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The Transference of Radioactive Calcium and Phosphorus from Hen to Chick
J. CLYDE DRIGGEBS, 1 RAY L. SHIRLEY, 2 GEORGE K. DAVIS 2 AND N. R.
MEHRHOF 1
Florida Agricultural Experiment Station, Gainesville, Florida (Received for publication August 15, 1950)
P
REVIOUS work at this station (Driggers and Comar, 1949) demonstrated that radioactive calcium (Ca45), when administered orally to a fasted hen reached the shell of an egg which was being formed within a very few minutes, and that by the end of 24 hours as much as one-third of the ingested dose was found in the entire contents of another egg which was laid. This work was confirmed by Spinks et al. (1949). O'Neil et al. (1948) showed also that radioactive phosphorus was deposited in the yolk and albumen of an egg soon after the oral administration of the isotope, and the eggs which were laid subsequent to that time showed increasing amounts up to the fifth day, after which time the total amount of P32 decreased. In this investigation, a study was made of (1) the transference of radioactive calcium and phosphorus, when administered separately and concurrently to previously mated hens, to chicks hatched 1
Department of Poultry Husbandry. Department of Animal Husbandry and Nutrition. Published with the permission of the Director of the Florida Agricultural Experiment Station. The Ca46 and P 32 were obtained from the Oak Ridge National Laboratory on allocation of the Isotopes Division of the U. S. Atomic Energy Commission. The authors express their appreciation to Francis L. Coune, Riley D. Owens, Jackson McClenny, John McCall, Marvin H. Hernden and Al Lovelace for technical assistance. 2
from eggs laid subsequent to the administration of the isotopes, and (2) the distribution of the isotopes in the various tissues of the chicks. EXPERIMENTAL Eight one-year-old S. C. White Leghorn hens weighing approximately 1750 grams each and having been previously mated were used in this experiment. They were kept in standard laying cages and fed a complete all-mash diet. The egg cycles of the hens had been estimated and the radioactive isotopes were administered at the expected time of ovulation. Using the technique employed in earlier work (Driggers and Comar, 1949), four of these hens received an oral dose of Ca45 as a solution of the chloride with an activity of approximately 70 microcuries. Two of the hens received 70 microcuries of the Ca45 plus 300 microcuries of carrier free P32 as H3PO4. One hen received 300 microcuries of P32 and the eighth hen received 200 microcuries of the P32 alone. Each egg laid thereafter was incubated and individually hatched to insure proper chick identification. On the day of hatching, the chicks were sacrificed and the following tissue samples were taken for analysis: yolk sac; femur; breast bone; brain; heart; liver; proventriculus; small intestine; breast muscle; and leg muscle. The samples were wet-ashed with concentrated HNO3 and evaporated to near-
199
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T h e Transference of Radioactive Calcium and Phosphorus from Hen to Chick
200
J. C. DRIGGERS, R. L. SHIRLEY, G. K. DAVIS AND N. T. MEHRHOF
nations were made using immersion-type Geiger-Mueller tubes in conjunction with commercial scalers. In the case of samples containing both Ca46 and P32 aliquots from the same solutions were taken for the respective determinations. The dipping-type GM tubes are insensitive to the weak beta emissions from the Ca46. Calcium oxalates prepared from solutions containing P32 and calcium which was free of Ca46 did not show any activity
TABLE 1.—Egg and chick production record of hens administered radioactive calcium and phosphorus
Hen number
Dose of the radioactive element
Days after administration x —egg laid on this day (x)—chick hatched from this egg
» 1
1 2 3 4
microcuries Ca«, 70
2
3
4
5
(x)
(x)
X
X
(x)
X
(x)
(x)
(x)
«
X
(x)
48
Ca , 70 45
Ca , 70 46
Ca , 70
X
X
6
7
X
X
(x)
X
8 X X
9 X
10
11
X
X
X
X
X X
X
12
13
14
IS
X
X
X
X
X
X
X
X
X
X X
46
S
Ca , 70 plus P32, 300
X
6
Ca45, 70 plus P32, 300
«
7
PS2, 300
8
32
P , 200
X
(x)
(x)
(x)
X
manner for radioactivity assay by means of a funnel and ring and disc assembly (Margnetti, 1949) which has been especially designed for this purpose. The Ca46 radioactivity assays were made on a commercial scaler using thin mica window (1.5-2.0 mg./cm.2) Geiger-Mueller tubes. Self-absorption and decay corrections were made in the usual manner (Kamen, 1947). Total calcium in the tissues was calculated from the weight of the calcium oxalate precipitate obtained from the aliquot prepared for radioactivity measurement. The phosphorus radioactivity determi-
(x)
(x)
(x)
(x)
X
(x)
(x) X
(x)
(x)
(x)
(x)
X
X
X
X
X
when the thin mica window GM tubes were used. Using the calcium oxalate method, no precipitation was found when 50 milligrams of phosphorus alone or 20 milligrams of phosphorus plus 20 milligrams of aluminum were used in synthetic solutions (Shirley, Owens and Davis, 1950). Total phosphorus was determined by the volumetric phosphomolybdate method (A.O.A.C, 1945). RESULTS
During this experiment, each of the hens receiving the Ca46 alone produced one or more chicks. One of the hens re-
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
dryness, diluted with distilled water, filtered, and made up to volume in volumetric flasks from which aliquots were taken for radioactivity -and total element determinations. The calcium was then precipitated as calcium oxalate (A.O.A.C., 1945) after 5 milligrams of inactive calcium had been added to each aliquot to insure precipitates weighing atleast 17 milligrams. The precipitate was prepared in a uniform
MATERNAL TRANSFER OF RADIOACTIVE CALCIUM AND PHOSPHORUS
ceiving concurrent doses of Ca46 and P322 did not produce any chicks, while thee other produced four. One hen receivingg the P32 alone produced nine chicks andi the other produced two. The egg produc-
201
the Ca45 dose was found in the tissues. Thereafter, there was a sharp decline until by the fourth day less than 0.05 percent of the dose could be found in any of the chicks. In Figure 2 the data ob-
100..
—
0.9Q. 050. 0.70.. m ^ 0.60| o u. 0.50J o uj 040.. §
1) E3 • •
YOLK SAC FEMUR BREAST BONE OTHER TISSUES BRAIN HEART LIVER PROVENTRICULUS INTESTINES BLOOD BREAST MUSCLE LEG MUSCLE
0.30.
J- 0.2Q. u o o.ioj
or a! 0.00.
I I 22 3 4 5 6 H 8 9 10 II 12 13 DAYS EGGS LAID AFTER ADMINISTRATION OF C A 4 5
14
15
FIG. 1. The percent of the dose of Ca46 administered which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: The chicks hatched from the first egg laid on the first day and the twelfth, fourteenth and fifteenth days after administration were those from the hens which received concurrent doses of Ca45 and P32.
tion and chicks hatched therefrom aree given in Table 1. Figure 1 shows graphically the percentt dose of Ca45 found in the various tissuess of the chicks hatched from eggs laid 1 to0 15 days after oral administration of thee isotope to the hens. The values obtainedi on the chicks from different hens were inti close agreement on the second and fourth•x days after administration and the valuess for the individual tissues in these casess were averaged. In the case of the chickss from the eggs laid on the first and second1 days after administration, approximatelyY 0.745 and 1.016 percent respectively off
tained for percent dose per milligram of total calcium in the various tissues are presented. The percent dose of P32 found in the chick tissues is presented in Figure 3. Chick tissue values obtained on the sixth and ninth days from different hens were averaged. In contrast to the above calcium data, very little of the P32 was found in the chicks hatched from eggs laid during the first two days after administration of the isotope. The P32 values increased rapidly from the second to the sixth day, at which time approximately 0.839 percent of the dose was found in all
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
I.IO
202
J. C. DRIGGERS, R. L. SHIRLEY, G. K. DAVIS AND N. R. MEHRHCTF
ffl
YOLK SAC FEMUR BREASTBONE BRAIN HEART LIVER PROVENTRICULUS INTESTINES BREAST MUSCLE LEG MUSCLE
O.OOJ
JBL I I 2 3 4 5 6 7 8 9 10 II 12 13 14 15 DAYS EGGS LAID AFTER ADMINISTRATION OF C A 4 5
FIG. 2. The percent of the dose of Ca45 administered to the hen per milligram of total calcium which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: Only traces of Ca46 per milligram of total calcium were found in chicks from the fourteenth and fifteenth days' eggs.
tissues analyzed. These values gradually decreased until the end of the fourteenth
D 0 D •
YOLK SAC FEMUR BREAST BONE BRAIN HEART LIVER D PROVENTRICULUS • INTESTINES m BREAST MUSCLE ID LEO MUSCLE
•
I 2 3 4 S 6 7 S 9 K) II 12 12 14 15 DAYS EGGS LAID AFTER ADMINISTRATION OF P32
day when less than 0.12 percent could be found in the newly hatched chicks. The data showing the percent dose of radioactive phosphorus per milligram of total phosphorus are in Figure 4. DISCUSSION A gross examination of the tissues indicated that the chicks hatched during this study were quite normal. The writers believe that infertility was the primary reason for the poor hatch. FIG. 3. The percent of the dose of P32 administered which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: The hen which laid the egg on the first day and the second egg on the twelfth day after administration received concurrent doses of Ca46 and P32. No Ca46 or P32 was found in the chick hatched from the egg laid on the first day after administration.
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H 0 • • S E D m H CD
MATERNAL TRANSFER OE RADIOACTIVE CALCIUM AND PHOSPHORUS
The greatest concentration of P32 was found in the chicks hatched from eggs laid 6 days after administration, allowing time for the entire yolk and white to receive the maximum dose of this element. Of particular interest is the gradual decrease in total P32 content of the entire chick after the peak has been reached on the sixth day as compared to the sharp decline in Ca45 content after the second day. In Figures 2 and 4 where the percent dose of Ca45 and P32 respectively per milligram of total element is reported, it
is evident that the radioactive isotope tends to be distributed uniformly throughout all the tissues with the stable isotope, According to these data, especially in the case of the phosphorus in Figure 4, all the tissues are essentially equivalent in concentration of the isotope per milligram of the total element. Such data as these
D
YOLK SAC 0 FEMUR D BREASTBONE BRAIN HEART 13 LIVER PROVENTRICULUS INTESTINES BREAST MUSCLE LEG MUSCLE
• s
a m n m
i ILii *
H
I 2 3 4 5 6 7 8 9 10 1! 12 12 W 15 DAYS EGOS LAID AFTER ADMINISTRATION OF P32
FIG. 4. The percent of the dose of P32 administered to the hen per milligram of total calcium which was found in the various tissues of the chicks hatched from eggs laid on the days indicated after administration.
might indicate abnormalities in isotope metabolism in certain tissues that would not be indicated where percent dose only is calculated. Each of the laying hens appeared healthy throughout the experiment. However, hens 5 and 6 which received approximately 7.23 rep's (roentgen equivalent physical) of radiation per day in the form of the combined dose of Ca46 and P32 molted and went out of production two days after the isotope administration but came back into production later.
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
Romanoff, as edited by Taylor (1949), reports that the newly hatched chick (exclusive of the yolk sac) contains about 150 milligrams more calcium than the entire contents of the unincubated egg. It is quite probable that the yolk of the egg which was laid one day after administration and subsequently hatched, contained no radioactive calcium as it was laid. However, on hatching, the yolk sac of the chick actually contained approximately 0.40 percent of the entire dose administered the hen. Since the yolk of the egg is formed 24 to 25 hours before laying, the only opportunity for the Ca45 to have reached the yolk sac which was drawn into the chick prior to hatching was by diffusion from the shell and to a much less extent from the white. This is substantiated by the fact that no P32 was found in the yolk sac of the chick hatched from the first egg laid by the hen after receiving the concurrent doses of radioactive calcium and phosphorus. It is difficult to estimate the amount of calcium which is actually translocated therein from the shell itself, but using the first and fourth days as a basis for calculation, approximately four times as much calcium is withdrawn from the shell for embryo formation as is derived from the yolk and white.
203
204
NEWS AND NOTES SUMMARY 45
32
REFERENCES Association Official Agricultural Chemists, 194S. Methods of analysis. 932 pages. Washington, D. C. Driggers, J. C , and C. L. Comar, 1949. The secretion of radioactive calcium (Ca46) in the hen's egg. Poultry Sci. 28: 420-424. Kamen, Martin D., 1947. Radioactive tracers in biology. Academic Press Inc., New York, N. Y., pp. 281. Margnetti, C , 1949. Precipitation techniques. Tracerlog 22: 9-11. (A commercial publication of Tracerlab, Inc., Boston 10, Mass.) O'Neil, J. IS., J. R. Jowsey, C. C. Lee, M. A. Reade and J. W. T. Spinks, 1948. Determination of the fate of phosphorus in the laying hen by means ot radiophosphorus (P32). Science 107: 295-296. Shirley, R. L., O. R. Deal and G. K. Davis, 19S0. Comparison of calcium oxalate and carbonate precipitates of radioactive assays. Anal. Chem. 22: 1003-1004. Spinks, J. W. T., M. R. Berlie and J. B. O'Neil, 1949. Determination of the fate of calcium in the laying hen by means of radiocalcium (Ca46). Science 110: 332-333. Taylor, Lewis W., 1949. Fertility and hatchability of chicken and turkey eggs. John Wiley and Sons, Inc., New York, N . Y., pp. 432.
News and Notes DELAWARE NOTES
Dr. A. T. Ringrose has joined the staff of the Department of Animal and Poultry Industry, Agricultural Experiment Station, University of Delaware, Newark. He will devote the major part of his work to the broiler industry in Delaware, determining the factors that go into the production of quality broilers. He was born in New York state and graduated from Cornell University with a
B.S. in 1930 and a Ph.D. in 1935. He was on the staff of the Poultry Department at Cornell from 1930 to 1935, becoming Assistant Director of Nutritional Research at National Oil Products Company, Harrison, N. J., in that year. In 1937 he joined the staff of the Department of Poultry Husbandry, Kentucky Agricultural Experiment Station, Lexington. During the war, Dr. Ringrose served with the Quartermaster Corps in nutrition work and
{Continued on page 212)
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1. Results of Ca and P analysis of chicks hatched from eggs laid subsequent to oral administration of these radioisotopes to the laying hens are presented. 2. Appreciable quantities (0.40 percent of the administered dose) of radioactive calcium were found in the absorbed yolk sac of chicks hatched from eggs laid one day after administration of the isotope. It is quite probable that all of this calcium was acquired from the egg shell. 3. The highest concentration of Ca45 (1.016 percent of dose); was found in chicks from eggs laid two days after administration. Thereafter, the amount dropped abruptly and then tended to decline gradually, indicating that the source of supply was mainly the yolk and white. 4. The concentration of P32 did not reach its peak (0.839 percent of the dose) in chicks until the eggs from which they were hatched were laid six days after administration of the isotope. 5. When the concentration of Ca46 and 32 P in the various chick tissues were calculated as percent dose per milligram of total element, respectively, all the
tissues were essentially equivalent in isotope concentration. This indicates that the isotopes are incorporated in the tissues in direct proportion to the total element assimilated.
MEHRHOF 1
Florida Agricultural Experiment Station, Gainesville, Florida (Received for publication August 15, 1950)
P
REVIOUS work at this station (Driggers and Comar, 1949) demonstrated that radioactive calcium (Ca45), when administered orally to a fasted hen reached the shell of an egg which was being formed within a very few minutes, and that by the end of 24 hours as much as one-third of the ingested dose was found in the entire contents of another egg which was laid. This work was confirmed by Spinks et al. (1949). O'Neil et al. (1948) showed also that radioactive phosphorus was deposited in the yolk and albumen of an egg soon after the oral administration of the isotope, and the eggs which were laid subsequent to that time showed increasing amounts up to the fifth day, after which time the total amount of P32 decreased. In this investigation, a study was made of (1) the transference of radioactive calcium and phosphorus, when administered separately and concurrently to previously mated hens, to chicks hatched 1
Department of Poultry Husbandry. Department of Animal Husbandry and Nutrition. Published with the permission of the Director of the Florida Agricultural Experiment Station. The Ca46 and P 32 were obtained from the Oak Ridge National Laboratory on allocation of the Isotopes Division of the U. S. Atomic Energy Commission. The authors express their appreciation to Francis L. Coune, Riley D. Owens, Jackson McClenny, John McCall, Marvin H. Hernden and Al Lovelace for technical assistance. 2
from eggs laid subsequent to the administration of the isotopes, and (2) the distribution of the isotopes in the various tissues of the chicks. EXPERIMENTAL Eight one-year-old S. C. White Leghorn hens weighing approximately 1750 grams each and having been previously mated were used in this experiment. They were kept in standard laying cages and fed a complete all-mash diet. The egg cycles of the hens had been estimated and the radioactive isotopes were administered at the expected time of ovulation. Using the technique employed in earlier work (Driggers and Comar, 1949), four of these hens received an oral dose of Ca45 as a solution of the chloride with an activity of approximately 70 microcuries. Two of the hens received 70 microcuries of the Ca45 plus 300 microcuries of carrier free P32 as H3PO4. One hen received 300 microcuries of P32 and the eighth hen received 200 microcuries of the P32 alone. Each egg laid thereafter was incubated and individually hatched to insure proper chick identification. On the day of hatching, the chicks were sacrificed and the following tissue samples were taken for analysis: yolk sac; femur; breast bone; brain; heart; liver; proventriculus; small intestine; breast muscle; and leg muscle. The samples were wet-ashed with concentrated HNO3 and evaporated to near-
199
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
T h e Transference of Radioactive Calcium and Phosphorus from Hen to Chick
200
J. C. DRIGGERS, R. L. SHIRLEY, G. K. DAVIS AND N. T. MEHRHOF
nations were made using immersion-type Geiger-Mueller tubes in conjunction with commercial scalers. In the case of samples containing both Ca46 and P32 aliquots from the same solutions were taken for the respective determinations. The dipping-type GM tubes are insensitive to the weak beta emissions from the Ca46. Calcium oxalates prepared from solutions containing P32 and calcium which was free of Ca46 did not show any activity
TABLE 1.—Egg and chick production record of hens administered radioactive calcium and phosphorus
Hen number
Dose of the radioactive element
Days after administration x —egg laid on this day (x)—chick hatched from this egg
» 1
1 2 3 4
microcuries Ca«, 70
2
3
4
5
(x)
(x)
X
X
(x)
X
(x)
(x)
(x)
«
X
(x)
48
Ca , 70 45
Ca , 70 46
Ca , 70
X
X
6
7
X
X
(x)
X
8 X X
9 X
10
11
X
X
X
X
X X
X
12
13
14
IS
X
X
X
X
X
X
X
X
X
X X
46
S
Ca , 70 plus P32, 300
X
6
Ca45, 70 plus P32, 300
«
7
PS2, 300
8
32
P , 200
X
(x)
(x)
(x)
X
manner for radioactivity assay by means of a funnel and ring and disc assembly (Margnetti, 1949) which has been especially designed for this purpose. The Ca46 radioactivity assays were made on a commercial scaler using thin mica window (1.5-2.0 mg./cm.2) Geiger-Mueller tubes. Self-absorption and decay corrections were made in the usual manner (Kamen, 1947). Total calcium in the tissues was calculated from the weight of the calcium oxalate precipitate obtained from the aliquot prepared for radioactivity measurement. The phosphorus radioactivity determi-
(x)
(x)
(x)
(x)
X
(x)
(x) X
(x)
(x)
(x)
(x)
X
X
X
X
X
when the thin mica window GM tubes were used. Using the calcium oxalate method, no precipitation was found when 50 milligrams of phosphorus alone or 20 milligrams of phosphorus plus 20 milligrams of aluminum were used in synthetic solutions (Shirley, Owens and Davis, 1950). Total phosphorus was determined by the volumetric phosphomolybdate method (A.O.A.C, 1945). RESULTS
During this experiment, each of the hens receiving the Ca46 alone produced one or more chicks. One of the hens re-
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
dryness, diluted with distilled water, filtered, and made up to volume in volumetric flasks from which aliquots were taken for radioactivity -and total element determinations. The calcium was then precipitated as calcium oxalate (A.O.A.C., 1945) after 5 milligrams of inactive calcium had been added to each aliquot to insure precipitates weighing atleast 17 milligrams. The precipitate was prepared in a uniform
MATERNAL TRANSFER OF RADIOACTIVE CALCIUM AND PHOSPHORUS
ceiving concurrent doses of Ca46 and P322 did not produce any chicks, while thee other produced four. One hen receivingg the P32 alone produced nine chicks andi the other produced two. The egg produc-
201
the Ca45 dose was found in the tissues. Thereafter, there was a sharp decline until by the fourth day less than 0.05 percent of the dose could be found in any of the chicks. In Figure 2 the data ob-
100..
—
0.9Q. 050. 0.70.. m ^ 0.60| o u. 0.50J o uj 040.. §
1) E3 • •
YOLK SAC FEMUR BREAST BONE OTHER TISSUES BRAIN HEART LIVER PROVENTRICULUS INTESTINES BLOOD BREAST MUSCLE LEG MUSCLE
0.30.
J- 0.2Q. u o o.ioj
or a! 0.00.
I I 22 3 4 5 6 H 8 9 10 II 12 13 DAYS EGGS LAID AFTER ADMINISTRATION OF C A 4 5
14
15
FIG. 1. The percent of the dose of Ca46 administered which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: The chicks hatched from the first egg laid on the first day and the twelfth, fourteenth and fifteenth days after administration were those from the hens which received concurrent doses of Ca45 and P32.
tion and chicks hatched therefrom aree given in Table 1. Figure 1 shows graphically the percentt dose of Ca45 found in the various tissuess of the chicks hatched from eggs laid 1 to0 15 days after oral administration of thee isotope to the hens. The values obtainedi on the chicks from different hens were inti close agreement on the second and fourth•x days after administration and the valuess for the individual tissues in these casess were averaged. In the case of the chickss from the eggs laid on the first and second1 days after administration, approximatelyY 0.745 and 1.016 percent respectively off
tained for percent dose per milligram of total calcium in the various tissues are presented. The percent dose of P32 found in the chick tissues is presented in Figure 3. Chick tissue values obtained on the sixth and ninth days from different hens were averaged. In contrast to the above calcium data, very little of the P32 was found in the chicks hatched from eggs laid during the first two days after administration of the isotope. The P32 values increased rapidly from the second to the sixth day, at which time approximately 0.839 percent of the dose was found in all
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
I.IO
202
J. C. DRIGGERS, R. L. SHIRLEY, G. K. DAVIS AND N. R. MEHRHCTF
ffl
YOLK SAC FEMUR BREASTBONE BRAIN HEART LIVER PROVENTRICULUS INTESTINES BREAST MUSCLE LEG MUSCLE
O.OOJ
JBL I I 2 3 4 5 6 7 8 9 10 II 12 13 14 15 DAYS EGGS LAID AFTER ADMINISTRATION OF C A 4 5
FIG. 2. The percent of the dose of Ca45 administered to the hen per milligram of total calcium which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: Only traces of Ca46 per milligram of total calcium were found in chicks from the fourteenth and fifteenth days' eggs.
tissues analyzed. These values gradually decreased until the end of the fourteenth
D 0 D •
YOLK SAC FEMUR BREAST BONE BRAIN HEART LIVER D PROVENTRICULUS • INTESTINES m BREAST MUSCLE ID LEO MUSCLE
•
I 2 3 4 S 6 7 S 9 K) II 12 12 14 15 DAYS EGGS LAID AFTER ADMINISTRATION OF P32
day when less than 0.12 percent could be found in the newly hatched chicks. The data showing the percent dose of radioactive phosphorus per milligram of total phosphorus are in Figure 4. DISCUSSION A gross examination of the tissues indicated that the chicks hatched during this study were quite normal. The writers believe that infertility was the primary reason for the poor hatch. FIG. 3. The percent of the dose of P32 administered which was found in the various tissues of chicks hatched from eggs laid on the days indicated after administration. Note: The hen which laid the egg on the first day and the second egg on the twelfth day after administration received concurrent doses of Ca46 and P32. No Ca46 or P32 was found in the chick hatched from the egg laid on the first day after administration.
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
H 0 • • S E D m H CD
MATERNAL TRANSFER OE RADIOACTIVE CALCIUM AND PHOSPHORUS
The greatest concentration of P32 was found in the chicks hatched from eggs laid 6 days after administration, allowing time for the entire yolk and white to receive the maximum dose of this element. Of particular interest is the gradual decrease in total P32 content of the entire chick after the peak has been reached on the sixth day as compared to the sharp decline in Ca45 content after the second day. In Figures 2 and 4 where the percent dose of Ca45 and P32 respectively per milligram of total element is reported, it
is evident that the radioactive isotope tends to be distributed uniformly throughout all the tissues with the stable isotope, According to these data, especially in the case of the phosphorus in Figure 4, all the tissues are essentially equivalent in concentration of the isotope per milligram of the total element. Such data as these
D
YOLK SAC 0 FEMUR D BREASTBONE BRAIN HEART 13 LIVER PROVENTRICULUS INTESTINES BREAST MUSCLE LEG MUSCLE
• s
a m n m
i ILii *
H
I 2 3 4 5 6 7 8 9 10 1! 12 12 W 15 DAYS EGOS LAID AFTER ADMINISTRATION OF P32
FIG. 4. The percent of the dose of P32 administered to the hen per milligram of total calcium which was found in the various tissues of the chicks hatched from eggs laid on the days indicated after administration.
might indicate abnormalities in isotope metabolism in certain tissues that would not be indicated where percent dose only is calculated. Each of the laying hens appeared healthy throughout the experiment. However, hens 5 and 6 which received approximately 7.23 rep's (roentgen equivalent physical) of radiation per day in the form of the combined dose of Ca46 and P32 molted and went out of production two days after the isotope administration but came back into production later.
Downloaded from http://ps.oxfordjournals.org/ at SUNY Health Science Center at Brooklyn - Medical Research Library on April 7, 2015
Romanoff, as edited by Taylor (1949), reports that the newly hatched chick (exclusive of the yolk sac) contains about 150 milligrams more calcium than the entire contents of the unincubated egg. It is quite probable that the yolk of the egg which was laid one day after administration and subsequently hatched, contained no radioactive calcium as it was laid. However, on hatching, the yolk sac of the chick actually contained approximately 0.40 percent of the entire dose administered the hen. Since the yolk of the egg is formed 24 to 25 hours before laying, the only opportunity for the Ca45 to have reached the yolk sac which was drawn into the chick prior to hatching was by diffusion from the shell and to a much less extent from the white. This is substantiated by the fact that no P32 was found in the yolk sac of the chick hatched from the first egg laid by the hen after receiving the concurrent doses of radioactive calcium and phosphorus. It is difficult to estimate the amount of calcium which is actually translocated therein from the shell itself, but using the first and fourth days as a basis for calculation, approximately four times as much calcium is withdrawn from the shell for embryo formation as is derived from the yolk and white.
203
204
NEWS AND NOTES SUMMARY 45
32
REFERENCES Association Official Agricultural Chemists, 194S. Methods of analysis. 932 pages. Washington, D. C. Driggers, J. C , and C. L. Comar, 1949. The secretion of radioactive calcium (Ca46) in the hen's egg. Poultry Sci. 28: 420-424. Kamen, Martin D., 1947. Radioactive tracers in biology. Academic Press Inc., New York, N. Y., pp. 281. Margnetti, C , 1949. Precipitation techniques. Tracerlog 22: 9-11. (A commercial publication of Tracerlab, Inc., Boston 10, Mass.) O'Neil, J. IS., J. R. Jowsey, C. C. Lee, M. A. Reade and J. W. T. Spinks, 1948. Determination of the fate of phosphorus in the laying hen by means ot radiophosphorus (P32). Science 107: 295-296. Shirley, R. L., O. R. Deal and G. K. Davis, 19S0. Comparison of calcium oxalate and carbonate precipitates of radioactive assays. Anal. Chem. 22: 1003-1004. Spinks, J. W. T., M. R. Berlie and J. B. O'Neil, 1949. Determination of the fate of calcium in the laying hen by means of radiocalcium (Ca46). Science 110: 332-333. Taylor, Lewis W., 1949. Fertility and hatchability of chicken and turkey eggs. John Wiley and Sons, Inc., New York, N . Y., pp. 432.
News and Notes DELAWARE NOTES
Dr. A. T. Ringrose has joined the staff of the Department of Animal and Poultry Industry, Agricultural Experiment Station, University of Delaware, Newark. He will devote the major part of his work to the broiler industry in Delaware, determining the factors that go into the production of quality broilers. He was born in New York state and graduated from Cornell University with a
B.S. in 1930 and a Ph.D. in 1935. He was on the staff of the Poultry Department at Cornell from 1930 to 1935, becoming Assistant Director of Nutritional Research at National Oil Products Company, Harrison, N. J., in that year. In 1937 he joined the staff of the Department of Poultry Husbandry, Kentucky Agricultural Experiment Station, Lexington. During the war, Dr. Ringrose served with the Quartermaster Corps in nutrition work and
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1. Results of Ca and P analysis of chicks hatched from eggs laid subsequent to oral administration of these radioisotopes to the laying hens are presented. 2. Appreciable quantities (0.40 percent of the administered dose) of radioactive calcium were found in the absorbed yolk sac of chicks hatched from eggs laid one day after administration of the isotope. It is quite probable that all of this calcium was acquired from the egg shell. 3. The highest concentration of Ca45 (1.016 percent of dose); was found in chicks from eggs laid two days after administration. Thereafter, the amount dropped abruptly and then tended to decline gradually, indicating that the source of supply was mainly the yolk and white. 4. The concentration of P32 did not reach its peak (0.839 percent of the dose) in chicks until the eggs from which they were hatched were laid six days after administration of the isotope. 5. When the concentration of Ca46 and 32 P in the various chick tissues were calculated as percent dose per milligram of total element, respectively, all the
tissues were essentially equivalent in isotope concentration. This indicates that the isotopes are incorporated in the tissues in direct proportion to the total element assimilated.