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Further Studies on Egg Yolk as a Source of Unidentified Growth Factors for Poults and Chicks1,2
PROTEIN REQUIREMENTS OF GROWING PULLETS
growth and feed efficiency. Poultry Sci. 34:1210. Roberts, R. E., 1929. Protein requirements of growing pullets. Poultry Sci. 9: 19-26. Schneider, A. J., B. B. Bohren and V. L. Anderson, 1955. The effect of restricted feeding on several genetically controlled characters in the fowl. Poultry Sci. 34: 691-702. Sunde, M. L., 1955. A relationship between protein level and energy level in chick rations. Federation Proc. 14: 451-452. Sunde, M. L., W. W. Cravens, H. R. Bird and J. G. Halpin, 1954. The effect of complete and incomplete growing diets on subsequent performance of the laying hen. Poultry Sci. 33: 779-784. Swift, R. W., A. Black, L. Voris and E. M. Funk, 1931. The optimum protein content of rations for growing chicks. Poultry Sci. 10: 288-298.
Further Studies on Egg Yolk as a Source of Unidentified Growth Factors for Poults and Chicks1,2 G. H. ARSCOTT AND J. A. HARPER Department of Poultry Husbandry AND J. R. SCHUBERT AND P. H. WESWIG Department of Agricultural Chemistry, Oregon State College, Corvallis, Oregon (Received for publication May 12, 1958)
D
ENTON et al. (1954) initially reported that egg yolk resulted in weight gains at 4 weeks of up to 18 percent when added to the diet of chicks. Several groups of investigators have since confirmed and extended this research (Hopper et al., 1956; Arscott, 1956; and Menge et al., 1957a, b). Complete agreement as to the nature of the response has not been possible. Hopper et al. (1956) have reported on the similarity of the fluid egg yolk activity to 1 Technical Paper No. 1121, Oregon Agricultural Experiment Station. 2 Reported in part at the 46th Annual Poultry Science Association meeting, Columbia, Mo., August 6-9, 1957.
the "fish factor" response. On the other hand Arscott (1956) and Menge et al. (1957a,b) in studies with dried egg yolk have indicated that factor(s) other than the "fish factor" may be involved. To substantiate the multiple nature of the growth response attributable to dried egg yolk, additional evidence from this station (Arscott et al., 1957) has also shown that fractions of dried egg yolk contain at least two separate unidentified growth factors. More recently Menge and associates (1957b) have elaborated further on the nature of the egg yolk response. Oleic and linoleic acids as well as lecithin and yolk ash were found inactive with activity concentrated in the nonphos-
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Combs, G. F., and G. L. Romoser, 1955. A new approach to poultry feed formulation. Feed Age, 5 (3): 50-58. Heady, E. 0., S. Balloun and R. McAlexander, 1956. Least-cost rations and optimum marketing weights for broilers. Res. Bui. 442, Agr. Exp. Station, Iowa State College, Ames, Iowa. Leong, K. C , M. L. Sunde, H. R. Bird and C. A. Elvehjem, 1955. Effect of energy: protein ration on growth rate, efficiency, feathering and fat deposition in chickens. Poultry Sci. 34: 1206. Milby, T. T., and D. H. Sherwood, 1956. The influence of feed intake during the growing period on the subsequent performance of laying hens. Poultry Sci. 35: 863-869. Potter, L. M., L. D. Matterson, D. Carlson and E. P. Singsen, 1955. Studies on the effect of varying protein levels in poultry rations on
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56
G. H. ARSCOTT, J. A. HARPER, J.. R. SCHUBERT AND P. H. WESWIG
pholipid and nonsaponifiable fractions of an ethanol extract of yolk. This paper reports further on egg yolk as a source of unidentified growth factors involving studies on (1) egg yolk components and certain reported growth factor supplements, (2) types and levels of yolk and (3) "carry-over" response from dam to progeny.
The design for each experiment is given in Tables 2 through 8. A range of 7 to 14 male chicks involving primarily a New Hampshire-Delaware cross was used in each group. A few experiments with New Hampshire chicks or a New HampshireWhite Rock cross were employed and will be indicated when necessary. Beltsville Small White turkeys of mixed sex were used in the poult study. All experiments were replicated either within a given trial and/or in subsequent trials. Management of battery raised chicks has previously been described (Arscott, 1956). The rat experiment involved a closed Sprague Dawley strain maintained within the Agricultural Chemistry Department for more than thirty years. The rats were weaned at three weeks and fed commercial chicken broiler crumbles until four weeks of age. This feed is routinely used as the stock diet for the rats in this laboratory. The basal rations used, unless otherwise noted, are shown in Table 1 and are designated as R-200, R-210 and R-300. Ration R-210, containing approximately 22.9 percent protein (calculated), differs from R-200 in that 4 percent soybean oil meal was added to the ration at the expense of sucrose. A number of unpublished experiments conducted at this laboratory has shown that the addition of 2, 4 or 8 percent soybean oil meal to R-200 resulted in a small but consistent increase
RESULTS AND DISCUSSION
Relation of egg yolk response to certain known components of egg yolk, reported growth factor supplements and energy: The summary data shown in Table 2 clearly show that the fatty acids (Series 1) palmitic, stearic and linoleic acids together with the sodium salt of oleic acid equivalent to that supplied by 8 percent yolk are without beneficial effect singly or in combination in promoting growth of chicks when compared to that obtained with dried egg yolk. Similarly a soybean lecithin product (Series 1) and the equivalent ash (550°C.) or cholesterol (Series 2 and 3) to 8 percent dried egg yolk appear without beneficial effect. These data concur with the findings of Menge et al. (1957a,b) with respect to oleic and linoleic acids, lecithin and the ash of egg yolk. Further evidence is presented with respect to palmitic and stearic acids and
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EXPERIMENTAL PROCEDURE
in growth that decreased the response obtained from 8 percent dried egg yolk from about 37 to 27 percent. The 8 percent level of soybean oil meal proved no better than the lower levels. Where R-200 is indicated it is generally accompanied by data involving R-210 with no apparent differences between treatments on the two diets. R-210 together with R-300, containing 32 percent protein, are considered adequate from the standpoint of all the known nutrients required by chicks and poults, respectively. All supplements__jygre added to the ba^sajjjationjitj^ The dried egg yolk employed was a commercially prepared product. Unless otherwise indicated the fluid yolks were obtained from chicken and/or turkey hens maintained on a commercial breeder ration. Individual body weights and feed consumption data were recorded at 4 weeks of age.
EGG YOLK AS A SOURCE OF UNIDENTIFIED FACTORS
57
/l?
TABLE 1.—Composition of basal rations
m'
Chick Ingredients
M}
v
J/
Poult
R-210
% 45.0 49.28 0.4 0.2 0.13 0.05 0.034 0.076
49.0 44.02 0.4 0.2 0.13 0.05 0.034 0.076
%
R-300 % 69.0 21.06
—
0.03 4.832
0.03 5.092
0.5 0.2 0.15 0.06 0.07 0.1 0.05 8.796
+
% of diet
+
+
—
4.0
—
—
— —
3.7 1.34 0.5
—
0.21
—
0.035 0.004 0.002 0.0001 0.0009 mg./100 gm. 3.3 1.2 1.0 0.4 0.4 0.4 0.2 0.1 0.075 0.010
0.5 0.29
0.035 0.004 0.002 0.0001 0.0009 3.3 1.2 1.0 0.4 0.4 0.4 0.2 0.1 0.075 0.010
—
—
—
7.0 1.0 0.5
—
0.25
—
0.035 0.006 0.003 0.00015 0.00135 7.5 3.3 2.0 0.9 l.S 1.5 0.3 0.5 0.4 0.03
,q-X.T<
cholesterol as not influencing growth. Carver and Johnson (1953), however, have reported linoleic and oleic acid concentrates as containing varying amounts of growth factor activity on more purified diets than those reported here. Under their conditions stearic acid appeared to depress growth as was the case in these experiments. Adenosine (Series 3) and orotic acid (Series 4) likewise do not appear involved in the expression of the egg yolk response. These data, therefore, would appear at variance with the report of Barnett et al. (1956) for adenosine and Combs et al.
(1954) and Fritz et al. (1954) for orotic acid until one relates these responses to "whey factor" supplements. Barnett et al. (1956) observed no response to orotic acid supplementation in the presence of whey while Huff et al. (1950) have isolated orotic acid from spray dried whey. The absence of the "whey factor" response in diets containing 44 percent solvent extracted soybean oil meal has been discussed elsewhere, Arscott (1956), and may serve to account for this apparent disagreement. Summary data on the role of energy supplied by corn oil in the expression of
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Soybean oil meal (44% protein) Sucrose Zein DL-methionine (98%) Vitamin A, dry (10,000 U.S.P.U./gm.) Vitamin D, dry (15,000 I.C.U./gm.) Vitamin E (14.71 gm./lb.) Choline chloride (86.9%) Inositol Iodinated casein ( 1 % thyroxine) Mineral mixture Vitamin mixture Mineral mixture Rock phosphate, defl. (34% Ca, 16.8% P) Di-calcium phosphate (23% Ca, 18% P) Limestone flour Salt, iodized Ferrous ammonium citrate (Fe(C6Hs06)2-6H20) Ferrous ammonium sulfate (Fe(NH4)2(SO.i)2' 6H 2 0) Manganese sulfate (65%) Manganese sulfate (70%) Potassium iodide Zinc chloride Cobalt sulfate (CoSCv 7H 2 0) Boric acid Vitamin mixture Niacin Ca-pantothenate (91.8%) Vitamin B« (1 mg./gm.) Pyridoxin HC1 Riboflavin Thiamine HC1 p-amino benzoic acid 2-methyl-l, 4-naphthoquinone Folacin Biotin
R-200
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G. H. ARSCOTT, J. A. HARPER, J. R. SCHUBERT AND P. H. WESWIG
TABLE 2.—Relation of egg yolk response to certain egg
yolk components and other known elements on chick growth Se- Grouf )
Av. 4week gain
Treatment
(gm.) 1 2 3 4 5 6 7 8
None 1 8 % Dried Egg Yolk 2.5% N a 0 1 e a t e = 8 % D.E.Y.3 1.3% Palmitic A c i d = 8 % D.E.Y. 3 0 . 3 % Stearic A c i d = 8 % D.E.Y.* 0.6% Linoleic Acid (60%) = 8 % D.E.Y.<1 As 3 + 4 + 5 + 6 1% Lecithin (Alcolec S)
375 (3)2 457 (3) 378(3) 387 (3) 358 (3) 385(3) 351(2) 367 (2)
2
1 2 3
None 1 8 % Dried Egg Yolk Ash = 8 % D . E . Y .
336 (6)5 427 (6) 332 (6)
3
1 2 3 4
None, R-210 8 % Dried Egg Yolk 3.12% Cholesterol = 8 % D.E.Y.» Adenosine (300 mg./kg.)
338 (2)« 432 (2) 328 (2) 322 (2)
4
1 2 3 4 5 6
None 1 8 % Dried Egg Yolk Orotic Acid (40 mg./kg.) As 2 + 3 Orotic Acid (80 mg./kg.) As 2 + 5
338 (3)' 420 (3) 302 (3) 400(3) 307 (3) 364 (3)
1 R-200 used in experiments 1 & 2; R-210 used in subsequent experiments. 2 Figure in parentheses indicate no. of experiments with following number of chicks per exp.: 12; 12; 14. 3 Non dry basis (Romanoff and Romanoff, 1949). 4 Moisture free basis (Romanoff and Romanoff, 1949). 6 Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 13; 13; 14X3; 10X2; 9-7; 12X2. 6 Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 10X2; 12X2. * Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 13; 12; 12.
the egg yolk response are given in Table 3. These results show a slight but consistent increase in growth when up to 4 percent corn oil is added to the basal ration, R-210. It is interesting to note, however, that the sensitivity of the experimental assay is not consistently or markedly re-
TABLE 3.—Summary data on influence of dried egg yolk on growth in presence or absence of corn oil Av. 4-week data Group
Treatment
Approx. fat content
% 1 2 3 4 5 6
None, R-210 8% Dried Egg Yolk 2% Corn Oil2 As 3 + 8 % D.E.Y. 4 % Corn Oil As 5 + 8 % D.E.Y.
0.5 4.5 2.5 6.5 4.5 8.5
Rel. gain (%)
Gain (gm.) 344 (7) 426 (7) 348 (3) 443(3) 360 (5) 440(2)
1
Figures in parentheses represent number of experiments. Mazola oil.
Group 1
Group 3
100 125 102 130 106 129
100 127 103 126
—
Group 5
— — —
100 122
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1
duced when relative gains are examined in the presence of 2 or 4 percent corn oil. Since energy might be considered a critical factor in interpreting these results, these data are of interest in view of the fact that the 4 percent level of corn oil approximates the amount of fat added to the diet by 8 percent dried egg yolk. Thus the response to energy by way of corn oil apparently accounts for no more than twelve percent of the response to egg yolk. Menge et al. (1957a,b) have likewise noted that 5 percent stabilized lard exerted little influence on the response to egg yolk. Relation of egg yolk response to amount and type of yolk: The summary data from numerous representative experiments presented in Table 4 clearly show the magnitude of the growth response obtained from the addition of 4, 8 and 12 percent dried egg yolk to the diet. These data show a significant increase (P = 0.01) in gain for each level of dried egg yolk when compared to the unsupplemented control. When comparing between supplements both the 8 and 12 percent levels proved highly significant (P = 0.01) over 4 percent dried egg yolk. Furthermore, the 12 percent level of dried egg yolk proved significant at the 5 percent level of probability when compared to the 8 percent level. When considering that dried egg yolk contains approximately twice the
59
EGG YOLK AS A SOURCE OF UNIDENTIFIED FACTORS TABLE 4.—Influence of dried egg yolk level on chick growth
TABLE 5.—Relation oj fluid turkey and chicken yolk to the dried egg yolk response for chicks and poults
Av. 4-week gain Treatment
Av. 4-week gain (gm.) (gm.)
None 4 % Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
344(15)' 378 2 (8) 4292>3 (15) 4552.3'4 (13)
solids (Romanoff and Romanoff, 1949) observed in fluid yolk, these data appear at variance with those of Hopper et al. (1956) who have reported optimal response with 5 percent fluid egg yolk which is equivalent to only 2.5 percent dried egg yolk. These data would suggest, therefore, that considerable destruction of activity occurs in processing egg yolk or that egg yolk varies considerably in its content of activity as influenced possibly by the diet of the dam. In view of this a number of experiments were conducted to compare the performance of chicks and poults to different levels and types of egg yolk. The results given in Table 5 reveal little difference in gain of chicks fed fluid turkey and chicken yolks with a suboptimal response indicated at the 4 and 8 percent levels. It would appear that the response from 12 percent fluid yolk is comparable to that from 8 percent dried egg yolk. It should be noted in these data that while the 8 percent level of dried egg yolk proved significantly greater (P = 0.05) than the 4 percent level, the 12 percent dried egg yolk addition did not prove significantly greater than the 8 percent level. On the other hand the data involving poults show a comparable response to either
Chicks 1 ' 2
Poults 3 ' 4
None, R-210
338
381
4 % Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
3911 422 419 443j
4221 433 425 420J
4 % Fluid Turkey Yolk 8% Fluid Turkey Yolk 12% Fluid Turkey Yolk
3841 398 403 427J
4491 391U24 433J
4 % Fluid Chicken Yolk 8% Fluid Chicken Yolk 12% Fluid Chicken Yolk
3781 405 404 430J
4181 429 425 428J
1 Av. data for 2 experiments with following no. of chicks per exp.: 9 X 2 ; 14X2. 2 Differences greater than 22 grams are statistically significant at P=0.05. 3 Av. data for 1 experiment with following no. of poults per exp.: 12X2. 4 Differences greater than 61 grams are statistically significant at P = 0.05.
type of fluid yolk or the dried egg yolk. Furthermore an optimal growth response appears evident at the 4 percent level, however, no significance was attained for individual treatments except for the group containing 4 percent fluid turkey yolk. While it must be recognized that poults of mixed sex were used in contrast to males in the chick experiments, it is evident that the magnitude of response is considerably less with more variability encountered than normally obtained with chicks. The results presented in Table 6 show the gain obtained with rats from 4 to 8 weeks as compared with male chicks fed identical rations. These data reveal that 8 percent dried egg yolk and an ether soluble extract of 8 percent dried egg yolk, while resulting in a slight improvement in growth over the basal control, failed to promote optimum growth when compared with commercial broiler crumbles. On the other hand, growth of chicks
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1 Figures in parentheses represent number of experiments. 2 Significantly greater than group 1 at one percent level of probability (t test). 3 Significantly greater than group 2 at one percent level of probability (t test). 4 Significantly greater than group 3 at five percent level of probability (t test).
Treatment
G. H. ARSCOTT, J. A. HARPER, J.. R. SCHUBERT AND P. H. WESWIG
60
TABLE 6.—Rat and chick response to egg yolk Group
1 2 3 4 1
TABLE 8.—Influence of maternal diet on egg yolk response
None, R-210 As 1 + 8 % Dried Egg Yolk As 1 + E t h e r Sol. Fract. = 8 % D . E . Y . Commercial Broiler Crumbles
Rats (4-8 wks.)
Chicks (0-4 wks.)
129(8)1 136 (8) 127(8) 147 (8)
339(16) 425 (15) 380(14) 376 (16)
Figures in parentheses represent number of survivors.
TABLE 7.—Influence of maternal diet on egg yolk response Av. 4-week gain 1
Treatment Chick'
(gm.)
None—control
None, R-200 8% Dried Egg Yolk
328 366
5 % Dried Egg Yolk
None, R-200 8% Dried Egg Yolk
342 384
Iodinated Casein (.1 gm./lb.)
None, R-200 8 % Dried Egg Yolk
343 375
As 2 + 3
None, R-200 8 % Dried Egg Yolk
327 380
1 Two-year data with 5 and 4 experiments with sexed males and2 mixed sexes, resp., per year. New Hampshires.
Chick'
Av. 4-week gain Hen2
(gm.)
None, R-210
344 (3)3
4% Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
381(3)1 432 (3) 427 469 (3) J
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(C-SM)« (C-SM)* (C-SM)*
382 (3)1 405 (3) U05 429 (3)J
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(C-SM-3%FM) (C-SM-3%FM) (C-SM-3%FM)
376(3)1 408(3) 411 450(3))
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(S-SM) (S-SM) (S-SM)
376(3)1 394 (3) 1395 416 (3) j
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(S-SM-3%FM) (S-SM-3%FM) (S-SM-3%FM)
394 (3) 1 405(2)5 1412 438 (3) J
1 2 3
New Hampshire XDelaware. New Hampshire. Figures in parentheses represent no. of experiments with 13 and 11 chicks per experiment. *C=corn; SM=soybean oil meal; FM=fish meal; S = sucrose. 6 Average of 2 groups only, water consumption restricted in second experiment.
dams housed on wire floors. It is evident in Table 7 that no "carry-over" of the unknown nutrient occurs in the presence of 5 percent dried egg yolk in the dam's diet (group 2) as indicated by gains of progeny to four weeks of age when fed R-200 with and without dried egg yolk. The increased response noted when egg yolk was fed in the presence of iodinated casein (group 4) was evident only in the first year's results and is therefore discounted. Likewise when fluid yolks from dams fed either a corn-soybean oil meal or sucrose-soybean oil meal with or without 3 percent fish meal were fed to chicks, Table 8, no differences in growth could be related to the diet fed the dams. These data concur with those of Keene et al. (1957) who have reported that egg yolks from hens raised on wire and fed a cornwheat-soybean oil meal type ration were as effective in improving chick growth as were yolks from hens reared on range and fed a complete ration. Lillie et al. (1957) in contrast have reported a significant
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compared favorably to that obtained previously with dried egg yolk or fat soluble extracts of dried egg yolk as reported by Arscott et al. (1957). The greater gain obtained with the egg yolk supplement when compared with the commercial broiler feed is of interest but cannot be accounted for at this time. Bentley (1954) has reported rat growth to be unaffected on corn-soybean oil meal or semi-synthetic type rations when egg yolk supplements were employed provided such rations contained no added iodinated casein. In the presence of 0.08 percent iodinated casein, however, eggs from hens fed antibiotics appeared to increase growth. The relation of egg yolk reponse to "carryover": The experiments in Tables 7 and 8 were designed to show the relation of "carry-over" of the unidentified nutrients) from the dam to the chick by using either chicks or eggs from treated
Hen!
Treatment
Av. 4-week gain (gm-)
Treatment
EGG YOLK AS A SOURCE or UNIDENTIFIED FACTORS
"carry-over" effect on progeny performance from the inclusion of 5 percent dried egg yolk in the maternal diet. SUMMARY
The authors wish to thank the following concerns for material kindly supplied during these investigations: Corn Products
Sales Co., New York, N. Y., for Argo Brand zein; Hoffmann-LaRoche, Inc., Nutley, N. J., for biotin; Lederle Laboratories Division, American Cyanamid Co., Pearl River, N. Y., for folacin and orotic acid; Merck & Co., Inc., Rahway, N. J., for other B-complex vitamins and 2-methyl-l, 4-napthoquinone; and Nopco Chemical Co., Richmond, Calf., for vitamin A (Nopcay 10). REFERENCES Arscott, G. H., 1956. Complexity of a chick growth response to egg yolk, animal fat and fish solubles additions to the diet. Poultry Sci. 35: 338-342. Arscott, G. H., P. H. Weswig and J. R. Schubert, 1957. Multiple nature of chick growth responses to fractions of dried egg yolk. Poultry Sci. 36: 513-516. Barnett, B. D., M. Lapidus, H. R. Bird and F. M. Strong, 1956. Adenosine as growth factor for chicks fed purified diet. Proc. Soc. Biol. Med. 92: 372-374. Bentley, 0. G., 1954. Rat growth response of egg yolks from hens fed antibiotics. Fed. Proc. 13 Pt. 1:181-182. Carver, D. S., and E. L. Johnson, 1953. Unidentified growth factors for the chick in vegetable oils and fatty acid concentrates. Poultry Sci. 32: 701-705. Combs, G. F., G. H. Arscott and H. L. Jones, 1954. Unidentified growth factors required by chicks and poults. 3. Chick studies involving practicaltype rations. Poultry Sci. 33: 71-79. Denton, C. A., R. J. Lillie and J. R. Sizemore, 1954. Effect of egg yolk, fat and fish solubles on growth of chicks. Fed. Proc. 13 Pt. 1: 455. Fritz, J. C , F. D. Wharton and R. M. Henley, 1954. Effect of orotic acid on growth of chicks and poults. Fed. Proc. 13 Pt. 1: 458. Hopper, J. H., H. M. Scott and B. C. Johnson, 1956. The growth promoting ability of fluid egg yolk. Poultry Sci. 35: 195-197. Huff, J. W., D. K. Basshardt, L. D. Wright, D. S. Spicer, K. A. Valetnik and H. R. Skeggs, 1950. A growth promoting substance for L. bulgaricus 09 in whey: Isolation and identification as orotic acid. Proc. Soc. Exp. Biol. Med. 75: 297-299. Keene, O. D., G. F. Combs and G. L. Romoser, 1957. Studies on carry-over and depletion of unidentified chick growth factors. Poultry Sci. 36: 11321133. Lillie, R. J., H. Menge and C. A. Denton, 1957. A breed difference with respect to carry-over effect
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Studies have been conducted involving (1) egg yolk components and reported growth factor supplements, (2) types and levels of yolk and (3) "carry-over" response from dam to progeny using a sucrose-soybean oil meal diet. Linoleic, palmitic and stearic acids, sodium oleate, cholesterol or an ashed yolk (550°C.) fed to chicks at levels equivalent to 8 percent dried yolk appeared to contain no growth stimulating effect. Soybean lecithin, orotic acid or adenosine were also without effect. Energy, supplied by corn oil, did not markedly reduce the response to egg yolk. Increased chick growth occurred when 4, 8 and 12 percent dried yolk was included in the diet. The 8 and 12 percent levels of dried egg yolk evoked significantly greater responses (P = 0.01) than the 4 percent level. In a summary involving thirteen experiments the 12 percent level of dried egg yolk proved significantly greater (P = 0.05) than the 8 percent level. Four, 8 and 12 percent fluid chicken or turkey yolk also responded in step-wise fashion with 12 percent fluid yolk appearing comparable to 8 percent dried egg yolk. Poults likewise responded to dried yolk or fluid chicken and turkey yolk with the 4 percent level of either appearing optimum. More variability and smaller differences were experienced with poults. Rats failed to show a marked response to dried yolk. No "carry-over" effect of the egg yolk factor was observed from dam to chick. ACKNOWLEDGMENTS
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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
H. B. FLUCKIGER 2 AND J. O. ANDERSON Poultry Department, Utah State University, Logan, Utah (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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I. EFFECT OF THYROXINE AND KIND OF PROTEIN ON THE ARGININE, METHIONINE AND GLYCINE REQUIREMENTS 1
growth and feed efficiency. Poultry Sci. 34:1210. Roberts, R. E., 1929. Protein requirements of growing pullets. Poultry Sci. 9: 19-26. Schneider, A. J., B. B. Bohren and V. L. Anderson, 1955. The effect of restricted feeding on several genetically controlled characters in the fowl. Poultry Sci. 34: 691-702. Sunde, M. L., 1955. A relationship between protein level and energy level in chick rations. Federation Proc. 14: 451-452. Sunde, M. L., W. W. Cravens, H. R. Bird and J. G. Halpin, 1954. The effect of complete and incomplete growing diets on subsequent performance of the laying hen. Poultry Sci. 33: 779-784. Swift, R. W., A. Black, L. Voris and E. M. Funk, 1931. The optimum protein content of rations for growing chicks. Poultry Sci. 10: 288-298.
Further Studies on Egg Yolk as a Source of Unidentified Growth Factors for Poults and Chicks1,2 G. H. ARSCOTT AND J. A. HARPER Department of Poultry Husbandry AND J. R. SCHUBERT AND P. H. WESWIG Department of Agricultural Chemistry, Oregon State College, Corvallis, Oregon (Received for publication May 12, 1958)
D
ENTON et al. (1954) initially reported that egg yolk resulted in weight gains at 4 weeks of up to 18 percent when added to the diet of chicks. Several groups of investigators have since confirmed and extended this research (Hopper et al., 1956; Arscott, 1956; and Menge et al., 1957a, b). Complete agreement as to the nature of the response has not been possible. Hopper et al. (1956) have reported on the similarity of the fluid egg yolk activity to 1 Technical Paper No. 1121, Oregon Agricultural Experiment Station. 2 Reported in part at the 46th Annual Poultry Science Association meeting, Columbia, Mo., August 6-9, 1957.
the "fish factor" response. On the other hand Arscott (1956) and Menge et al. (1957a,b) in studies with dried egg yolk have indicated that factor(s) other than the "fish factor" may be involved. To substantiate the multiple nature of the growth response attributable to dried egg yolk, additional evidence from this station (Arscott et al., 1957) has also shown that fractions of dried egg yolk contain at least two separate unidentified growth factors. More recently Menge and associates (1957b) have elaborated further on the nature of the egg yolk response. Oleic and linoleic acids as well as lecithin and yolk ash were found inactive with activity concentrated in the nonphos-
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Combs, G. F., and G. L. Romoser, 1955. A new approach to poultry feed formulation. Feed Age, 5 (3): 50-58. Heady, E. 0., S. Balloun and R. McAlexander, 1956. Least-cost rations and optimum marketing weights for broilers. Res. Bui. 442, Agr. Exp. Station, Iowa State College, Ames, Iowa. Leong, K. C , M. L. Sunde, H. R. Bird and C. A. Elvehjem, 1955. Effect of energy: protein ration on growth rate, efficiency, feathering and fat deposition in chickens. Poultry Sci. 34: 1206. Milby, T. T., and D. H. Sherwood, 1956. The influence of feed intake during the growing period on the subsequent performance of laying hens. Poultry Sci. 35: 863-869. Potter, L. M., L. D. Matterson, D. Carlson and E. P. Singsen, 1955. Studies on the effect of varying protein levels in poultry rations on
55
56
G. H. ARSCOTT, J. A. HARPER, J.. R. SCHUBERT AND P. H. WESWIG
pholipid and nonsaponifiable fractions of an ethanol extract of yolk. This paper reports further on egg yolk as a source of unidentified growth factors involving studies on (1) egg yolk components and certain reported growth factor supplements, (2) types and levels of yolk and (3) "carry-over" response from dam to progeny.
The design for each experiment is given in Tables 2 through 8. A range of 7 to 14 male chicks involving primarily a New Hampshire-Delaware cross was used in each group. A few experiments with New Hampshire chicks or a New HampshireWhite Rock cross were employed and will be indicated when necessary. Beltsville Small White turkeys of mixed sex were used in the poult study. All experiments were replicated either within a given trial and/or in subsequent trials. Management of battery raised chicks has previously been described (Arscott, 1956). The rat experiment involved a closed Sprague Dawley strain maintained within the Agricultural Chemistry Department for more than thirty years. The rats were weaned at three weeks and fed commercial chicken broiler crumbles until four weeks of age. This feed is routinely used as the stock diet for the rats in this laboratory. The basal rations used, unless otherwise noted, are shown in Table 1 and are designated as R-200, R-210 and R-300. Ration R-210, containing approximately 22.9 percent protein (calculated), differs from R-200 in that 4 percent soybean oil meal was added to the ration at the expense of sucrose. A number of unpublished experiments conducted at this laboratory has shown that the addition of 2, 4 or 8 percent soybean oil meal to R-200 resulted in a small but consistent increase
RESULTS AND DISCUSSION
Relation of egg yolk response to certain known components of egg yolk, reported growth factor supplements and energy: The summary data shown in Table 2 clearly show that the fatty acids (Series 1) palmitic, stearic and linoleic acids together with the sodium salt of oleic acid equivalent to that supplied by 8 percent yolk are without beneficial effect singly or in combination in promoting growth of chicks when compared to that obtained with dried egg yolk. Similarly a soybean lecithin product (Series 1) and the equivalent ash (550°C.) or cholesterol (Series 2 and 3) to 8 percent dried egg yolk appear without beneficial effect. These data concur with the findings of Menge et al. (1957a,b) with respect to oleic and linoleic acids, lecithin and the ash of egg yolk. Further evidence is presented with respect to palmitic and stearic acids and
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EXPERIMENTAL PROCEDURE
in growth that decreased the response obtained from 8 percent dried egg yolk from about 37 to 27 percent. The 8 percent level of soybean oil meal proved no better than the lower levels. Where R-200 is indicated it is generally accompanied by data involving R-210 with no apparent differences between treatments on the two diets. R-210 together with R-300, containing 32 percent protein, are considered adequate from the standpoint of all the known nutrients required by chicks and poults, respectively. All supplements__jygre added to the ba^sajjjationjitj^ The dried egg yolk employed was a commercially prepared product. Unless otherwise indicated the fluid yolks were obtained from chicken and/or turkey hens maintained on a commercial breeder ration. Individual body weights and feed consumption data were recorded at 4 weeks of age.
EGG YOLK AS A SOURCE OF UNIDENTIFIED FACTORS
57
/l?
TABLE 1.—Composition of basal rations
m'
Chick Ingredients
M}
v
J/
Poult
R-210
% 45.0 49.28 0.4 0.2 0.13 0.05 0.034 0.076
49.0 44.02 0.4 0.2 0.13 0.05 0.034 0.076
%
R-300 % 69.0 21.06
—
0.03 4.832
0.03 5.092
0.5 0.2 0.15 0.06 0.07 0.1 0.05 8.796
+
% of diet
+
+
—
4.0
—
—
— —
3.7 1.34 0.5
—
0.21
—
0.035 0.004 0.002 0.0001 0.0009 mg./100 gm. 3.3 1.2 1.0 0.4 0.4 0.4 0.2 0.1 0.075 0.010
0.5 0.29
0.035 0.004 0.002 0.0001 0.0009 3.3 1.2 1.0 0.4 0.4 0.4 0.2 0.1 0.075 0.010
—
—
—
7.0 1.0 0.5
—
0.25
—
0.035 0.006 0.003 0.00015 0.00135 7.5 3.3 2.0 0.9 l.S 1.5 0.3 0.5 0.4 0.03
,q-X.T<
cholesterol as not influencing growth. Carver and Johnson (1953), however, have reported linoleic and oleic acid concentrates as containing varying amounts of growth factor activity on more purified diets than those reported here. Under their conditions stearic acid appeared to depress growth as was the case in these experiments. Adenosine (Series 3) and orotic acid (Series 4) likewise do not appear involved in the expression of the egg yolk response. These data, therefore, would appear at variance with the report of Barnett et al. (1956) for adenosine and Combs et al.
(1954) and Fritz et al. (1954) for orotic acid until one relates these responses to "whey factor" supplements. Barnett et al. (1956) observed no response to orotic acid supplementation in the presence of whey while Huff et al. (1950) have isolated orotic acid from spray dried whey. The absence of the "whey factor" response in diets containing 44 percent solvent extracted soybean oil meal has been discussed elsewhere, Arscott (1956), and may serve to account for this apparent disagreement. Summary data on the role of energy supplied by corn oil in the expression of
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Soybean oil meal (44% protein) Sucrose Zein DL-methionine (98%) Vitamin A, dry (10,000 U.S.P.U./gm.) Vitamin D, dry (15,000 I.C.U./gm.) Vitamin E (14.71 gm./lb.) Choline chloride (86.9%) Inositol Iodinated casein ( 1 % thyroxine) Mineral mixture Vitamin mixture Mineral mixture Rock phosphate, defl. (34% Ca, 16.8% P) Di-calcium phosphate (23% Ca, 18% P) Limestone flour Salt, iodized Ferrous ammonium citrate (Fe(C6Hs06)2-6H20) Ferrous ammonium sulfate (Fe(NH4)2(SO.i)2' 6H 2 0) Manganese sulfate (65%) Manganese sulfate (70%) Potassium iodide Zinc chloride Cobalt sulfate (CoSCv 7H 2 0) Boric acid Vitamin mixture Niacin Ca-pantothenate (91.8%) Vitamin B« (1 mg./gm.) Pyridoxin HC1 Riboflavin Thiamine HC1 p-amino benzoic acid 2-methyl-l, 4-naphthoquinone Folacin Biotin
R-200
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G. H. ARSCOTT, J. A. HARPER, J. R. SCHUBERT AND P. H. WESWIG
TABLE 2.—Relation of egg yolk response to certain egg
yolk components and other known elements on chick growth Se- Grouf )
Av. 4week gain
Treatment
(gm.) 1 2 3 4 5 6 7 8
None 1 8 % Dried Egg Yolk 2.5% N a 0 1 e a t e = 8 % D.E.Y.3 1.3% Palmitic A c i d = 8 % D.E.Y. 3 0 . 3 % Stearic A c i d = 8 % D.E.Y.* 0.6% Linoleic Acid (60%) = 8 % D.E.Y.<1 As 3 + 4 + 5 + 6 1% Lecithin (Alcolec S)
375 (3)2 457 (3) 378(3) 387 (3) 358 (3) 385(3) 351(2) 367 (2)
2
1 2 3
None 1 8 % Dried Egg Yolk Ash = 8 % D . E . Y .
336 (6)5 427 (6) 332 (6)
3
1 2 3 4
None, R-210 8 % Dried Egg Yolk 3.12% Cholesterol = 8 % D.E.Y.» Adenosine (300 mg./kg.)
338 (2)« 432 (2) 328 (2) 322 (2)
4
1 2 3 4 5 6
None 1 8 % Dried Egg Yolk Orotic Acid (40 mg./kg.) As 2 + 3 Orotic Acid (80 mg./kg.) As 2 + 5
338 (3)' 420 (3) 302 (3) 400(3) 307 (3) 364 (3)
1 R-200 used in experiments 1 & 2; R-210 used in subsequent experiments. 2 Figure in parentheses indicate no. of experiments with following number of chicks per exp.: 12; 12; 14. 3 Non dry basis (Romanoff and Romanoff, 1949). 4 Moisture free basis (Romanoff and Romanoff, 1949). 6 Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 13; 13; 14X3; 10X2; 9-7; 12X2. 6 Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 10X2; 12X2. * Figure in parentheses indicate no. of experiments with following no. of chicks per experiment: 13; 12; 12.
the egg yolk response are given in Table 3. These results show a slight but consistent increase in growth when up to 4 percent corn oil is added to the basal ration, R-210. It is interesting to note, however, that the sensitivity of the experimental assay is not consistently or markedly re-
TABLE 3.—Summary data on influence of dried egg yolk on growth in presence or absence of corn oil Av. 4-week data Group
Treatment
Approx. fat content
% 1 2 3 4 5 6
None, R-210 8% Dried Egg Yolk 2% Corn Oil2 As 3 + 8 % D.E.Y. 4 % Corn Oil As 5 + 8 % D.E.Y.
0.5 4.5 2.5 6.5 4.5 8.5
Rel. gain (%)
Gain (gm.) 344 (7) 426 (7) 348 (3) 443(3) 360 (5) 440(2)
1
Figures in parentheses represent number of experiments. Mazola oil.
Group 1
Group 3
100 125 102 130 106 129
100 127 103 126
—
Group 5
— — —
100 122
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1
duced when relative gains are examined in the presence of 2 or 4 percent corn oil. Since energy might be considered a critical factor in interpreting these results, these data are of interest in view of the fact that the 4 percent level of corn oil approximates the amount of fat added to the diet by 8 percent dried egg yolk. Thus the response to energy by way of corn oil apparently accounts for no more than twelve percent of the response to egg yolk. Menge et al. (1957a,b) have likewise noted that 5 percent stabilized lard exerted little influence on the response to egg yolk. Relation of egg yolk response to amount and type of yolk: The summary data from numerous representative experiments presented in Table 4 clearly show the magnitude of the growth response obtained from the addition of 4, 8 and 12 percent dried egg yolk to the diet. These data show a significant increase (P = 0.01) in gain for each level of dried egg yolk when compared to the unsupplemented control. When comparing between supplements both the 8 and 12 percent levels proved highly significant (P = 0.01) over 4 percent dried egg yolk. Furthermore, the 12 percent level of dried egg yolk proved significant at the 5 percent level of probability when compared to the 8 percent level. When considering that dried egg yolk contains approximately twice the
59
EGG YOLK AS A SOURCE OF UNIDENTIFIED FACTORS TABLE 4.—Influence of dried egg yolk level on chick growth
TABLE 5.—Relation oj fluid turkey and chicken yolk to the dried egg yolk response for chicks and poults
Av. 4-week gain Treatment
Av. 4-week gain (gm.) (gm.)
None 4 % Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
344(15)' 378 2 (8) 4292>3 (15) 4552.3'4 (13)
solids (Romanoff and Romanoff, 1949) observed in fluid yolk, these data appear at variance with those of Hopper et al. (1956) who have reported optimal response with 5 percent fluid egg yolk which is equivalent to only 2.5 percent dried egg yolk. These data would suggest, therefore, that considerable destruction of activity occurs in processing egg yolk or that egg yolk varies considerably in its content of activity as influenced possibly by the diet of the dam. In view of this a number of experiments were conducted to compare the performance of chicks and poults to different levels and types of egg yolk. The results given in Table 5 reveal little difference in gain of chicks fed fluid turkey and chicken yolks with a suboptimal response indicated at the 4 and 8 percent levels. It would appear that the response from 12 percent fluid yolk is comparable to that from 8 percent dried egg yolk. It should be noted in these data that while the 8 percent level of dried egg yolk proved significantly greater (P = 0.05) than the 4 percent level, the 12 percent dried egg yolk addition did not prove significantly greater than the 8 percent level. On the other hand the data involving poults show a comparable response to either
Chicks 1 ' 2
Poults 3 ' 4
None, R-210
338
381
4 % Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
3911 422 419 443j
4221 433 425 420J
4 % Fluid Turkey Yolk 8% Fluid Turkey Yolk 12% Fluid Turkey Yolk
3841 398 403 427J
4491 391U24 433J
4 % Fluid Chicken Yolk 8% Fluid Chicken Yolk 12% Fluid Chicken Yolk
3781 405 404 430J
4181 429 425 428J
1 Av. data for 2 experiments with following no. of chicks per exp.: 9 X 2 ; 14X2. 2 Differences greater than 22 grams are statistically significant at P=0.05. 3 Av. data for 1 experiment with following no. of poults per exp.: 12X2. 4 Differences greater than 61 grams are statistically significant at P = 0.05.
type of fluid yolk or the dried egg yolk. Furthermore an optimal growth response appears evident at the 4 percent level, however, no significance was attained for individual treatments except for the group containing 4 percent fluid turkey yolk. While it must be recognized that poults of mixed sex were used in contrast to males in the chick experiments, it is evident that the magnitude of response is considerably less with more variability encountered than normally obtained with chicks. The results presented in Table 6 show the gain obtained with rats from 4 to 8 weeks as compared with male chicks fed identical rations. These data reveal that 8 percent dried egg yolk and an ether soluble extract of 8 percent dried egg yolk, while resulting in a slight improvement in growth over the basal control, failed to promote optimum growth when compared with commercial broiler crumbles. On the other hand, growth of chicks
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1 Figures in parentheses represent number of experiments. 2 Significantly greater than group 1 at one percent level of probability (t test). 3 Significantly greater than group 2 at one percent level of probability (t test). 4 Significantly greater than group 3 at five percent level of probability (t test).
Treatment
G. H. ARSCOTT, J. A. HARPER, J.. R. SCHUBERT AND P. H. WESWIG
60
TABLE 6.—Rat and chick response to egg yolk Group
1 2 3 4 1
TABLE 8.—Influence of maternal diet on egg yolk response
None, R-210 As 1 + 8 % Dried Egg Yolk As 1 + E t h e r Sol. Fract. = 8 % D . E . Y . Commercial Broiler Crumbles
Rats (4-8 wks.)
Chicks (0-4 wks.)
129(8)1 136 (8) 127(8) 147 (8)
339(16) 425 (15) 380(14) 376 (16)
Figures in parentheses represent number of survivors.
TABLE 7.—Influence of maternal diet on egg yolk response Av. 4-week gain 1
Treatment Chick'
(gm.)
None—control
None, R-200 8% Dried Egg Yolk
328 366
5 % Dried Egg Yolk
None, R-200 8% Dried Egg Yolk
342 384
Iodinated Casein (.1 gm./lb.)
None, R-200 8 % Dried Egg Yolk
343 375
As 2 + 3
None, R-200 8 % Dried Egg Yolk
327 380
1 Two-year data with 5 and 4 experiments with sexed males and2 mixed sexes, resp., per year. New Hampshires.
Chick'
Av. 4-week gain Hen2
(gm.)
None, R-210
344 (3)3
4% Dried Egg Yolk 8% Dried Egg Yolk 12% Dried Egg Yolk
381(3)1 432 (3) 427 469 (3) J
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(C-SM)« (C-SM)* (C-SM)*
382 (3)1 405 (3) U05 429 (3)J
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(C-SM-3%FM) (C-SM-3%FM) (C-SM-3%FM)
376(3)1 408(3) 411 450(3))
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(S-SM) (S-SM) (S-SM)
376(3)1 394 (3) 1395 416 (3) j
4% Fluid Egg Yolk 8% Fluid Egg Yolk 12% Fluid Egg Yolk
(S-SM-3%FM) (S-SM-3%FM) (S-SM-3%FM)
394 (3) 1 405(2)5 1412 438 (3) J
1 2 3
New Hampshire XDelaware. New Hampshire. Figures in parentheses represent no. of experiments with 13 and 11 chicks per experiment. *C=corn; SM=soybean oil meal; FM=fish meal; S = sucrose. 6 Average of 2 groups only, water consumption restricted in second experiment.
dams housed on wire floors. It is evident in Table 7 that no "carry-over" of the unknown nutrient occurs in the presence of 5 percent dried egg yolk in the dam's diet (group 2) as indicated by gains of progeny to four weeks of age when fed R-200 with and without dried egg yolk. The increased response noted when egg yolk was fed in the presence of iodinated casein (group 4) was evident only in the first year's results and is therefore discounted. Likewise when fluid yolks from dams fed either a corn-soybean oil meal or sucrose-soybean oil meal with or without 3 percent fish meal were fed to chicks, Table 8, no differences in growth could be related to the diet fed the dams. These data concur with those of Keene et al. (1957) who have reported that egg yolks from hens raised on wire and fed a cornwheat-soybean oil meal type ration were as effective in improving chick growth as were yolks from hens reared on range and fed a complete ration. Lillie et al. (1957) in contrast have reported a significant
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compared favorably to that obtained previously with dried egg yolk or fat soluble extracts of dried egg yolk as reported by Arscott et al. (1957). The greater gain obtained with the egg yolk supplement when compared with the commercial broiler feed is of interest but cannot be accounted for at this time. Bentley (1954) has reported rat growth to be unaffected on corn-soybean oil meal or semi-synthetic type rations when egg yolk supplements were employed provided such rations contained no added iodinated casein. In the presence of 0.08 percent iodinated casein, however, eggs from hens fed antibiotics appeared to increase growth. The relation of egg yolk reponse to "carryover": The experiments in Tables 7 and 8 were designed to show the relation of "carry-over" of the unidentified nutrients) from the dam to the chick by using either chicks or eggs from treated
Hen!
Treatment
Av. 4-week gain (gm-)
Treatment
EGG YOLK AS A SOURCE or UNIDENTIFIED FACTORS
"carry-over" effect on progeny performance from the inclusion of 5 percent dried egg yolk in the maternal diet. SUMMARY
The authors wish to thank the following concerns for material kindly supplied during these investigations: Corn Products
Sales Co., New York, N. Y., for Argo Brand zein; Hoffmann-LaRoche, Inc., Nutley, N. J., for biotin; Lederle Laboratories Division, American Cyanamid Co., Pearl River, N. Y., for folacin and orotic acid; Merck & Co., Inc., Rahway, N. J., for other B-complex vitamins and 2-methyl-l, 4-napthoquinone; and Nopco Chemical Co., Richmond, Calf., for vitamin A (Nopcay 10). REFERENCES Arscott, G. H., 1956. Complexity of a chick growth response to egg yolk, animal fat and fish solubles additions to the diet. Poultry Sci. 35: 338-342. Arscott, G. H., P. H. Weswig and J. R. Schubert, 1957. Multiple nature of chick growth responses to fractions of dried egg yolk. Poultry Sci. 36: 513-516. Barnett, B. D., M. Lapidus, H. R. Bird and F. M. Strong, 1956. Adenosine as growth factor for chicks fed purified diet. Proc. Soc. Biol. Med. 92: 372-374. Bentley, 0. G., 1954. Rat growth response of egg yolks from hens fed antibiotics. Fed. Proc. 13 Pt. 1:181-182. Carver, D. S., and E. L. Johnson, 1953. Unidentified growth factors for the chick in vegetable oils and fatty acid concentrates. Poultry Sci. 32: 701-705. Combs, G. F., G. H. Arscott and H. L. Jones, 1954. Unidentified growth factors required by chicks and poults. 3. Chick studies involving practicaltype rations. Poultry Sci. 33: 71-79. Denton, C. A., R. J. Lillie and J. R. Sizemore, 1954. Effect of egg yolk, fat and fish solubles on growth of chicks. Fed. Proc. 13 Pt. 1: 455. Fritz, J. C , F. D. Wharton and R. M. Henley, 1954. Effect of orotic acid on growth of chicks and poults. Fed. Proc. 13 Pt. 1: 458. Hopper, J. H., H. M. Scott and B. C. Johnson, 1956. The growth promoting ability of fluid egg yolk. Poultry Sci. 35: 195-197. Huff, J. W., D. K. Basshardt, L. D. Wright, D. S. Spicer, K. A. Valetnik and H. R. Skeggs, 1950. A growth promoting substance for L. bulgaricus 09 in whey: Isolation and identification as orotic acid. Proc. Soc. Exp. Biol. Med. 75: 297-299. Keene, O. D., G. F. Combs and G. L. Romoser, 1957. Studies on carry-over and depletion of unidentified chick growth factors. Poultry Sci. 36: 11321133. Lillie, R. J., H. Menge and C. A. Denton, 1957. A breed difference with respect to carry-over effect
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Studies have been conducted involving (1) egg yolk components and reported growth factor supplements, (2) types and levels of yolk and (3) "carry-over" response from dam to progeny using a sucrose-soybean oil meal diet. Linoleic, palmitic and stearic acids, sodium oleate, cholesterol or an ashed yolk (550°C.) fed to chicks at levels equivalent to 8 percent dried yolk appeared to contain no growth stimulating effect. Soybean lecithin, orotic acid or adenosine were also without effect. Energy, supplied by corn oil, did not markedly reduce the response to egg yolk. Increased chick growth occurred when 4, 8 and 12 percent dried yolk was included in the diet. The 8 and 12 percent levels of dried egg yolk evoked significantly greater responses (P = 0.01) than the 4 percent level. In a summary involving thirteen experiments the 12 percent level of dried egg yolk proved significantly greater (P = 0.05) than the 8 percent level. Four, 8 and 12 percent fluid chicken or turkey yolk also responded in step-wise fashion with 12 percent fluid yolk appearing comparable to 8 percent dried egg yolk. Poults likewise responded to dried yolk or fluid chicken and turkey yolk with the 4 percent level of either appearing optimum. More variability and smaller differences were experienced with poults. Rats failed to show a marked response to dried yolk. No "carry-over" effect of the egg yolk factor was observed from dam to chick. ACKNOWLEDGMENTS
61
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
H. B. FLUCKIGER 2 AND J. O. ANDERSON Poultry Department, Utah State University, Logan, Utah (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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I. EFFECT OF THYROXINE AND KIND OF PROTEIN ON THE ARGININE, METHIONINE AND GLYCINE REQUIREMENTS 1