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Effect of Ration on Yolk Color*
Effect of Ration on Yolk Color* E. W. HENDERSON AND H. L. WILCKE Iowa State College, Ames (Presented at Annual Meeting, August 17-20, 1932)
I
* Journal Paper No. B66 of the Iowa Agricultural Experiment Station.
t Project financed in part by funds from Swift and Company, Chicago, Illinois.
[266]
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NTEREST in the factors which influ-i- color from the exterior of the hen in terms ence the color of the yolk of hens'i' of the number of eggs produced rather than eggs is being revived. There are at least >t in terms of the length of time the hen had two reasons for renewed interest in this is been in egg production. A poultry extenproblem. One is, the illogical discriminationn sion specialist was recently credited with in certain markets against rich yellow yolks,;, the statement that a hen which had comand the other is, the close association be-;- pletely exhausted the yellow color from her tween vitamin A and yellow pigment-pro-i- skin and shanks would of necessity produce ducing feeds. Evidence has been accumulat-:- a pale or colorless egg yolk. If the foregoing ing to show that carotin is the precursorr opinions are correct it might be inferred of vitamin A. This work recently has beenn that the hen possesses the ability to store reviewed by Mattikow (1932). Karrer, vona yellow pigment and possibly vitamin A in Euler, and Rydbom (1930), Russell andd her body fat for later deposition in the egg Weber (1931), and Kline, Schultze, andd yolks produced. There is little experimental Hart (1932) have shown that xanthophyll] evidence to support the above inference, from plant tissue does not have vitamin A^ and the work of Palmer and Kempster producing properties. Palmer and Kempsterr (1919) definitely suggests that as far as (1919) showed that the yellow color of eggg yellow pigment is concerned, once it is deyolk is produced almost exclusively by/ posited in the body it does not find its xanthophyll instead of carotin, but egg yolks way into the egg yolk, was one of the first substances in whichi vitamin A was found by McCollum and * EXPERIMENTAL! Davis (1913). The object of this work was to deterThe Maine Agricultural Experiment Stamine whether or not factors other than the tion (1914) and Blakeslee and Warnerr feed of the hen influence the color of the (1915), according to Palmer and Kempsterr egg yolks she produces. We were especially (1919), advanced the hypothesis "thatt interested in the possible influence that the the growth of the egg abstracts the pig, abdominal fat might have on the color of ments from the body tissues." Probably as the yolk, since it was shown by Palmer a result of this explanation some poultrymen seem to be of the opinion that the> and Kempster (1919) that this region rehen has the power to withdraw yellow pig- tained its yellow color after the skin had ment from the body fat and the skin and[ faded and that a fat stain (Sudan III) deposit it in the yolk of the egg. Certaint would be deposited in the abdominal fat production judges are in the habit of ex- independently of the skin. It seemed logical that, if the body fat pressing the degree of fading of yellowj
7. 8. 9. PLATE I. Yolks of hen 3937; Exp. II (see p. 268).
1.
8.
2..
3.
lo. 11. PLATE II. Yolks of hen 3868; Exp. I l l (see p. 271).
A.
It.
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267
contributed to the color of the egg yolk independently of the feed, a fat stain such as Sudan III would follow the same path. A procedure for staining the body fat by injection without feeding the dye was suggested, but Professor H. L. Kempster pointed out that, if the dye were fed to hens that were not in egg production, the adipose tissue could be more uniformly stained. The latter procedure was followed.
the number of eggs produced is fully as important as the factor of time. Hen 3911 laid six eggs which were unmistakably colored, and five which were recorded as doubtful. It is entirely possible that some of these five may have contained no Sudan III. It is equally possible that these yolks may have been developing in the hen's ovary at the time of the dye feeding, but because of their small size or slow rate of growth the amount of dye deposited in the yolks may have been so small as to be almost invisible to the eye. The eggs laid after the sixteenth day were free from dye. This seemed to be evidence that either the dye had disappeared from the fat or that it was not exerting any influence on the color of the egg yolks. On March 19 all the hens were killed and an examination of the abdominal fat revealed positive evidence of the presence of Sudan III dye. There was evidence of the dye on the surface of the shanks and feet of the birds, but this was easily accounted for by the fact that a considerable quantity of the feed was knocked out of the containers onto the floor of the cages so that the feet were probably colored by direct contact. The record of the appearance of the egg yolks produced by the hens in Experiment I is shown in Table 1. Presence of the pink dye is indicated by plus signs, absence by the minus sign. Doubtful eggs are indicated by the plus and minus sign. Autopsy record, March 19, 1932.—Hen 3893 was killed and her fat in the abdominal and mesenteric regions was distinctly stained a pink color as compared to normal fat. The external fat in the dorsal region was not stained. One fully developed egg was taken from the oviduct and several yolks were taken from the ovary. None of these egg yolks showed evidence of Sudan III dye. Hen 3911 was au-
1933
. VOL.
XII,
No.
Experiment I For a preliminary trial five White Plymouth Rock hens which were not laying were selected on February 12, 1932, and isolated in exhibition cages. They were fed a standard laying mash which contained enough Sudan III so that each hen consumed approximately 35 milligrams of dye per day. The dye feeding was continued for 14 days. One hen (3938) was killed on February 23. There was no positive evidence of the dye in the body fat. She was affected by leukemia and evidently had not eaten the dyed feed. A second hen (3909) was autopsied and her abdominal and mesenteric fat were colored distinctly pink as compared to the normal fat. No trace of pink color was evident in the fat of the dorsal region. The first egg was laid by this group of hens on February 17 (S days after feeding began). The yolk was distinctly colored by the dye. The yolks of tae eggs laid by the three remaining hens continued to show evidence of the Sudan III dye until March 13, 16 days after the dye feeding stopped. This is not in complete agreement with the results of Riddle (1911) and of Gage and Fish (1924), who found that the last dyed egg was laid 5 to 8 days after Sudan III was fed. However, no hen, with the exception of 3911, laid more than eight colored eggs after the cessation of dye feeding. It would appear from these results that
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4
JULY,
268
POULTRY TABLE 1. Evidence of dye in yolks Experiment I
Hen No.
3911
3948
Date Feb. 17
3893
++ ++
18 19 20 21
+++
23 24
++ +
25 26* 27
++ + +++
.+ + +
28 29 Mar. 1
+++
++ + ++ +
+++
2 3 4 5
++ + +
++ +
6 7
+
8
±
9
+
10
±
, + + •
11
+
12 13
+
14
-
15 16
-
19
-
* Dye feeding stopped.
-
-
topsied with the same results yielded by 3893. An autopsy study of hen 3948 likewise yielded the same results as hen 3893. Experiment II Since the body fat of the hens in Experiment I did not show as much dye as seemed possible, the dosage for the hens used in a second experiment was increased to 100 milligrams per hen per day. Twelve White Plymouth Rock hens were selected for this experiment and the dye feeding began on March 24, 1932, and continued to April 4, 1932. The eggs produced by the different hens were studied in three different ways. (1) Cross sections of hard boiled yolks were drawn and tinted by an artist with the aid of color charts. (2) Photographs were made of cross sections of hard boiled yolks. (3) The yolks were analyzed by a Razek Mulder color analyzer. It was not possible to keep all the hens in continuous production, probably because of the change in rations and quarters. Hen 3937 laid more consistently than the others. Her yolks are illustrated in Plate I. The dates the eggs were laid by hen 3937 are as follows: Date: 3/31 4/3 4/5 4/6 4/7 4/10 4/24 4/25 4/26 Egg No. 1 2 3 4 5 6 7 8 9 The eggs laid by hen 3868 in Experiment II were not illustrated, but a record of the changing amount of dye with the age of the eggs as determined by ocular observation is shown in Table 2. While samples of eggs for each day of the period of dye feeding were not available the samples are fairly well distributed over the period. Hens 3874 and 3895 laid eggs without traces of dye on April 26 and April 22 respectively. Traces of dye were still visible
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22
++ ++ +++ ++ +++
SCIENCE
JULY,
1933.
VOL.
XII,
No.
4
269
TABLE 2. Evidence of dye in yolks Experiment II Hen No.
3895
Date Mar. 27 28
3964
3874
3937
2932 3860 3891 3913f
3868
+
-
+
3929* 3901
+
+
29
3883 Floor
+ + ++
30
Apr.
++ ++ +++ +++ ++++
1
+
.3
+++
4t
+ + +++ + +++++ + ++++
5 6 7
+ +++ +++
8 9
+
10
++++
11
++
12 13
+
14
+++
15 16
++
+
++ +
±
17 18
+
19 20
++ ~
-
21 22
+
+
25
-
++
+
-
-
+
-
+
-
+
-
-
23 24
+
-
-
* 3929 Raw yolks used on color analyzer—no results available, f 3913 Killed for autopsy Apr. 6th—3 ova showed dye. t Dye feeding stopped.
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+
31
270
POULTRY
SCIENCE
TABLE 1 . {Continued)
Hen No. 3895
3964
3937
3874
-
Apr. 26
-
+
27
-
-
3929* 3901
2932 3860 3891 3913f
-
-
30
-
-
-
-
-
-
-
3 4
-
5 * 3929 Raw yolks used on color analyzer—no results available. f3913 Killed for autopsy Apr. 6th—3 ova showed dye.
in the yolks of the eggs laid by hen 3937 up to April 26 although the traces were very slight from April 24 to April 26 inclusive. Hen 3883 laid one eggs free from dye on April 24 but the egg she produced on April 25 showed slight traces of the dye. It is possible that there was an error made in dating these eggs but it is also possible that the yolk of the egg laid on April 25 could have been older than the one laid on April 24. In all cases the dye appeared first on the outer margin of the yolk. In the case of the egg laid by 3883 the dye appeared on the outer margin of the yolk on March 27, 3 days after the dye feeding started, but the eggs laid by 3895 on March 25 and 26 showed no trace of dye. Dye was evident in the egg of 3895 on March 28. The most highly colored yolk was laid by hen 3886 on April 7, 14 days after the dye feeding had started and 3 days after it had stopped. The dye began to recede on April 7 as shown by the yolks laid by number 3937 on that date. In two cases it had entirely disappeared by April 24, one month after the dye feeding began and 20 days after it
stopped. At this time the rations of the hens were reduced to % normal with the idea of noting whether the fat reserve might be utilized to a greater extent and possibly increase the dye in the egg yolks. The hens promptly stopped laying, which was not unexpected. Only one hen was killed to determine the extent of the dye in the abdominal fat for it had been shown to persist in this region after it ceased appearing in the yolks in Experiment I. Autopsy record.—Hen 3913 was killed for autopsy on April 6. All body and mesenteric fat was colored a distinct pink. The bird was in fair flesh. The back fat, and side fat was colored deeply enough to be seen through the skin. A small egg (smaller than a pheasant egg) was taken from the oviduct. Three large ova in the ovary were distinctly colored. Experiment III The same group of hens was continued on a ration designed to improve their condition. They were again fed the Sudan III dye from May 15 to May 22. Three days after the dye feeding began it appeared
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2
-
-
29
1
3883 Floor
-
-
28
May
3868
JULY,
1933.
VOL.
XII,
No.
ell
4
Time Dye Remains in Body Fat While it was necessary to demonstrate that the dye fed in this experiment was present in the body fat after it ceased to be deposited in the eggs, the length of time it might remain after feeding was of minor interest. There were only a few hens for use in determining this point; therefore, the results can be considered only an indication of the possibilities. Hens 3893, 3948, and 3911 were killed 22 days after dye
feeding ceased and no visible dye color was seen in the body fat. Some time between 22 and 26 days after dye feedings stopped, fat stained by our method loses its dye color. CONCLUSIONS
The pigment in the ration of the hen is primarily responsible for the color of the egg yolk. RAZEK -MULDER COLOR REFLECTION O t a
BURGESS-PARR
COMPANY
ANALYZER
FACTOR 01 Vl
C HICAGO
B
ILLINOI
FIG. 1. Color reflection curves (hen 3937).
The hen does not withdraw Sudan III pigment from her body fat for deposit in the egg yolks. The time required for the feed to exert the initial visible influence on the color of the egg yolk is between 3 and S days, depending upon how soon eggs are laid after feeding begins. The influence of dyes in the feed may remain evident for as long as 20 days after the dye feeding stops, depend-
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in the yolk of an egg laid by hen 3868. Fourteen days after the dye feeding stopped it had been reduced to a mere trace in the center of the yolk which was laid on June 5 by 3868. The eggs laid by this hen from May 18 to June 5 are typical of the lot. Cross sections of the yolks are illustrated in Plate II. This hen stopped laying on June 5, but the eggs laid by hen 3937 beginning June 6 continued to show dye until June 14, after which all eggs laid to June 20 were free from dye. Hen 3937 was killed on June 20 and the color of her abdominal fat was compared to that of a normal hen by use of a color analyzer. The fat from hen 3937 was distinctly colored a deep pink. The color reflection curves of the two fat samples are shown in Figure 1. There is no question but that the dye remained visible in the body fat of hen 3937 after she ceased to produce dye in her egg yolk. We suggest that the reason hen 3937 continued to produce dye in her eggs for a longer period of time than any of the others was because she had a large number of partially developed yolks which had accumulated during the period of dye feeding. She failed to come into production until after the dye feeding stopped, but the accumulated yolks were produced with the dye acquired during the dye feeding. As soon as the accumulated yolks were exhausted, she produced eggs without dye.
272
POUL TRY
SCIENCE
TABLE 3. Evidence of dye in yolks Experiment I I I Hen No.
3868
3929
3937
3874
Date May 16
3922
3901
3869
Floor
-
17 18 19 20
+ + ++
21 22* 23
+++ +++ +
24
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++ +++ ++++ ++ +++
25 26
+++++
27 28 29
++ + + ++
30 31 June
1 2
+' + +
+++ ++
+
3
+
4 5
+
6
++
7 8 9
+ +
-
10 11
+
12 13 14 * Dye feeding stopped.
+
—
JULY,
1933.
VOL. XII,
No.
273
4
TABLE 3. (Continued)
Hen No.
3868
3929
3937
3874
-
-
3922
3901
3869
Floor
June 15
16 17 18
-
REFERENCES
Blakeslee, A. F., and D. E. Warner, 1915. Correlation between egg-laying activity and yellow pigment in the domestic fowl. Science, 41:432. Gage, S. H., and P. A. Fish, 1924. Fat digestion, absorption, and assimilation in man and animals as determined by the dark-field microscope, and a fat-soluble dye. Amer. Jour, of Anat. 34:1-86. Karrer, P., H. v. Euler, and M. Rydbom, 1930. Neue Versuche uber die physiologische Wirkung des Xantophylls. Helvetica Chimica Acta, 13 :• 1059-1062. Kline, O. L., M. O. Schultze, and E. G. Hart, 1932. Carotene and xanthophyll as sources of vitamin A for the growing chick. Jour. Biol. Chem. 97:83-91.
Maine Agr. Exp. Sta., Press Letter 144, 1914. Cited by Palmer and Kempster (1919). Mattikow, M., 1932. A critical review of the literature on the coloring matter in egg yolk. Poultry Sci., 11:83-93. McCollum, E. V. and M. Davis, 1913. The necessity of certain lipins in the diet during growth. Jour. Biol. Chem., 15 :167-175. Palmer, L. S., and H. L. Kempster, 1919. Relation of plant carotinoids to growth, fecundity, and reproduction of fowls. Jour. Biol. Chem., 39:299-312. , 1919. The physiological relation between fecundity and the natural yellow pigmentation of certain breeds of fowls. Jour. Biol. Chem., 39:313-330. , 1919. The influence of specific feeds and certain pigments on the color of the egg yolk and body fat of fowls. Jour. Biol. Chem., 39:331-337. Riddle, O. A., 1911. On the formation, significance, and chemistry of the white and yellow yolk of eggs. Jour. Morph., 22:455-490. Russell, W. C. and A. L. Weber, 1931. Plant pigments in the nutrition of the chicken. Proc. Soc. Exp. Biol, and Med., 29:297-298.
Downloaded from http://ps.oxfordjournals.org/ at New York University on May 22, 2015
ing upon the rate and time of production. The maximum influence of dye feeding is obtained in approximately 14 days but this depends upon the rate of production. The length of time required for the development and production of an egg varies with the individual hens.
I
* Journal Paper No. B66 of the Iowa Agricultural Experiment Station.
t Project financed in part by funds from Swift and Company, Chicago, Illinois.
[266]
Downloaded from http://ps.oxfordjournals.org/ at New York University on May 22, 2015
NTEREST in the factors which influ-i- color from the exterior of the hen in terms ence the color of the yolk of hens'i' of the number of eggs produced rather than eggs is being revived. There are at least >t in terms of the length of time the hen had two reasons for renewed interest in this is been in egg production. A poultry extenproblem. One is, the illogical discriminationn sion specialist was recently credited with in certain markets against rich yellow yolks,;, the statement that a hen which had comand the other is, the close association be-;- pletely exhausted the yellow color from her tween vitamin A and yellow pigment-pro-i- skin and shanks would of necessity produce ducing feeds. Evidence has been accumulat-:- a pale or colorless egg yolk. If the foregoing ing to show that carotin is the precursorr opinions are correct it might be inferred of vitamin A. This work recently has beenn that the hen possesses the ability to store reviewed by Mattikow (1932). Karrer, vona yellow pigment and possibly vitamin A in Euler, and Rydbom (1930), Russell andd her body fat for later deposition in the egg Weber (1931), and Kline, Schultze, andd yolks produced. There is little experimental Hart (1932) have shown that xanthophyll] evidence to support the above inference, from plant tissue does not have vitamin A^ and the work of Palmer and Kempster producing properties. Palmer and Kempsterr (1919) definitely suggests that as far as (1919) showed that the yellow color of eggg yellow pigment is concerned, once it is deyolk is produced almost exclusively by/ posited in the body it does not find its xanthophyll instead of carotin, but egg yolks way into the egg yolk, was one of the first substances in whichi vitamin A was found by McCollum and * EXPERIMENTAL! Davis (1913). The object of this work was to deterThe Maine Agricultural Experiment Stamine whether or not factors other than the tion (1914) and Blakeslee and Warnerr feed of the hen influence the color of the (1915), according to Palmer and Kempsterr egg yolks she produces. We were especially (1919), advanced the hypothesis "thatt interested in the possible influence that the the growth of the egg abstracts the pig, abdominal fat might have on the color of ments from the body tissues." Probably as the yolk, since it was shown by Palmer a result of this explanation some poultrymen seem to be of the opinion that the> and Kempster (1919) that this region rehen has the power to withdraw yellow pig- tained its yellow color after the skin had ment from the body fat and the skin and[ faded and that a fat stain (Sudan III) deposit it in the yolk of the egg. Certaint would be deposited in the abdominal fat production judges are in the habit of ex- independently of the skin. It seemed logical that, if the body fat pressing the degree of fading of yellowj
7. 8. 9. PLATE I. Yolks of hen 3937; Exp. II (see p. 268).
1.
8.
2..
3.
lo. 11. PLATE II. Yolks of hen 3868; Exp. I l l (see p. 271).
A.
It.
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oo oooo ooo oooo
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267
contributed to the color of the egg yolk independently of the feed, a fat stain such as Sudan III would follow the same path. A procedure for staining the body fat by injection without feeding the dye was suggested, but Professor H. L. Kempster pointed out that, if the dye were fed to hens that were not in egg production, the adipose tissue could be more uniformly stained. The latter procedure was followed.
the number of eggs produced is fully as important as the factor of time. Hen 3911 laid six eggs which were unmistakably colored, and five which were recorded as doubtful. It is entirely possible that some of these five may have contained no Sudan III. It is equally possible that these yolks may have been developing in the hen's ovary at the time of the dye feeding, but because of their small size or slow rate of growth the amount of dye deposited in the yolks may have been so small as to be almost invisible to the eye. The eggs laid after the sixteenth day were free from dye. This seemed to be evidence that either the dye had disappeared from the fat or that it was not exerting any influence on the color of the egg yolks. On March 19 all the hens were killed and an examination of the abdominal fat revealed positive evidence of the presence of Sudan III dye. There was evidence of the dye on the surface of the shanks and feet of the birds, but this was easily accounted for by the fact that a considerable quantity of the feed was knocked out of the containers onto the floor of the cages so that the feet were probably colored by direct contact. The record of the appearance of the egg yolks produced by the hens in Experiment I is shown in Table 1. Presence of the pink dye is indicated by plus signs, absence by the minus sign. Doubtful eggs are indicated by the plus and minus sign. Autopsy record, March 19, 1932.—Hen 3893 was killed and her fat in the abdominal and mesenteric regions was distinctly stained a pink color as compared to normal fat. The external fat in the dorsal region was not stained. One fully developed egg was taken from the oviduct and several yolks were taken from the ovary. None of these egg yolks showed evidence of Sudan III dye. Hen 3911 was au-
1933
. VOL.
XII,
No.
Experiment I For a preliminary trial five White Plymouth Rock hens which were not laying were selected on February 12, 1932, and isolated in exhibition cages. They were fed a standard laying mash which contained enough Sudan III so that each hen consumed approximately 35 milligrams of dye per day. The dye feeding was continued for 14 days. One hen (3938) was killed on February 23. There was no positive evidence of the dye in the body fat. She was affected by leukemia and evidently had not eaten the dyed feed. A second hen (3909) was autopsied and her abdominal and mesenteric fat were colored distinctly pink as compared to the normal fat. No trace of pink color was evident in the fat of the dorsal region. The first egg was laid by this group of hens on February 17 (S days after feeding began). The yolk was distinctly colored by the dye. The yolks of tae eggs laid by the three remaining hens continued to show evidence of the Sudan III dye until March 13, 16 days after the dye feeding stopped. This is not in complete agreement with the results of Riddle (1911) and of Gage and Fish (1924), who found that the last dyed egg was laid 5 to 8 days after Sudan III was fed. However, no hen, with the exception of 3911, laid more than eight colored eggs after the cessation of dye feeding. It would appear from these results that
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4
JULY,
268
POULTRY TABLE 1. Evidence of dye in yolks Experiment I
Hen No.
3911
3948
Date Feb. 17
3893
++ ++
18 19 20 21
+++
23 24
++ +
25 26* 27
++ + +++
.+ + +
28 29 Mar. 1
+++
++ + ++ +
+++
2 3 4 5
++ + +
++ +
6 7
+
8
±
9
+
10
±
, + + •
11
+
12 13
+
14
-
15 16
-
19
-
* Dye feeding stopped.
-
-
topsied with the same results yielded by 3893. An autopsy study of hen 3948 likewise yielded the same results as hen 3893. Experiment II Since the body fat of the hens in Experiment I did not show as much dye as seemed possible, the dosage for the hens used in a second experiment was increased to 100 milligrams per hen per day. Twelve White Plymouth Rock hens were selected for this experiment and the dye feeding began on March 24, 1932, and continued to April 4, 1932. The eggs produced by the different hens were studied in three different ways. (1) Cross sections of hard boiled yolks were drawn and tinted by an artist with the aid of color charts. (2) Photographs were made of cross sections of hard boiled yolks. (3) The yolks were analyzed by a Razek Mulder color analyzer. It was not possible to keep all the hens in continuous production, probably because of the change in rations and quarters. Hen 3937 laid more consistently than the others. Her yolks are illustrated in Plate I. The dates the eggs were laid by hen 3937 are as follows: Date: 3/31 4/3 4/5 4/6 4/7 4/10 4/24 4/25 4/26 Egg No. 1 2 3 4 5 6 7 8 9 The eggs laid by hen 3868 in Experiment II were not illustrated, but a record of the changing amount of dye with the age of the eggs as determined by ocular observation is shown in Table 2. While samples of eggs for each day of the period of dye feeding were not available the samples are fairly well distributed over the period. Hens 3874 and 3895 laid eggs without traces of dye on April 26 and April 22 respectively. Traces of dye were still visible
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22
++ ++ +++ ++ +++
SCIENCE
JULY,
1933.
VOL.
XII,
No.
4
269
TABLE 2. Evidence of dye in yolks Experiment II Hen No.
3895
Date Mar. 27 28
3964
3874
3937
2932 3860 3891 3913f
3868
+
-
+
3929* 3901
+
+
29
3883 Floor
+ + ++
30
Apr.
++ ++ +++ +++ ++++
1
+
.3
+++
4t
+ + +++ + +++++ + ++++
5 6 7
+ +++ +++
8 9
+
10
++++
11
++
12 13
+
14
+++
15 16
++
+
++ +
±
17 18
+
19 20
++ ~
-
21 22
+
+
25
-
++
+
-
-
+
-
+
-
+
-
-
23 24
+
-
-
* 3929 Raw yolks used on color analyzer—no results available, f 3913 Killed for autopsy Apr. 6th—3 ova showed dye. t Dye feeding stopped.
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+
31
270
POULTRY
SCIENCE
TABLE 1 . {Continued)
Hen No. 3895
3964
3937
3874
-
Apr. 26
-
+
27
-
-
3929* 3901
2932 3860 3891 3913f
-
-
30
-
-
-
-
-
-
-
3 4
-
5 * 3929 Raw yolks used on color analyzer—no results available. f3913 Killed for autopsy Apr. 6th—3 ova showed dye.
in the yolks of the eggs laid by hen 3937 up to April 26 although the traces were very slight from April 24 to April 26 inclusive. Hen 3883 laid one eggs free from dye on April 24 but the egg she produced on April 25 showed slight traces of the dye. It is possible that there was an error made in dating these eggs but it is also possible that the yolk of the egg laid on April 25 could have been older than the one laid on April 24. In all cases the dye appeared first on the outer margin of the yolk. In the case of the egg laid by 3883 the dye appeared on the outer margin of the yolk on March 27, 3 days after the dye feeding started, but the eggs laid by 3895 on March 25 and 26 showed no trace of dye. Dye was evident in the egg of 3895 on March 28. The most highly colored yolk was laid by hen 3886 on April 7, 14 days after the dye feeding had started and 3 days after it had stopped. The dye began to recede on April 7 as shown by the yolks laid by number 3937 on that date. In two cases it had entirely disappeared by April 24, one month after the dye feeding began and 20 days after it
stopped. At this time the rations of the hens were reduced to % normal with the idea of noting whether the fat reserve might be utilized to a greater extent and possibly increase the dye in the egg yolks. The hens promptly stopped laying, which was not unexpected. Only one hen was killed to determine the extent of the dye in the abdominal fat for it had been shown to persist in this region after it ceased appearing in the yolks in Experiment I. Autopsy record.—Hen 3913 was killed for autopsy on April 6. All body and mesenteric fat was colored a distinct pink. The bird was in fair flesh. The back fat, and side fat was colored deeply enough to be seen through the skin. A small egg (smaller than a pheasant egg) was taken from the oviduct. Three large ova in the ovary were distinctly colored. Experiment III The same group of hens was continued on a ration designed to improve their condition. They were again fed the Sudan III dye from May 15 to May 22. Three days after the dye feeding began it appeared
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2
-
-
29
1
3883 Floor
-
-
28
May
3868
JULY,
1933.
VOL.
XII,
No.
ell
4
Time Dye Remains in Body Fat While it was necessary to demonstrate that the dye fed in this experiment was present in the body fat after it ceased to be deposited in the eggs, the length of time it might remain after feeding was of minor interest. There were only a few hens for use in determining this point; therefore, the results can be considered only an indication of the possibilities. Hens 3893, 3948, and 3911 were killed 22 days after dye
feeding ceased and no visible dye color was seen in the body fat. Some time between 22 and 26 days after dye feedings stopped, fat stained by our method loses its dye color. CONCLUSIONS
The pigment in the ration of the hen is primarily responsible for the color of the egg yolk. RAZEK -MULDER COLOR REFLECTION O t a
BURGESS-PARR
COMPANY
ANALYZER
FACTOR 01 Vl
C HICAGO
B
ILLINOI
FIG. 1. Color reflection curves (hen 3937).
The hen does not withdraw Sudan III pigment from her body fat for deposit in the egg yolks. The time required for the feed to exert the initial visible influence on the color of the egg yolk is between 3 and S days, depending upon how soon eggs are laid after feeding begins. The influence of dyes in the feed may remain evident for as long as 20 days after the dye feeding stops, depend-
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in the yolk of an egg laid by hen 3868. Fourteen days after the dye feeding stopped it had been reduced to a mere trace in the center of the yolk which was laid on June 5 by 3868. The eggs laid by this hen from May 18 to June 5 are typical of the lot. Cross sections of the yolks are illustrated in Plate II. This hen stopped laying on June 5, but the eggs laid by hen 3937 beginning June 6 continued to show dye until June 14, after which all eggs laid to June 20 were free from dye. Hen 3937 was killed on June 20 and the color of her abdominal fat was compared to that of a normal hen by use of a color analyzer. The fat from hen 3937 was distinctly colored a deep pink. The color reflection curves of the two fat samples are shown in Figure 1. There is no question but that the dye remained visible in the body fat of hen 3937 after she ceased to produce dye in her egg yolk. We suggest that the reason hen 3937 continued to produce dye in her eggs for a longer period of time than any of the others was because she had a large number of partially developed yolks which had accumulated during the period of dye feeding. She failed to come into production until after the dye feeding stopped, but the accumulated yolks were produced with the dye acquired during the dye feeding. As soon as the accumulated yolks were exhausted, she produced eggs without dye.
272
POUL TRY
SCIENCE
TABLE 3. Evidence of dye in yolks Experiment I I I Hen No.
3868
3929
3937
3874
Date May 16
3922
3901
3869
Floor
-
17 18 19 20
+ + ++
21 22* 23
+++ +++ +
24
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++ +++ ++++ ++ +++
25 26
+++++
27 28 29
++ + + ++
30 31 June
1 2
+' + +
+++ ++
+
3
+
4 5
+
6
++
7 8 9
+ +
-
10 11
+
12 13 14 * Dye feeding stopped.
+
—
JULY,
1933.
VOL. XII,
No.
273
4
TABLE 3. (Continued)
Hen No.
3868
3929
3937
3874
-
-
3922
3901
3869
Floor
June 15
16 17 18
-
REFERENCES
Blakeslee, A. F., and D. E. Warner, 1915. Correlation between egg-laying activity and yellow pigment in the domestic fowl. Science, 41:432. Gage, S. H., and P. A. Fish, 1924. Fat digestion, absorption, and assimilation in man and animals as determined by the dark-field microscope, and a fat-soluble dye. Amer. Jour, of Anat. 34:1-86. Karrer, P., H. v. Euler, and M. Rydbom, 1930. Neue Versuche uber die physiologische Wirkung des Xantophylls. Helvetica Chimica Acta, 13 :• 1059-1062. Kline, O. L., M. O. Schultze, and E. G. Hart, 1932. Carotene and xanthophyll as sources of vitamin A for the growing chick. Jour. Biol. Chem. 97:83-91.
Maine Agr. Exp. Sta., Press Letter 144, 1914. Cited by Palmer and Kempster (1919). Mattikow, M., 1932. A critical review of the literature on the coloring matter in egg yolk. Poultry Sci., 11:83-93. McCollum, E. V. and M. Davis, 1913. The necessity of certain lipins in the diet during growth. Jour. Biol. Chem., 15 :167-175. Palmer, L. S., and H. L. Kempster, 1919. Relation of plant carotinoids to growth, fecundity, and reproduction of fowls. Jour. Biol. Chem., 39:299-312. , 1919. The physiological relation between fecundity and the natural yellow pigmentation of certain breeds of fowls. Jour. Biol. Chem., 39:313-330. , 1919. The influence of specific feeds and certain pigments on the color of the egg yolk and body fat of fowls. Jour. Biol. Chem., 39:331-337. Riddle, O. A., 1911. On the formation, significance, and chemistry of the white and yellow yolk of eggs. Jour. Morph., 22:455-490. Russell, W. C. and A. L. Weber, 1931. Plant pigments in the nutrition of the chicken. Proc. Soc. Exp. Biol, and Med., 29:297-298.
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ing upon the rate and time of production. The maximum influence of dye feeding is obtained in approximately 14 days but this depends upon the rate of production. The length of time required for the development and production of an egg varies with the individual hens.