- Email: [email protected]
The Effect of Amphenone in the Chicken1
TOLERANCE OF CHICKENS FOR ZINC
of rats fed high levels of 2inc. J. Nutrition, 72: 233-242. Mehring, A. L., Jr., J. H. Brumbaugh and H. W. Titus, 1956. A comparison of the growth of chicks fed diets containing different quantities of zinc. Poultry Sci. 35: 956-958. Roberson, R. H., and P. J. Schaible, 1960. The tolerance of growing chicks for high levels of different forms of zinc. Poultry Sci. 39: 893-896. Sandell, E. B., 1959. Colorimetric Determination of Traces of Metals, 3rd ed. Interscience Publishers, Inc., New York. Shirley, R. L., E. J. Benne and E. J. Miller, 1949. Report on zinc in plants. J. Assoc. Off. Agric. Chem. 32: 276-280.
The Effect of Amphenone in the Chicken 1 BRUCE GLICK
Department of Poultry Husbandry, Mississippi State University, State College, Miss. (Efeceived for publication May 10, 1961)
PROCEDURE
INTRODUCTION
T
HE growth, regression, and relationship of the bursa of Fabricius to other glands have been of primary concern in our studies, Glick (1960). The adrenal cortex appears to play a prominent part in the regression of the bursa, Selye (1943), Bannister (1951), Garren and Shaffner (1956), and Glick (1959). Inactivation of the adrenal cortex might be expected to prevent bursa regression. Amphenone [3,3-bis (p-aminophenyl)-butanone-2-dihydrochloride] is a drug capable of inhibiting adrenal cortical function in rats, Manrique et al. (1958), but not in the golden hamster, Marks et al. (1958), It was of interest to determine if Amphenone was capable of inhibiting adrenal cortical function in chickens.
1 Mississippi Agricultural journal article no. 936.
Experiment
Station
All New Hampshire chicks were raised in battery brooders and fed a basal ration. In all trials, Amphenone was taken up in sterile distilled water and injected intramuscularly. The sterile distilled water employed as a control injection was adjusted to the approximate pH of the Amphenone solution. The adrenocorticotrophic hormone (ACTH) was in a gelatin carrier. In trial 1 all injections were administered three times, twelve hours apart, to 3-week-old male chicks. Total and differential white blood cell counts were made two and six hours after the first injection and twenty hours after the last injection. Immediately after the last blood cell determination, all birds were killed with an overdose of Nembutal and body, bursa, and adrenal weights recorded. In the second trial, four daily injections
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Brink, M. F., D. E. Becker, S. W. Terrill and A. H. Jensen, 1959. Zinc toxicity in the weanling pig. J. Animal Sci. 18: 836-842. Grant-Frost, D. R., and E. J. Underwood, 1958. Zinc toxicity in the rat and its interrelation with copper. Australian J. Exp. Biol. Med. Sci. 36: 339-346. Insko, W. M., Jr., M. Lyons and J. H. Martin, 1938. The effect of manganese, zinc, aluminum, and iron salts on the incidence of perosis in chicks. Poultry Sci. 17: 264-269. Klussendorf, R. D., and J. Pensack, 1958. Newer aspects of zinc metabolism. J. Am. Vet. Med. Assoc. 132: 446-450. Magee, A. C , and G. Matrone, 1960. Studies on growth, copper metabolism and iron metabolism
317
318
B. GLICK
We employed 5-month-old pullets that were laying to test Amphenone since Perek and Eckstein (1959) have indicated that the adrenals of laying pullets are
more sensitive to ACTH than the adrenals of non-layers. Total white blood cell counts were determined by the Natt-Herrick technique (1946) and differential stains were made by the method of Wright. Blood cell counts and body weight differences and gland weights were analysed by the analysis of variance and covariance, respectively, Snedecor (1946). Significant mean difference comparisons were made by the method of Duncan (1955). All means are accompanied by their standard deviations. RESULTS
The smallest percentage of lymphocytes and highest percentage of heterophils were found after a single injection of 50 mg. of Amphenone, Table 1. In only one case was a significant mean difference noted: heterophil count two hours after a water injection and 50 mg. Amphenone injection. The injections did not significantly affect body, bursa, or adrenal weights of 3-week-old males, Table 2. The injection of 1-week-old chicks with 20 mg. of
TABLE 1.—The influence of Amphenone on the white blood cells of 3-week-old New Hampshire males1 Amphenone
Total white blood cells, cells/ mm.'X 1,000 A B C Lymphocytes, % A B C Heterophil, % A B C
Control
Water, .2 ml./inj.
19 + 6 20 + 8 20 + 4
.1 ml./inj. (10 mg./inj.)
.2 ml./inj. (20 mg./inj.)
.5 ml./inj. (50 mg./inj.)
20+10 23+ 7 19+ 7
19+ 5 26+11 16+ 7
15 + 3 19 + 5 18 + 7
15 + 3 33 + 4 21 + 6
92 + 5 87 + 7 85 + 8
88+ 8 84+10 85+ 7
89+ 2 81+ 9 90+ 8
91 + 2 81+9 86 + 8
80 + 7 71 + 7 87+3
6+ 4 9+ 7 12 + 8
12*+ 7 13 + 9 13+5
9+ 2 14± 9 7+ 6
7+ 2 17 + 9 11 + 8
17* + 6 23 + 8 11 + 3
1 All means represented by 5 birds. A—2 hours after 1st injection. B—6 hours after 1st injection. C—20 hours after 3rd injection. *P<.05.
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
of Amphenone were administered to 1week-old males. Ten hours after the last injection, all birds were killed and the body, bursae, and adrenals weighed. In trial 3, twenty-five 5-month-old pullets were equally divided into five groups. One group of pullets were controls and received no injections. A second group received two 1 ml. injections of water separated by twenty hours. A third group received two 1 ml. (20 mg.) injections of Amphenone separated by twenty hours. The fourth group of pullets received three 1 ml. (20 mg.) injections of Amphenone. Ten hours separated each injection. The last two Amphenone injections were accompanied by an injection of .25 ml. (10 I.U.) of ACTH. The final group received two .25 ml. (10 I.U.) injections of ACTH. All initial injections were administered at the same time. All blood cell determinations were made three hours after the first and last injections.
319
EFFECTS OF AMPHENONE TABLE 2.—The effect of Amphenone on the weight of the bursa of Fabricius and adrenal glands of 3-itieek-old chicks1 Amphenone Water, .2 ml./inj.
Control
Gain in body weight, gm. Bursa of Fabricius, gm. Adrenal, mg. 1
36±12 1.57±.50 56±14
38+10 1.99±.59 52 + 4
.1 ml./inj. (10 mg./inj.)
.2 ml./inj. (20 mg./inj.)
.5 ml./inj. (50 mg./inj.)
37 ±15 1.71±.43 59 ±12
36 ±12 1.61+ .5 52 ±13
37 ± 1 5 1.42+.69 48±9
Each mean represented by 8 birds.
DISCUSSION
It is known that the injections of cortisone acetate and ACTH into birds will decrease the relative lymphocyte count and increase the relative heterophil count, Bannister (1951), Huble, (1955), and Glick (1959). It is generally agreed that the release of ACTH occurs normally in mammals in the presence of certain agents (called stressor agents) and that such TABLE 3.-—The
release will stimulate the production of glucocorticoids which are thymolytic and lymphopenic, Selye and Heuser (1956). A similar phenomena appears to occur in birds in the presence of stressor agents, Garren and Shaffner (1956), Newcomer (1958), and Chancellor and Glick (1960). The increase in percentage of heterophils and decrease in percentage of lymphocytes that occurred in the presence of Amphenone or Amphenone plus ACTH in this study would indicate that the pituitaryadrenal axis was stimulated. At least, the data support the argument that Amphenone under the conditions of this test did not inactivate the adrenal cortex of the chicken. The consistent reduction in bursa size, although not significant, tends to confirm the inability of Amphenone to inactivate the adrenal cortex since we know the bursa to regress in the presence
effect of Amphenone on the weight of the bursa of Fabricius, adrenals, and body of 1-week-old chicks1^ Amphenone Water
Control
. 0 5 ml./inj. (10 mg./inj.)
.10 ml./inj. (20 mg./inj.)
Initial body weight, gm.
44± 5
40± 5
45+ 4
45+ 6
Final body weight, gm.
61+7
57 ± 1 3
54+ 6
45± 9
Bursa of Fabricius, mg.
110 + 28
92 ± 3 4
76 + 31
65 ± 2 0
11+ 2
13± 3
12+ 2
11± 4
Adrenal, mg. 1 2
Each mean represented by 8 birds. All means not underscored by the same line are significantly different at the 5% level, Duncan (1956).
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Amphenone (Table 3) significantly reduced body weight, but did not significantly affect the bursa or adrenals. The apparent reduction in bursa weight was a reflection of the inhibition of body growth. It is apparent that Amphenone alone and ACTH alone will decrease the relative lymphocyte count and increase the relative heterophil counts of 5-month-old pullets, Table 4.
320
B. GLICK TABLE 4.—The influence of Amphenone and ACTH on the white blood cell counts of 5-month-old New Hampshire pullets1 Amphenone, -m--iK' ACTH, .25 ml. Al »phenone, inj.+ACTH, ( 1 Q j ft y . . 1m v .25 ml. " ' (20 mg.)/mj. (10 I.U.)/inj.
l
Control
Total white blood cells, celIs/mm. 3 X 1,000 3 hrs. after 1st inj. 3 hrs. after last inj.
27+ 82 26+ 5
37+ 4 29+ 5
23+ 92 29 + 15
34+ 5 31+5
31 + 4 21+4
Lymphocyte, % 3 hrs. after 1st inj.
70+ 6
70 ±10
51 + 11
45+ 8
38±6
3 hrs. after last inj.
75 + 10
74+11
54+12
59+10
59 + 8
Heterophil, % 3 hrs. after 1st inj.
26+10
28 + 11
45+ 9
50+ 7
58 + 5
3 hrs. after last inj.
20+ 9
22 + 11
43 + 12
38 + 10
38 + 7
1 2
All means not underscored by the same line significantly different at the 5% level. These means were significantly smaller than the control mean.
of glucocorticoids. The reaction of the week-old chicks to Amphenone should be noted. About 10 seconds after administering 20 mg. of Amphenone, the chicks closed their eyes and appeared drowsy. Usually the chicks exhibited a gasping reaction after receiving Amphenone. Within 20 seconds, the chicks lost their balance and in less than a minute were prostrate. We lost two chicks after the first Amphenone injection. SUMMARY
Single and multiple injections of Amphenone were administered to male chicks and 5-month-old pullets. White blood cell counts and weights of the body, bursa of Fabricius, and adrenals were the variables studied. The decrease in percent lymphocytes and increase in percent heterophils, coupled with a consistent reduction in bursa size in the presence of Amphenone or Amphenone plus ACTH, support the conclusion that Amphenone is incapable of inactivating adrenal cortical function in the chicken.
ACKNOWLEDGMENT
Appreciation is expressed to Dr. E. W. Young of the Upjohn Co., Kalamazoo, Michigan, for making the Amphenone available and to Mrs. Bobbie Brashear, Laboratory Technician, for her assistance in the collection of the data. REFERENCES Bannister, G. L., 1951. Avian blood changes following injections of cortisone. Canad. J. Comp. Med. 15: 169-171. Chancellor, L., and B. Glick, 1960. Effect of temperature as a stresser on white blood cells, adrenals, and bursae of Fabricius of chicks. Am. J. Physiol. 198: 1346-1348. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Garren, H., and C. S. Shaffner, 1956. How the period of exposure to different stress stimuli affects the endocrine and lymphatic gland weights of young chickens. Poultry Sci. 34: 266. Glick, B., 1959. The experimental production of the stress picture with cortisone and the effect of penicillin in young chickens. Ohio J. Sci. 59: 8 1 86. Glick, B., 1960. Growth of the bursa of Fabricius and its relationship to the adrenal gland in the White Pekin duck, White Leghorn, outbred and inbred New Hampshire. Poultry Sci. 39: 130-139.
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Water, 1 ml./inj.
EFFECTS OF AMPHENONE Huble, J., 1955. Haematological changes in cockerels after ACTH and cortisone acetate treatment. Poultry Sci. 34: 1357-1360. Manrique, J., R. Paredes, J. Arabehety and S. J. Gray, 1958. Effect of Amphenone on gastric secretory activity. Am. J. Physiol. 195: 221-228. Marks, B. H., M. Alpert and F. A. Krueger, 1958. Effect of Amphenone upon steroidogenesis in the adrenal cortex of the goldem hamster. Endocrinology, 63: 75-81. Natt, M. R., and C. A. Herrick, 1946. A new blood diluent for counting the erythrocytes and leukocytes of the chicken. Poultry Sci. 31: 735-738. Newcomer, W. S., 1958. Physiologic factors which
321
influence acidophilia induced by stressors in the chicken. Am. J. Physiol. 194: 251. Perek, M., and B. Eckstein, 1959. The adrenal ascorbic acid content of molting hens and the effect of ACTH on the adrenal ascorbic acid content of laying hens. Poultry Sci. 38: 996-999. Selye, H., and G. Heuser, 1956. The Fifth Annual Report on Stress. MD Publications, New York, pp. 815. Selye, H., 1943. Morphological changes in the fowl following chronic overdosages with various steroids. J. Morphol. 73: 401-241. Snedecor, G. W., 1946. Statistical Methods. The Iowa State College Press, Ames, Iowa.
2. CHANGES IN ADRENAL CHOLESTEROL AND BLOOD CONSTITUENT LEVELS H. S. SIEGEL Poultry Department, Virginia Polytechnic Institute, Blacksburg (Received for publication May 10, 1961)
R
ESISTANCE to general systemic stress in mammals has been characterized by adrenal hyperfunction as manifested by adrenal hypertrophy, adrenal cholesterol and ascorbic acid depletion, general lymphatic tissue involution and associated blood element changes (Selye, 1950). It is generally accepted that these responses are mediated through increased hypophysial corticotropin output. Similarly, a variety of stress stimuli have been shown to induce adrenal hypertrophy and lymphatic involution in birds (Garren and Shaffner, 1956; Siegel, 1960; Siegel and Siegel, 1961). Chronic or prolonged doses of exogenous corticotropin (ACTH) induce increases in adrenal weight and involution of lymphatic tissue in birds (Bates et al, 1940; Jailer and Boas, 1950; Zarrow and Baldini, 1952; Garren and Hill, 1958). Garren et al. (1961) have also
observed that route and vehicle of application may modify responses. However, possibly as a result of greater functional independence of avian cortical tissue from the anterior pituitary (Brown et al., 1958; Newcomer, 1959), ACTH may not be as efficacious in producing these changes in birds as in laboratory mammals (Dulin, 1953; Conner, 1959; Flickinger, 1959). Furthermore, it has been shown that age and sex may also act to modify such responses (Siegel, 1961). In a comparative study, Elton et al. (1959) concluded that adrenal cholesterol and ascorbic acid levels were not changed in chickens 1 to 4 hours after ACTH administration. Siegel and Beane (1961) have reported, however, that maximum and significant reductions in adrenal cholesterol levels occurred in male chickens, 46 days of age, 12 hours after ACTH
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Age and Sex Modification of Responses to Adrenocorticotropin in Young Chickens
of rats fed high levels of 2inc. J. Nutrition, 72: 233-242. Mehring, A. L., Jr., J. H. Brumbaugh and H. W. Titus, 1956. A comparison of the growth of chicks fed diets containing different quantities of zinc. Poultry Sci. 35: 956-958. Roberson, R. H., and P. J. Schaible, 1960. The tolerance of growing chicks for high levels of different forms of zinc. Poultry Sci. 39: 893-896. Sandell, E. B., 1959. Colorimetric Determination of Traces of Metals, 3rd ed. Interscience Publishers, Inc., New York. Shirley, R. L., E. J. Benne and E. J. Miller, 1949. Report on zinc in plants. J. Assoc. Off. Agric. Chem. 32: 276-280.
The Effect of Amphenone in the Chicken 1 BRUCE GLICK
Department of Poultry Husbandry, Mississippi State University, State College, Miss. (Efeceived for publication May 10, 1961)
PROCEDURE
INTRODUCTION
T
HE growth, regression, and relationship of the bursa of Fabricius to other glands have been of primary concern in our studies, Glick (1960). The adrenal cortex appears to play a prominent part in the regression of the bursa, Selye (1943), Bannister (1951), Garren and Shaffner (1956), and Glick (1959). Inactivation of the adrenal cortex might be expected to prevent bursa regression. Amphenone [3,3-bis (p-aminophenyl)-butanone-2-dihydrochloride] is a drug capable of inhibiting adrenal cortical function in rats, Manrique et al. (1958), but not in the golden hamster, Marks et al. (1958), It was of interest to determine if Amphenone was capable of inhibiting adrenal cortical function in chickens.
1 Mississippi Agricultural journal article no. 936.
Experiment
Station
All New Hampshire chicks were raised in battery brooders and fed a basal ration. In all trials, Amphenone was taken up in sterile distilled water and injected intramuscularly. The sterile distilled water employed as a control injection was adjusted to the approximate pH of the Amphenone solution. The adrenocorticotrophic hormone (ACTH) was in a gelatin carrier. In trial 1 all injections were administered three times, twelve hours apart, to 3-week-old male chicks. Total and differential white blood cell counts were made two and six hours after the first injection and twenty hours after the last injection. Immediately after the last blood cell determination, all birds were killed with an overdose of Nembutal and body, bursa, and adrenal weights recorded. In the second trial, four daily injections
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Brink, M. F., D. E. Becker, S. W. Terrill and A. H. Jensen, 1959. Zinc toxicity in the weanling pig. J. Animal Sci. 18: 836-842. Grant-Frost, D. R., and E. J. Underwood, 1958. Zinc toxicity in the rat and its interrelation with copper. Australian J. Exp. Biol. Med. Sci. 36: 339-346. Insko, W. M., Jr., M. Lyons and J. H. Martin, 1938. The effect of manganese, zinc, aluminum, and iron salts on the incidence of perosis in chicks. Poultry Sci. 17: 264-269. Klussendorf, R. D., and J. Pensack, 1958. Newer aspects of zinc metabolism. J. Am. Vet. Med. Assoc. 132: 446-450. Magee, A. C , and G. Matrone, 1960. Studies on growth, copper metabolism and iron metabolism
317
318
B. GLICK
We employed 5-month-old pullets that were laying to test Amphenone since Perek and Eckstein (1959) have indicated that the adrenals of laying pullets are
more sensitive to ACTH than the adrenals of non-layers. Total white blood cell counts were determined by the Natt-Herrick technique (1946) and differential stains were made by the method of Wright. Blood cell counts and body weight differences and gland weights were analysed by the analysis of variance and covariance, respectively, Snedecor (1946). Significant mean difference comparisons were made by the method of Duncan (1955). All means are accompanied by their standard deviations. RESULTS
The smallest percentage of lymphocytes and highest percentage of heterophils were found after a single injection of 50 mg. of Amphenone, Table 1. In only one case was a significant mean difference noted: heterophil count two hours after a water injection and 50 mg. Amphenone injection. The injections did not significantly affect body, bursa, or adrenal weights of 3-week-old males, Table 2. The injection of 1-week-old chicks with 20 mg. of
TABLE 1.—The influence of Amphenone on the white blood cells of 3-week-old New Hampshire males1 Amphenone
Total white blood cells, cells/ mm.'X 1,000 A B C Lymphocytes, % A B C Heterophil, % A B C
Control
Water, .2 ml./inj.
19 + 6 20 + 8 20 + 4
.1 ml./inj. (10 mg./inj.)
.2 ml./inj. (20 mg./inj.)
.5 ml./inj. (50 mg./inj.)
20+10 23+ 7 19+ 7
19+ 5 26+11 16+ 7
15 + 3 19 + 5 18 + 7
15 + 3 33 + 4 21 + 6
92 + 5 87 + 7 85 + 8
88+ 8 84+10 85+ 7
89+ 2 81+ 9 90+ 8
91 + 2 81+9 86 + 8
80 + 7 71 + 7 87+3
6+ 4 9+ 7 12 + 8
12*+ 7 13 + 9 13+5
9+ 2 14± 9 7+ 6
7+ 2 17 + 9 11 + 8
17* + 6 23 + 8 11 + 3
1 All means represented by 5 birds. A—2 hours after 1st injection. B—6 hours after 1st injection. C—20 hours after 3rd injection. *P<.05.
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
of Amphenone were administered to 1week-old males. Ten hours after the last injection, all birds were killed and the body, bursae, and adrenals weighed. In trial 3, twenty-five 5-month-old pullets were equally divided into five groups. One group of pullets were controls and received no injections. A second group received two 1 ml. injections of water separated by twenty hours. A third group received two 1 ml. (20 mg.) injections of Amphenone separated by twenty hours. The fourth group of pullets received three 1 ml. (20 mg.) injections of Amphenone. Ten hours separated each injection. The last two Amphenone injections were accompanied by an injection of .25 ml. (10 I.U.) of ACTH. The final group received two .25 ml. (10 I.U.) injections of ACTH. All initial injections were administered at the same time. All blood cell determinations were made three hours after the first and last injections.
319
EFFECTS OF AMPHENONE TABLE 2.—The effect of Amphenone on the weight of the bursa of Fabricius and adrenal glands of 3-itieek-old chicks1 Amphenone Water, .2 ml./inj.
Control
Gain in body weight, gm. Bursa of Fabricius, gm. Adrenal, mg. 1
36±12 1.57±.50 56±14
38+10 1.99±.59 52 + 4
.1 ml./inj. (10 mg./inj.)
.2 ml./inj. (20 mg./inj.)
.5 ml./inj. (50 mg./inj.)
37 ±15 1.71±.43 59 ±12
36 ±12 1.61+ .5 52 ±13
37 ± 1 5 1.42+.69 48±9
Each mean represented by 8 birds.
DISCUSSION
It is known that the injections of cortisone acetate and ACTH into birds will decrease the relative lymphocyte count and increase the relative heterophil count, Bannister (1951), Huble, (1955), and Glick (1959). It is generally agreed that the release of ACTH occurs normally in mammals in the presence of certain agents (called stressor agents) and that such TABLE 3.-—The
release will stimulate the production of glucocorticoids which are thymolytic and lymphopenic, Selye and Heuser (1956). A similar phenomena appears to occur in birds in the presence of stressor agents, Garren and Shaffner (1956), Newcomer (1958), and Chancellor and Glick (1960). The increase in percentage of heterophils and decrease in percentage of lymphocytes that occurred in the presence of Amphenone or Amphenone plus ACTH in this study would indicate that the pituitaryadrenal axis was stimulated. At least, the data support the argument that Amphenone under the conditions of this test did not inactivate the adrenal cortex of the chicken. The consistent reduction in bursa size, although not significant, tends to confirm the inability of Amphenone to inactivate the adrenal cortex since we know the bursa to regress in the presence
effect of Amphenone on the weight of the bursa of Fabricius, adrenals, and body of 1-week-old chicks1^ Amphenone Water
Control
. 0 5 ml./inj. (10 mg./inj.)
.10 ml./inj. (20 mg./inj.)
Initial body weight, gm.
44± 5
40± 5
45+ 4
45+ 6
Final body weight, gm.
61+7
57 ± 1 3
54+ 6
45± 9
Bursa of Fabricius, mg.
110 + 28
92 ± 3 4
76 + 31
65 ± 2 0
11+ 2
13± 3
12+ 2
11± 4
Adrenal, mg. 1 2
Each mean represented by 8 birds. All means not underscored by the same line are significantly different at the 5% level, Duncan (1956).
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Amphenone (Table 3) significantly reduced body weight, but did not significantly affect the bursa or adrenals. The apparent reduction in bursa weight was a reflection of the inhibition of body growth. It is apparent that Amphenone alone and ACTH alone will decrease the relative lymphocyte count and increase the relative heterophil counts of 5-month-old pullets, Table 4.
320
B. GLICK TABLE 4.—The influence of Amphenone and ACTH on the white blood cell counts of 5-month-old New Hampshire pullets1 Amphenone, -m--iK' ACTH, .25 ml. Al »phenone, inj.+ACTH, ( 1 Q j ft y . . 1m v .25 ml. " ' (20 mg.)/mj. (10 I.U.)/inj.
l
Control
Total white blood cells, celIs/mm. 3 X 1,000 3 hrs. after 1st inj. 3 hrs. after last inj.
27+ 82 26+ 5
37+ 4 29+ 5
23+ 92 29 + 15
34+ 5 31+5
31 + 4 21+4
Lymphocyte, % 3 hrs. after 1st inj.
70+ 6
70 ±10
51 + 11
45+ 8
38±6
3 hrs. after last inj.
75 + 10
74+11
54+12
59+10
59 + 8
Heterophil, % 3 hrs. after 1st inj.
26+10
28 + 11
45+ 9
50+ 7
58 + 5
3 hrs. after last inj.
20+ 9
22 + 11
43 + 12
38 + 10
38 + 7
1 2
All means not underscored by the same line significantly different at the 5% level. These means were significantly smaller than the control mean.
of glucocorticoids. The reaction of the week-old chicks to Amphenone should be noted. About 10 seconds after administering 20 mg. of Amphenone, the chicks closed their eyes and appeared drowsy. Usually the chicks exhibited a gasping reaction after receiving Amphenone. Within 20 seconds, the chicks lost their balance and in less than a minute were prostrate. We lost two chicks after the first Amphenone injection. SUMMARY
Single and multiple injections of Amphenone were administered to male chicks and 5-month-old pullets. White blood cell counts and weights of the body, bursa of Fabricius, and adrenals were the variables studied. The decrease in percent lymphocytes and increase in percent heterophils, coupled with a consistent reduction in bursa size in the presence of Amphenone or Amphenone plus ACTH, support the conclusion that Amphenone is incapable of inactivating adrenal cortical function in the chicken.
ACKNOWLEDGMENT
Appreciation is expressed to Dr. E. W. Young of the Upjohn Co., Kalamazoo, Michigan, for making the Amphenone available and to Mrs. Bobbie Brashear, Laboratory Technician, for her assistance in the collection of the data. REFERENCES Bannister, G. L., 1951. Avian blood changes following injections of cortisone. Canad. J. Comp. Med. 15: 169-171. Chancellor, L., and B. Glick, 1960. Effect of temperature as a stresser on white blood cells, adrenals, and bursae of Fabricius of chicks. Am. J. Physiol. 198: 1346-1348. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Garren, H., and C. S. Shaffner, 1956. How the period of exposure to different stress stimuli affects the endocrine and lymphatic gland weights of young chickens. Poultry Sci. 34: 266. Glick, B., 1959. The experimental production of the stress picture with cortisone and the effect of penicillin in young chickens. Ohio J. Sci. 59: 8 1 86. Glick, B., 1960. Growth of the bursa of Fabricius and its relationship to the adrenal gland in the White Pekin duck, White Leghorn, outbred and inbred New Hampshire. Poultry Sci. 39: 130-139.
Downloaded from http://ps.oxfordjournals.org/ at University of Leeds on May 17, 2015
Water, 1 ml./inj.
EFFECTS OF AMPHENONE Huble, J., 1955. Haematological changes in cockerels after ACTH and cortisone acetate treatment. Poultry Sci. 34: 1357-1360. Manrique, J., R. Paredes, J. Arabehety and S. J. Gray, 1958. Effect of Amphenone on gastric secretory activity. Am. J. Physiol. 195: 221-228. Marks, B. H., M. Alpert and F. A. Krueger, 1958. Effect of Amphenone upon steroidogenesis in the adrenal cortex of the goldem hamster. Endocrinology, 63: 75-81. Natt, M. R., and C. A. Herrick, 1946. A new blood diluent for counting the erythrocytes and leukocytes of the chicken. Poultry Sci. 31: 735-738. Newcomer, W. S., 1958. Physiologic factors which
321
influence acidophilia induced by stressors in the chicken. Am. J. Physiol. 194: 251. Perek, M., and B. Eckstein, 1959. The adrenal ascorbic acid content of molting hens and the effect of ACTH on the adrenal ascorbic acid content of laying hens. Poultry Sci. 38: 996-999. Selye, H., and G. Heuser, 1956. The Fifth Annual Report on Stress. MD Publications, New York, pp. 815. Selye, H., 1943. Morphological changes in the fowl following chronic overdosages with various steroids. J. Morphol. 73: 401-241. Snedecor, G. W., 1946. Statistical Methods. The Iowa State College Press, Ames, Iowa.
2. CHANGES IN ADRENAL CHOLESTEROL AND BLOOD CONSTITUENT LEVELS H. S. SIEGEL Poultry Department, Virginia Polytechnic Institute, Blacksburg (Received for publication May 10, 1961)
R
ESISTANCE to general systemic stress in mammals has been characterized by adrenal hyperfunction as manifested by adrenal hypertrophy, adrenal cholesterol and ascorbic acid depletion, general lymphatic tissue involution and associated blood element changes (Selye, 1950). It is generally accepted that these responses are mediated through increased hypophysial corticotropin output. Similarly, a variety of stress stimuli have been shown to induce adrenal hypertrophy and lymphatic involution in birds (Garren and Shaffner, 1956; Siegel, 1960; Siegel and Siegel, 1961). Chronic or prolonged doses of exogenous corticotropin (ACTH) induce increases in adrenal weight and involution of lymphatic tissue in birds (Bates et al, 1940; Jailer and Boas, 1950; Zarrow and Baldini, 1952; Garren and Hill, 1958). Garren et al. (1961) have also
observed that route and vehicle of application may modify responses. However, possibly as a result of greater functional independence of avian cortical tissue from the anterior pituitary (Brown et al., 1958; Newcomer, 1959), ACTH may not be as efficacious in producing these changes in birds as in laboratory mammals (Dulin, 1953; Conner, 1959; Flickinger, 1959). Furthermore, it has been shown that age and sex may also act to modify such responses (Siegel, 1961). In a comparative study, Elton et al. (1959) concluded that adrenal cholesterol and ascorbic acid levels were not changed in chickens 1 to 4 hours after ACTH administration. Siegel and Beane (1961) have reported, however, that maximum and significant reductions in adrenal cholesterol levels occurred in male chickens, 46 days of age, 12 hours after ACTH
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Age and Sex Modification of Responses to Adrenocorticotropin in Young Chickens