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The Variation in the Calcium Level of the Blood of the Domestic Fowl*
T h e Variation in the Calcium Level of the Blood of the Domestic Fowl* H. R. KNOWLES, E. B. HART, AND J. G. HALPIN University of Wisconsin, Madison (Received for Publication May 24, 1934)
* Published with the permission of the Director of the Wisconsin Agricultural Experiment Station.
from 9 to 27 mg. Ca per 100 cc. serum. The increase in the level of calcium occurred approximately 108 hours before the first egg was laid and a decrease from the high level set in after the second egg was laid. Hughes, Titus, and Smits ((1927) published data dealing with the calcium level of the blood of chickens. Their work showed that the level of calcium was much higher in laying hens—even up to three times—than in immature pullets. Russell, Howard, and Hess (1930) stated that the serum calcium of hens lay between 13.0 and 26.7 mg. Ca per 100 cc. serum if an ovum greater than 1 cm. in diameter were present, even though the birds were on a ration low in the antirachitic factor. Buckner, Martin, and Hull (1930) showed that the laying hen absorbed calcium at a rapid rate from the intestines, since the calcium content of the blood in the anterior mesenteric vein was 2 mg. per 100 cc. serum higher than that in the anterior mesenteric artery. This difference was not found in the case of the non-laying hen. That this was not due to the lack of a readily available source of calcium was evident, since a post-mortem examination revealed the presence of calcium containing grit in the gizzard and intestines of the non-laying hen. Sun and MacOwan (1930) found that the microscopical structure of the parathyroid glands in the laying hen differed from that of the non-laying hen. In the former the cells were larger and not as closely packed,
[83]
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AS Bell (1908) knew of no method for l \ determining the level of calcium in the blood, he developed a method which he called the calcium index. He precipitated the calcium from the blood as the oxalate and counted the crystals of calcium oxalate on a hemocytometer plate. In the case of a non-laying hen he found the calcium index to vary between 0.9 to 1.3 on successive days, whereas in the laying hen the variation was from 0.S to 3.1. Pearl and Surface (1909) suggested that the causal factor for the deposition of calcium on the egg was a mechanical one. They sutured the large intestine to the oviduct anterior to the shell gland and found that the faeces were then covered with a calcareous deposit on their passage through the gland. Buckner, Martin, and Peter (1925) showed that there was not enough calcium in the shell gland to account for all the calcium in the shell. The gland contained only 0.000S to 0.0071 gram CaO whereas the egg contained approximately 2.5 grams. They then postulated that the transfer of calcium during shell formation was by a direct removal from the circulation. Riddle and Reinhart (1926) while studying the variation in the calcium level of the blood of pigeons found that the level in male pigeons did not vary more than 1.2 mg. Ca per 100 cc. serum. However, in the case of the hen pigeon the variation was
84
POULTRY
SCIENCE
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and frequently exhibited a degree of vacuo- was as follows: The large cephalic vein was lation. Collip (1931) stated that he was un- exposed in its sheath by an incision through able to demonstrate any effects of injections the skin, and an extremely finely pointed of parathormone upon non-laying hens. instrument—a number 11 Bard Parker knife MacOwan (1932) reaffirmed her previous blade—was carefully inserted into the vein work on the structure of the parathyroid until blood just flowed out when slight presgland during the period of ovulation. She sure was applied. By careful manipulation also injected chickens with parathormone. such a puncture could be used for drawing By taking another sample of blood approxi- as many as a dozen samples of blood. At mately seventeen hours later, she was able each subsequent bleeding the small clot in to show an increase of 1.5 mg. per 100 cc. the wound was wiped away after pressure serum in the case of pullets, a varied in- had been applied. The blood was collected crease and decrease in the case of non-laying into the special tube described by Elvehjem hens, and no response in cocks. and Kline (1933) and its calcium content Charles and Hogben (1933) attempted to determined as prescribed by them. correlate the level of calcium in the blood The method of restraint used was that with the period in the ovarian cycle, by kill- of simply tying the legs of the bird above ing off the hens at varying intervals after the hock joint with a narrow strip of cloth laying. They found that as long as there and laying the bird on its side on a table. was no egg in the oviduct the serum calcium Samples of blood from the shell gland durremained about 17 mg. per 100 cc. serum ing the period of shell secretion were re(14.7-19.6). On the other hand when the quired for another phase of this work. The egg was in the oviduct the range was hen was anaesthetized with ether. An inci10.5-28.5 mg. Ca. per 100 cc. serum. They sion was made from the point of the breaststate that it is not surprising that exception- bone to the cloaca and around one side of ally low values should occur while the rapid the base of the tail. Then a transverse inelimination of calcium is in progress due to cision was made from midway along the first shell deposition; but what is striking is the incision toward the right shoulder, care fact that the rapid elimination of calcium being taken to stop before puncturing the by the secretory activity of the shell gland thoracic cavity. A third incision, similar to may be associated with a blood calcium level the second, was made on the left side. By which is much higher than that which is this means the abdominal cavity was exfound when shell secretion is not in progress. posed and a minimum of large blood vessels As there were no data available in the severed. In locating the uterine blood vesliterature showing the daily variation in the sels the intestines were pushed to one side level of blood calcium of the laying hen, it .and a very small hypodermic needle inserted was thought necessary to study this phase into the vein or artery and the sample withdrawn. The latter operation was not always of the problem. successful. EXPERIMENTAL In the studies on the response to parathormone,* the parathormone was injected Two difficulties presented themselves— under the skin on the bare site above the obtaining small samples of blood and the femoral feather tract. The birds were bled analysis of such small samples for their before and after the injection. The results calcium content. Various methods of bleeding were at* In the experiments dealing with the action of tempted but the one yielding the best results parathormone, Mr. Bernard Kline co-operated.
MARCH,
1935.
VOL.
XIV,
No.
85
2
TABLE 1.—Variations in level of blood calcium independent of intervals Time Hen No.
Date Bled
9573
9566
12/1/33 12/2/33
12/6/33 12/7/33
5:15 p.m. 10:40 a.m. 4:30 p.m. 10:45 p.m. 9:00 3:15 9:40 8:45 3:40
a.m. p.m. p.m. a.m. p.m.
12/1/33 12/2/33
5:30p.m. 9:30 a.m. 5:00 p.m. 11:15 p.m.
9572
12/6/33
9:00 a.m. 3:30 p.m. 10:00 p.m. 9:00 a.m. 3:10 p.m.
12/7/33
obtained in this study are tabulated in Tables 1 to 7. There are variations in the level of blood calcium independent of intervals between bleeding. This is shown in Table 1. In Table 2 data are presented showing the relation of calcium level in the blood to the time of shell formation. DISCUSSION The frequent bleeding is believed to have had no deleterious effect upon the hens, since no decrease in egg production was noticed during the experimental period. The hens that were bled most frequently laid as follows: Repeated Bleeding without Effect upon Egg Production Hen No. 9S70 9572 9574
Total times Production bled in 2 months December January 43 21 22 36 21 21 42 23 22
By taking samples of blood with long intervals between bleedings (see Table 1) a
10:30 a.m.
12:30p.m.
9:00 a.m.
Grams blood in aliquot
Mg. Caper 100 grams blood (mean)
1.06 0.61 0.55 1.41
17.7 19.0 21.1 19.1
0.57 0.83 0.57 1.15 0.80
21.7 23.1 17.9 17.9 17.7
0.49 0.98 0.48 0.52
20.0 13.8 20.2 20.1
0.76 0.65 0.49 1.23 0.74
18.1 20.3 14.3 13.5 14.1
variation in the level of blood calcium can be demonstrated. Since the sample withdrawn was of the size of 1-3 cc. its withdrawal could not have been responsible for the changes which occurred. A study of Tables 2 to 5 shows that after an egg is laid the calcium level of the blood rises and then decreases. Further, they show that the calcium level of the blood of the same hen does not always rise to the same high levels, cf. Tables 2 and 4. Having established the nature of the variation, the causes came under consideration. Since the available data all pointed to a direct removal of calcium from the blood, attempts were made to secure samples of uterine and venous blood during shell deposition. Only one complete sample was obtained. Calcium Content of Blood of Shell Gland During Period of Shell Formation (Hen 9569)
Arterial blood Venous blood
Mg. C a / 1 0 0 grams Blood 1 2 Mean 18.0 18.6 18.3 10.6 10.6 10.6
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9579
Laid
bleedings
86
POULTRY -Variation in the level of blood calcium Hen no. 9570
TABLE 2.-
Mg. Ca per 100 grams blood (Mean)
Bleeding 10:50 a.m. 2:25 p.m.
0.77 0.65
13.4 23". 8
12/19/1933
10:00 a.m. 12:00 a.m. 2:25 p.m. 4:20 p.m. 6:20 p.m. 8:55 p.m. 11:00p.m.
0.51 1.52 1.18 0.96 1.20 0.82 1.80
8.7 16.5 15.6 16.7 11.0 15.6 16.5
12/20/1933
9:05 a.m.
Laid
0.47
14.5
1/ 3/1934
9:55 a.m. 12:00 a.m. 2:00 p.m. 4:15 p.m. 6:10 p.m. 8:30 p.m. 10:20 p.m.
No egg
1.23 0.87 0.71 1.28 1.70 0.78 0.81
31.6 37.0 30.4 27.7 32.0 26.5 23.0
1/ 4/1934
12:20 a.m. 8:50 a.m.
0.86 1.51
22.8 23.0
1/27/1934
1.76 1.74 1.18 1.60
23.8 27.2 26.8 16.5
0.81 0.92 1.43 1.40
27.0 26.4 .24.7 22.8
Laying No egg 8:00 a.m.
8:00 a.m. No egg
1/28/1934
9:25 a.m. 11:55 a.m. 2:36 p.m. 8:40 p.m.
1/29/1934
8:40 a.m. 11:00 a.m. 5:00 p.m. 9:50 p.m. (under ether) 10:10p.m.
9:40 a.m.
1:45 p.m. Died—no egg in oviduct
Thus one of the explanations for the decrease in the level of calcium is thought to be the following. The removal of calcium from the blood for the shell causes a lowering in the calcium level in the uterine circulation. This blood on being mixed with the other systemic blood causes the calcium level in the systemic blood to be decreased, provided the amount picked up in the intestinal circulation or elsewhere does not exceed the amount removed for shell formation. When shell deposition is completed the level will increase again.
In Tables 6 and 7 the effect of injections of parathormone on the level of calcium in the blood of chickens is shown. The remarkable phenomenon is the rapid response, and in the majority of cases the rapid decrease. The data obtained bring out the fact that the chicken responds to injections of parathormone, but in order to ascertain the rise the samples must be taken early after injection. The same bird, however, does not always give a response of the same degree, cf. Table 7, hen 9SS8. Since the chicken responds to injections of parathormone and
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Grams of blood in aliquot
Time of Date 12/18/1933
SCIENCE
MARCH,
1935.
VOL.
XIV,
No.
2
87
TABLE 3.—Variation in the level of blood calcium
Hen no. 9572 Grams of blood in aliquot
Mg. Ca per 100 grams blood (Mean)
0.88 1.19 0.61 1.04 1.05 0.88 0.56 0.65
17.4 17.4 16.8 14.1 17.8 17.2 17.6 18.0
0.84 1.02
15.3 14.1
No egg
1.09 1.23 1.51 1.03 1.33 0.94
14.8 14.4 13.0 16.8 11.8 12.3
11:00 a.m.
1.40 0.68 1.14
15.7 16.7 15.9
Time of Date Bleeding 12/27/1933 12/28/1933
9:00 a.m. 9:15 11:15 1:15 3:30 5:20 7:10 9:15 11:00
a.m. a.m. p.m. p.m. p.m. p.m. p.m. p.m.
1:00 a.m. 9:00 a.m.
1/27/1934
8:30 10:34 12:25 2:40 4:50 9:40
1/28/1934
11:20 a.m. 2:32 p.m. 8:55 p.m.
a.m. a.m. p.m. p.m. p.m. p.m.
11:00 a.m.
4:00 p.m.
the rate of response and duration of effect are sudden in character, it is felt that the variation in the high levels of blood calcium during the laying cycle are due to the activity of the parathyroid gland. Hence the following explanation is offered: (1) When a pullet comes into lay, the rise in the level of blood calcium is due primarily to the increased activity of the parathyroid gland. (2) When an eggshell is being formed, the calcium level of the blood decreases because of the rapid removal of calcium. When the shell is fully formed, the level rises again because either no more calcium is being removed, or a new stimulus is acting on the parathyroid. SUMMARY
There is a variation in the level of the calcium in the blood of laying hens from one oviposition to another. The maxima and minima are not constant even for the same
The period of low level corresponds to the time taken for shell deposition. The period of high level corresponds to the absence of a fully formed egg in the shell gland. In a single case a sample of blood taken TABLE 6.—Effect
of injection of parathormone on
level of blood calcium of hen. 2 cc. parathormone (40 Units) injected Hen No. 9562 Time after injection in hours 0 1 2 3 4 5 6 7 9 11 13 31 35
Mg. Ca per 100 grams blood 18.9 21.4 22.0 23.4 25.3 28.7 37.6 48.4 29.0 12.9 11.9 7.8 Died
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12/29/1933
Laying
POULTRY
SCIENCE
TABLE 4.—Variation in the level of blood calcium Hen no. 9574 Time of Date Bleeding
Laying
12/18/1933
Grams of blood in aliquot
Mg. Ca per 100 grams blood (Mean)
No egg 9:40 a.m. 11:50 a.m. 2:20 a.m. 4:00 p.m. 6:10 p.m. 8:45 p.m. 10:50 p.m.
9:40 a.m.
1.20 1.14 0.61 1.24 0.88 1.41 1.08
15.7 12.1 13.1 13.2 11.6 12.6 12.0
12/20/1933
8:30 a.m.
2:55 p.m.
0.56
12.8
1/ 2/1934
No egg
1/ 3/1934
9:45 a.m. 11:50 a.m. 2:10 p.m. 4:00 p.m. 6:05 p.m. 8:20 p.m. 10:10 p.m.
1.34 1.60 1.38 1.19 0.74 0.71 0.93
21.1 17.2 23.2 24.4 21.1 21.8 21.8
1/ 4/1934
12:15 a.m. 8:45 a.m. 10:30 a.m.
0.91 0.55 1.11
18.9 18.9 17.9
1.04 1.46 0.72 0.85
18.4 15.8 17.9 15.7
1.42 0.90 1.27 1.24
14.9 17.5 17.7 15.0
10:30 a.m.
1/26/1934
No egg
1/27/1934
9:00 12:20 2:25 4:40
a.m. p.m. p.m. p.m.
1/28/1934
9:30 a.m. 12:00 a.m. 2:41p.m. 9:00p.m.
10:15 a.m.
2:00 p.m.
1/29/1934
No egg TABLE 5.—Variation in the level of blood calcium Hen no. 9568 Time of
Date Bleeding 1/26/1934 1/27/1934
1/28/1934
Laying
Grams of blood in aliquot
Mg. Caper 100 grams blood (Mean)
0.80 1.23 1.02 1.00 0.73
17.0 15.3 16.6 16.0 14.6
0.97 0.89 0.91 0.57
12.0 15.7 15.3 24.4
No egg 8:45 a.m. 10:30 a.m. 12:30 p.m. 2:30 p.m. 4:45 p.m. 9:20 a.m. 11:50 a.m. 2:30 p.m. 8:50 p.m.
10:00 a.m.
2:00 p.m.
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12/19/1933
MARCH,
1935.
TABLE 7.—Effects
Bird
4 6 8 9 10 12 13 22 25
No.
89
2
of injections of parathormone on level of blood calcium mg. Ca per 100 grams blood
3cc.
8.0 9.1
12.3
11.5
20.0
B.P. B.P. Rock hen no. Rock 9558. hen N.B. Not laying
B.P. Rock hen no. 9554. Not laying
B.P. Rock hen no. 9578. Not laying
1.5 cc.
1.0 cc.
1.5 cc.
1.5 cc.
11.9
9.4
10.4
10.6
14.5
11.6
14.4
14.3 14.9 12.9
11.6 11.7 11.2
11.9
11.2 11.2
lcc.
B.P. B.P. Rock S.C.W. Rock capon Leghorn cock no. capon 8556
B.P. Rock cock no. 8584
1 cc.
1 cc.
2cc.
1 cc.
14.5
7.6
6.5
9.2
6.7
12.2
22.5
9.5
10.7
25.2 16.3 11.8
12.9 13.1 13.6
16.6 14.3 14.2
8.0 7.8
7.4 6.8
9.0
8.5
8.8 7.9
10.6 10.8
14.1 11.8
8.3
6.9
12.4
7.8
7.0
7.7
13.4 14.8 10.3
13.3
from the uterine artery and uterine vein, during shell deposition, showed a marked difference in the level of calcium. Chickens have been shown to respond to subcutaneous injections of parathormone. The response is rapid and sudden. REFERENCES
Bell, B., 1908. Menstruation and its relationship to the calcium metabolism. Proc. Royal Soc. Med., Vol. 1, 291. Buckner, G. Davis, J. H. Martin and A. M. Peter, 1925. Chemical studies of the oviduct of the hen. Amer. Jour. Physiol. 71:349-3S4. Buckner, G. D., J. H. Martin and F. E. Hull, 1930. The distribution of blood calcium in the circulation of laying hens. Amer. Jour. Physiol. 93, 86. Charles, Enid and Lancelot Hogben, 1933. The serum calcium and magnesium level in the ovarian cycle of the laying hen. Quart. Jour, of Exp. Physiol. 23, 343. Collip, J. B., 1931. The physiology of the parathyroid glands. Can. Med. Assn. Jour. 24, 651.
Elvehjem, C. A. and B. E. Kline, 1933. Calcium and phosphorus studies in the chick. Jour. Biol. Chem. 103, 733. Hughes, J. S., R. W. Titus and B. L. Smits, 1927. The increase in the calcium of hen's blood accompanying egg production. Science, vol. lxv, 1680, p. 264. MacOwan, Marion, 1932. Observations on the ductless glands, the serum calcium, and egg laying in the fowl. Quart. Jour. Exp. Physiol. 21, 383. Pearl, Raymond and F. M. Surface, 1909. The nature of the stimulus which causes a shell to be formed on a bird's egg. Science (N. Ser. xxix) 741:428-429. Riddle, Oscar and W. F. Reinhart, 1926. Blood calcium changes in the reproductive cycle. Amer. Jour. Physiol. 76, 660-676. Russell, W. C , C. H. Howard and A. F . Hess, 1930. The relationship in the hen between the development of ova, blood calcium and the antirachitic factor. Science (vol. Ixxii) 1872: 506. Sun, T. P. and Marion MacOwan, 1930. Changes in parathyroid and adrenal glands and in blood calcium in relationship to egg formation in fowls. Jour. Physiol. 70: pp. iv-v.
Downloaded from http://ps.oxfordjournals.org/ at Ritsumeikan University on June 9, 2015
3i
XIV,
B.P. S.C.W. B.P. Rock Leghorn Rock pullet hen no. hen no. 9560. (10 9555. Not wks.) Not laying laying
Dose of Parathormone. . 3cc. Hours after injection 0 1 2
VOL.
* Published with the permission of the Director of the Wisconsin Agricultural Experiment Station.
from 9 to 27 mg. Ca per 100 cc. serum. The increase in the level of calcium occurred approximately 108 hours before the first egg was laid and a decrease from the high level set in after the second egg was laid. Hughes, Titus, and Smits ((1927) published data dealing with the calcium level of the blood of chickens. Their work showed that the level of calcium was much higher in laying hens—even up to three times—than in immature pullets. Russell, Howard, and Hess (1930) stated that the serum calcium of hens lay between 13.0 and 26.7 mg. Ca per 100 cc. serum if an ovum greater than 1 cm. in diameter were present, even though the birds were on a ration low in the antirachitic factor. Buckner, Martin, and Hull (1930) showed that the laying hen absorbed calcium at a rapid rate from the intestines, since the calcium content of the blood in the anterior mesenteric vein was 2 mg. per 100 cc. serum higher than that in the anterior mesenteric artery. This difference was not found in the case of the non-laying hen. That this was not due to the lack of a readily available source of calcium was evident, since a post-mortem examination revealed the presence of calcium containing grit in the gizzard and intestines of the non-laying hen. Sun and MacOwan (1930) found that the microscopical structure of the parathyroid glands in the laying hen differed from that of the non-laying hen. In the former the cells were larger and not as closely packed,
[83]
Downloaded from http://ps.oxfordjournals.org/ at Ritsumeikan University on June 9, 2015
AS Bell (1908) knew of no method for l \ determining the level of calcium in the blood, he developed a method which he called the calcium index. He precipitated the calcium from the blood as the oxalate and counted the crystals of calcium oxalate on a hemocytometer plate. In the case of a non-laying hen he found the calcium index to vary between 0.9 to 1.3 on successive days, whereas in the laying hen the variation was from 0.S to 3.1. Pearl and Surface (1909) suggested that the causal factor for the deposition of calcium on the egg was a mechanical one. They sutured the large intestine to the oviduct anterior to the shell gland and found that the faeces were then covered with a calcareous deposit on their passage through the gland. Buckner, Martin, and Peter (1925) showed that there was not enough calcium in the shell gland to account for all the calcium in the shell. The gland contained only 0.000S to 0.0071 gram CaO whereas the egg contained approximately 2.5 grams. They then postulated that the transfer of calcium during shell formation was by a direct removal from the circulation. Riddle and Reinhart (1926) while studying the variation in the calcium level of the blood of pigeons found that the level in male pigeons did not vary more than 1.2 mg. Ca per 100 cc. serum. However, in the case of the hen pigeon the variation was
84
POULTRY
SCIENCE
Downloaded from http://ps.oxfordjournals.org/ at Ritsumeikan University on June 9, 2015
and frequently exhibited a degree of vacuo- was as follows: The large cephalic vein was lation. Collip (1931) stated that he was un- exposed in its sheath by an incision through able to demonstrate any effects of injections the skin, and an extremely finely pointed of parathormone upon non-laying hens. instrument—a number 11 Bard Parker knife MacOwan (1932) reaffirmed her previous blade—was carefully inserted into the vein work on the structure of the parathyroid until blood just flowed out when slight presgland during the period of ovulation. She sure was applied. By careful manipulation also injected chickens with parathormone. such a puncture could be used for drawing By taking another sample of blood approxi- as many as a dozen samples of blood. At mately seventeen hours later, she was able each subsequent bleeding the small clot in to show an increase of 1.5 mg. per 100 cc. the wound was wiped away after pressure serum in the case of pullets, a varied in- had been applied. The blood was collected crease and decrease in the case of non-laying into the special tube described by Elvehjem hens, and no response in cocks. and Kline (1933) and its calcium content Charles and Hogben (1933) attempted to determined as prescribed by them. correlate the level of calcium in the blood The method of restraint used was that with the period in the ovarian cycle, by kill- of simply tying the legs of the bird above ing off the hens at varying intervals after the hock joint with a narrow strip of cloth laying. They found that as long as there and laying the bird on its side on a table. was no egg in the oviduct the serum calcium Samples of blood from the shell gland durremained about 17 mg. per 100 cc. serum ing the period of shell secretion were re(14.7-19.6). On the other hand when the quired for another phase of this work. The egg was in the oviduct the range was hen was anaesthetized with ether. An inci10.5-28.5 mg. Ca. per 100 cc. serum. They sion was made from the point of the breaststate that it is not surprising that exception- bone to the cloaca and around one side of ally low values should occur while the rapid the base of the tail. Then a transverse inelimination of calcium is in progress due to cision was made from midway along the first shell deposition; but what is striking is the incision toward the right shoulder, care fact that the rapid elimination of calcium being taken to stop before puncturing the by the secretory activity of the shell gland thoracic cavity. A third incision, similar to may be associated with a blood calcium level the second, was made on the left side. By which is much higher than that which is this means the abdominal cavity was exfound when shell secretion is not in progress. posed and a minimum of large blood vessels As there were no data available in the severed. In locating the uterine blood vesliterature showing the daily variation in the sels the intestines were pushed to one side level of blood calcium of the laying hen, it .and a very small hypodermic needle inserted was thought necessary to study this phase into the vein or artery and the sample withdrawn. The latter operation was not always of the problem. successful. EXPERIMENTAL In the studies on the response to parathormone,* the parathormone was injected Two difficulties presented themselves— under the skin on the bare site above the obtaining small samples of blood and the femoral feather tract. The birds were bled analysis of such small samples for their before and after the injection. The results calcium content. Various methods of bleeding were at* In the experiments dealing with the action of tempted but the one yielding the best results parathormone, Mr. Bernard Kline co-operated.
MARCH,
1935.
VOL.
XIV,
No.
85
2
TABLE 1.—Variations in level of blood calcium independent of intervals Time Hen No.
Date Bled
9573
9566
12/1/33 12/2/33
12/6/33 12/7/33
5:15 p.m. 10:40 a.m. 4:30 p.m. 10:45 p.m. 9:00 3:15 9:40 8:45 3:40
a.m. p.m. p.m. a.m. p.m.
12/1/33 12/2/33
5:30p.m. 9:30 a.m. 5:00 p.m. 11:15 p.m.
9572
12/6/33
9:00 a.m. 3:30 p.m. 10:00 p.m. 9:00 a.m. 3:10 p.m.
12/7/33
obtained in this study are tabulated in Tables 1 to 7. There are variations in the level of blood calcium independent of intervals between bleeding. This is shown in Table 1. In Table 2 data are presented showing the relation of calcium level in the blood to the time of shell formation. DISCUSSION The frequent bleeding is believed to have had no deleterious effect upon the hens, since no decrease in egg production was noticed during the experimental period. The hens that were bled most frequently laid as follows: Repeated Bleeding without Effect upon Egg Production Hen No. 9S70 9572 9574
Total times Production bled in 2 months December January 43 21 22 36 21 21 42 23 22
By taking samples of blood with long intervals between bleedings (see Table 1) a
10:30 a.m.
12:30p.m.
9:00 a.m.
Grams blood in aliquot
Mg. Caper 100 grams blood (mean)
1.06 0.61 0.55 1.41
17.7 19.0 21.1 19.1
0.57 0.83 0.57 1.15 0.80
21.7 23.1 17.9 17.9 17.7
0.49 0.98 0.48 0.52
20.0 13.8 20.2 20.1
0.76 0.65 0.49 1.23 0.74
18.1 20.3 14.3 13.5 14.1
variation in the level of blood calcium can be demonstrated. Since the sample withdrawn was of the size of 1-3 cc. its withdrawal could not have been responsible for the changes which occurred. A study of Tables 2 to 5 shows that after an egg is laid the calcium level of the blood rises and then decreases. Further, they show that the calcium level of the blood of the same hen does not always rise to the same high levels, cf. Tables 2 and 4. Having established the nature of the variation, the causes came under consideration. Since the available data all pointed to a direct removal of calcium from the blood, attempts were made to secure samples of uterine and venous blood during shell deposition. Only one complete sample was obtained. Calcium Content of Blood of Shell Gland During Period of Shell Formation (Hen 9569)
Arterial blood Venous blood
Mg. C a / 1 0 0 grams Blood 1 2 Mean 18.0 18.6 18.3 10.6 10.6 10.6
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9579
Laid
bleedings
86
POULTRY -Variation in the level of blood calcium Hen no. 9570
TABLE 2.-
Mg. Ca per 100 grams blood (Mean)
Bleeding 10:50 a.m. 2:25 p.m.
0.77 0.65
13.4 23". 8
12/19/1933
10:00 a.m. 12:00 a.m. 2:25 p.m. 4:20 p.m. 6:20 p.m. 8:55 p.m. 11:00p.m.
0.51 1.52 1.18 0.96 1.20 0.82 1.80
8.7 16.5 15.6 16.7 11.0 15.6 16.5
12/20/1933
9:05 a.m.
Laid
0.47
14.5
1/ 3/1934
9:55 a.m. 12:00 a.m. 2:00 p.m. 4:15 p.m. 6:10 p.m. 8:30 p.m. 10:20 p.m.
No egg
1.23 0.87 0.71 1.28 1.70 0.78 0.81
31.6 37.0 30.4 27.7 32.0 26.5 23.0
1/ 4/1934
12:20 a.m. 8:50 a.m.
0.86 1.51
22.8 23.0
1/27/1934
1.76 1.74 1.18 1.60
23.8 27.2 26.8 16.5
0.81 0.92 1.43 1.40
27.0 26.4 .24.7 22.8
Laying No egg 8:00 a.m.
8:00 a.m. No egg
1/28/1934
9:25 a.m. 11:55 a.m. 2:36 p.m. 8:40 p.m.
1/29/1934
8:40 a.m. 11:00 a.m. 5:00 p.m. 9:50 p.m. (under ether) 10:10p.m.
9:40 a.m.
1:45 p.m. Died—no egg in oviduct
Thus one of the explanations for the decrease in the level of calcium is thought to be the following. The removal of calcium from the blood for the shell causes a lowering in the calcium level in the uterine circulation. This blood on being mixed with the other systemic blood causes the calcium level in the systemic blood to be decreased, provided the amount picked up in the intestinal circulation or elsewhere does not exceed the amount removed for shell formation. When shell deposition is completed the level will increase again.
In Tables 6 and 7 the effect of injections of parathormone on the level of calcium in the blood of chickens is shown. The remarkable phenomenon is the rapid response, and in the majority of cases the rapid decrease. The data obtained bring out the fact that the chicken responds to injections of parathormone, but in order to ascertain the rise the samples must be taken early after injection. The same bird, however, does not always give a response of the same degree, cf. Table 7, hen 9SS8. Since the chicken responds to injections of parathormone and
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Grams of blood in aliquot
Time of Date 12/18/1933
SCIENCE
MARCH,
1935.
VOL.
XIV,
No.
2
87
TABLE 3.—Variation in the level of blood calcium
Hen no. 9572 Grams of blood in aliquot
Mg. Ca per 100 grams blood (Mean)
0.88 1.19 0.61 1.04 1.05 0.88 0.56 0.65
17.4 17.4 16.8 14.1 17.8 17.2 17.6 18.0
0.84 1.02
15.3 14.1
No egg
1.09 1.23 1.51 1.03 1.33 0.94
14.8 14.4 13.0 16.8 11.8 12.3
11:00 a.m.
1.40 0.68 1.14
15.7 16.7 15.9
Time of Date Bleeding 12/27/1933 12/28/1933
9:00 a.m. 9:15 11:15 1:15 3:30 5:20 7:10 9:15 11:00
a.m. a.m. p.m. p.m. p.m. p.m. p.m. p.m.
1:00 a.m. 9:00 a.m.
1/27/1934
8:30 10:34 12:25 2:40 4:50 9:40
1/28/1934
11:20 a.m. 2:32 p.m. 8:55 p.m.
a.m. a.m. p.m. p.m. p.m. p.m.
11:00 a.m.
4:00 p.m.
the rate of response and duration of effect are sudden in character, it is felt that the variation in the high levels of blood calcium during the laying cycle are due to the activity of the parathyroid gland. Hence the following explanation is offered: (1) When a pullet comes into lay, the rise in the level of blood calcium is due primarily to the increased activity of the parathyroid gland. (2) When an eggshell is being formed, the calcium level of the blood decreases because of the rapid removal of calcium. When the shell is fully formed, the level rises again because either no more calcium is being removed, or a new stimulus is acting on the parathyroid. SUMMARY
There is a variation in the level of the calcium in the blood of laying hens from one oviposition to another. The maxima and minima are not constant even for the same
The period of low level corresponds to the time taken for shell deposition. The period of high level corresponds to the absence of a fully formed egg in the shell gland. In a single case a sample of blood taken TABLE 6.—Effect
of injection of parathormone on
level of blood calcium of hen. 2 cc. parathormone (40 Units) injected Hen No. 9562 Time after injection in hours 0 1 2 3 4 5 6 7 9 11 13 31 35
Mg. Ca per 100 grams blood 18.9 21.4 22.0 23.4 25.3 28.7 37.6 48.4 29.0 12.9 11.9 7.8 Died
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12/29/1933
Laying
POULTRY
SCIENCE
TABLE 4.—Variation in the level of blood calcium Hen no. 9574 Time of Date Bleeding
Laying
12/18/1933
Grams of blood in aliquot
Mg. Ca per 100 grams blood (Mean)
No egg 9:40 a.m. 11:50 a.m. 2:20 a.m. 4:00 p.m. 6:10 p.m. 8:45 p.m. 10:50 p.m.
9:40 a.m.
1.20 1.14 0.61 1.24 0.88 1.41 1.08
15.7 12.1 13.1 13.2 11.6 12.6 12.0
12/20/1933
8:30 a.m.
2:55 p.m.
0.56
12.8
1/ 2/1934
No egg
1/ 3/1934
9:45 a.m. 11:50 a.m. 2:10 p.m. 4:00 p.m. 6:05 p.m. 8:20 p.m. 10:10 p.m.
1.34 1.60 1.38 1.19 0.74 0.71 0.93
21.1 17.2 23.2 24.4 21.1 21.8 21.8
1/ 4/1934
12:15 a.m. 8:45 a.m. 10:30 a.m.
0.91 0.55 1.11
18.9 18.9 17.9
1.04 1.46 0.72 0.85
18.4 15.8 17.9 15.7
1.42 0.90 1.27 1.24
14.9 17.5 17.7 15.0
10:30 a.m.
1/26/1934
No egg
1/27/1934
9:00 12:20 2:25 4:40
a.m. p.m. p.m. p.m.
1/28/1934
9:30 a.m. 12:00 a.m. 2:41p.m. 9:00p.m.
10:15 a.m.
2:00 p.m.
1/29/1934
No egg TABLE 5.—Variation in the level of blood calcium Hen no. 9568 Time of
Date Bleeding 1/26/1934 1/27/1934
1/28/1934
Laying
Grams of blood in aliquot
Mg. Caper 100 grams blood (Mean)
0.80 1.23 1.02 1.00 0.73
17.0 15.3 16.6 16.0 14.6
0.97 0.89 0.91 0.57
12.0 15.7 15.3 24.4
No egg 8:45 a.m. 10:30 a.m. 12:30 p.m. 2:30 p.m. 4:45 p.m. 9:20 a.m. 11:50 a.m. 2:30 p.m. 8:50 p.m.
10:00 a.m.
2:00 p.m.
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12/19/1933
MARCH,
1935.
TABLE 7.—Effects
Bird
4 6 8 9 10 12 13 22 25
No.
89
2
of injections of parathormone on level of blood calcium mg. Ca per 100 grams blood
3cc.
8.0 9.1
12.3
11.5
20.0
B.P. B.P. Rock hen no. Rock 9558. hen N.B. Not laying
B.P. Rock hen no. 9554. Not laying
B.P. Rock hen no. 9578. Not laying
1.5 cc.
1.0 cc.
1.5 cc.
1.5 cc.
11.9
9.4
10.4
10.6
14.5
11.6
14.4
14.3 14.9 12.9
11.6 11.7 11.2
11.9
11.2 11.2
lcc.
B.P. B.P. Rock S.C.W. Rock capon Leghorn cock no. capon 8556
B.P. Rock cock no. 8584
1 cc.
1 cc.
2cc.
1 cc.
14.5
7.6
6.5
9.2
6.7
12.2
22.5
9.5
10.7
25.2 16.3 11.8
12.9 13.1 13.6
16.6 14.3 14.2
8.0 7.8
7.4 6.8
9.0
8.5
8.8 7.9
10.6 10.8
14.1 11.8
8.3
6.9
12.4
7.8
7.0
7.7
13.4 14.8 10.3
13.3
from the uterine artery and uterine vein, during shell deposition, showed a marked difference in the level of calcium. Chickens have been shown to respond to subcutaneous injections of parathormone. The response is rapid and sudden. REFERENCES
Bell, B., 1908. Menstruation and its relationship to the calcium metabolism. Proc. Royal Soc. Med., Vol. 1, 291. Buckner, G. Davis, J. H. Martin and A. M. Peter, 1925. Chemical studies of the oviduct of the hen. Amer. Jour. Physiol. 71:349-3S4. Buckner, G. D., J. H. Martin and F. E. Hull, 1930. The distribution of blood calcium in the circulation of laying hens. Amer. Jour. Physiol. 93, 86. Charles, Enid and Lancelot Hogben, 1933. The serum calcium and magnesium level in the ovarian cycle of the laying hen. Quart. Jour, of Exp. Physiol. 23, 343. Collip, J. B., 1931. The physiology of the parathyroid glands. Can. Med. Assn. Jour. 24, 651.
Elvehjem, C. A. and B. E. Kline, 1933. Calcium and phosphorus studies in the chick. Jour. Biol. Chem. 103, 733. Hughes, J. S., R. W. Titus and B. L. Smits, 1927. The increase in the calcium of hen's blood accompanying egg production. Science, vol. lxv, 1680, p. 264. MacOwan, Marion, 1932. Observations on the ductless glands, the serum calcium, and egg laying in the fowl. Quart. Jour. Exp. Physiol. 21, 383. Pearl, Raymond and F. M. Surface, 1909. The nature of the stimulus which causes a shell to be formed on a bird's egg. Science (N. Ser. xxix) 741:428-429. Riddle, Oscar and W. F. Reinhart, 1926. Blood calcium changes in the reproductive cycle. Amer. Jour. Physiol. 76, 660-676. Russell, W. C , C. H. Howard and A. F . Hess, 1930. The relationship in the hen between the development of ova, blood calcium and the antirachitic factor. Science (vol. Ixxii) 1872: 506. Sun, T. P. and Marion MacOwan, 1930. Changes in parathyroid and adrenal glands and in blood calcium in relationship to egg formation in fowls. Jour. Physiol. 70: pp. iv-v.
Downloaded from http://ps.oxfordjournals.org/ at Ritsumeikan University on June 9, 2015
3i
XIV,
B.P. S.C.W. B.P. Rock Leghorn Rock pullet hen no. hen no. 9560. (10 9555. Not wks.) Not laying laying
Dose of Parathormone. . 3cc. Hours after injection 0 1 2
VOL.