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Investigations Concerning the Effects of Cortisone in the Domestic Fowl1
Investigations Concerning the Effects of Cortisone in the Domestic Fowl1 J. J. KUDZIA AND L. R. CHAMPION Department of Poultry Husbandry, Michigan State College, East Lansing (Received for publication October 3, 1952)
T
HE isolation of cortisone, principal hormone of the adrenal cortex, by Kendall (1938) has resulted in highly significant advances in medical science. Numerous clinical experiments have been conducted with cortisone on human patients for the treatment of various diseases. Most of the emphasis in the investigations has been directed toward the therapeutic value of cortisone. However, data are recorded in the literature indicating the possibility of deleterious effects occurring in laboratory animals receiving cortisone.
Hart and Reese (1950) reported an increase in the susceptibility of mice receiving cortisone to experimental tuberculosis. Glaubach, Antopol and Groff (1951) found that cortisone administered to pregnant mice had a growth-inhibition effect on the developing embryos. Cortisone administered to rats resulted in increased urinary nitrogen excretion and decreased protein synthesis, Clark (1950). Follis (1951) injected cortisone subcutaneously into young rats and observed a plateauing of the growth curves followed by a gradual loss of body weight. A retardation of wound healing in rats and rabbits receiving cortisone was ob-
served by Howes, Platz, Blunt and Rogan (1950). Courrier and Collange (1951) working with pregnant rabbits receiving cortisone, found that this extract caused abortions, hemorrhages, pale placentas, and resorbed or macerated fetuses greatly reduced in size. Landauer (1947) injected an adrenal cortex extract into the yolk sac of early developing chick embryos and observed growth retardation. Karnofsky, Patterson and Ridgeway (1949) and Karnofsky, Ridgeway and Patterson (1951) confirmed Landauer's (1947) growth retardation observation in chick embryos. Further, these researchers found that a selective inhibition of feather follicle formation occurred in the embryos. Mushett, Porter and Silber (1941) found that the route of administration of cortisone had an influence on the effects produced. When dogs were administered cortisone subcutaneously, polyurea, polydipsia and a slight anemia resulted. Oral administration of cortisone did not produce these effects. This investigation was initiated to determine the effects of cortisone acetate* on the body weight, egg production, egg size, fertility, hatchability, and feather formation of mature normal chickens.
1
Published with the approval of the Director of the Michigan Agricultural Experiment Station as Journal Article No. 1412. The data reported herein are taken from a thesis submitted by the senior author to the School of Graduate Studies, Michigan State College, in partial fulfillment of the requirements for the Master of Science degree in Poultry Husbandry. 476
MATERIALS AND METHODS
Three experiments designated herein * We are indebted to Merck and Company, Inc., Rahway, New Jersey for the cortisone acetate used in these studies.
CORTISONE EFFECTS IN FOWL
as Experiment 1, 2 and 3 were conducted using yearling male and female New Hampshire chickens. The cortisone acetate was injected subcutaneously at various dosage levels. The females were housed in individual cages in laying batteries equipped with raised wire bottoms. The males were housed in special male cages in the same building. Both the experimental and control birds received the station basal ration ad libitum and fresh water was constantly available. All weekly matings were accomplished by artificial insemination. Prior to the start of the experimental period, all individuals were observed for a five-week interval to obtain a "normal picture" for the factors previously enumerated. Body weights to the nearest ten gram unit were obtained for all birds three times per week, with the weighing procedure performed at a set time each day. Weekly measurements were made of all feathers removed (pectoral, cushion and primary) from date of removal until full development had been reached. All eggs were pedigreed and weighed to the nearest gram daily. Weekly settings of eggs were made. Upon hatching the chicks were pedigree wing-banded and day-old and two-week chick weights were taken. The chicks were fed the station starting ration. Experiment 1.—Six males divided into two equal groups comprised the experimental and control groups. The experimental males received subcutaneous injections of cortisone acetate at the 1 mg./lb. of body weight level. The control males were administered a .1 normal saline solution in proportions based on 1 mg./lb. of body weight to eliminate the possibility of any effects which could be attributed to the carrier of the cortisone. All injections were given on four consecutive days followed by a lag period of three days and then injections were
477
resumed for four additional consecutive days. Primaries I and II were removed from the wings of the males one day prior to the start of injections. Experiment 2.—Twenty-four females selected for uniformity of body weight and egg production and six males selected for body weight were equally divided into an experimental and control group. The experimental females received subcutaneous injections of cortisone acetate at the 1 mg./lb. of body weight level as in Experiment 1. The control females received the standard saline solution in appropriate proportions. The anterior pectoral tract feathers were removed from all females when injections were initiated. Experiment 3.—-Twelve selected females and four selected males made up the experimental and control populations. The experimental females received 3 mg./lb. of cortisone acetate whereas the control females were injected with the saline solution as in Experiments 1 and 2. Primaries I and II were removed from each wing of all females and in addition, an area of approximately four square inches of cushion feathers were removed. Analysis of variance and covariance (Fisher, 1930) were employed in the statistical treatment of these data. Covariance was used to correct the final body weights by adjusting for differences between individual birds in initial weights. Further, these data were analyzed for the pre-injection, injection and post-injection periods. RESULTS AND DISCUSSION Body Weight and Egg Production.—In Experiment 1, the experimental males lost an average of 243 grams in body weight during the injection period. This body weight loss was highly significant (Table 1). After the discontinuance of cortisone injections the males began to recover their
478
J. J. KUDZIA AND L. R. CHAMPION
TABLE 1.—The results of an analysis of covariance of initial (x) and final (y) body weights for experimental males during the injection period. Initial body weights observed, three days prior to injections, final weights four days post-injection Experiment 1. Males, 1 mg./lb. Sums of squares and products
Source of variation
d.f.
Total Between Treat. Within Treat.
5 1 4
Errors of estimate
Sx*
Sxy
Sf
d.f.
S.S.
M.S.
2,312 11 2,301
1,706 -83 1,789
2,059 640 1,419
1 3
771 29
771f 9.7
t Highly significant value of F.
body weight loss but their original body weight was not regained at the conclusion of this experiment. It is postulated that the loss in body weight resulted from alterations in carbohydrate and protein metabolism. The average body weight loss of 90 grams for the experimental females (Experiment 2) during the injection period was non-significant. The females did not lay as well as they had during the preinjection period, but the reduction in egg numbers was not significant. In Experiment 3, the experimental females lost an average of 20 grams in body weight during the injection period. This body weight loss was not significant. Four weeks after injections had ceased, the females had gained an average of 100 grams in body weight. The gain in body weight over and above their initial weight was highly significant (Table 2). Individual body weight changes for the experimental males and females (Experiments 1 and 3) are presented in Table 3. The increased TABLE 2.-
dosage level of cortisone (3 mg./lb.) caused egg production to cease completely within four days after the initiation of injections in five of the six females. The reduced egg production during the injection period was highly significant (selfevident). The females were out of production for seventeen days. It is suggested that the gain in body weight by the experimental females over and above their initial body weight was due to the cessation of egg production and consequently an increased deposition of fat. Further, it is hypothesized that cortisone administered in large doses to mature female chickens has an inhibitory effect (directly or indirectly) on the secretion of luteinizing hormone by the anterior pituitary. Egg Weight.—Cortisone acetate administered at the 1 mg./lb. of body weight level did not significantly affect egg size during or post injection. Fertility and Hatchability.—Cortisone injected into the male birds at the 1 mg./lb. of body weight level resulted in a
•The results of an analysis of covariance of initial (x) and final (y) body weights for experimental females for the post-injection period Experiment 3. Females, 3 mg./lb. Sums of squares and products
Source of variation
d.f.
Total Between Treat. Within Treat.
11 1 10
t Highly significant value of F.
Errors of estimate
Sx*
Sxy
sy
d.f.
S.S.
M.S.
8,240 1,102 7,138
8,347 355 7,992
9,536 114 9,422
1 9
607 474
607f 53
479
CORTISONE EFFECTS IN FOWL TABLE 3.—Body weights {gms.) of mature male and female chickens receiving cortisone acetate Experiment 1. Males, 1 mg./lb. Experiment
Control
Injection period
Bird No.
Pre
During
37 40 45
4,160 4,320 3,880
Ave.
4,090
TABLE 5.—The results of an analysis of variance of fertility for the post-injection period Experiment 2. Females, 1 mg./lb. Source of variation
d.f.
S.S.
M.S.
Injection period
Post
Bird No.
Pre
During
Post
3,880 4,060 3,610
3,980 3,990 3,660
42 43 44
4,060 3,280 3,860
4,050 4,200 3,920
4,040 4,140 3,940
3,850
3,880
Ave.
4,070
4,060
4,040
Experiment 3. Females, 3 mg ./lb. 3 5 7 9 16 18
3,010 2,860 3,000 2,510 2,700 2,520
3,080 2,870 3,020 2,390 2,740 2,420
3,120 3,030 3,220 2,610 2,740 2,500
1 2 12 25 30 35
2,760 2,490 2,930 3,210 3,040 3,320
2,830 2,420 2,960 3,060 2,930 3,140
2,690 2,440 2,980 3,250 3,040 3,220
Ave.
2,770
2,750
2,870
Ave.
2,960
2,890
2,940
TABLE 4.—The results of analysis of variance of fertility during the injection period Experiment 1. Males, 1 mg./lb. Source of variation
d.f.
S.S.
Total Between Treat. Within Treat.
21 1 20
29,291 7,287 22,004
M.S. 7,287* 1,100
* Significant value of F.
significant decrease in fertility during the injection period (Table 4). Differences in fertility during the post-injection period were not significant. Also, a significant loss in fertility was found to exist during the post-injection period when the females received cortisone at the 1 mg./lb. of body weight level (Table 5). No significant differences were found in hatchability in Experiments 1 and 2 for the preinjection, injection and post-injection periods.
Total Between Treat. Between Weeks Interaction
1,727 1,145 460 122
1,145* 230 61
* Significant value of F.
Feather Growth.—-As shown in Table 6, a highly significant retardation of primary feather growth in the males was found during the first week after injections of cortisone started. Differences in rate of feather growth were not significant thereafter. Inhibition of cushion feather growth (Experiment 3) was highly significant for the first three weeks after injections started, significant for the fourth week and non-significant thereafter (Table 7). An interesting observation was that the rejuvenating primary and cushion feathers were exceedingly stringy in appearance and had abnormally long shafts. These data confirm and extend to mature chickens the observations of Karnofsky el al. (1949, 1951) that cortisone retards feather formation in chick embryos. It is suggested that tissues vary greatly in their susceptibility to the action of cortisone, the physiological mode of action being complicated. The effects observed may be the result of alterations in the metabolic chain of events. Chick Weight.—Cortisone administered to the parental individuals had no sig-
TABLE 6.—The results of an analysis of variance of primary feather growth Experiment 1. Males, 1 mg./lb. Source of variation Total Between Treat Within Treat
Week d.f.
S.S.
5 1 4
3.6 3.1 .5
t Highly significant value of F.
2
1 M.S. 3.If 0.1
4
3
S.S.
M.S.
S.S.
M.S.
S.S.
M.S.
5.6 3.2 2.4
3.2 0.6
8.3 3.0 5.3
3.0 1.3
10.3 5.8 4.5
5.8 1.1
480
J. J. KUDZIA AND L. R. CHAMPION
TABLE 7.—The results of analysis of variance of cushion feather growth Experiment 3. Females, 3 mg./lb. Source of variance Total Between Treat. Within Treat.
Week
1
2
d.f.
s.s.
M.S.
S.S.
M.S.
S.S.
M.S.
11 1 10
1.0 0.5 0.5
0.5f 0.05
1.5 0.8 0.7
0.8f 0.07
3.8 2.6 1.2
2.6f 0.12
4 Total Between Treat. Within Treat.
3
11 1 10
3.0 1.4 1.6
5 1.4* 0.16
6.0 1.0 5.0
6 1.0 0.5
1.4 0.02 1.38
0.02 0.14
* Significant value of F. t Highly significant value of F. nificant effect on day-old or two-week chick weight. SUMMARY Two different levels of cortisone acetate were administered subcutaneously to mature New Hampshire male and female chickens. T h e suspensions were injected into the birds for four consecutive days followed by a lag period of three days, after which time injections were resumed for four additional consecutive days. The following results were secured: 1. Cortisone acetate caused a highly significant body weight loss in mature males during the injection period. 2. The body weight loss and reduced egg production observed by the females administered cortisone a t the 1 m g . / l b . of body weight level was not significant. 3. T h e body weight loss in females receiving 3 mg./lb. of cortisone per pound of body weight was not significant. However, the gain in body weight over and above initial body weight which was found to exist four weeks after injections ceased was highly significant. 4. A highly significant reduction in egg production was found to occur in females injected with 3 mg. of cortisone per pound of body weight.
5. A highly significant retardation in primary feather growth occurred in the males during the first week of injections of cortisone. 6. Inhibition of cushion feather growth in the females was highly significant for the first three weeks after initiation of injections, significant for the fourth week and non-significant thereafter. 7. Cortisone administered a t the 1 mg./lb. level caused a significant decrease in fertility, whereas at the high level (3 mg./lb.) significant reductions in fertility were not found. 8. No significant differences were found in egg weight, hatchability, day-old or two-week chick weight. REFERENCES Clark, I., 1950. Effect of cortisone on protein metabolism in the rat as studied with isotopic glycine. Federation Proc. 9: 161. Courrier, R., and A. Collonge, 1951. Cortisone and gestation. J. Am. Med. Assoc. 146: 493. Fisher, R. A., 1930. Statistical Methods for Research Workers. Oliver and Boyd, London. Follis, R. H., Jr., 1951. Effect of cortisone on growing bones of the rat. Proc. Soc. Exp. Biol. Med. 76: 722-724. Glaubach, S., W. Antopol and S. Graff, 1951. Excessive doses of cortisone in pregnant mice: effect on development and survival of the fetus and newborn and on the maternal breast tissue. Bui. New York Acad. Med. 27:398.
NORMAL OVULATION IN NON-LAYING HENS Hart, P. D'A., and R. J. W. Rees, 1950. Enhancing effect of cortisone on tuberculosis in the mouse. Lancet, 2: 391-395. Howes, E. L., C. M. Plotz, J. W. Blunt and C. Rogan, 1950. Retardation of wound healing by cortisone. Surgery, 28: 177-181. Kamofsky, D. A., P. A. Patterson and L. P. Ridgeway, 1949. Effect of folic acid, "4-amino" folic acids and related substances on growth of chick embryos. Proc. Soc. Exp. Biol. Med. 71:447-452. Kamofsky, D . A., L. P. Ridgeway and P. A. Patterson, 1951. Growth-inhibiting effect of cortisone
481
acetate on the chick embryo. Endocrinology, 48: 596-616. Kendall, E. C , 1938. Isolation in crystalline form of the hormone of the adrenal cortex. Proc. Staff Meeting, Mayo Clinic 11: 351. Landauer, W. L., 1947. Potentiating effects of adrenal cortical extract on insulin-induced abnormalities of chick development. Endocrinology, 41:489-493. Mushett, C. W., C. C. Porter and R. H. Silber, 1951. Effects of cortisone administered subcutaneously and orally to dogs. Federation Proc. 10: 366.
Normal Ovulation in Non-Laying Hens R. K. COLE AND F. B. HUTT Department of Poultry Husbandry, Cornell University Agricultural Experiment Station, Ithaca, New York (Received for publication October 6, 1952)
f \ V U L A T I O N and the subsequent lay^* ing of a normal egg, usually within 26 hours/ are preceded and accompanied by certain changes in the body of the hen. The external characters that are associated with an active state of egg production are well known and are used as the basis for the periodic culling of commercial laying flocks. By the absence of external changes considered indicative of egg production, one is able to identify nonlayers and to remove them from the flock, if so desired. I t is customary to consider as layers those birds which show the external characteristics associated with egg production. We have learned, however, that some birds behave as though they were laying, and show appropriate changes in body condition, without actually producing an egg. This paper presents the pertinent findings when groups of such birds were subjected to study. SOURCES OF DATA
Among large populations of pedigreed
White Leghorns that were being trapnested there were a number of birds which had only occasionally been credited with an egg, and in most cases, none at all. The non-laying hens from four populations, S, T, U, and V, hatched in the four years from 1948 to 1951, respectively, have been segregated and studied, although under somewhat different conditions of management each year. 5 population. In order to distinguish between non-layers and the floor-layers, the 39 birds in the S population for which there were no eggs recorded, other than sporadic ones, were transferred early in May to individual laying cages and kept there for a period of one month to ascertain their egg-producing status. The ten birds found to be laying at regular intervals were returned to their original pens and thereafter considered as floorlayers. The 29 proven non-layers were transferred to a small pen and kept there until early in October, at the end of the first laying year, at which time the reproductive systems of the 14 survivors
T
HE isolation of cortisone, principal hormone of the adrenal cortex, by Kendall (1938) has resulted in highly significant advances in medical science. Numerous clinical experiments have been conducted with cortisone on human patients for the treatment of various diseases. Most of the emphasis in the investigations has been directed toward the therapeutic value of cortisone. However, data are recorded in the literature indicating the possibility of deleterious effects occurring in laboratory animals receiving cortisone.
Hart and Reese (1950) reported an increase in the susceptibility of mice receiving cortisone to experimental tuberculosis. Glaubach, Antopol and Groff (1951) found that cortisone administered to pregnant mice had a growth-inhibition effect on the developing embryos. Cortisone administered to rats resulted in increased urinary nitrogen excretion and decreased protein synthesis, Clark (1950). Follis (1951) injected cortisone subcutaneously into young rats and observed a plateauing of the growth curves followed by a gradual loss of body weight. A retardation of wound healing in rats and rabbits receiving cortisone was ob-
served by Howes, Platz, Blunt and Rogan (1950). Courrier and Collange (1951) working with pregnant rabbits receiving cortisone, found that this extract caused abortions, hemorrhages, pale placentas, and resorbed or macerated fetuses greatly reduced in size. Landauer (1947) injected an adrenal cortex extract into the yolk sac of early developing chick embryos and observed growth retardation. Karnofsky, Patterson and Ridgeway (1949) and Karnofsky, Ridgeway and Patterson (1951) confirmed Landauer's (1947) growth retardation observation in chick embryos. Further, these researchers found that a selective inhibition of feather follicle formation occurred in the embryos. Mushett, Porter and Silber (1941) found that the route of administration of cortisone had an influence on the effects produced. When dogs were administered cortisone subcutaneously, polyurea, polydipsia and a slight anemia resulted. Oral administration of cortisone did not produce these effects. This investigation was initiated to determine the effects of cortisone acetate* on the body weight, egg production, egg size, fertility, hatchability, and feather formation of mature normal chickens.
1
Published with the approval of the Director of the Michigan Agricultural Experiment Station as Journal Article No. 1412. The data reported herein are taken from a thesis submitted by the senior author to the School of Graduate Studies, Michigan State College, in partial fulfillment of the requirements for the Master of Science degree in Poultry Husbandry. 476
MATERIALS AND METHODS
Three experiments designated herein * We are indebted to Merck and Company, Inc., Rahway, New Jersey for the cortisone acetate used in these studies.
CORTISONE EFFECTS IN FOWL
as Experiment 1, 2 and 3 were conducted using yearling male and female New Hampshire chickens. The cortisone acetate was injected subcutaneously at various dosage levels. The females were housed in individual cages in laying batteries equipped with raised wire bottoms. The males were housed in special male cages in the same building. Both the experimental and control birds received the station basal ration ad libitum and fresh water was constantly available. All weekly matings were accomplished by artificial insemination. Prior to the start of the experimental period, all individuals were observed for a five-week interval to obtain a "normal picture" for the factors previously enumerated. Body weights to the nearest ten gram unit were obtained for all birds three times per week, with the weighing procedure performed at a set time each day. Weekly measurements were made of all feathers removed (pectoral, cushion and primary) from date of removal until full development had been reached. All eggs were pedigreed and weighed to the nearest gram daily. Weekly settings of eggs were made. Upon hatching the chicks were pedigree wing-banded and day-old and two-week chick weights were taken. The chicks were fed the station starting ration. Experiment 1.—Six males divided into two equal groups comprised the experimental and control groups. The experimental males received subcutaneous injections of cortisone acetate at the 1 mg./lb. of body weight level. The control males were administered a .1 normal saline solution in proportions based on 1 mg./lb. of body weight to eliminate the possibility of any effects which could be attributed to the carrier of the cortisone. All injections were given on four consecutive days followed by a lag period of three days and then injections were
477
resumed for four additional consecutive days. Primaries I and II were removed from the wings of the males one day prior to the start of injections. Experiment 2.—Twenty-four females selected for uniformity of body weight and egg production and six males selected for body weight were equally divided into an experimental and control group. The experimental females received subcutaneous injections of cortisone acetate at the 1 mg./lb. of body weight level as in Experiment 1. The control females received the standard saline solution in appropriate proportions. The anterior pectoral tract feathers were removed from all females when injections were initiated. Experiment 3.—-Twelve selected females and four selected males made up the experimental and control populations. The experimental females received 3 mg./lb. of cortisone acetate whereas the control females were injected with the saline solution as in Experiments 1 and 2. Primaries I and II were removed from each wing of all females and in addition, an area of approximately four square inches of cushion feathers were removed. Analysis of variance and covariance (Fisher, 1930) were employed in the statistical treatment of these data. Covariance was used to correct the final body weights by adjusting for differences between individual birds in initial weights. Further, these data were analyzed for the pre-injection, injection and post-injection periods. RESULTS AND DISCUSSION Body Weight and Egg Production.—In Experiment 1, the experimental males lost an average of 243 grams in body weight during the injection period. This body weight loss was highly significant (Table 1). After the discontinuance of cortisone injections the males began to recover their
478
J. J. KUDZIA AND L. R. CHAMPION
TABLE 1.—The results of an analysis of covariance of initial (x) and final (y) body weights for experimental males during the injection period. Initial body weights observed, three days prior to injections, final weights four days post-injection Experiment 1. Males, 1 mg./lb. Sums of squares and products
Source of variation
d.f.
Total Between Treat. Within Treat.
5 1 4
Errors of estimate
Sx*
Sxy
Sf
d.f.
S.S.
M.S.
2,312 11 2,301
1,706 -83 1,789
2,059 640 1,419
1 3
771 29
771f 9.7
t Highly significant value of F.
body weight loss but their original body weight was not regained at the conclusion of this experiment. It is postulated that the loss in body weight resulted from alterations in carbohydrate and protein metabolism. The average body weight loss of 90 grams for the experimental females (Experiment 2) during the injection period was non-significant. The females did not lay as well as they had during the preinjection period, but the reduction in egg numbers was not significant. In Experiment 3, the experimental females lost an average of 20 grams in body weight during the injection period. This body weight loss was not significant. Four weeks after injections had ceased, the females had gained an average of 100 grams in body weight. The gain in body weight over and above their initial weight was highly significant (Table 2). Individual body weight changes for the experimental males and females (Experiments 1 and 3) are presented in Table 3. The increased TABLE 2.-
dosage level of cortisone (3 mg./lb.) caused egg production to cease completely within four days after the initiation of injections in five of the six females. The reduced egg production during the injection period was highly significant (selfevident). The females were out of production for seventeen days. It is suggested that the gain in body weight by the experimental females over and above their initial body weight was due to the cessation of egg production and consequently an increased deposition of fat. Further, it is hypothesized that cortisone administered in large doses to mature female chickens has an inhibitory effect (directly or indirectly) on the secretion of luteinizing hormone by the anterior pituitary. Egg Weight.—Cortisone acetate administered at the 1 mg./lb. of body weight level did not significantly affect egg size during or post injection. Fertility and Hatchability.—Cortisone injected into the male birds at the 1 mg./lb. of body weight level resulted in a
•The results of an analysis of covariance of initial (x) and final (y) body weights for experimental females for the post-injection period Experiment 3. Females, 3 mg./lb. Sums of squares and products
Source of variation
d.f.
Total Between Treat. Within Treat.
11 1 10
t Highly significant value of F.
Errors of estimate
Sx*
Sxy
sy
d.f.
S.S.
M.S.
8,240 1,102 7,138
8,347 355 7,992
9,536 114 9,422
1 9
607 474
607f 53
479
CORTISONE EFFECTS IN FOWL TABLE 3.—Body weights {gms.) of mature male and female chickens receiving cortisone acetate Experiment 1. Males, 1 mg./lb. Experiment
Control
Injection period
Bird No.
Pre
During
37 40 45
4,160 4,320 3,880
Ave.
4,090
TABLE 5.—The results of an analysis of variance of fertility for the post-injection period Experiment 2. Females, 1 mg./lb. Source of variation
d.f.
S.S.
M.S.
Injection period
Post
Bird No.
Pre
During
Post
3,880 4,060 3,610
3,980 3,990 3,660
42 43 44
4,060 3,280 3,860
4,050 4,200 3,920
4,040 4,140 3,940
3,850
3,880
Ave.
4,070
4,060
4,040
Experiment 3. Females, 3 mg ./lb. 3 5 7 9 16 18
3,010 2,860 3,000 2,510 2,700 2,520
3,080 2,870 3,020 2,390 2,740 2,420
3,120 3,030 3,220 2,610 2,740 2,500
1 2 12 25 30 35
2,760 2,490 2,930 3,210 3,040 3,320
2,830 2,420 2,960 3,060 2,930 3,140
2,690 2,440 2,980 3,250 3,040 3,220
Ave.
2,770
2,750
2,870
Ave.
2,960
2,890
2,940
TABLE 4.—The results of analysis of variance of fertility during the injection period Experiment 1. Males, 1 mg./lb. Source of variation
d.f.
S.S.
Total Between Treat. Within Treat.
21 1 20
29,291 7,287 22,004
M.S. 7,287* 1,100
* Significant value of F.
significant decrease in fertility during the injection period (Table 4). Differences in fertility during the post-injection period were not significant. Also, a significant loss in fertility was found to exist during the post-injection period when the females received cortisone at the 1 mg./lb. of body weight level (Table 5). No significant differences were found in hatchability in Experiments 1 and 2 for the preinjection, injection and post-injection periods.
Total Between Treat. Between Weeks Interaction
1,727 1,145 460 122
1,145* 230 61
* Significant value of F.
Feather Growth.—-As shown in Table 6, a highly significant retardation of primary feather growth in the males was found during the first week after injections of cortisone started. Differences in rate of feather growth were not significant thereafter. Inhibition of cushion feather growth (Experiment 3) was highly significant for the first three weeks after injections started, significant for the fourth week and non-significant thereafter (Table 7). An interesting observation was that the rejuvenating primary and cushion feathers were exceedingly stringy in appearance and had abnormally long shafts. These data confirm and extend to mature chickens the observations of Karnofsky el al. (1949, 1951) that cortisone retards feather formation in chick embryos. It is suggested that tissues vary greatly in their susceptibility to the action of cortisone, the physiological mode of action being complicated. The effects observed may be the result of alterations in the metabolic chain of events. Chick Weight.—Cortisone administered to the parental individuals had no sig-
TABLE 6.—The results of an analysis of variance of primary feather growth Experiment 1. Males, 1 mg./lb. Source of variation Total Between Treat Within Treat
Week d.f.
S.S.
5 1 4
3.6 3.1 .5
t Highly significant value of F.
2
1 M.S. 3.If 0.1
4
3
S.S.
M.S.
S.S.
M.S.
S.S.
M.S.
5.6 3.2 2.4
3.2 0.6
8.3 3.0 5.3
3.0 1.3
10.3 5.8 4.5
5.8 1.1
480
J. J. KUDZIA AND L. R. CHAMPION
TABLE 7.—The results of analysis of variance of cushion feather growth Experiment 3. Females, 3 mg./lb. Source of variance Total Between Treat. Within Treat.
Week
1
2
d.f.
s.s.
M.S.
S.S.
M.S.
S.S.
M.S.
11 1 10
1.0 0.5 0.5
0.5f 0.05
1.5 0.8 0.7
0.8f 0.07
3.8 2.6 1.2
2.6f 0.12
4 Total Between Treat. Within Treat.
3
11 1 10
3.0 1.4 1.6
5 1.4* 0.16
6.0 1.0 5.0
6 1.0 0.5
1.4 0.02 1.38
0.02 0.14
* Significant value of F. t Highly significant value of F. nificant effect on day-old or two-week chick weight. SUMMARY Two different levels of cortisone acetate were administered subcutaneously to mature New Hampshire male and female chickens. T h e suspensions were injected into the birds for four consecutive days followed by a lag period of three days, after which time injections were resumed for four additional consecutive days. The following results were secured: 1. Cortisone acetate caused a highly significant body weight loss in mature males during the injection period. 2. The body weight loss and reduced egg production observed by the females administered cortisone a t the 1 m g . / l b . of body weight level was not significant. 3. T h e body weight loss in females receiving 3 mg./lb. of cortisone per pound of body weight was not significant. However, the gain in body weight over and above initial body weight which was found to exist four weeks after injections ceased was highly significant. 4. A highly significant reduction in egg production was found to occur in females injected with 3 mg. of cortisone per pound of body weight.
5. A highly significant retardation in primary feather growth occurred in the males during the first week of injections of cortisone. 6. Inhibition of cushion feather growth in the females was highly significant for the first three weeks after initiation of injections, significant for the fourth week and non-significant thereafter. 7. Cortisone administered a t the 1 mg./lb. level caused a significant decrease in fertility, whereas at the high level (3 mg./lb.) significant reductions in fertility were not found. 8. No significant differences were found in egg weight, hatchability, day-old or two-week chick weight. REFERENCES Clark, I., 1950. Effect of cortisone on protein metabolism in the rat as studied with isotopic glycine. Federation Proc. 9: 161. Courrier, R., and A. Collonge, 1951. Cortisone and gestation. J. Am. Med. Assoc. 146: 493. Fisher, R. A., 1930. Statistical Methods for Research Workers. Oliver and Boyd, London. Follis, R. H., Jr., 1951. Effect of cortisone on growing bones of the rat. Proc. Soc. Exp. Biol. Med. 76: 722-724. Glaubach, S., W. Antopol and S. Graff, 1951. Excessive doses of cortisone in pregnant mice: effect on development and survival of the fetus and newborn and on the maternal breast tissue. Bui. New York Acad. Med. 27:398.
NORMAL OVULATION IN NON-LAYING HENS Hart, P. D'A., and R. J. W. Rees, 1950. Enhancing effect of cortisone on tuberculosis in the mouse. Lancet, 2: 391-395. Howes, E. L., C. M. Plotz, J. W. Blunt and C. Rogan, 1950. Retardation of wound healing by cortisone. Surgery, 28: 177-181. Kamofsky, D. A., P. A. Patterson and L. P. Ridgeway, 1949. Effect of folic acid, "4-amino" folic acids and related substances on growth of chick embryos. Proc. Soc. Exp. Biol. Med. 71:447-452. Kamofsky, D . A., L. P. Ridgeway and P. A. Patterson, 1951. Growth-inhibiting effect of cortisone
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acetate on the chick embryo. Endocrinology, 48: 596-616. Kendall, E. C , 1938. Isolation in crystalline form of the hormone of the adrenal cortex. Proc. Staff Meeting, Mayo Clinic 11: 351. Landauer, W. L., 1947. Potentiating effects of adrenal cortical extract on insulin-induced abnormalities of chick development. Endocrinology, 41:489-493. Mushett, C. W., C. C. Porter and R. H. Silber, 1951. Effects of cortisone administered subcutaneously and orally to dogs. Federation Proc. 10: 366.
Normal Ovulation in Non-Laying Hens R. K. COLE AND F. B. HUTT Department of Poultry Husbandry, Cornell University Agricultural Experiment Station, Ithaca, New York (Received for publication October 6, 1952)
f \ V U L A T I O N and the subsequent lay^* ing of a normal egg, usually within 26 hours/ are preceded and accompanied by certain changes in the body of the hen. The external characters that are associated with an active state of egg production are well known and are used as the basis for the periodic culling of commercial laying flocks. By the absence of external changes considered indicative of egg production, one is able to identify nonlayers and to remove them from the flock, if so desired. I t is customary to consider as layers those birds which show the external characteristics associated with egg production. We have learned, however, that some birds behave as though they were laying, and show appropriate changes in body condition, without actually producing an egg. This paper presents the pertinent findings when groups of such birds were subjected to study. SOURCES OF DATA
Among large populations of pedigreed
White Leghorns that were being trapnested there were a number of birds which had only occasionally been credited with an egg, and in most cases, none at all. The non-laying hens from four populations, S, T, U, and V, hatched in the four years from 1948 to 1951, respectively, have been segregated and studied, although under somewhat different conditions of management each year. 5 population. In order to distinguish between non-layers and the floor-layers, the 39 birds in the S population for which there were no eggs recorded, other than sporadic ones, were transferred early in May to individual laying cages and kept there for a period of one month to ascertain their egg-producing status. The ten birds found to be laying at regular intervals were returned to their original pens and thereafter considered as floorlayers. The 29 proven non-layers were transferred to a small pen and kept there until early in October, at the end of the first laying year, at which time the reproductive systems of the 14 survivors