on the arginine requirement of the chick. J. Nutrition, 59: 491-500. Griminger, P., H. Fisher and H. M. Scott, 1955. On the arginine requirement of the growing chick. Poultry Sci. 34: 1247-1249. Hegsted, D. M., G. M. Briggs, C. A. Elvehjem and
71